Day 1 :
Keynote Forum
Aaron Lerner
AESKU.KIPP Institute, Germany
Keynote: New serological markers for celiac disease diagnosis: anti-neo-epitope human and microbial transglutaminases antibodies
Time : 10:00-10:30
Biography:
Professor Aaron Lerner after receiving his MD from the Sakler school of medicine, Tel-Aviv University (1976), Professor Lerner specialized in Pediatrics ( 1982 ), Pediatric Gastroenterology and Nutrition ( 1984 ) and Adult Gastroenterology ( 1987). Took several senior positions as head of Department of paediatrics (1995-2005) and head of Pediatric Gastroenterology and Nutrition unit, at the Carmel Medical Center, Haifa, Israel. Finished his Medical Management degree M.H.A, at Ben- Gurion University, Beer-Sheba, Israel (1999), spent research sabbaticals in Hahnemann University, Philadelphia, PA, USA (1991), State University of North Carolina, Chapel Hill, N.C,U.S.A (2005) and currently, in an extended scientific sabbatical in Aesku. Kipp Institute, Wendelsheim, Germany.(2014-16).Prof. Lerner presented in numerous international congresses, mainly of pediatrics, nutrition and autoimmunity, published 250 manuscripts in peer reviewed journals and is on the editorial board of 14 international journal.
Abstract:
The guidelines of ESPGHAN for the diagnosis of pediatric celiac disease (PCD) rely on anti-human tissue transglutaminase (tTg) as the prime and unique antibody for screening PCD population. Microbial Tg (mTg)is a family member of human tissue transglutaminase (tTg). Both enzymes,tTg and mTg,complexed to gliadin present neo-epitopes and antibodies against these complexes are called tTgneo-epitope andmTg neo-epitope. mTg is capable of cross-linking numerous molecules. Despite declarations of mTg safety, direct evidence for immunogenicity of the enzyme is lacking. The reliability of those antibodies in CD was evaluated.rnMaterials & Methods: The serological activity of mTg, tTg, mTg neo-epitope and tTg neo-epitope were studied in: 95 pediatric celiac patients (CD), 99 normal children (NC) and 79 normal adults (NA). Sera were tested by ELISAs, detecting IgA, IgG or both IgA and IgG: AESKULISA® tTg (tTg, RUO), AESKULISA® tTg New Generation (tTg neo-epitope (tTg-neo)), AESKULISA® mTg(RUO) and AESKULISA® mTg neo-epitope (mTg-neo, RUO). Marsh criteria were used for the degree of intestinal injury.rnResults: Comparing pediatric CD patients with the 2 normal groups: mTg-neo IgA, IgG and IgA+IgG antibody activities exceed the comparable mTg ones (p<0.0001). All mTg-neo and tTg-neo levels were higher (p<0.001). tTg IgA and IgG+IgA were higher than mTg IgA and IgA+IgG (p<0.0001). The levels of tTg-neo IgA/IgG were higher than tTg IgA/IgG (p<0.0001). The sequential antibody activities reflecting best the increased intestinal damage were: tTg-neo IgG ≥ mTg-neo IgG > mTg-neo IgA+IgG > tTg-neo IgA. Taken together, mTg-neo IgG and tTg-neo IgG correlated best with intestinal pathology (r2=0.989, r2=0.989, p<0.0001, p<0.0001, respectively).rnConclusion: mTg is immunogenic in children with CD and by complexing to gliadin its immunogenicity is enhanced. It represents a new serological marker for CD and matches the performance of the tTg neo-epitope. Both antibodies correlate with intestinal damage to the same degree. The neo-epitope tTg and mTg are new powerful serological markers for CD diagnosis. Further studies are needed to explore the pathogenic potential of those antibodies in CD.rn
Keynote Forum
Jurgen Bernhagen
Ludwig-Maximilians University of Munich, Germany
Keynote: MIF proteins as prototypical innate chemokines in inflammatory and cardiovascular disease
Time : 10:30-11:00
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.
