International Conference on Innate Immunity July 20-21, 2015 Barcelona, Spain

Alireza Badiei is a trained veterinarian and he worked as a veterinarian before he started his Postgraduate studies in Immunobiology from the School of Medicine, Putra University of Malaysia in 2008. He was appointed as a Lecturer at the University of Malaysia Kelantan, Faculty of Veterinary Medicine. Currently, he is finalizing his PhD in the Department of Pathology, University of Otago, Christ Church (age 39 years). He has published more than 10 papers from his Master’s and PhD projects.

Hydrogen sulfide (H2S) plays an important role in inflammation. It is produced by the activity of the enzyme cystathionine γ-lyase (CSE). Research on the role of CSE is confounded by the lack of specificity of pharmacological agents that inhibit CSE. The use of small interfering RNA (siRNA) to inhibit the production of the CSE is more a specific approach. Using a mouse model of caerulein-induced acute pancreatitis, we have observed that higher levels of pro-inflammatory cytokines including TNF-α, IL-6, MCP-1 and MIP-2, as well as increased activity of pancreatic and lung MPO and also plasma amylase levels. We used siRNA to silence gene for expression CSE in monocytes in an in vivo model of mouse acute pancreatitis. Our results show that siRNA treatment attenuates inflammation in the pancreas and lungs of mice following acute pancreatitis and reduces levels of pro-inflammatory mediators including IL-6, TNF-α, MCP-1, andMIP-2. Pancreatic and lung MPO activities as well as plasma amylase levels were reduced following in vivo silencing of CSE with siRNA. These findings show a crucial pro-inflammatory role for H2S synthesized by CSE in acute pancreatitis.

Despite permanent improvement of congenital heart disease surgical technique and perioperative care still significant number of procedures remain complicated or unsuccessful. The research continues into the impact of patient’s individual susceptibility to complications as a consequence of genetically determined possibility to react to wide spectrum of stressful stimuli occurring during surgery: cardiopulmonary bypass (CPB), infection, hypothermia and others. The attention has been paid to the complement system which is strongly involved in response to special conditions generated during cardiac surgery procedure. The problem was intensely investigated in adult population but has been up till now not elucidated in pediatric patients.We investigated the association between MBL serum concentrations, corresponding MBL2 genotypes and the postoperative course of children in whom cardiac surgery with the use of CPB was performed, in a prospective study. Blood/serum samples were collected from 195 patients. The obtained results suggest protective, beneficial effect of low MBL serum levels. The incidence of SIRS was lower in children with preoperative MBL concentrations<1000 ng/ml and point mutations within exon 1 of the MBL2 gene (A/O and O/O genotypes). MBL deficient patients (<100 ng/ml) with very low MBL levels had generally shorter ICU length of stay and lower inotropic score. In contrast, high MBL-producing genotypes (YA/YA) were more common among children who developed SIRS. This work was supported by the Polish National Science Centre (grant UMO-2011/03/B/NZ6/00052).

Jiandong Huang has completed his PhD at the age of 29 years from University of California, Los Angleles and postdoctoral studies from NCI, NIH. He is the director of Center of Synthetic Biology and Engineering, Shenzhen Institute of Advanced Integration Technology, Shenzhen, China. He has published more than 100 papers in reputed journals and has been serving as Professor at the University of Hong Kong

Metastasis is the most lethal feature of cancer. More than 90% of cancer patients die from secondary cancerous growths. Long-term dormancy, drug resistance and multiple growths all contribute to deadly side of metastasis. We have been engineering Salmonella typhimurium for cancer therapy and successfully developed a tumor-hypoxia targeting strain YB1, whose growth is strictly regulated by oxygen. Administration of YB1 to tumor-bearing mice partially inhibits the growth of primary tumor, but impressively, largely reduces lung metastasis. To investigate how YB1 treatment affects the process of metastasis, we set up orthotopic and experimental metastasis models on either immunocompetent or immunodeficient mice. The overall process of metastasis includes localized invasion, intravasation, circulation, extravasation and colonization. By implanting mouse breast cancer cells to mice and treating with YB1 at different stages of metastasis, we find that only treatments taken at early stages could reduce lung metastasis. Experimental metastasis model demonstrates further that YB1 treatment interferes with the colonization process of cancer cells to lung, thus reducing metastasis. Direct cytotoxicity of Salmonella to cancer cells is excluded as the anti-metastasis mechanism. The proinflammatory cytokine storms induced in tumor and blood circulation by YB1 contribute to the reduction of metastasis. After cytokine storms are induced, intravenously injected 4T1 cells are not able to colonize and form lung metastasis. The same phenomenon can be observed even on immunodeficient NOD SCID mice, indicating that the innate immunity is sufficient to reduce lung metastasis. These results demonstrate the potential of using bacteria as immunotherapy against cancer metastasis.

