2nd International Conference on Innate Immunity and Immune System Diseases July 21-22, 2016 Berlin, Germany

Biography:

Jurgen Bernhagen has studied Biochemistry and Immunology at the University of Tübingen, Germany and at Queen Mary College, London, UK. He has performed a sandwich PhD thesis at the University of Tübingen and at the Picower Institute for Medical Research, Manhasset, NY, USA and trained as a Post doctorate at the Picower. He is currently a full Professor of Biochemistry and Molecular Cell Biology at RWTH Aachen University, Germany, and the Chair and Director of the homonymous institute. His main research interest has been on cytokines, chemokines and their role in inflammation with a focus on MIF and the biochemical and structural features and mechanisms of such inflammatory mediators. He has also studies the COP9 signalosome, disease models encompass rodent model of atherosclerosis, sepsis, liver and kidney disease as well as colitis and colorectal cancer. He has authored more than 120 peer-reviewed papers in these areas, several of them published in leading journals such as Nature, Nature Medicine, or PNAS. He serves on the Editorial Board of several journals and serves on several Review committees for extramural funding and various fellowship organizations.

Abstract:

Inflammatory processes such as those promoting atherosclerotic lesion formations are pivotally driven by components of the innate and adaptive immune axis. Chemokines and their receptors are particularly prominent part of the innate immune arm. While the role of classical chemokines, i.e., belonging to the CC or CXC families is increasingly well understood, an emerging family of chemokine like inflammatory mediators termed ‘innate chemokines’, CLF chemokines or micro-chemokines, which additionally struc¬turally and functionally overlaps with the mediator class of alarmins, has been identified, but it yet has to be comprehensively characterized regarding its molecular mechanism and role in disease. For example, innate chemokines modulate inflammatory reactions in the atherogenic arterial wall and numerous other inflamed tissues, but the precise receptor signaling mechanisms are still only poorly understood. What is known is that many innate chemokines share functional homology with classical chemokines and signal through classical chemokine receptors, whereas they do not exhibit conserved structural features such as N-terminal tandem cysteine residues or the chemokine fold. Thus, important receptor binding motifs yet have to be characterized. This lecture will give an overview of the mechanisms underlying “molecular hijacking” of classical chemokine receptors by innate chemokines, featuring their pathophysiological role. Examples will encompass high mobility group binding protein-1 (HMGB1), macrophage migration inhibitory factor (MIF), MIF-2/D-DT and certain β-defensins. Receptor usage, binding domains, signaling, innate immune cell regulation and involvement in various inflammatory conditions, including atherosclerosis will be discussed. The lecture will outline strategies to target such mediators in disease either in conjunction or explicit exclusion of the co-targeting of classical chemokines. Finally, a cross kingdom analysis will be shared offering more general understanding of some of these mediators.

Biography:

Thomas C G Bosch has studied Biology at the University of Munich, Germany and Swansea, United Kingdom and earned his Doctorate in 1986. Since 2000, he has been a Professor of General Zoology at Kiel University. From 2010 to 2013, he has served as Vice President of Kiel University. Since 2013 he is heading the interdisciplinary research center “Kiel Life Science” (KLS). He is also Coordinator of a new DFG-funded Collaborative Research Center on “Origin and Function of Metaorganisms”. He is the recipient of numerous awards including an honorary Doctorate degree from St. Petersburg State University, Russia.

Abstract:

Characterization of the innate immune repertoire of evolutionary old organisms is of both fundamental and applied interest; it not only provides insights into the basic immunological "tool kit" of the common ancestor of all animals but is also likely to be important in understanding human barrier disorders by describing ancient mechanisms of host/microbial interactions and the resulting evolutionary selection processes. Here, I evaluate our current understanding of the evolution of epithelial-based innate immunity. I highlight growing evidence that the innate immune system with its host-specific antimicrobial peptides and rich repertoire of pattern recognition receptors has evolved in response to the need for controlling resident beneficial microbes rather than to defend against invasive pathogens.

Biography:

John H White has obtained his PhD from Harvard University and has been working in the vitamin-D field since 2000. His lab was the first to perform comprehensive gene expression profiling studies on genes regulated by the hormonal form of vitamin-D. These studies provided multiple insights into novel areas of vitamin-D physiology, including its role as a regulator in innate immunity in humans.

Abstract:

Abstract Hormonal 1,25-dihydroxyvitamin D (1,25D) signals through the nuclear vitamin-D receptor (VDR), a ligand-regulated transcription factor. Vitamin-D was discovered as the cure to nutritional rickets and 1,25D is a critical regulator of calcium homeostasis during development and in the adult. However, the VDR is expressed in several tissues not implicated in calcium homeostasis, including throughout the immune system and a wide range of studies over recent years has revealed that hormonal vitamin-D is an important regulator of innate immunity. In humans, the 1,25D-bound VDR directly induces the transcription of genes encoding antimicrobial peptides (AMPs), pattern recognition receptors and key cytokines implicated in innate immune responses. These findings provide a molecular basis for a number of clinical studies providing a correlation between vitamin-D deficiency and an increased risk of infection. Notably, while global mechanisms of innate immune responses to pathogen threat have been conserved during evolution, the details of those responses and their regulation are species-specific. We find that the cognate binding sites for the VDR (vitamin D response elements; VDREs) present in a number of human genes encoding key components of innate immune responses are highly conserved in primates, but not present in rodent genes. Similarly, 1,25D-induced production of AMPs appears to be absent in mice. Given that other work has shown that 1,25D does control innate immune responses in rodent models of disease, the similarities and differences in 1,25D-regulated innate immune responses between species will be discussed.

Biography:

icancer agents that infect and effectively

Abstract:

Abstract Oncolytic viruses (OVs) are novel anticancer agents that infect and effectively kill cancer cells but not normal cells. Although tumor growth is delayed or eliminated in numerous animal models following treatment with OVs, several cancer models remain partially or completely resistant to viral oncolysis. To overcome this resistance, experimental strategies are now combining OVs with different cytotoxic compounds to improve OV efficacy. Our laboratory has previously demonstrated that OV replication can be bolstered by co-administration of other chemical agents such as Triptolide, a natural molecule derived from the medicinal herb. In the current study, we investigated the capacity of sulforaphane (SFN); an anti-cancer compound naturally occurred in cruciferous vegetables with demonstrated potent antioxidant and possible anti-inflammatory actions to enhance vesicular stomatitis virus (VSV) oncolysis in OV-resistant cancer cells. We ultimately demonstrate that the resistant PC3 prostate cancer cell line can be sensitized to VSV by addition of SFN. Indeed, SFN dose-dependently enhances the replication of VSV. Neither VSV (MOI 0.1) nor SFN (20 uM) alone are toxic against PC3 cells in vitro; however, in combination they greatly increased the oncolytic capacity of VSV by reducing cancer cell viability and promoting apoptosis-mediated cell death. Furthermore, the potentiation of VSV oncolysis by SFN is dependent on the production of ROS and is associated with the induction of autophagy. SFN is known to induce phase II antioxidant genes via Nrf2 activation, which regulates ROS levels and stimulates autophagy in prostate cancer cells. Mechanistically, SFN inhibited the innate antiviral response by blocking the type-1 interferon (IFN) signaling pathway, through the activationof the Nrf2 transcription factor. Exogenous Nrf2 expression inhibits Interferon-Stimulated Response Element (ISRE) promoter activity in a dose dependent manner following virus infection or IFN treatment. Taken together, these results demonstrate for the first time the synergic effect of SFN and VSV and indicate that SFN treatment increases VSV replication and the subsequent apoptosis of tumor cells by inhibiting IFN signaling. We are currently investigating the molecular mechanism involved in VSV-induced oncolysis by Nrf2 activators and evaluating the therapeutic potential of the combination of OV and Nrf2 activators in a mouse model of prostate cancer.

Biography:

Nayef Jarrous is currently working at “The Hebrew University of Jerusalem, Israel”. His research interest is based on “Human nuclear RNase P ribonucleoprotein in tRNA processing”. He has published many articles in reputed journals.

Abstract:

Abstract RNA polymerase III (Pol III) is a key player in innate immunity, as it serves as a sensor of viral and bacterial DNA of infected cells. This sensing asset is based on promoter independent recognition of foreign DNA templates in the cytoplasm and transcription via nonspecific initiation mechanism. The resulting 5’-triphosphate RNA transcripts activate the retinoic acid induced gene I (RIG-I), thus leading to induction of type-I interferon. We have previously shown that the human catalytic ribonucleoprotein RNase P is implicated in formation of proficient initiation complexes of nuclear Pol III on 5S rRNA and tRNA genes. However, it was unknown if this ribonucleoprotein is also implicated in nonspecific initiation of gene transcription by cytoplasmic Pol III. We will present preliminary results that show that the H1 RNA subunit of human RNase P is implicated in promoter independent initiation of transcription of synthetic circular DNA templates (COLIGOs) by cytoplasmic Pol III. The regulatory role of H1 RNA in this transcription system may explain the existence of RNase P like RNA genes in DNA viruses and the possible roles of their transcripts in evading antiviral innate immune responses.

Biography:

Many bacterial pathogens utilize a type-III secretion system (T3SS) to inject virulence proteins (effectors) into host cells to subv

Abstract:

Many bacterial pathogens utilize a type-III secretion system (T3SS) to inject virulence proteins (effectors) into host cells to subvert various biological functions. Effector subversion of pro-inflammatory host responses is well studied, but less attention has been given to the potential inhibition of host interferon (IFN) signaling. Type-I IFNs are important both to maintaining intestinal homeostasis and to responding to pathogen infection. Pathogens have evolved strategies to interfere with host type-I IFN production. A recent study found both that IFN-β is induced by enteropathogenic E. coli (EPEC) infection and that the EPEC T3SS effector NleD inhibits IFN-β induction. IFN expression is known to be important to limiting Citrobacter rodentium infection but whether C. rodentium T3SS effectors inhibit host IFN-β induction is unclear. We screened C. rodentium strains bearing deletions in individual T3SS effectors to determine the extent to which this pathogen might inhibit the host IFN-β response. To determine if C. rodentium T3SS effectors inhibit the host type-I IFN response, we monitored the survival of a recombinant vesicular stomatitis virus (VSV). Since TRAF3 is critical to IFN signaling, we also monitored effector mediated inhibition of the TNF receptor (TNFR) associated factor 3 (TRAF3) ubiquitination in RAW264.7 cells. Supernatants from cells infected with C. rodentium escN inhibited VSV to levels similar to those induced by LPS treatment. By contrast, supernatants from cells infected with WT C. rodentium did not inhibit VSV-GFP growth. These data suggested that a T3SS-effector inhibits the production of a host factor involved in virus inhibition. We then infected HeLa cells with C. rodentium strains lacking individual T3SS effector genes and screened the cell supernatants for anti-viral activity. nleB inhibited virus replication most significantly. By monitoring TRAF3 activity in C. rodentium infected cells, we also revealed the selective impact of NleB on K63 linked TRAF3 ubiquitination. We conclude that the T3SS effector NleB inhibits host IFN-β production by reducing the extent of the activation associated K63 linked TRAF3 ubiquitination.

Biography:

Eva E Avila is a Professor at the Universidad de Guanajuato, Mexico. Her research interest is in innate immunity defense, especially antimicrobial peptides and some virulence mechanisms of the parasites Trichomonas vaginalis and Entamoeba histolytica. She also enjoys teaching in bachelor and postgraduate levels.

Abstract:

Trichomonas vaginalis is a flagellated parasite that causes human trichomoniasis; this is the non-viral most common sexually transmitted disease worldwide. Trichomoniasis is associated with the premature birth of newborns, with infertility and with increased susceptibility to human immunodeficiency virus and papillomavirus infections. This infection is characterized by a heavy inflammatory response with abundant number of neutrophils. Neutrophils, the most abundant cells in the bloodstream, have a main function to eliminate the pathogenic microorganisms through phagocytosis, degranulation and the formation of neutrophil extracellular traps (NETs). NETs are DNA fibers associated with histones and antimicrobial peptides that trap and prevent the spread of pathogens. The purpose of this research was to characterize the interaction between Trichomonas vaginalis and human neutrophils in vitro. The formation of NETs was activated by trophozoites and by its surface lipophosphoglycan (LPG), which was reduced in the presence of an antibody to TLR-4, suggesting the participation of this receptor in NETs formation induced by T. vaginalis. NETs trapped trophozoites, as observed by confocal microscopy and after a 3-hour interaction, the viability of T. vaginalis decreased significantly. These results suggest that neutrophil extracellular traps are effective against T. vaginalis; however, the presence of excessive number of neutrophils during infection may also contribute to the damage of epithelial mucosa.

Biography:

Cecile Oury is a PhD in Biology and has studied Platelet Biology for about 15 years. In 2001, she was the Laureate of the Prize Boerhinger Ingelheim for her research on Thrombosis and Haemostasis. She is currently the Senior Research Associate at the Belgium Fund for Scientific Research and she is the Head of the Laboratory of Thrombosis and Haemostasis, part of the Unit GIGA-Cardiovascular Sciences at the University of Liege, Belgium. Her research focuses on the elucidation of the mechanisms of thrombosis using in vitro studies and animal models. She has h-index of 19, more than 55 international peer-reviewed papers and 1595 citations. She is a Council Member of the Belgian Society on Thrombosis and Haemostasis and also a Member of the International Society on Thrombosis and Haemostasis.