Keynote Forum
Andreas Weinhausel
Austrian Institute of Technology GmbH, Austria
Keynote: Immunomics using protein and peptide microarrays for (cancer) biomarker development
Time : 11:15-11:45
Biography:
Andreas Weinhausel is a Biotechnologist and Specialist in Human Genetics. He has more than 20 years experience in molecular diagnostics. He has worked at the Children’s Cancer Research Institute, Vienna (1995-2004); he specialized in human molecular genetics diagnostics of syndromal and hereditary neoplastic disease. Since 2004, he has been working in the “Molecular Diagnostics” unit at the AIT-Austrian Institute of Technology and his focus is on DNA-methylation and protein biomarker development for cancerous and other systemic human disease using omics discovery and high throughput validation technologies. He is also an Associate Professor for Molecular Biology at the University of Natural Resources and Applied Life Sciences, Vienna.
Abstract:
An estimated 2.7 million new cancer cases and 7.6 million cancer related deaths were reported worldwide in 2008 and incidences are increasing. It is well accepted that early cancer diagnosis can improve survival, thus there is a great need and anticipation to identify novel biomarkers for cancer diagnosis at the earliest possible stage, which can ideally be integrated in minimal-invasive diagnostic assays. Cancer onset and progression produces mutated or aberrantly expressed proteins generally also termed as tumor associated antigens (TAAs) which are able to act as antigens and evoke an immune response which results in the production of autoantibodies. These autoantibodies can be detected months or years before the clinical diagnosis of cancer and can therefore be used as biomarkers for the early diagnosis of cancer. We have setup immunomics discovery technologies using high density protein and peptide microarrays for elucidation of novel biomarkers. By microarray discovery we have defined cancer specific classifiers with high diagnostic performance, obtaining AUCs>0.9 for the big 4 cancer entities. Autoantibody based strategies outperform the current clinical diagnostic methods and would be of high value for improving cancer diagnostics and patient management. To transfer assays onto clinical applicable formats our current developments of different technological variant settings using medium scaled multiplexed assays in microarray and bead array formats will be presented.
Keynote Forum
Walter Schubert
ToposNomos Ltd., Germany
Keynote: The human toponome project: Translating the spatial protein network code into efficient therapies
Time : 11:45-12:15
Biography:
Walter Schubert is founder and director of the international human toponome Project, and he is the head of the Molecular Pattern Recognition Research (MPRR) group at the Medical Faculty of the Otto-von-Guericke-University, Magdeburg, Germany, and guest Professor for Toponomics at the Max-Planck-CAS (CAS-MPG) partner Institute for computational biology, Shanghai, China. Walter Schubert has studied medicine at the universities of Aachen and Bonn, Germany, and has led the neuromuscular diagnostic center at the university of Bonn. He has invented and developed during his clinical work the toponome imaging robot technology MELC/TIS in 1990 (to decipher the protein networks in humans), is co-initiator of the course of studies in computervisualistics (CV) at Magdeburg university (MDU), and teaches human biology for students in CV and medicine at the MDU. Walter Schubert holds many patents, has received several national and international awards and honours, such as the american ISAC best paper award 2008 (for the three symbol code of organized proteomes, \"toponome\"), and has launched the human toponome project aiming at the functional decoding of the protein networks in humans.rnrnHe is keynote speaker at international congresses (presenting the human toponome project), invited distinguished lecturer at the Case Western Reserve University (2010), and member of several american and european editorial and scientific advisory boards. WS chaired three interdisciplinary national joint projects of the Deutsche Forschungsgemeinschaft and the BMBF (at MDU, and transnational) connecting engineering, informatics, medicine and cell biology in the field of toponomics. The work of the his Magdeburg group and coworkers has been acknowledged by a Research Highlight \"Mapping togetherness\" (Nature 443, p 609, 2006).rnrn
Abstract:
Imaging cycler® technology (IC®M) is presented as key thenology for (i) the spatial resolution of large protein networks at the target sites of disese with a discriminatory power for an unlimited number or proteins at a time (dimension unlimited imaging); (ii) for the in situ detection of thousands of distinct multi protein complexes; (iii) for the construction of machines able to decode the mechanism of cell invasion into organs, such as the invasion of autoimmune cells and cancer cells, and (iv) application of this technology for the efficient finding of therapies selectively blocking these invasions. The example of amyotrophic lateral sclerosis (ALS) is presented showing that (i) “ALS cells” were seen by IC® for the first time in the blood, (ii) the mechanism of CNS invasion and pathogenic neuronal axotomy of these cells was completely decoded by IC®, and (iv) these “ALS cells” were efficiently depleted in blood of patients. This ALS example can be translated for other diseases based on cell invasion. The IC® detection of somatotopic coding in the innate immune system is key.