Magdalena Materniak works in Department of Biochemistry of National Veterinary Research Institute in Pulawy, Poland. During her PhD studies she developed the diagnostic tools for the detection of bovine foamy virus infections in cows and determined the prevalence of this virus in dairy cattle in Poland. Afterwards she continued her study in the field of veterinary retrovirology and diagnostics, especially on identification of foamy viruses in livestock and companion animals as well as zoonotic potential of these viruses. She has been contributed in several grants carried out in NVRI and papers published in reputed journals.

Porcine reproductive and respiratory syndrome virus (PRRSV) is a causative agent of an important infectious disease causing serious economic losses to swine industry. The clinical signs of the PRRS indcude respiratory disorders, abortions and variable mortality in piglets. The experimental study was performed to compare the virulence of highly diverse East European strains belonging to subtype 2 (Russian strain ILI and Belarusian strain BOR), as well as Danish strain from classical subtype 1 (DAN). The infection with strain BOR resulted in the most severe clinical picture, while the infection with remaining two strains was milder. To gain insight into transcriptome changes after different PRRSV strains infection, gene expression profiles of peripheral blood mononuclear cells lymphocytes were analysed in piglets infected with three PRRSV strains vs control piglets by microarray analysis. The genes were determined to be differentially expressed if the fold change (FC) was greater than 1.5 in up or down-regulation. Validation of microarray results was performed using RT-qPCR and showed high reliability of FC values determined by both methods. Microarray data were analysed using the Ingenuity Systems Pathway Analysis program to determine most significantly affected immunological pathways and regulators. Surprisingly, despite the differences in clinical picture of infection, ILI and BOR infected groups showed similar patterns of activation score for particular immunological pathways analysed, as well as, comparable upstream regulators activity (IFNG, IFNB1, Ifnar, IFNL1), when compared to DAN group. Therefore further analysis is required to deterimine genes which may be crucial for the course of PRRSV infection.

Seong Ji Woo has completed her PhD at 2014 from Chonbuk National University Medical School. She is a post-doc in the same scool and has a interest in the the role of macrophage in the obesity-related inflammation and insulin resistance.

Macrophage infiltration and inflammation in adipose tissue are well established to cause obesity-linked insulin resistance and type 2 diabetes. AMP-activated protein kinase (AMPK) has pleiotropic effects for energy metabolism, but its role for adipocyte-promoted macrophage chemotaxis has not been explored. Here, we report that an AMPK inhibitor, compound C (CC), significantly inhibited adipocyte conditioned medium (CM)-induced macrophage chemotaxis in RAW 264.7 cells in a concentration dependent manner, and this inhibitory effect was accompanied by the inhibition of focal adhesion kinase (FAK), AKT and inhibitory κB kinase (IKK) in macrophages, which all are important regulators of cell migration. Inhibition of macrophage chemotaxis by CC was not prevented but potentiated by co-treatment with AMPK activator 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), indicating that CC inhibition of AMPK is not involved in its suppression of chemotaxis. Instead, FAK phosphorylation was additively reduced by combined treatment of CC and AICAR, suggesting that CC and AICAR prevent adipocyte macrophage chemotaxis through targeting FAK pathway. CC treatment also prevented the expression of proinflammatory genes in RAW 264.7 macrophages when stimulated with either CM or lipopolysaccharide (LPS) and this effect was mediated by inhibition of IKK/NFB pathway. Lastly, we demonstrated that CC functioned as a repressor of macrophage-mediated insulin resistance in adipocytes. Our results suggest that CC might serve as a useful molecule in research on adipocyte-mediated macrophage chemotaxis and as a potential lead compound for the treatment of obesity-linked insulin resistance.