Abstract:

Short nuclease-resistant phosphorothioate synthetic CpG motif-bearing oligonucleotides (CpG ODNs) mimicking bacterial DNA display potent immunostimulatory activity. Pre-clinical studies and human clinical trials have demonstrated that CpG ODNs significantly improve vaccine-specific antibody responses. CpG ODN recognition by B-cells and plasmacytoid dendritic cells depends on the interaction of CpG ODNs with the intracellular Toll-like receptor 9 (TLR9). Besides immune cells, a recent study indicates that phosphorothioate modified CpG ODNs activate platelets and, therefore, may cause dangerous thrombotic events. In this work, we used pharmacological and genetic approaches on human and mouse platelets to investigate the mechanisms of platelet activation by CpG ODNs in order to identify potential protective strategies. We showed that CpG ODNs bound on platelet surface and were internalized. They caused P-selectin exposure and binding of fibrinogen and fibronectin on platelet surface and induced their aggregation. In vivo, intravenous injection of CpG ODN in mice promoted thrombus growth and fibrin generation at site of endothelial injury. Surprisingly, the use of TLR9-deficient platelets indicated that TLR9 signaling was dispensable for CpG ODN induced platelet activation. We found that CpG ODNs stimulated platelet tyrosine kinase-dependent pathway and Syk phosphorylation, similar to ITAM and αIIbβ3 integrin outside-in signaling. Mouse platelets deficient for the hemITAM C-type lectin-like receptor CLEC-2 were unable to capture and internalize CpG ODN. CLEC-2 deficiency abolished CpG ODN-induced platelet activation and aggregation. Accordingly, CpG ODN stimulated CLEC-2 dimerization in human platelets and CpG ODN-induced platelet aggregation was inhibited by an anti-CLEC-2 blocking antibody. Furthermore, β3-deficient platelets could not be activated by CpG ODNs. In contrast, mouse platelets deficient for the collagen ITAM receptor GPVI responded normally to CpG ODNs. Pre-treating platelets with the ATP/ADP scavenger apyrase or with the P2Y12 receptor antagonist cangrelor fully inhibited CpG ODN uptake and subsequent platelet activation, while the cyclooxygenase inhibitor indomethacin and the αIIbβ3 antagonists tirofiban and eptifibatide conferred partial protection. In agreement with these results, CpG ODN failed to activate platelets from clopidogrel-treated patients or from a patient with Glanzmann thrombasthenia. In conclusion, CpG ODN-induced platelet activation depends on multiple platelet receptors including CLEC-2, P2Y12 and αIIbβ3. Importantly, inhibiting P2Y12 receptors with clinically available antiplatelet drugs may confer full protection against possible adverse pro-thrombotic effects of CpG ODN vaccine adjuvants.

Biography:

TLRs), RIG-I-like receptors (RLRs) and cytoplasmic DNA receptors (CDRs). While esse

Abstract:

Key to the activation of the innate immune system is the pattern-recognition receptors (PRRs) including Toll-like receptors (TLRs), RIG-I-like receptors (RLRs) and cytoplasmic DNA receptors (CDRs). While essential for protection against infections, activation of PRRs requires tight control to avert inflammatory diseases. The mechanisms underlying this strict regulation are unclear. The reversible attachment to and removal of ubiquitins from proteins by ubiquitin ligases and deubiquitinases respectively is a versatile system that regulates diverse aspects of biology including the immune system. The H2A deubiquitinase MYSM1 (H2A-DUB) MYSM1 is a nuclear metalloprotease previously described as a key component of epigenetic signaling machinery. We now show that MYSM1 is a master negative regulator of PRR pathways. In response to infections or inflammation, we found that MYSM1 rapidly accumulates in the cytoplasm. In the cytoplasm MYSM1 interacts with and inactivates key signaling complexes for the TLR, RLR and CDR pathways via the removal of K63 linked polyubiquitin chains. Hence mice deficient in MYSM1 are hyper-responsive to various innate immune stimuli, exhibit resistance to viral infections but are more susceptible to inflammatory disease such as sepsis. These results highlight MYSM1 as a key negative regulator of the innate immune system that protects against an overzealous self-destructive immune response.

Biography:

Prof. Mira Barda-Saad, a returning scientist from the National Cancer Institute at NIH in Maryland, is a Senior Lecturer at the Mina and Everard Goodman Faculty of Life Sciences. Barda-Saad is currently examining the molecular signaling mechanisms controlling immune cell response with the primary goal of relating this knowledge to pathophysiological conditions of the immune system. She believes that understanding the dynamic behavior of signaling and cytoskeletal molecules that control immune cell activation is essential for identification of targets relevant for the treatment of cancer, autoimmune diseases and immunodeficiencies

Abstract:

Natural killer (NK) cells represent a powerful weapon of immune defense against viral infections and tumor growth via the cytotoxicity of target cells and the production of cytokines. NK cell function is regulated by a balance between activating and inhibitory signals. Cancer cells or viruses often perturb this balance by expressing ligands for activating NK cell receptors and by down-regulating ligands for the inhibitory receptors, i.e., MHC class I molecules, resulting in target cell killing. Engagement of inhibitory receptors, including the killer cell immunoglobulin-like receptor (KIR), antagonizes activating pathways through the recruitment and activation of the SH2-containing protein tyrosine phosphatase-1 (SHP-1) to the NK immunological synapse (NKIS). To date, only the signaling molecule VAV1 was clearly demonstrated as a direct substrate of SHP-1 in human NK cells. Since SHP-1 activity is the major mechanism that prevents NK cell autoimmune response, it is of great importance to determine whether additional substrates of SHP-1 exist and whether additional molecular mechanisms down-regulate NK cell activation. Moreover, the mechanisms that control SHP-1 activity remain to be unraveled. In the present study, we demonstrate that in response to KIR receptor engagement, SHP-1 and the E3 ubiquitin ligases Cbls negatively regulate the linker for the activation of T cells (LAT) and phospholipase Cγ (PLCγ) 1/2. LAT dephosphorylation by SHP-1 abrogated PLCγ 1/2 recruitment to NKIS and decreased calcium flux and degranulation, thus abolishing NK cell cytotoxicity. Furthermore, LAT ubiquitylation via c-Cbl and Cbl-b following NK cell inhibition leads to its degradation and to the down-regulation of NK cell activation. Using a cutting-edge microscope system, we follow this cellular signaling cascade from the moment of encounter through target-cell killing. Our data suggest that LAT phosphorylation triggers its ubiquitylation, implying a collateral inhibitory mechanism in which a pool of phosphorylated LAT that escapes SHP-1 dephosphorylation is targeted to proteasomal degradation. These mechanisms serve as a key checkpoint in tuning NK cell activation threshold and the immune response.

Biography:

Ludovica F Butto has completed her Master’s degree from the University of Verona, Italy, investigating the production and oral delivery of vaccines for diabetes type-I in plant. She won a fellowship position funded by the European Food Safety Authority (EFSA) at the Centre of Competence for the innovation in the agro-environmental field, Agroinnova (Turin, Italy). She carried out her PhD at the Alimentary Pharmabiotic Centre (University College Cork, Cork, Ireland) focused on TLR signaling in bacteria-host immune cells interaction. She is currently a Postdoctoral Researcher funded by Helmsley Trust (NY) at the Chair of Nutrition and Immunology, Technische Universität Munchen (Weihenstephan, Germany), investigating the impact of the human intestinal microbiota from highly refractory Crohn’s disease patients who underwent autologous hematopoietic stem cell transplantation inducing drug-free remission, in germfree mouse models of Crohn’s disease and ulcerative colitis.

Abstract:

Recognition of microbes by toll-like receptors (TLRs) is critical for initiation of appropriate innate and adaptive immune responses. How crosstalk between TLRs situated in various cellular locations contributes to host-microbe dialogue and immunoregulation is presently unclear. Here we report a dual role for TLR2 in regulating the response of macrophages to internalized bacteria. Cell surface TLR2 initiated an inflammatory response while sub cellular TLR2 negatively regulated a TLR4-mediated hyper-inflammatory response that was phagocytosis dependent and driven by a type-I interferon autocrine loop. TLR2 deficient macrophages were hyper responsive to commensal and pathogenic bacteria and developed an M1 like phenotype, which was absent in TLR2/TLR4 deficient macrophages. Our findings identify sub cellular TLR2 as being important for the negative regulation of inappropriate TLR4 mediated inflammatory responses to bacteria.

Biography:

peutic option for a subset of asthmatics who continue to be symptomatic despite high dose glucocorticoid

Abstract:

Rationale: Bronchial thermoplasty (BT) is a therapeutic option for a subset of asthmatics who continue to be symptomatic despite high dose glucocorticoid therapy. Mechanisms underlying the impact of BT on airway inflammation are largely unknown. Myeloid-derived regulatory cells (MDRCs) are important regulators of chronic inflammation in human asthma. There is also increasing appreciation for a relationship between airway microbiome and respiratory inflammation. We hypothesized that the longitudinal changes in diversity and or abundance of the airway microbiome may modulate immune regulation by MDRCs following BT and contribute to the beneficial outcome of BT. Methods: Bronchial washings (BW) and peripheral blood samples were collected at each of the three bronchoscopic procedures for BT from 5 patients with severe asthma. Multi-parameter flow cytometry was performed to enumerate MDRC subsets and regulatory T-cells (Treg). Microbial genomic DNA was isolated, 16S rDNA genes were amplified using V4 primers and PCR products were sequenced using the Illumina MiSeq platform. Sequences were processed, integrated, analyzed and reported using QIIME and in-house software. Statistical significance of longitudinal changes in microbial signatures and cell proportions were determined by linear mixed model regression analysis. Correlation between microbiome composition and time points was determined by PERMANOVA. Microbial phyla and cell MDRC subsets were correlated by linear mixed model regressing one variable to the other adjusting time points as a fixed effect and sample ID as a random effect. All analyses were conducted in the lme4 R package. Results: Significant longitudinal changes were noted at the level of phyla in comparisons of operational taxonomic unit and time points of bronchoscopies. Significant reduction in both proteobacteria (P=0.04) and verrucomicrobia (P=0.020) was observed in the BW. Correlation analyses revealed significant correlations between microbial alterations and proportions of MDRC subsets in the BW following BT. Alterations in phyla tenericutes and verrucomicrobia significantly correlated (P=0.039,P=0.0131) with the reduction in proportions of immunosuppressive CD14+CD16-HLA-DR- MDRCs in BW. Changes in phyla proteobacteria and fusobacteria correlated (P=0.002, P=0.017) with the enhancement of CD14+CD16+HLA-DR- MDRCs in the BW. Importantly, changes in kingdom Archaea; phylum euryarchaeota correlated with the progressive reduction of pro-inflammatory CD163+HLA-DR+CD11b+CD33+MDRCs (P=0.00003) and enhancement of the CD4+CD25+CD127loFoxp3+Tregs (P=0.0135) and CD14+CD16+HLA-DR- MDRCs (P=0.044) in the BW. Conclusions: Correlation of progressive modulation of airway microbiome and myeloid cell dynamics suggest a functional relationship/interaction between these components and an important contribution to immune regulation in the airways that can account for some of the benefits observed in patients following BT.

Biography:

Abstract:

Rationale: Myeloid-derived suppressor cells (MDSCs) have been well established as regulators of anti-tumor immunity. MDSCs modulate amino acid metabolism in the tumor microenvironment and suppress T-cell function. However, it is less clear whether MDSCs regulate B-cell responses during tumor progression. Methods: Using a syngeneic orthotopic model for lung cancer with murine Lewis Lung Carcinoma cells, we evaluated B-cell subsets in tumor bearing mice by multi parameter flow cytometry. The amount of serum IgG or IL-7 was determined by ELSIA. Phospho-STAT5 and total STAT5 were detected by immunoblotting. To investigate MDSC-mediated suppression of B cell lymphopoiesis, we adoptively transferred MDSCs derived from bone marrow of CD45.2+ tumor bearing mice intratibially into congenic CD45.1+ mice. B-cell subsets in recipient mice at day 7 post MDSC transfer were enumerated as above. In vitro B-cell inhibitory assay was performed by co-culturing CFSE-labeled pre-activated splenocytes with MDSCs purified from bone marrow of tumor-bearing mice at a ratio of 1:1 in the absence or presence of arginase inhibitor nor-NOHA (20 M), iNOS inhibitor 1400W (500 nM) or IDO inhibitor 1-D-MT (1 mM) for 48 hours. The percentage of CD19+CFSElow cells (proliferating cells) was determined by FACS analysis. Results: Percentages and absolute numbers of Pro-, Pre- and mature B-cells were reduced in bone marrow (BM) of tumor bearing mice. Moreover, percentage and absolute number of follicular B cells were reduced, while immature B-cells increased in the spleen of tumor bearing mice. Levels of serum IgG were reduced in tumor-bearing mice. Furthermore, IL-7 and downstream STAT-5 signaling were impaired in tumor bearing mice. Transfer of BM-derived MDSCs from tumor bearing mice into congenic recipients resulted in significant reduction in both percentages and absolute numbers of immature and mature B-cells in peripheral blood of recipient mice. Pre-B cells and immature B-cells also decreased in BM of MDSC transferred recipients. Additionally, MDSCs suppress B-cell proliferation and IgG production by B-cells in an arginase and iNOS dependent but IDO independent manner. Conclusions: In the present study, we demonstrate that B-cell differentiation in vivo is impaired in the BM and spleen of mice with lung cancer. Adoptive transfer studies with congenic mice demonstrate that MDSCs derived from Lewis Lung Carcinoma bearing mice may suppress B-cell differentiation in tumor naive mice. These results together suggest that tumor-related MDSCs may potentially regulate humoral immune responses to promote tumor survival.