- Cellular Immunology Inflammatory/Autoimmune Diseases Infectious Diseases and Immune System Innate Immunity Innate Immune Evasion Other Species Immunity Innate Molecular Immunology
Chair
Andreas Weinhausel
Austrian Institute of Technology GmbH, Austria
Co-Chair
Jessy S Deshane
University of Alabama at Birmingham, USA
Session Introduction
Philip R Hardwidge
Kansas State University, USA
Title: Citrobacter rodentium NleB blocks TRAF3 K63-linked ubiquitination to inhibit interferon-β production
Time : 13:15-13:35
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. ï„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.
Bozena Futoma-Koloch
University of Wroclaw, Poland
Title: C3 component deposition on Salmonella O48 cells characterized by sialylated lipopolysaccharide and different pattern of outer membrane proteins
Time : 13:35-13:55
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.
Jessy S Deshane
University of Alabama at Birmingham, USA
Title: Longitudinal changes in airway microbiome signatures and immunoregulatory cell dynamics following bronchial thermoplasty
Time : 13:55-14:15
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.
Hanan Al-Khalifa
Kuwait Institute for Scientific Research, Kuwait
Title: Lymphocyte subpopulations as affected by dietary fatty acids
Time : 14:15-14:35
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.
Nelson Gekara
Laboratory for Molecular Infection Medicine Sweden, Sweden
Title: The functions of the Histone H2A deubiquitinase (H2A-DUB/MYSM1) in innate immune regulation
Time : 14:35-14:55
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.
Un-Hwan Ha
Korea University, South Korea
Title: Pseudomonas aeruginosa mediated modulation of host defense responses
Time : 14:55-15:15
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.
Eva E Avila
Universidad de Guanajuato, Mexico
Title: Interaction of the parasite Trichomonas vaginalis with human neutrophil extracellular traps
Time : 15:15-15:35
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.
Tewodros Firdissa Duressa
KU Leuven, Belgium
Title: Locust innate immunity: Hemocyte replenishment, cytokine identification and involvement of an angiotensin converting ortholog
Time : 15:35-15:55
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.
Tzung-Yan Lee
Chang Gung University, Taiwan
Title: Electroacupuncture prevents white adipose tissue inflammation through modulation of hypoxia-inducible factors 1α-dependent pathway in obese animals
Time : 16:10-16:30
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.
Mira Barda-Saad
Bar-Ilan University, Israel
Title: Signaling cascades regulating natural killer cell activation threshold
Time : 16:30-16:50
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.
Nayef Jarrous
The Hebrew University of Jerusalem, Israel
Title: Non specific transcription initiation by RNA polymerase III and its possible implication in innate immunity
Time : 16:50-17:10
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.
Sumru Savas
Ege University, Turkey
Title: No relationship between lipoprotein associated phospholipase A2, proinflammatory cytokines and neopterin in Alzheimer’s disease
Time : 17:10-17:30
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.
Andreia Marques Ribeiro
National University of Ireland, Ireland
Title: A rapid and quantifiable flow cytometry-based potency assay to measure the immunomodulatory properties of mesenchymal stromal cells
Time : 17:30-17:50
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.