Beatriz Basso completed her PhD at School of Biochemistry, National Cordoba University, Argentina. She is Professor of Medicine School at the same University and Director of the Parasitology Laboratory, National Service of Chagas disease. She was Visiting Researcher in Universities of USA and Belgium, Director of graduate students and PhD. She has published more than 100 papers in national and international journals, over 200 presentations at congress, with several awards. She is evaluator of national and international research project, Editorial Board Member and Reviewer of international research journal, among other academic and research activities

Chagas disease, caused by Trypanosoma cruzi, is a major health problem in Latin America. Immune response to T cruzi infection is highly complex and involves many components, regulators and effectors. This parasite is able to survive and replicate in its host because it is adapted to resist host defences. A model for vaccinating mice with Trypanosoma rangeli, a parasite closely related to T. cruzi, but non-pathogenic to humans, has been designed in our laboratory, showing protection against T. cruzi infection. The aim of this work was to analyse some mechanisms of the early innate immune response in this vaccination model. In the first hours of infection, vaccinated mice showed a strong innate immune response. The adherent cells revealed important phagocytic activity, and some soluble mediators showed modifications: Respiratory Burst: significantly increase; NO: adequate levels; Arginasa: significantly increase. Furthermore, they showed an increase of macrophages, NK, granulocytes, and regulation of IL6, IFNγ, TNFα and IL10, with an increase of IL12, with respect to only infected mice. Conclusion: In this model some cellular populations and mediators soluble, involved in the innate immune response, plays an important role in vaccinated mice for the early elimination of the parasites. These results also suggest that in mice vaccinated with T. rangeli innate response could develop some kind of immunological memory, equivalent to adaptative immune response, recognizing shared antigens between T. rangeli and T. cruzi. These results could contribute to the knowledge of new mechanisms which would have an important role in the immunonoprevention of Chagas disease.

Marbel Torres has completed his PhD at the age of 35 years from Universite Francois Rabelais d Tours - France.

BACKGROUND: Toxoplasmosis is one of the most common parasitic infections in humans. It can establish chronic infection and is characterized by the formation of tissue cysts in the brain. The cysts remain largely quiescent for the life of the host, but can reactivate and cause life-threatening toxoplasmic encephalitis in immunocompromised patients, such as those with AIDS, neoplastic diseases and organ transplants. Toll-like receptor (TLR) adaptor MyD88 activation is required for the innate sensing of Toxoplasma gondii. Mice deficient in MyD88 have defective IL-12 and Th1 effector responses, and are highly susceptible to the acute phase of T. gondii infection. However, the role of this signaling pathway during cerebral infection is poorly understood and requires examination. METHOD: MyD88-deficient mice and control mice were orally infected with T. gondii cysts. Cellular and parasite infiltration in the peripheral organs and in the brain were determined by histology and immunohistochemistry. Cytokine levels were determined by ELISA and chemokine mRNA levels were quantified by real-time PCR (qPCR). RESULTS: Thirteen days after infection, a higher parasite burden was observed but there was no histological change in the liver, heart, lungs and small intestine of MyD88⁻/⁻ and MyD88⁺/⁺ mice. However, MyD88⁻/⁻ mice compared to MyD88⁺/⁺ mice were highly susceptible to cerebral infection, displayed high parasite migration to the brain, severe neuropathological signs of encephalitis and succumbed within 2 weeks of oral infection. Susceptibility was primarily associated with lower expression of Th1 cytokines, especially IL-12, IFN-γ and TNF-α, significant decrease in the expression of CCL3, CCL5, CCL7 and CCL19 chemokines, marked defect of CD8⁺ T cells, and infiltration of CD11b⁺ and F4/80⁺ cells in the brain. CONCLUSION: MyD88 is essential for the protection of mice during the cerebral installation of T. gondii infection. These results establish a role for MyD88 in T cell-mediated control of T. gondii in the central nervous system (CNS).

Youngyi Lee has completed her master from Woosuk University. Currently, she is a PhD student of Chonbuk National University Medical School. She has an interest in the pathogenesis of rheumatoid arthritis.