Biography:

Oziniel Ruzvidzo is a full Professor of Plant Biotechnology at the North-West University, South Africa. He has completed his Doctoral degree in Plant Biotechnology in 2009 at the University of the Western Cape, South Africa. He also undertook some Postdoctoral studies in the same subject area in 2010 at the University of Cape Town in South Africa. Currently, he is the Head of Department and Principal Investigator of Plant Biotechnology in the Department of Biological Sciences at the North-West University, Mafikeng Campus, South Africa, where he has already extensively published high impact factor papers in this subject area of his research specialization. He is an active Member of several university committees including the senate as well as an Editorial Member of various academic and research boards locally, regionally and internationally.

Abstract:

The need to develop agricultural crops with improved stress resistance has led plant biotechnologists to strategically focus on those plant molecules involved in the maintenance of homeostasis. One such molecule that is typically involved in signal transduction is the cyclic adenosine 3′,5′-monophosphate (cAMP), which is generated by adenylate cyclases (AC). Even though ACs have previously been experimentally proven to be centrally involved in numerous stress response systems in animals, prokaryotes and lower eukaryotes, their existence and or functional properties in higher plants have until recently been a very serious matter of debate and controversy. To date, only four ACs have since been confirmed in higher plants, specifically in the Arabidopsis thaliana, the Zea mays, the Nicotiana benthamiana and the Hippeastrum hybridum plants. Since it is inconceivable that a single AC per plant can essentially account for all the known and or currently reported cAMP-dependent processes in plants, we then set out to practically identify and functionally characterize a second probable AC candidate from A. thaliana in the form of a putative clathrin assembly protein (AtCAP: At1g68110), with a view of elucidating its exact physiological and biological roles in higher plants. Firstly, we conducted a preliminary bioinformatic analysis of this protein, which then showed us that this putative protein candidate is actually a multi-domain, multi-functional molecule with a possible role in AC-dependent stress response and adaptation mechanisms. We then proceeded to clone and recombinantly express its AC-containing fragment domain (AtCAP-AC) in chemically competent E. cloni BL21 (DE3) pLysS DUOs cells and then showed its ability to induce the generation of endogenous cAMP in this prokaryotic expression system. Furthermore, we also demonstrated the complementation of a mutant non-lactose fermenting cyaA SP850 Escherichia coli strain by this recombinant AtCAP-AC protein, to eventually ferment lactose as a result of this protein’s ability to generate the most required cAMP for this process. In addition, we also undertook the purification process of this truncated AtCAP-AC protein followed by the functional characterization of its in vitro AC activities, which in turn revealed that this recombinant is indeed a bona fide soluble adenylate cyclase (sAC) physiological and biochemical roles may be mediated by cAMP and possibly, via a calmodulin-dependent signaling system. Lastly and in order to gain more insights into the possible physiological roles of this AtCAP-AC protein in higher plants, we then bioinformatically analyzed its expressional profiles using the various computational and web-based bioinformatic tools and found out that the protein is centrally involved in responses to biotic stress factors, whose systems are both cAMP and SORLIP1AT core motif-dependent.

Biography:

Francesco Peri is a Professor of Organic and Medicinal Chemistry at the University of Milano Bicocca. He also has a permanent Professorship in Medicinal Chemistry at the Ecole Normale Superieure (ENS) in Lyon, France. He has published more than 87 papers in reputed journals, H-index: 24; H-index from 2010: 20.

Abstract:

Toll like Receptors (TLRs) activation by pathogen associated molecular patterns (PAMPs) is a pivotal molecular event in inflammation and innate immunity and TLRs and their agonists are responsible for the efficacy of almost every vaccine. Conversely, TLRs hyper activation by endogenous factors such as oxidized phopsholipids or heat shock proteins is the main cause of many inflammatory and autoimmune diseases. Activating or inhibiting specifically TLRs provides access to a new generation of therapeutics. We developed synthetic molecules able to modulate TLR4 activation and signaling and we studied the mechanism of action (MOA) of these non toxic and drug like compounds. Positively or negatively charged synthetic glycolipids are active in blocking TLR4 activation by specifically targeting the CD14 co receptor. These molecules are very efficient in inhibiting TLR4 activation in cells and in contrasting diseases related to TLR4 hyper activation by infectious and endogenous agents in animal models. We investigated at a molecular level the MOA of these molecules by binding experiments with purified CD14, MD-2 and TLR4 receptors and experiments on dendritic cells, macrophages and HEK-TLR4 cells. The unique MOA of these molecules is based on the capacity to dissociate CD14 and TLR4 endocytosis, thus creating an inducible CD14 deficiency at the cell surface. These conditions are expected to antagonize TLR4 signaling more effectively than simply competing with LPS for CD14 and TLR4. Very promising results have been obtained at a preclinical level using these drug hits and recent (2015) data on animal models of atherosclerosis, neuroinflammation and amyotrophic lateral sclerosis will be reported.

Biography:

Denis Soulet has completed his PhD in Neuroimmunology from Laval University, Canada and Postdoctoral studies from Universitet , Sweden. He is an Associate Professor at the Medicine Faculty of Laval University, Canada. He has published more than 40 papers in reputed journals and has been serving as an Editorial Board Member of SM Journal of Gastroenterology and Hepathology. He is leading a research team dedicated to study the role of peripheral inflammation in the enteric nervous system and its contribution to Parkinson’s disease. The ultimate goal of his research program is to design immunomodulatory based disease modifying drugs for PD.

Abstract:

Parkinson’s disease (PD) is a neurological disorder characterized by motor symptoms which are often preceded by non-motor symptoms, including gastrointestinal dysfunctions. Common treatments are only symptomatic; there is still no disease modifying drug available to cure patients. Since numerous pro-inflammatory markers have been measured in the central and peripheral nervous system, this deleterious immune response seems to be a potential target to develop new therapeutic strategies. Therefore, a better understanding of the role of the immune response in the etiology and progression of PD is essential. During my talk, I will present original data about the impact of the innate immune response on enteric neuronal damage in PD models. At first, we characterized the immune response induced by the neurotoxin MPTP in the enteric nervous system of partially immunodeficient mice. We demonstrated the timeline of inflammatory events occurring prior to the neuronal demise and the critical role of monocytes and macrophages in the gut. Thereafter, we tested various estrogenic compounds for their immunomodulatory and neuroprotective properties in PD models both in vivo and in vitro, delineating the major contribution of various estrogenic receptors, mainly the G Protein-coupled Estrogen Receptor 1 (GPER1). More recently, we successfully explored the therapeutic potential of a clinically approved selective estrogen receptor modulator, Raloxifene, for drug repurposing in PD. In conclusion, our data highlight the critical role of the immune response at early stages of PD and the immunomodulatory and neuroprotective potential of estrogen-based hormonotherapy at the pre-clinical level.

Biography:

Philip R Hardwidge is an Associate Professor at Kansas State University. His laboratory is interested in understanding, treating and preventing diarrheal disease caused by bacterial pathogens. His research team has discovered several mechanisms by which bacterial proteins subvert the host innate immune system to promote bacterial colonization and transmission. He is directing his knowledge of these proteins and their mammalian targets to innovative studies of metabolic syndromes, autoimmune disorders and cancer. He is also developing proteomic techniques to identify vaccine targets in other organisms.

Abstract:

Many bacterial pathogens utilize a type-III secretion system (T3SS) to inject virulence proteins (effectors) into host cells to subvert various biological functions. Effector subversion of pro-inflammatory host responses is well studied, but less attention has been given to the potential inhibition of host interferon (IFN) signaling. Type-I IFNs are important both to maintaining intestinal homeostasis and to responding to pathogen infection. Pathogens have evolved strategies to interfere with host type-I IFN production. A recent study found both that IFN-β is induced by enteropathogenic E. coli (EPEC) infection and that the EPEC T3SS effector NleD inhibits IFN-β induction. IFN expression is known to be important to limiting Citrobacter rodentium infection but whether C. rodentium T3SS effectors inhibit host IFN-β induction is unclear. We screened C. rodentium strains bearing deletions in individual T3SS effectors to determine the extent to which this pathogen might inhibit the host IFN-β response. To determine if C. rodentium T3SS effectors inhibit the host type-I IFN response, we monitored the survival of a recombinant vesicular stomatitis virus (VSV). Since TRAF3 is critical to IFN signaling, we also monitored effector mediated inhibition of the TNF receptor (TNFR) associated factor 3 (TRAF3) ubiquitination in RAW264.7 cells. Supernatants from cells infected with C. rodentium escN inhibited VSV to levels similar to those induced by LPS treatment. By contrast, supernatants from cells infected with WT C. rodentium did not inhibit VSV-GFP growth. These data suggested that a T3SS-effector inhibits the production of a host factor involved in virus inhibition. We then infected HeLa cells with C. rodentium strains lacking individual T3SS effector genes and screened the cell supernatants for anti-viral activity. nleB inhibited virus replication most significantly. By monitoring TRAF3 activity in C. rodentium infected cells, we also revealed the selective impact of NleB on K63 linked TRAF3 ubiquitination. We conclude that the T3SS effector NleB inhibits host IFN-β production by reducing the extent of the activation associated K63 linked TRAF3 ubiquitination.

Biography:

Bozena Futoma-Koloch was graduated from University of Wrocław in 2004. She has received her PhD in Microbiology from the University of Wrocław in 2008. After that, she has held an academic position of Assistant Professor in the Department of Microbiology in the Institute of Genetics and Microbiology at University of Wroclaw. She is a Member of Polish Society of Microbiologists, European Society of Clinical Microbiology and Infectious Diseases and International Complement Society. She has published more than 50 papers in national and international journals, 30 presentations at congresses with several awards. Her research program focuses on bacterial surface antigens as molecular targets of the protective immune response and their role in bacterial resistance to disinfectants.

Abstract:

The mechanisms used by bacteria to avoid host immunological defenses are not entirely understood. Microorganisms getting into contact with human blood or plasma have developed a variety of strategies to evade complement attack. One strategy is the incorporation of the sialic acid into the bacterial surface glycoconjugates that usually results in an increase in serum resistance. Nothing is known about the influence of sialylated bacterial surface structures on C3 fixation in serum. The role of the outer membrane proteins (OMP) in Salmonella susceptibility to serum has not also been investigated thoroughly. Therefore, C3 deposition on the O48 group of Salmonella bacteria has been studied. The tested microorganisms Salmonella O48 are characterized by sialylated lipopolysaccharide (LPS) and different patterns of OMP. Our investigations showed that bacteria were sensitive to human serum (HS) although they possessed sialylated LPS. We found that the greatest C3 deposition occurred on Salmonella sv. Isaszeg cells with low content of sialic acid in LPS. A weaker C3 deposition ratio was noted to Salmonella vs. Ngozi and Salmonella subsp. arizonae with the high contents of sialic acid in LPS. Immunoblotting revealed that C3 complement protein bound to OMP common to three tested strains. We suggest that the differential sensitivity of tested bacteria to HS may be due to a weaker C3 activation on strongly sialylated LPS and a binding of C3 components to the OMP.

Biography:

Jessy S Deshane is a pulmonary Immunologist with expertise in immune regulation in asthma. She investigates myeloid-derived regulatory cell biology and free radical mechanisms that regulate their differentiation and function. She pioneered these investigations both in mouse models and human asthma. She has authored 46 peer-reviewed publications, including high impact journals like Journal of Experimental Medicine, Journal of Clinical Investigations, Journal of Allergy and Clinical Immunology, Immunity and Cancer Research. She serves on the Editorial Boards for the journals Allergy and American Journal of Respiratory Cell and Molecular Biology and serves on grant review committees.

Abstract:

Rationale: Bronchial thermoplasty (BT) is a therapeutic option for a subset of asthmatics who continue to be symptomatic despite high dose glucocorticoid therapy. Mechanisms underlying the impact of BT on airway inflammation are largely unknown. Myeloid-derived regulatory cells (MDRCs) are important regulators of chronic inflammation in human asthma. There is also increasing appreciation for a relationship between airway microbiome and respiratory inflammation. We hypothesized that the longitudinal changes in diversity and or abundance of the airway microbiome may modulate immune regulation by MDRCs following BT and contribute to the beneficial outcome of BT. Methods: Bronchial washings (BW) and peripheral blood samples were collected at each of the three bronchoscopic procedures for BT from 5 patients with severe asthma. Multi-parameter flow cytometry was performed to enumerate MDRC subsets and regulatory T-cells (Treg). Microbial genomic DNA was isolated, 16S rDNA genes were amplified using V4 primers and PCR products were sequenced using the Illumina MiSeq platform. Sequences were processed, integrated, analyzed and reported using QIIME and in-house software. Statistical significance of longitudinal changes in microbial signatures and cell proportions were determined by linear mixed model regression analysis. Correlation between microbiome composition and time points was determined by PERMANOVA. Microbial phyla and cell MDRC subsets were correlated by linear mixed model regressing one variable to the other adjusting time points as a fixed effect and sample ID as a random effect. All analyses were conducted in the lme4 R package. Results: Significant longitudinal changes were noted at the level of phyla in comparisons of operational taxonomic unit and time points of bronchoscopies. Significant reduction in both proteobacteria (P=0.04) and verrucomicrobia (P=0.020) was observed in the BW. Correlation analyses revealed significant correlations between microbial alterations and proportions of MDRC subsets in the BW following BT. Alterations in phyla tenericutes and verrucomicrobia significantly correlated (P=0.039,P=0.0131) with the reduction in proportions of immunosuppressive CD14+CD16-HLA-DR- MDRCs in BW. Changes in phyla proteobacteria and fusobacteria correlated (P=0.002, P=0.017) with the enhancement of CD14+CD16+HLA-DR- MDRCs in the BW. Importantly, changes in kingdom Archaea; phylum euryarchaeota correlated with the progressive reduction of pro-inflammatory CD163+HLA-DR+CD11b+CD33+MDRCs (P=0.00003) and enhancement of the CD4+CD25+CD127loFoxp3+Tregs (P=0.0135) and CD14+CD16+HLA-DR- MDRCs (P=0.044) in the BW. Conclusions: Correlation of progressive modulation of airway microbiome and myeloid cell dynamics suggest a functional relationship/interaction between these components and an important contribution to immune regulation in the airways that can account for some of the benefits observed in patients following BT.

Biography:

Hanan Al-Khalifa has obtained her Master’s degree in Parasitological Diseases and Immunology at University of Manchester and completed her PhD in 2007 in the University of Reading, UK, investigating the effect of n-3 fatty acids on the immune response and general health status. Her interests include but are not limited to immunological techniques, parasitological diseases, effect of nutrition, espicially fatty acids, on the immune status in both humans and expermental animals. She executed many research projects that focused on the effect of nutrition on immunology. She has attended many scientific events and published more than 60 papers in refereed journals and conference proceedings.

Abstract:

Abnormal numbers of specific types of leukocytes may indicate immuno-suppression or immunocompetence in response to an immunomodulator. The objective of this study is to investigate the effect of feeding broilers on diet containing flaxseed on splenocyte T and B-cells. Upon hatching, all chicks were given the same basal diet for 13 days. Dietary supplementation of flaxseed started at 14 days of ages until the end of the cycle at 35 days of age. At slaughter, samples of spleen were collected. Spleen cells were harvested in cell suspensions. Immune cells were then enumerated using the flow haemocytometer in the Physiology Laboratory. The overall differences between the dietary treatments were analyzed using one-way analysis of variance (ANOVA) and the general linear model procedure of Minitab. Statistically, there was no significant effect of flaxseed on the percentage positive or mean fluorescence intensity of the leukocyte subsets under investigation. However, there was a trend towards an increase in the proportion of B-cells in the spleen after feeding 15% of flaxseed, which approached significance (P=0.058). There was also a trend towards a decrease in the mean fluorescence intensity of CD8+ subsets in the spleen, which was close to statistical significance (P=0.054). This trend is particularly interesting, given the fact that the bursa in these chickens were significantly observed to be smaller and thinner. It suggests that flaxseed may either prevent the homing of B-lymphocytes to the bursa or encourage the release of B-lymphocytes from the bursa into the circulation and then in the spleen.

Biography:

Laboratory for Molecular Infection Medicine Sweden (MIMS), Sweden

Abstract:

Key to the activation of the innate immune system are the pattern-recognition receptors (PRRs) including Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), and cytoplasmic DNA receptors (CDRs). While essential for protection against infections, activation of PRRs require tight control to avert inflammatory diseases. The mechanisms underlying this strict regulation are unclear. The reversible attachment to and removal of ubiquitins from proteins by ubiquitin ligases and deubiquitinases respectively is a versatile system that regulates diverse aspects of biology including the immune system. The H2A deubiquitinase MYSM1 (H2A-DUB) MYSM1 is a nuclear metalloprotease previously described as a key component of epigenetic signaling machinery. We now show that MYSM1 is a master negative regulator of PRR pathways. In response to infections or inflammation, we found that MYSM1 rapidly accumulates in the cytoplasm. In the cytoplasm MYSM1 interacts with and inactivates key signaling complexes for the TLR, RLR and CDR pathways via the removal of K63 linked polyubiquitin chains. Hence mice deficient in MYSM1 are hyper-responsive to various innate immune stimuli, exhibit resistance to viral infections but are more susceptible to inflammatory disease such as sepsis. These results highlight MYSM1 as a key negative regulator of the innate immune system that protects against an overzealous self-destructive immune response.

Biography:

Un-Hwan Ha has completed his PhD in Microbiology from the University of Florida in 2002 and Postdoctoral studies in Cellular Microbiology from House Ear Institute and University of Rochester Medical Center. In 2008, he has begun to serve as an Assistant Professor at the Department of Biotechnology and Bioinformatics, Korea University and is currently positioned as a Professor. He has published more than 20 papers in reputed journals since 2008.

Abstract:

Pseudomonas aeruginosa is a Gram negative opportunistic bacterial pathogen that has a notorious reputation about drug resistance against commonly used antibiotics as well as an infectious agent to the respiratory tract of immunocompromised patients along with other microbial invaders. P. aeruginosa possesses diverse secretory systems, which play critical parts in releasing a number of virulence factors that are involved in causing acute and chronic infections. The potential effects of these factors on the modulation of host defense responses have been proposed. However, the resulting modulation effects against competitive bacteria, such as Staphylococcus aureus, are unknown since the clinical impact of polymicrobial diseases caused by combinations of pathogens has received much attention from the medical community. Here, we report that components secreted from P. aeruginosa enhance the expression of bradykinin receptors which act as important host defense responses against invading microbes by interacting with a ligand, bradykinin. In addition to this, LPS as a well-known membrane associated molecule pattern of P. aeruginosa induces the expression of TLR2, which plays a dominant role in sensing PAMPs typically expressed by Gram positive bacteria. Up-regulation of TLR2 influences the magnitude of proinflammatory responses to the secondary S. aureus infection. Moreover, P. aeruginosa Ndk with the aid of flagellin, increases the expression of interleukin-1 which is an important pro-inflammatory cytokine via NF-κB/inflammasome pathways. Taken together, the results of this study demonstrate that P. aeruginosa is capable of modulating host defense responses through the actions of associated or released virulence factors and this may have impacts on against a secondary microbial infection.

Biography:

Eva E Avila is a Professor at the Universidad de Guanajuato, Mexico. Her research interest is in innate immunity defense, especially antimicrobial peptides and some virulence mechanisms of the parasites Trichomonas vaginalis and Entamoeba histolytica. She also enjoys teaching in bachelor and postgraduate levels.

Abstract:

Trichomonas vaginalis is a flagellated parasite that causes human trichomoniasis; this is the non-viral most common sexually transmitted disease worldwide. Trichomoniasis is associated with the premature birth of newborns, with infertility and with increased susceptibility to human immunodeficiency virus and papillomavirus infections. This infection is characterized by a heavy inflammatory response with abundant number of neutrophils. Neutrophils, the most abundant cells in the bloodstream, have a main function to eliminate the pathogenic microorganisms through phagocytosis, degranulation and the formation of neutrophil extracellular traps (NETs). NETs are DNA fibers associated with histones and antimicrobial peptides that trap and prevent the spread of pathogens. The purpose of this research was to characterize the interaction between Trichomonas vaginalis and human neutrophils in vitro. The formation of NETs was activated by trophozoites and by its surface lipophosphoglycan (LPG), which was reduced in the presence of an antibody to TLR-4, suggesting the participation of this receptor in NETs formation induced by T. vaginalis. NETs trapped trophozoites, as observed by confocal microscopy and after a 3-hour interaction, the viability of T. vaginalis decreased significantly. These results suggest that neutrophil extracellular traps are effective against T. vaginalis; however, the presence of excessive number of neutrophils during infection may also contribute to the damage of epithelial mucosa.

Biography:

Roger Huybrechts has obtained his PhD at KU Leuven in 2015. His research focuses Locust cellular innate immunity.

Abstract:

Within the order Insecta, knowledge about the innate immune response is mainly based on studies regarding holometabolous model organisms: In both fruit fly and moths the cellular immune response of phagocytosis, nodule formation and encapsulation is supported by a strong humoral pro-inflammatory response including coagulation, prophenoloxidase activation and antimicrobial peptide synthesis. In hemimetabolous locusts, genes encoding the classical antimicrobial peptide precursors are missing and they mainly depend upon their efficient cellular immune response. This elaborated cellular immune response has its trade off as it results in a steep decrease in the number of circulating hemocytes. Immune challenge makes a GBP hemocyte spreading peptide activating the hemocytes. Most tissues except gut and hemocytes display a high basal transcription of this cytokine. Spreading and increased adhesive character of activated hemocytes partly explains their rapid disappearance and reappearance following infection. It turned out that in adult locusts the persisting “hematopoietic organ” is not involved in replenishment of lost hemocytes but rather has a prophylactic function as main phagocytotic organ at later age. The instant hemocyte replenishment by circulating prohemocyte stem cells supports this hypothesis, an immune challenge by either Gram positive or Gram negative bacteria but not by a challenge with fungi results in a significant selective increase in expression of insect angiotensin converting enzyme in Locusta migratoria hemocytes. Knockdown of Locusta ACE by both RNAi and captopril inhibitor elucidated the involvement of ACE, either direct or indirect, in the appearance of LPS induced hemolymph peptides of which most have a so far unidentified function.

Biography:

Tzung-Yan Lee has completed his PhD degree from National Yang-Ming University and Postdoctoral studies from Institute of Biological Chemistry, Academia Sinica. He has published more than 50 papers in reputed journals and has been serving as an Editorial Board Member of repute.

Abstract:

An important initiator of the inflammatory response to obesity is adipose tissue, which is involved in obesity induced insulin resistance and chronic inflammation. Electroacupuncture (EA) shows anti-inflammation and several pleiotropic effects that interact with metabolic pathways. Numerous studies have demonstrated the clinical efficacy of acupuncture in weight loss. However, the precise mechanism of its potential effect related to adipose tissue remains poorly understood. Obese animals treated with EA showed significantly reduced body weight. EA decreased the number of F4/80 and CD11b positive macrophages in epididymal adipose tissue. We found that EA at Zusanli (ST36) acupoints significantly alleviated macrophage recruitment and then improved the obesity associated factors of sterol regulatory element binding protein (SREBP)-1 and target genes expression in obese animals. Adipose tissue tumor necrosis factor-α (TNF-α), interleukin-1 (IL-6), monocyte chemotactic protein-1 (MCP-1) and CD68 mRNA expression were significantly reduced by EA treatment in obese animals. On the other hand, EA significantly down-regulated HIF-1α level in a time course dependent manner in ob/ob mice. The expression level of hypoxia related genes (VEGFA, Slc2al, GPX1) and inflammation related genes (TNF-α, IL-6, MCP-1) were also poorly expressed in adipose tissue after EA treatment. This phenomenon was paralleled by the levels of inflammatory cytokines, such as TNF-α, IL-6 and IL-1β in obese mice. We conclude that EA offers a beneficial effect on adipose tissue mass in obese animals, at least partly, via attenuation of lipogenesis signaling, thus resulting in improved inflammatory response. Therefore, EA prevents weight gain through modulation of HIF-1α-dependent pathways and inflammatory response in obese adipose tissues.

Biography:

Mira Barda-Saad is a returning Scientist from the National Cancer Institute at NIH in Maryland, Senior Lecturer at the Mina and Everard Goodman Faculty of Life Sciences. She is currently examining the molecular signaling mechanisms controlling immune cell response with the primary goal of relating this knowledge to pathophysiological conditions of the immune system. She believes that understanding the dynamic behavior of signaling and cytoskeletal molecules that control immune cell activation is essential for identification of targets relevant for the treatment of cancer, autoimmune diseases and immunodeficiencies.