The type III histone deacetylase SIRT1 is an enzyme critical in the modulation of immune and inflammatory responses. However, data on its role in rheumatoid arthritis (RA) are limited and controversial. To better understand how SIRT1 regulates adaptive immune responses in RA, we evaluated collagen-induced arthritis (CIA) in myeloid cell-specific SIRT1 knockout (mSIRT1 KO) mice. CIA was induced in mSIRT1 KO and wild-type (WT) mice. Arthritis severity was gauged by clinical, radiographic, and pathologic scores. Levels of cytokines and transcription factors were determined by RT-PCR and ELISA. Flow cytometry was used to quantify T cells and dendritic cells (DCs). T-cell proliferation was also investigated when co-cultured with antigen-pulsed DCs. Compared with WT counterparts, arthritis in mSIRT1 KO mice was less severe and proved less destructive to joints. Expression levels of inflammatory cytokines, matrix metalloproteinases, and ROR-γT in mSIRT1 KO mice were also reduced by comparison, paralleled by reductions in Th1 and Th17 cells and CD80- or CD86-positive DCs. In addition, impaired maturation of DCs and resultant declines in Th1/Th17 immune response were observed in mSIRT1 KO mice. In co-culture, DCs of SIRT1 KO mice were less adept at driving T-cell proliferation and Th1/Th17 immune response. Myeloid cell-specific deletion of SIRT1 appears to suppress CIA through modulation of DC maturation. Our data suggest that careful investigation of DC-specific SIRT1 downregulation is needed to weigh the therapeutic utility of agents targeting SIRT1 in RA.

Laurence O’Neill is completing a Masters Research at the Institute of Technology, Carlow (IT Carlow). The project is funded by the Presidents’ Fellowship Award Scholarship Scheme at IT Carlow. He completed his undergraduate degree in biosciences with biopharmaceuticals (Honours) at IT Carlow.

Matrix metalloproteinase-2 and -9 are produced in arthritis conditions such as osteoarthritis and rheumatoid arthritis. Pro-inflammatory cytokines such as Interleukin-6 (IL-6) and Tumor necrosis factor-alpha (TNF-α) are known to be elevated, whereas anti-inflammatory cytokines such as Interleukin-10 (IL-10) and Interleukin-17 (IL-17) are lower. The balance between pro- and anti-inflammatory cytokines determines the severity of arthritis. This is due to the pro-inflammatory cytokines initiating a sequence of events which ultimately leads to joint destruction. Our research seeks to compare the levels of matrix metalloproteinases found in blood samples from arthritis patients to the levels obtained in blood from healthy volunteers. Using these results to then determine if a diagnosis can be made from the cytokine profile present in a patients’ samples. Gelatin zymography was carried out to determine the levels of Matrix metalloproteinases (MMPs) in blood and synovial fluid samples and the results were analysed using Image J. Enzyme-linked immunosorbent assay (ELISA) was also used to determine the levels of cytokines IL-1, IL-6, TNF-α and IL-17 in all the blood specimens.

Jae Young Kim has completed his PhD at the age of 32 years from Dongguk University and postdoctoral studies from Seoul National University College of Medicine, Seoul, Korea. He had served as a chairman of the Department of Biological Science, Gachon University, Incheon, Korea. He has written more than 25 papers in immunology and has been serving as a senior editor of Korean Society for Transplantation.

Toll-like receptors (TLR) are essential components of innate immune system. TLR5 is the receptor for flagellin, the principal protein component of bacterial flagella. TLR5 gene is known to have 6 exons. However, previous transcriptome data of TLR5 suggest the possible existence of an additional exon derived from the fourth intron. In this study, we have identified and sequenced the extra exon from the human TLR5 transcripts. The new human TLR5 exon consists of 94 nucleotides and is located between known exons 4 and 5 in the 5’-untranslated region (5’-UTR). Real-time PCR analysis of the two alternatively spliced variants of the human TLR5 transcripts in various immune and non-immune cells showed that the transcripts with the extra exon abundantly express in non-immune cells. Ratios of the transcripts with the extra exon to those without the extra exon of human non-immune cells, such as epithelial cells, glial cells, keratinocytes and mesenchymal stem cells, were in the range of 1.3 to 4.5. In contrast, those of immune cells including human monocytes and T cells were around 0.9. These ratios of human monocytes were decreased by the treatment of IFN-γ in a concentration dependent manner, while those were not changed by the IL-4 treatment. The ratio began to decline at 3 h, reach 60% of normal ratio at 6 h, and then return to normal at 24 h after the IFN-γ treatment. Our results reveal that expression levels of new splicing variant of TLR5 gene in monocytes were lower than those in non-immune cells and were significantly decreased by the treatment of IFN-γ, suggesting possible negative regulatory role of this extra exon of 5’-UTR in TLR5 protein expression of human monocytes.