Abstract:

Natural killer (NK) cells represent a powerful weapon of immune defense against viral infections and tumor growth via the cytotoxicity of target cells and the production of cytokines. NK cell function is regulated by a balance between activating and inhibitory signals. Cancer cells or viruses often perturb this balance by expressing ligands for activating NK cell receptors and by down-regulating ligands for the inhibitory receptors, i.e., MHC class I molecules, resulting in target cell killing. Engagement of inhibitory receptors, including the killer cell immunoglobulin-like receptor (KIR), antagonizes activating pathways through the recruitment and activation of the SH2-containing protein tyrosine phosphatase-1 (SHP-1) to the NK immunological synapse (NKIS). To date, only the signaling molecule VAV1 was clearly demonstrated as a direct substrate of SHP-1 in human NK cells. Since SHP-1 activity is the major mechanism that prevents NK cell autoimmune response, it is of great importance to determine whether additional substrates of SHP-1 exist and whether additional molecular mechanisms down-regulate NK cell activation. Moreover, the mechanisms that control SHP-1 activity remain to be unraveled. In the present study, we demonstrate that in response to KIR receptor engagement, SHP-1 and the E3 ubiquitin ligases Cbls negatively regulate the linker for the activation of T cells (LAT) and phospholipase Cγ (PLCγ) 1/2. LAT dephosphorylation by SHP-1 abrogated PLCγ 1/2 recruitment to NKIS and decreased calcium flux and degranulation, thus abolishing NK cell cytotoxicity. Furthermore, LAT ubiquitylation via c-Cbl and Cbl-b following NK cell inhibition leads to its degradation and to the down-regulation of NK cell activation. Using a cutting-edge microscope system, we follow this cellular signaling cascade from the moment of encounter through target-cell killing. Our data suggest that LAT phosphorylation triggers its ubiquitylation, implying a collateral inhibitory mechanism in which a pool of phosphorylated LAT that escapes SHP-1 dephosphorylation is targeted to proteasomal degradation. These mechanisms serve as a key checkpoint in tuning NK cell activation threshold and the immune response.

Biography:

Nayef Jarrous is currently working at “The Hebrew University of Jerusalem, Israel”. His research interest is based on “Human nuclear RNase P ribonucleoprotein in tRNA processing”. He has published many articles in reputed journals.

Abstract:

RNA polymerase III (Pol III) is a key player in innate immunity, as it serves as a sensor of viral and bacterial DNA of infected cells. This sensing asset is based on promoter independent recognition of foreign DNA templates in the cytoplasm and transcription via nonspecific initiation mechanism. The resulting 5’-triphosphate RNA transcripts activate the retinoic acid induced gene I (RIG-I), thus leading to induction of type-I interferon. We have previously shown that the human catalytic ribonucleoprotein RNase P is implicated in formation of proficient initiation complexes of nuclear Pol III on 5S rRNA and tRNA genes. However, it was unknown if this ribonucleoprotein is also implicated in nonspecific initiation of gene transcription by cytoplasmic Pol III. We will present preliminary results that show that the H1 RNA subunit of human RNase P is implicated in promoter independent initiation of transcription of synthetic circular DNA templates (COLIGOs) by cytoplasmic Pol III. The regulatory role of H1 RNA in this transcription system may explain the existence of RNase P like RNA genes in DNA viruses and the possible roles of their transcripts in evading antiviral innate immune responses.

Biography:

Sumru Savas is an Internal Medicine Specialist since 1999, Graduate of European Academy for Medicine of Ageing (2015). She is currently a PhD student in Elderly Health-Gerontology (From 2011) at Ege University Health Sciences Institute. She is an Internist and Lecturer at Geriatrics Section of Internal Medicine Department.

Abstract:

Lipoprotein associated phospholipase A2 (Lp-PLA2) is a reported risk factor for dementia. However, the relationship between Alzheimer’s disease (AD) and Lp-PLA2 is still debatable and to the best of our knowledge, no study has evaluated the associations between levels of Lp-PLA2, proinflammatory cytokines and neopterin in AD. In total, 59 patients with AD and 38 non-demented individuals were included in the case control study. Fasting serum concentrations of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), neopterin and Lp-PLA2 were determined using ELISA. The associations between AD and each of the variables were analyzed by logistic regression. The median Lp-PLA2 levels in AD and controls were similar (P=0.29, not significant). Median serum neopterin and IL-6 levels were significantly higher in patients with AD than in controls (P=0.0001 and P=0.03, respectively). In regression analyses, median neopterin levels, a lower level of education and female gender were significantly associated with AD when compared with controls (OR, 31.44, 95% CI 3.59-275.28, P=0.002; OR, 4.35, 95% CI 1.13-16.61, P=0.032; OR, 7.25, 95% CI 1.88-28.00, P=0.004, respectively). In contrast to previous evidence suggesting its role in dementia and AD, Lp-PLA2 enzyme levels were higher in the controls and no relationship between Lp-PLA2 and either proinflammatory cytokines or neopterin was identified in AD. Elevated neopterin levels may be considered inflammatory markers of AD.

Biography:

Andreia Marques Ribeiro has completed her graduate degree in Biology and Master’s degree in Microbiology from Aveiro University in Portugal. Since 2010, she has been working as a Research Assistant in Flow Cytometry and Immunology groups in Portugal and in Ireland. In 2013, she started her PhD in the DECIDE consortium and completed in 2016 from National University of Ireland, Galway.

Abstract:

Mesenchymal Stromal Cells (MSC) possesses immunomodulatory and anti-inflammatory properties, having several effects on immune cells. For this reason MSC have been proposed as a potential therapeutic modality for osteoarthritis (OA) and rheumatoid arthritis (RA). However, there is a commercial need to have a reliable, rapid, quantifiable assay to assess the potency of allogeneic human MSC. The aims of this assay were to determine by flow cytometry the effects of MSC from bone marrow (BM MSC) and adipose tissue (ASC) on TNF-α and IL-6 production by LPS stimulated monocytes of healthy and patient samples. A number of factors were considered prior to optimizing the assay, including: Brefeldin A concentration, type of anticoagulant (heparin; K2EDTA or citrate), LPS concentration; blood dilution and incubation time. MSC numbers were then titrated and co-cultured with whole blood of healthy donors. All results are expressed as intracellular expression of TNF-α and IL-6, on gated monocytes identified as CD45+CD14+ cells using an Accuri four color flow cytometer. Result show that BM MSC and ASC significantly reduced TNF-α and IL-6 expression by monocytes from healthy donors and in OA and RA patients. Thus, we have established a rapid, reliable and quantifiable screening assay to determinate the effects of MSC on LPS activated monocytes. Such an assay could be used to screen the recipients’ monocytes for inhibition by the MSC preparation that will be injected, thereby contributing to personalized medicine. For this reason this assay is being used in the ADIPOA2 clinical trial.

Biography:

Jessy S Deshane is a pulmonary Immunologist with expertise in immune regulation in asthma. She investigates myeloid-derived regulatory cell biology and free radical mechanisms that regulate their differentiation and function. She pioneered these investigations both in mouse models and human asthma. She has authored 46 peer-reviewed publications, including high impact journals like Journal of Experimental Medicine, Journal of Clinical Investigations, Journal of Allergy and Clinical Immunology, Immunity and Cancer Research. She serves on the Editorial Boards for the journals Allergy and American Journal of Respiratory Cell and Molecular Biology and serves on grant review committees.

Abstract:

Rationale: Myeloid-derived suppressor cells (MDSCs) have been well established as regulators of anti-tumor immunity. MDSCs modulate amino acid metabolism in the tumor microenvironment and suppress T-cell function. However, it is less clear whether MDSCs regulate B-cell responses during tumor progression. Methods: Using a syngeneic orthotopic model for lung cancer with murine Lewis Lung Carcinoma cells, we evaluated B-cell subsets in tumor bearing mice by multi parameter flow cytometry. The amount of serum IgG or IL-7 was determined by ELSIA. Phospho-STAT5 and total STAT5 were detected by immunoblotting. To investigate MDSC-mediated suppression of B cell lymphopoiesis, we adoptively transferred MDSCs derived from bone marrow of CD45.2+ tumor bearing mice intratibially into congenic CD45.1+ mice. B-cell subsets in recipient mice at day 7 post MDSC transfer were enumerated as above. In vitro B-cell inhibitory assay was performed by co-culturing CFSE-labeled pre-activated splenocytes with MDSCs purified from bone marrow of tumor-bearing mice at a ratio of 1:1 in the absence or presence of arginase inhibitor nor-NOHA (20 M), iNOS inhibitor 1400W (500 nM) or IDO inhibitor 1-D-MT (1 mM) for 48 hours. The percentage of CD19+CFSElow cells (proliferating cells) was determined by FACS analysis. Results: Percentages and absolute numbers of Pro-, Pre- and mature B-cells were reduced in bone marrow (BM) of tumor bearing mice. Moreover, percentage and absolute number of follicular B cells were reduced, while immature B-cells increased in the spleen of tumor bearing mice. Levels of serum IgG were reduced in tumor-bearing mice. Furthermore, IL-7 and downstream STAT-5 signaling were impaired in tumor bearing mice. Transfer of BM-derived MDSCs from tumor bearing mice into congenic recipients resulted in significant reduction in both percentages and absolute numbers of immature and mature B-cells in peripheral blood of recipient mice. Pre-B cells and immature B-cells also decreased in BM of MDSC transferred recipients. Additionally, MDSCs suppress B-cell proliferation and IgG production by B-cells in an arginase and iNOS dependent but IDO independent manner. Conclusions: In the present study, we demonstrate that B-cell differentiation in vivo is impaired in the BM and spleen of mice with lung cancer. Adoptive transfer studies with congenic mice demonstrate that MDSCs derived from Lewis Lung Carcinoma bearing mice may suppress B-cell differentiation in tumor naive mice. These results together suggest that tumor-related MDSCs may potentially regulate humoral immune responses to promote tumor survival.

Biography:

Ildiko Molnar has completed her PhD in the special field of Graves’ Ophthalmopathy at the Hungarian Academy of Science. Her work and research connected her to Kenezy County and Teaching Hospital from 1977 to 2008. Her research activities are on field of internal medicine, endocrinology, immunology and allergology. Currently she is the Chief of EndoMed, Immunoendocrinology and Osteoporosis Centre, Private Outpatient Clinic from 2008. She is an expert in laboratory methods (ELISA, blotting, allergy testing) and DXA measurement. She has published more than 53 papers in reputed journals, 16 chapters and 2 books.

Abstract:

Type II 5’-deiodinase enzyme (DII) activity is responsible for T4 conversion to T3 resulting in the majority of intracellular T3 concentration. DII is a membrane-anchored protein characterized by tissue-specificity; highly expressed in thyroid, pituitary, skeletal, eye and cardiac muscles, brain, adipose tissue and bone. Decreased DII activity leads to hypothyroidism in euthyroid sick syndrome. We demonstrated DII expression in thyroid, eye and skeletal muscle tissues by immunohistochemistry using immunized guinea pig and patients sera with Graves’ orbitopathy. Decreased DII activities were measured after adding proinflammatory cytokines and patients sera with hyperthyroid Graves’ orbitopathy and systemic sclerosis. Antibodies to DII inhibited the mitogen-activated protein kinase (MAPK) activation in thyroid tissue. Proinflammatory cytokines (IL-6, TNFα, IFNγ) inhibited thyroid DII activities in dose-dependent manner (Vmax: 4.1×10-3 pmol/mg/min for IL-6; 0.18 pmol/mg/min for TNFα; 0.23 pmol/mg/min for IFNγ). Hyperthyroid patient sera with Graves’ orbitopathy decreased better thyroid DII activities than eye muscle DII ones (3.99±5.79 vs. 7.66±10.49 pmol/mg/min, P<0.05, n=26). Patient sera with systemic sclerosis (SSc, n=19) decreased DII activities compared to those in controls (n=16) (4.99±1.04 vs. 2.88±0.61 pmol/mg/min, P<0.0001). Immunized guinea pig and Graves’ patient sera with anti-DII antibodies resulted in relevant inhibition of MAPK activation. In conclusion, DII protein can be a new autoantigen in thyroid autoimmunity, particularly in Graves’ orbitopathy. DII activity blocking cytokines could be responsible for low FT3 levels causing euthyroid sick syndrome in systemic sclerosis. The difference in tissue-specific DII activities could be implicated in the development of orbitopathy in hyperthyroid Graves’ disease.

Biography:

Rongtuan Lin is an Associate Professor in the Department of Medicine at McGill University and a Project Director of the Molecular Oncology Group at the Lady Davis Institute for Medical Research. He has received his PhD from Concordia University and completed Post-doctoral training at the Lady Davis Institute for Medical Research. He made important contributions in the fields of interferon signaling and innate antiviral immunity. He has a highly successful laboratory research program with 100 scientific publications, which have been cited more than 5,500 times. He was a recipient of a Chercheur-boursier Senior and Junior 2 from Fonds de la Recherche en Sante du Quebec. In 1996 and 1998, he has received the Milstein Young Investigator Award from the International Society for Interferon and Cytokine Research.

Abstract:

Oncolytic viruses (OVs) are novel anticancer agents that infect and effectively kill cancer cells but not normal cells. Although tumor growth is delayed or eliminated in numerous animal models following treatment with OVs, several cancer models remain partially or completely resistant to viral oncolysis. To overcome this resistance, experimental strategies are now combining OVs with different cytotoxic compounds to improve OV efficacy. Our laboratory has previously demonstrated that OV replication can be bolstered by co-administration of other chemical agents such as Triptolide, a natural molecule derived from the medicinal herb. In the current study, we investigated the capacity of sulforaphane (SFN); an anti-cancer compound naturally occurred in cruciferous vegetables with demonstrated potent antioxidant and possible anti-inflammatory actions to enhance vesicular stomatitis virus (VSV) oncolysis in OV-resistant cancer cells. We ultimately demonstrate that the resistant PC3 prostate cancer cell line can be sensitized to VSV by addition of SFN. Indeed, SFN dose-dependently enhances the replication of VSV. Neither VSV (MOI 0.1) nor SFN (20 uM) alone are toxic against PC3 cells in vitro; however, in combination they greatly increased the oncolytic capacity of VSV by reducing cancer cell viability and promoting apoptosis-mediated cell death. Furthermore, the potentiation of VSV oncolysis by SFN is dependent on the production of ROS and is associated with the induction of autophagy. SFN is known to induce phase II antioxidant genes via Nrf2 activation, which regulates ROS levels and stimulates autophagy in prostate cancer cells. Mechanistically, SFN inhibited the innate antiviral response by blocking the type-1 interferon (IFN) signaling pathway, through the activation of the Nrf2 transcription factor. Exogenous Nrf2 expression inhibits Interferon-Stimulated Response Element (ISRE) promoter activity in a dose dependent manner following virus infection or IFN treatment. Taken together, these results demonstrate for the first time the synergic effect of SFN and VSV and indicate that SFN treatment increases VSV replication and the subsequent apoptosis of tumor cells by inhibiting IFN signaling. We are currently investigating the molecular mechanism involved in VSV-induced oncolysis by Nrf2 activators and evaluating the therapeutic potential of the combination of OV and Nrf2 activators in a mouse model of prostate cancer.

Biography:

Meltem Elitas is a Faculty Member in Mechatronics Program at Sabanci University. Her background is in Electrical and Mechatronics Engineering. She has obtained her Doctorate from Bioengineering and Biotechnology Department at Ecole Polytechnique Federale de Lausanne. She has performed her Postdoctoral studies at Yale University Biomedical Engineering Department. She has published more than 25 papers in reputed journals. Her research interests are biomechatronics, cellular heterogeneity, cellular interactions, tumor microenvironment, live cell imaging and development of microfabricated tools for quantitative biology.

Abstract:

Understanding the interactions between tumor cells and immune cells in a quantitative manner will provide valuable information to reveal the mechanism of diseases, immune defense and development of new treatment reagents and strategies for the diseases. Today one of the biggest limitations relies on the traditional methods and tools that we use to investigate the rare cells and specific events in biology particularly in immunology. Since these techniques are not adequate enough to be selective, specific and quantitative, the rare cells such as the metastatic or drug resistant ones or the events such as onset symptoms of tumors and infections are being masked by majority of the cells or events in the population. Therefore, we cannot diagnosis on time or provide successful strategies. As a consequence, our approaches might not target the right cells at the right time in the right place. To overcome these limitations, we might profit from engineering approaches and tools. We can develop quantitative, accurate, reproducible and precise methods and use microfabricated tools to understand the nature and behavior of rare cells and events. The improvements from microfabricated tools in conjunction with microscopy might provide statistics from large numbers of single cells, short assay time, less sample consumption, less waste production, quantitative and reproducible data, single-cell resolution images, high-throughput, spatio-temporal tracking and real-time assays, etc. This talk will present recently developed microfabricated tools to understand the immune cell-tumor cell interactions. I will present our microfluidic applications and their preliminary data from my research group.

Biography:

Amina Dahmani is currently a PhD candidate in Microbiology-Immunology at Université de Montréal, Canada. She has completed her Master degree in Immunology at Univérsité Laval, Canada, in Cellular Therapy Lab directed by Dr Jacques P Tremblay, where she studied the development of immunological tolerance to allogeneic myoblasts transplantation as potential therapy for Duchenne Muscular Dystrophy. Later she has joined Dr Jean-Sébastien Delisle team's, dedicated to cancer and viral adoptive immunotherapy to complete her PhD. She is currently working to improve adoptive immunotherapy protocols.

Abstract:

Adoptive immunotherapy (AI) has emerged as a potentially curative therapy for advanced cancer and infections. Recent findings suggest that the transfer of T-cells with “early” memory features may improve the therapeutic potential of AI. TGF-β is a pleiotropic cytokine that controls a large spectrum of biological and pathological processes. In T-cell biology, TGF-β is mostly known for its immunoregulatory properties, but recent evidence has revealed a novel role of TGF-β in T-cell memory differentiation and maintenance. Thus, we investigated whether TGF-β could promote features of memory in ex vivo stimulated human T-cells to further improve the efficacy of clinical protocols for AI. Here we show that agonistic TGF-β stimulation leads to the expression of central memory markers without significantly altering T-cell expansion or polyfunctional cytokine secretion following stimulation. Furthermore, TGF-β exposure decreased expression of transcription factors responsible for effector differentiation (T-BET, GATA3 and BLIMP1) and increased those associated with memory differentiation, notably ID3. The knock-down of ID3 by specific siRNA revealed that TGF-β-driven T-cell memory differentiation largely depends on ID3. Moreover, TGF-β-exposed T-cells showed enhanced persistence, expansion and alloreactivity after adoptive transfer into NSG mice. Finally, using clinically relevant culture methods to generate T-cell lines against viral and tumor antigens, we found that TGF-β programmed the expression of early memory markers without significantly curtailing T-cell expansion or antigen-specificity. This finding provides a rationale for clinical use of TGF-β to optimize memory phenotype of ex vivo pathogen/antigen-specific T-cells expanded for AI.

Biography:

He completed his Phd and working as a Group Leader at Klinikum der Universität München, Germany

Abstract:

Macrophage migration inhibitory factor (MIF) proteins (MIF and MIF-2) are chemokine-like inflammatory mediators with unique structural properties distinct from classical chemokines. MIF proteins play a role in the control of both physiological and pathophysiological immune responses. With MIF-2 only very recently identified, most evidence currently is available for MIF. In fact, MIF is a pivotal upstream mediator of innate immunity, while some contribution to the adaptive response has been reported. When dysregulated, MIF is an exacerbating promoter of several inflammatory diseases including atherosclerosis, a chronic inflammatory condition of large and medium-sized arteries and the major underlying cause of cardiovascular morbidity and mortality worldwide. MIF orchestrates the atherogenic recruitment of monocytes/macrophages and T lymphocytes through non-cognate interaction with the CXC chemokine receptors CXCR2 and CXCR4, respectively, and contributes to the inflammation and destabilization in atherosclerotic lesions. These processes have been considered as the effects of an innate chemokine on innate inflammatory cells in the atherosclerotic lesion area. Here we show that MIF also controls adaptive immune cells in atherosclerotic pathogenesis. We present data that MIF is a novel B cell chemokine that promotes B cell migration and proliferation via the chemokine receptors CXCR4 and CXCR7 as well as CD74, the surface form of MHC class II invariant chain. MIF-driven B cell responses are mediated through the ZAP-70 and ERK1/2 signaling pathways and encompass the activation of calcium transients. We also studied the impact of Mif gene deletion in the pro-atherogenic ApoE-/- genetic background in mice and have unraveled a surprising atherogenic phenotype with a previously unrecognized link between MIF and B cell pathobiology. This suggests that MIF could be a potential therapeutic target to induce protective B cell responses in such diseases.

Biography:

Denis Soulet has completed his PhD in Neuroimmunology from Laval University, Canada and Postdoctoral studies from Ycee Claude Bernard, Sweden. He is an Associate Professor at the Medicine Faculty of Laval University, Canada. He has published more than 40 papers in reputed journals and has been serving as an Editorial Board Member of SM Journal of Gastroenterology and Hepathology. He is leading a research team dedicated to study the role of peripheral inflammation in the enteric nervous system and its contribution to Parkinson’s disease. The ultimate goal of his research program is to design immunomodulatory based disease modifying drugs for PD.

Abstract:

Parkinson’s disease (PD) is a neurological disorder characterized by motor symptoms which are often preceded by non-motor symptoms, including gastrointestinal dysfunctions. Common treatments are only symptomatic; there is still no disease modifying drug available to cure patients. Since numerous pro-inflammatory markers have been measured in the central and peripheral nervous system, this deleterious immune response seems to be a potential target to develop new therapeutic strategies. Therefore, a better understanding of the role of the immune response in the etiology and progression of PD is essential. During my talk, I will present original data about the impact of the innate immune response on enteric neuronal damage in PD models. At first, we characterized the immune response induced by the neurotoxin MPTP in the enteric nervous system of partially immunodeficient mice. We demonstrated the timeline of inflammatory events occurring prior to the neuronal demise and the critical role of monocytes and macrophages in the gut. Thereafter, we tested various estrogenic compounds for their immunomodulatory and neuroprotective properties in PD models both in vivo and in vitro, delineating the major contribution of various estrogenic receptors, mainly the G Protein-coupled Estrogen Receptor 1 (GPER1). More recently, we successfully explored the therapeutic potential of a clinically approved selective estrogen receptor modulator, Raloxifene, for drug repurposing in PD. In conclusion, our data highlight the critical role of the immune response at early stages of PD and the immunomodulatory and neuroprotective potential of estrogen-based hormonotherapy at the pre-clinical level.

Biography:

Ganapathi Bhat Mugulthimoole is a Senior Consultant Medical Oncologist & Stem Cell Transplant Physician at Jaslok Hospital & Research Centre. He has completed his graduation in Medicine in 1993 and Post graduation in General Medicine in 2000. He has further trained in the field of oncology in various institutions in India and later gained expertise in stem cell transplant while working in Kuwait Cancer Control Centre (2002-2006). He has gained specialized training in stem cell transplantation as part of the ESH-EBMT (2007), 2011 (Labaule, France) and ICAS training program (2009) from Ulm University, Germany. He has numerous academic articles published in Indian, international journals and textbooks to his credit.

Abstract:

The tumor microenvironment is a principal feature of cancer biology that supports the initiation and progression of the tumor and response to therapy. Cells and molecules of the immune system are the essential elements of the tumor microenvironment. Therapeutic stratagems can harness the immune system to precisely target tumor cells by possibly stimulating tumor-specific immunological memory, which may lead to long term regression. Understanding the complexity of immunomodulation by tumors is important for the expansion and promotion of immunotherapy. Several approaches are being carried out to augment the anti-tumor immune responses. Among them are immunotherapeutic vaccines, adoptive cell transfer therapies and checkpoint blocking drugs. However, cancer cells try to evade the immune watchdogs by reducing surface tumor antigen or inducing cells that express certain proteins that affect immune cell inactivation or by promoting tumor proliferation and survival. Gaining a deeper understanding of the tumor immunogenicity by employing advanced techniques such as sequencing of the tumor DNA, will help to better address the challenges and gain an appreciation of the delicate association between cancer and our immune systems.

Biography:

Moshe Giladi has joined Novocure in 2005 and served as the Head of the NovoBiotic project until 2008. He was then promoted to Head of Novocure's Preclinical Research leading a team of experts of various fields: Cancer, immunology, cell biology and also responsible for research collaboration with academic institutes. He leads research activities studying tumor treating fields mechanism of action. He has received his PhD in Life Sciences from the Department of Molecular Microbiology and Biotechnology, Faculty of Life Sciences and his MBA from the Leon Recanati Graduate School of Business Administration both at the Tel Aviv University, Israel.

Abstract:

Tumor treating fields (TTFields) are an effective anti-neoplastic treatment modality delivered via non-invasive application of low intensity, intermediate frequency, alternating electric fields. This therapy is approved for the treatment of patients with glioblastoma. Previous investigations have shown that TTFields disrupt microtubules and septin filaments, both of which govern key roles in mitosis. Some of the outcomes of mitosis under TTFields application include abnormal chromosome segregation and ER stress which trigger different forms of cell death. The goal of this study was to evaluate whether TTFields induced cancer cell death can be perceived as immunogenic by the immune system and to explore the possibility of combining TTFields with immune-modulating drugs. Murine Lewis lung carcinoma (LLC) and ovarian surface epithelial (MOSE) cells were treated with TTFields using the inovitro system. The exposure of calreticulin (CRT) on the surface of treated cells was evaluated using flow cytometry. High-mobility group box 1 protein (HMGB1) secretion was measured using ELISA assay. For in vivo experiments, immunocompetent C57BL/6 mice were orthotopically implanted with LLC cells and treated with TTFields, anti-PD-1 or combination of the two modalities. Changes in tumor volume were monitored and flow cytometry analysis was performed for phenotypic characterization of tumor infiltrating immune cells. We demonstrate that application of TTFields leads to the exposure of CRT on the cell surface and also promotes release of HMGB1 from cancer cells in vitro. In vivo, the combined treatment of TTFields and anti-PD-1 led to a significant decrease in tumor volume compared to control group and to animals treated with anti-PD-1 alone. An increase in CD45+ tumor infiltrating cells was observed in both anti-PD-1 and TTFields+anti-PD-1 groups although statistical significance was reached only in the combination treatment group. Interestingly, CD45+ cells from the combination treatment group also demonstrated a significant upregulation of PD-L1 expression on the cell surface. Specifically, this upregulation in the PD-L1 expression was observed in both F4/80+CD11+ cells (macrophages) and CD11c+ cells (dendritic cells) whereas no significant effect on the infiltration pattern of these immune cell populations was noted. Taken together, our results demonstrate that TTFields application potentiates immunogenic cell death in cancer cells and that combining TTFields with specific immunotherapies such as anti-PD-1 might achieve tumor control by further enhancing antitumor immunity.

Biography:

Tali Feferman has completed her PhD in Molecular Biology at the Macquarie University and Postdoctoral Training at The Heart Research Institute in Sydney. She has then joined the Weizmann Institute of Science in 2000. Her initial interest involved exploring the mechanism of action and immunomodulation of Myasthenia Gravis (MG) and its model experimental autoimmune MG (EAMG). In recent years her interest is mainly focused on addressing questions important for optimizing cancer immunotherapy using cutting-edge imaging microscopic techniques in live mice.

Abstract:

Poor tumor vascularization is an obstacle to immunotherapy by CTLs. It impairs tumor infiltration but also introduces hypoxia, known to interfere with T-cell migration. It is yet unknown how suboptimal vascularization affects CTL migration and function within tumors. To study this question, we combined immunohistochemistry of human melanoma samples with two-photon imaging in live mice. Orthotopically implanted B16-OVA tumors were studied after adoptive transfer of in vitro matured antigen-specific OT-I CTLs. In patients, CD8 T-cells concentrated around peripheral vessels in the tumor and sparsely infiltrated avascular areas. In mice, CTLs crawled rapidly in oxygenated areas within 50 µm of flowing blood vessels. Occluding intratumoral blood vessels triggered immediate arrest of CTL motility, which was quickly reversed when flow was resumed. Immunohistology indicated that CTLs avoided hypoxic tumor areas. Live CTL imaging in vitro showed deceleration under hypoxic conditions and when oxidative phosphorylation was blocked. To circumvent intratumoral CTL dysfunction we attempted to increase vascular density by implanting tumors in matrices containing bFGF. bFGF-laced tumors were more easily rejected after transfer of CTLs and displayed delayed growth in untreated mice but were not affected in mice deficient in CD8 T-cells. CTLs infiltrated such tumors in normal numbers but displayed enhanced motility in highly vascular tumors, suggesting that enhanced rejection resulted from improved intratumoral CTL migration. Taken together, the results suggest that hypoxia limits CTL function away from blood vessels and that alleviating it may synergize with immunotherapy.

Biography:

Hisham Abd El Dayem is presently working in Ain Shams University, Egypt.

Abstract:

Purpose: To compare expression of multidrug-resistant protein 1/P-glycoprotein (MDR1/Pgp) in retinoblastoma in eyes treated by primary enucleation due to advanced tumor at initial presentation and those enucleated after being resistant to chemotherapy. Methods: This study was a prospective study. Twenty retinoblastoma patients presented to Retinoblastoma Clinic at Ophthalmology Department, Ain Shams University Hospitals. All patients had enucleation and were divided into 2 groups. Patients in group-1 underwent primary enucleation due to advanced tumor at presentation. Patients in group-2 underwent secondary enucleation after failure of conservative treatment. Immunohistochemical studies were performed searching for expression of multidrug-resistant protein 1/P-glycoprotein (MDR1/Pgp) in the two groups. Results: Analysis of the primary enucleation group showed high positive, low positive and negative expression in 1 (10%), 2 (20%) and 6 cases (70%) respectively. In secondary enucleation group: 5 cases (50%), 3 cases (30%) and 2 cases (20%) showed high positive, low positive and negative expression respectively. Conclusions: This pilot study though, not being able to demonstrate statistical significance in MDR1 expression in primary enucleated vs. secondary enucleated resistant cases, demonstrated p-value low enough to indicate a trend for more MDR1 expression in resistant cases (P=0.068). Further study with a larger sample size is warranted.

Biography:

Nan-Shan Chang is currently the Professor and Director of the Molecular Medicine Institute, National Cheng Kung University (NCKU) in Taiwan, and the Adjunct Professor with the SUNY Upstate Medical University and the NYS Institute for Basic Research in Developmental Disabilities, New York. Dr. Chang is most noted for his discovery of tumor suppressor WWOX in 2000. Recent Awards: Breast cancer and neurofibromatosis research awards from the Department of Defense, USA, in 2008 and 2010; NCKU Distinguished Professor Award 2010, 2013 and 2016; Distinguished Scientist Award 2011 from the Society of Experimental Biology & Medicine, USA.

Abstract:

Whether tumor suppressor WWOX stimulates immune cell maturation is largely unknown. Here, we determined that Tyr33-phosphorylated WWOX physically binds non-phosphorylated ERK and IκBα in immature acute lymphoblastic leukemia MOLT-4 T cell and in the naïve mouse spleen. The IκBα/ERK/WWOX complex was shown to localize, in part, in the mitochondria. WWOX prevents IκBα from proteosomal degradation. Upon stimulating MOLT-4 with ionophore A23187/phorbol myristate acetate (IoP), endogenous IκBα and ERK undergo rapid phosphorylation in less than 5 min, and subsequently WWOX is Tyr33 and Tyr287 de-phosphorylated and Ser14 phosphorylated. Three hr later, IκBα starts to degrade and ERK returns to basal or non-phosphorylation, and this lasts in the next 12 hr. Finally, expression of CD3 and CD8 occurs in MOLT-4, along with re-appearance of the IκBα/ERK/WWOX complex near 24 hr. Inhibition of ERK phosphorylation by U0126 or IκBα degradation by MG132 prevents MOLT-4 maturation. By time-lapse FRET microscopy, IκBα/ERK/WWOX complex exhibits an increased binding strength by 1-2 fold after exposure to IoP for 15-24 hrs. Meanwhile, a portion of ERK and WWOX relocate to the nucleus, suggesting their role in the induction of CD3 and CD8 expression in MOLT-4. (Supported by MOST and NHRI, Taiwan, and DoD, USA)

Biography:

Miao Dong is currently pursuing PhD in City University of Hong Kong. She has obtained her Bachelor degree from Liaoning University, China, majoring in Ecology and completed her Master degree from Shandong Agricultural University. During three years study, she investigated antioxidant defense system in zebrafish and has 6 publications (3 of 6 are as first author). During her PhD study, she has done research on interaction between innate immune proteins in fish blood and bacteria, also the collection of natural occurring antimicrobial peptides from medaka fish blood. The related manuscript has been submitted to JBC.

Abstract:

The excessive use of antibiotics in aquaculture contributes to the uprising of antibiotic resistance that threatens human health. We explore the innate immunity of fish for naturally occurring antimicrobial factors that can be developed into potential antibiotic agents. Antimicrobial peptides (AMPs) have been studied in many organisms but efforts on the systematic identification of AMPs in fish have been lacking. In this study, we systematically identified naturally occurring peptides in medaka plasma. Blood collected from medaka of different gender, age and infection status were combined and thereby providing a resource of plasma macromolecules under various possible physiological conditions. Peptides under the molecular weight of 3 kDa were fractionated and purified followed by mass spectrometry analysis. In total, 6483 unique peptides were identified against the medaka genome, constituting a database of circulating peptides in this organism. After evaluation with a combination of web based prediction tool and conserved physicochemical properties of AMPs, 83 potential antimicrobial peptides were predicted. One of them, a 13-residue peptide named VPS13D3241-3253, showed broad spectrum toxicity on fish and human pathogenic bacteria (Gram positive or Gram negative) without significant cytotoxicity on mammalian cell lines. Scanning electron microscopy indicated that VPS13D3241-3253 disrupted the cell wall of both Gram positive and negative bacteria. SOS response assay showed that this peptide efficiently induced DNA damage in bacteria. The identification of VPS13D3241-3253 illustrates the feasibility of the proteomic approach in the discovery of potentially novel AMPs from fish. These AMPs will form an important basis for the development of new antibacterial agents in the fishery.

Biography:

Anne Stinn is currently a PhD student at the Max Planck Institute for Infection Biology in Berlin, Germany. From 2008 to 2013 she studied Biology at the Justus Liebig University in Giessen, Germany. After completing her study she went to London as an Intern in the Research Group of Cell Death, Cancer and Inflammation (CCI) at the UCL Cancer Institute. She has started her PhD in the year 2014.

Abstract:

Acute myeloid leukemia (AML) is a highly malignant cancer of the myeloid cell lineage that is characterized by the rapid growth of abnormal white blood cells. Although AML is a relatively rare disease accounting for about 1% of all cancer cases (US), it is the type of leukemia showing the lowest survival rate. In the majority of all AML cases mutations in the kinase domain of the FMS-like tyrosine kinase III receptor (FLT3; CD135) are reported. Besides treatment based on chemo and radiation therapy as well as bone marrow transplantation, bispecific antibodies are studied for the use in immunotherapy against AML. These antibodies recognize tumor-associated antigens (TAAs) as well as the agonistic T-cell receptor/CD3 complex (TCR/CD3) and should thereby lead to a tumor cell-restricted activation of immune cells and specific lysis of cancer cells. In this study a structural analysis of the bispecific antibody NF-CU, the first known to date was performed. In addition to the structure of the NF-CU itself, the crystallographic structure of the antibody bound to FLT3 and CD3 was investigated. Deciphering the crystal structure of the antibody-antigens complex should give an inside into epitope recognition as well as the molecular mechanism leading to T-cell activation and tumor cell death.

Biography:

Maryam Golshani was graduated from the Pasteur Institute with a degree in Medical Bacteriology. Presently, she is a Post doctorate fellow and Junior Research Group Leader at IPI working on new Brucella vaccine candidates. Her research mainly focuses on in silico investing the immunogenicity of new vaccine targets and in vivo evaluating their protective efficacy against Brucella infection. She has involved in more than 11 projects and published 11 papers in reputed journals.

Abstract:

Objectives: Brucellosis is the most common bacterial zoonosis worldwide and no safe and effective vaccine is available for the prevention of human brucellosis. In humans, brucellosis is mostly caused by Brucella melitensis and Brucella abortus. According to our in silico studies, Omp2b is predicted to be potentially immunogenic antigen conserved in main Brucella pathogens. The aim of this study was to design truncated form of Omp2b and to evaluate the immunogenicity and protective efficacy of a recombinant protein vaccine encoding tOmp2b. Methods: Bioinformatics tools were used to design the truncated protein based on conserved domains and regions of epitopes with strong affinity for MHC molecules. The humoral/cellular immune response and protection levels against challenge with wild B. melitensis and B. abortus infections were evaluated in rtOmp2b+ adjuvants immunized mice and control groups. Results: Vaccination of BALB/c mice rtOmp2b provided the significant protection level against both B. melitenisis and B. abortus. Moreover, rtOmp2b elicited a strong specific IgG response (higher IgG2a titers) and significant IFN-γ/IL2 production. Conclusion: According to the results, rtOmp2b is able to induce cross-protection against B. melitensis and B. abortus infections. Therefore, it could be a new potential candidate for the development of Brucella subunit vaccines.

Biography:

Jalil Mehrzad has completed his PhD at the age of 32 years from Ghent University, Faculty of Veterinary Medicine, and postdoctoral studies from McGill University. He is an assciate professor of Immunology in Ferdowsi University of Mashhad (next year will move to Tehran University as full-time scientific member of departmet of Microbiology and Immunology). With H-index and citations of 16 and 1315, respectively, he has published more than 45 papers in reputed journals and has been serving as regular reviewer for many journals in the area of immunobiology, molecular biotechnology and medicine.

Abstract:

Caspases-mediated apoptosis/cell death activation is key regulatory response in many physiopathological conditions. Application of bioluminescence and the reaction of luciferase would provide a powerfully novel in vitro/vivo assay for apoptosis detecion. As key brain immune cells, astrocytes and microglials, are vital part of the central nervous system (CNS); they are the main responder to inflammation in CNS; any disruption on their function would lead to CNS damage. Aflatoxin B1 (AFB1) is commonly found in foodstuffs, and can be the cause of many diseases including cancer. AFB1 and its metabolites cause oxidative stress in especially the CNS-derived cells, adversely affecting their normal activities, thus leading to the neurodegenerative diseases including multiple sclerosis (MS), Alzheimer’s and Huntington’s diseases. Considering the importance of astrocytes and the inevitable existence of AFB1 in the feed/foods, worldwide, the study of astrocytes-AFB1 interactions is valuable. We therefore investigated the impact of AFB1 on the apoptosis of one of the key accessory supportive CNS, astrocytes, using several biochemical experimentations including intracellular ATP and caspases 3/7 measured by bioluminescence and luciferase reactions. The release of cytochrome c and apoptosis/necrosis of AFB1-treated astrocytes with various concentration of AFB1 and exposure time was also tested using Western blotting and flow cytometry techniques, respectively. Bioluminescence results revealed decreased intracellular ATP, increased caspases 3/7 activities, cytochrome-c release and apoptotic/necrotic of astrocytes particularly at higher timepoints and doses of AFB1. Considering the broad roles of astrocytes in CNS, this finding deepens our understanding of the molecular mechanisms and functional consequences of the neural cells damage neurotoxicity triggered by AFB1 exposure in mammals.

Biography:

Thomas Boldicke has completed his PhD at the Max Planck Institute for Molecular Genetics in Berlin. He has been working for 25 years in the field of recombinant antibodies particularly intrabodies. He has published more than 20 papers in reputed journals.

Abstract:

Intracellular antibodies (intrabodies) are targeted into a cell expressing the corresponding antigen, binding of the intrabody to the antigen results in inhibition of protein function. The advantages of high specificity, no off target effects and targeting of post translational modifications are the reasons that such molecules are very valuable in functional genomics. Two developments will boost the intrabody technology in the future: Cytoplasmic intrabodies can be stable expressed as single domain antibodies, mostly from camels. Alternatively, the construction of human VL and VH domains is ongoing. The single domain antibody approach is an effective alternative to other approaches for selection of stable cytoplasmic intrabodies such as the Intracellular Antibody Capture Technology (IACT) based on the yeast two hybrid system and Complementarity Determining Region (CDR) grafting or introduction of synthetic CDRs in stable frameworks. ER intrabodies: Selection of recombinant antibody fragments by in vitro display systems mainly phage and yeast display. One cloning step is sufficient to express scFv fragments as ER intrabodies. Most promising are these intrabodies retaining proteins passing the ER. Recently we demonstrated in mice a delay of metastasis of rhabdomyosarcoma tumor cells mediated by two specific intrabodies retaining two polysialyltransferases inside the ER. Finally transgenic ER intrabody mice have been generated. An intrabody mouse expressing an anti-VCAM intrabody is not lethal in comparison to the genetic knockout counterpart. 30% of genetic knockouts are lethal; therefore intrabody knockdown mice will be very useful in case the genetic knockdown is embryonically lethal.

Biography:

Hanan Al-Khalifa has obtained her Master’s degree in Parasitological Diseases and Immunology at University of Manchester and completed her PhD in 2007 in the University of Reading, UK, investigating the effect of n-3 fatty acids on the immune response and general health status. Her interests include but are not limited to immunological techniques, parasitological diseases, effect of nutrition, espicially fatty acids, on the immune status in both humans and expermental animals. She executed many research projects that focused on the effect of nutrition on immunology. She has attended many scientific events and published more than 60 papers in refereed journals and conference proceedings.

Abstract:

The phagocytosis assay allows quantitative measurement of the percentage of phagocytes and the enzymatic activity of each phagocyte. Immunomodulation of fatty acids in flaxseed may alter phagocytosis activity. The objective of this work was to compare the effect of feeding normal broiler chickens 15% of dietary flaxseed on phagocytic activity of monocytes and heterophils in the peripheral blood. One day old broilers were used. Upon hatching, all chicks were given the same basal diet for 13 days. Following this, dietary supplementation of flaxseed started at 14 days of ages until the end of the cycle at 35 days of age. At slaughter, samples of blood were collected from each bird. The quantitative analysis of the phagocytic activity of peripheral blood mononuclear phagocytes in whole blood was performed using PHAGOTEST commercial kits. Results were expressed as percentage of fluorescent cells (% phagocytosing cells) and mean fluorescence intensity (MFI). Feeding flaxseed at 15% did not affect either the percentage of cells participating in phagocytosis or the Mean Fluorescence Intensity (MFI). However, there was a trend towards a decrease in the percentage of monocytes involved in phagocytosis in birds fed diets containing 15% flaxseed. Also, there was a trend towards a decreased MFI (p=0.056) for monocytes. In general, results of the current study showed no effect of flaxseed on phagocytosis of peripheral blood cells.

Biography:

Laurence Macia is a Group Leader at the Charles Perkins Centre, School of Medicine of the University of Sydney. She has published over 30 articles in journals such as Nature Communication and Nature Reviews Immunology. Her research interest is the impact of diet on gut microbiota and development of inflammatory diseases. She has obtained her PhD in 2006 at the Pasteur Institute of Lille in France where she studied the inter-relation between metabolism and immunity. She has then worked at the Garvan Institute and at Monash University in Professor Mackay’s Lab to investigate the impact of diet on the development of Western diseases.

Abstract:

Incidence of food allergy has increased dramatically in recent decades particularly in Western countries. The diet hypothesis states that western diet enriched in fat and sugars while deprived in fibre contributes to development of western diseases such as allergy. Dietary fibre is potent prebiotic, reshaping beneficially gut microbiota. It is also fermented in the colon by anaerobic bacteria into short chain fatty acids (SCFA) that bind specific G-protein coupled receptors widely expressed in the host. The aim of this study was to determine the impact of diet enriched in dietary fibre and SCFA on the development of food allergy in mice. Mice were fed on diets either enriched or deprived in fibre in models of oral tolerance to peanut and of peanut allergy. In both models, dietary fibre was beneficial as enhanced oral tolerance and protection from food allergy were observed under high fibre feeding conditions. SCFA were behind these benefits as both acetate and butyrate protected from development of peanut allergy while propionate had no effects. Accordingly, mice knockout for GPR43 or for GPR109A, respectively preferential receptor for acetate and butyrate were not protected from food allergy development under high fiber feeding conditions. To determine the role of gut microbiota, germ free mice were reconstituted with microbiota isolated from high fibre vs. zero fibre fed mice and we found that the first was protective in food allergy. In conclusion, high fibre feeding protects from food allergy development by reshaping of gut microbiota and through the SCFA acetate and butyrate.

Biography:

Sundeep Kumar Upadhyaya is a Senior Consultant at New Delhi’s Indraprastha Apollo Hospitals. He is involved in the treatment of autoimmune disorders like Arthritides, Vasculitides, Lupus and Spondyloarthritis since 16 years. He is an Associate Professor and Teaching Faculty at the Teaching Program at the AHERF and National Boards of the Apollo Group of Hospitals. He is involved with cutting edge clinical work on these disorders and has developed treatment algorithms for lupus nephritis, early and established rheumatoid arthritis and has been on various national bodies and involved in management protocols for autoimmune disorders.

Abstract:

Rheumatoid arthritis (RA) is conventionally treated with cDMARDs (like Methotrexate, Leflunomide etc) but more severe, cDMARD resistant RA, is treated with a combinations of cDMARDs and Biologics (example Anti-TNF Biologics). Despite close monitoring and follow-up only a fraction of treated patients achieve complete remission and attain a drug free state. A combination of cDMARDs and Biologics in the treatment of early RA has been able to achieve drug free remissions in only a small minority of patients. Complete remissions are rare when patients are treated with cDMARDs alone. Presented here is the clinical description of patients with RA who achieved long-term drug free clinical remissions (CR) with cDMARDs alone at a single rheumatology clinic in northern India and remained in CR over a long period (at least 6 months). The immune-biology of such states and the clinical factors leading to the prolonged CR will be discussed next.

Biography:

Li Yu-Jung is currently a Lecturer at St. Mary’s Medicine, Nursing and Management College. He has completed his training program of Oral and Maxillofacial Surgery in Veterans General Hospital-Taipei, Taiwan during 2002-2006. He is also a Doctoral candidate majored in Mechanical and Electrical Engineering from National Taipei University of Technology. He has received his MS degree of Clinical Dental Science from Institute of Clinical Dentistry, National Yang-Ming University. He has also received MS degrees of Chemistry and Biophysics from Graduate Institute of Biophysics, National Central University and Institute of Chemistry, Tamkang University during 2006-2010 and Bachelor’s degree of Dentistry from Department of Dentistry, Chung Shan Medical University in 2002.

Abstract:

Due to lack of pulp structure and periodontal ligament (PDL), which are regarded as the mainly pain origins of tooth painful sensation, dental implant have the opportunity for painless molecular releasing and blood monitoring with long-term steady and continuous properties. The new pathway may allow functional peptides and even the bigger molecular releasing and monitoring, which is impossible to absorb throughout the gastrointestinal (GI) tract. Therefore we design the replaceable drug delivery and bio-sensing modules above the titanium dental implant fixture which is immobile inside the maxillary bone marrow. The drug delivery module contains the piezoelectric micro-pump, the drug container and the power supply inside, while the bio-sensing module is constructed by the integrated circuit (IC), the Bluetooth module and the power supply. The total loading volume of the drug delivery module is around 0.5-1 ml and the drug is polymerized due to safety concern. Therefore the drug releasing type throughout this module is slowly and continuously diffused into the surrounding blood pool inside the bone marrow. The released drug type also needs to be carefully selected to avoid surrounding bony destructions. In contrast, the biosensor may provide various molecular types of continuous blood monitoring within five minute intervals and lasting for about 1 month within current technology. With standard dental and medical protocol establishments, the device may provide more useful applications in clinical practice.

Biography:

Andreia Ribeiro has completed her Biology Degree and Microbiology Master in Aveiro University in Portugal. Since 2010 she has been working as Research Assistant in Flow Cytometry and Immunology Groups in Portugal and in Ireland. In 2013, she has started her PhD in the DECIDE consortium and completed in 2016 from National University of Ireland, Galway.

Abstract:

Extracellular matrix (ECM) plays an important role in the tumor microenvironment and in biologic processes such as hematopoiesis. ECM contributes to regulation of cell survival, proliferation and cell differentiation. The aims of this project were to study the quality, quantity and biological role of ECM produced by a cloned mouse mesenchymal stromal cell line (MS5) on cell differentiation and to study the role of hypoxia on cell differentiation. To carry out these studies, we have used two methods of producing ECM in vitro. In both methods ECM is produced in normoxia and hypoxia. In method 1, cells are lysed by osmotic shock with a Tris/EDTA buffer, the standard way of preparing ECM in many studies. In method 2, MS5 were transduced with a caspase 9 vector, allowing induction of apoptosis in the cells following ECM production. Balb/c bone marrow mesenchymal stromal cells (MSC) were then seeded either in uncoated plastic dishes or in dishes covered with ECM and differentiation assays were performed, again either in normoxia or hypoxia. Results show that the two methods produce qualitatively different ECM and that hypoxia plays a role in ECM composition. Moreover, compared with hypoxia, normoxia is a better condition for adipogenic differentiation of fresh MSC. In contracts, osteogenic differentiation is better on ECM in hypoxia. In conclusion, different methods of preparing ECM in vitro lead to different protein composition and different outcomes in cell differentiation. Hypoxia also makes a difference in ECM composition and cell differentiation.

Biography:

Roghaye Arezumand has completed her PhD from Pasteur Institute of Iran in 2015. She has published about 10 papers in scientific journals.

Abstract:

Nanobody is smallest antigen binding domain derived from camelids family. The small size and other evolutionary property could introduce it as novel drug candidate especially against cancer. Over expression of angiogenesis is a highlighted character of tumor tissues. Many angiogenic factors like VEGF family involved in new vessels formation in cancerous tissues. Placenta growth factor (PLGF) is highly expressed in pathologic condition of angiogenesis. Targeting of PLGF could have inhibitory effect on angiogenesis in cell and animal model. The aim of this study is targeting of PLGF by developed novel nanobody. We constructed a PLGF nanobody library in pHEN-4 phagemid vector. This library specify by biopanning on immobilized recombinant PLGF. After screening of individual colonies, we selected different nanobody for soluble expression. Affinity of this nanobody was done by ELISA based method and the effect of this nanobody on angiogenesis were assessment by proliferation, migration, invasion, 3D capillary formation and chorioallantoic membrane assay (CAM). This nanobody could inhibit the proliferation, migration and tube formation of HUVEC cells and invasion of MDA-MB231 breast cancer cells. In addition of in vitro assays, this nanobody could inhibit the neo-vascular formation of fertilized eggs. A novel and high affinity specific-Nb against PLGF with anti-angiogenesis effects on endothelial, breast cancer cells and on in vivo model was developed in this investigation.

Biography:

University of Baghdad, Iraq

Abstract:

Background: Hepatitis C virus (HCV) is a serious infectious disease that can cause lifelong infection. Infection with chronic hepatitis C virus (HCV) can lead to autoimmune hepatitis (AIH) in a minority of patients. Viral infection induces tumor necrosis factor (TNF-alpha) production in hepatocytes. On the other hand prolactin which is an endocrinal hormone acts as a cytokine and is also involved in immune responses. These findings suggest that both parameters may have an important role in the patho-physiology of human liver diseases induced by viruses. Aim: The aim of the presents study was evaluate the role of the immunoendocrine system in the pathogenesis of the disease, by measuring serum prolactin and tumor necrosis factor-alpha. Subject and Methods: Sixty- one chronic hepatitis C patients were consequently selected from the Medical city, Gastrointestinal Hospital in Baghdad, Iraq, during the period from July 2014 to September 2014, their median age was 34.8 year, 29 of them were males and 32 were females. All patients were diagnosed having positive for HCV RNA by means of polymerase chain reaction. The study also included twenty apparently healthy adult age and sex matched considered as controls, which were negatively screened with hepatitis C virus. Peripheral blood sample of 2 ml was aspirated using disposal syringes. Samples were collected between (9.00a.m-12.00p.m). The blood was allowed to clot in plain tube for 30-45 minutes at room temperature. Sera were obtained by centrifugation of the collected blood and then stored in plain tubes at -20 c. ELISA method was used to measure (TNF and Prolactin) Results: The results of this study showed an increase in mean value of both TNF and prolactin hormone in chronic hepatitis C patients. However no significant correlations were found between both parameters studied. Conclusions: Chronic hepatitis C is associated with an immunological abnormality mainly represented by tumor necrosis factor-alpha and prolactine. This might shed a light of the type of therapy and drug of choice when managing the disease.