Yangsheng Yu

Microbial Infection and Rheumatoid Arthritis

Rheumatoid arthritis (RA) is a complex autoimmune disease affecting 1–2% of general worldwide population. The etiopathogenesis of RA involves the interplay of multiple genetic risk factors and environmental triggers. Microbial infections are believed to play an important role in the initiation and perpetuation of RA. Recent clinical studies have shown the association of microbial infections with RA. Accumulated studies using animal models have also found that microbial infections can induce and/or exaggerate the symptoms of experimental arthritis. In this review, we have identified the most common microbial infections associated with RA in the literature and summarized the current evidence supporting their pathogenic role in RA. We also discussed the potential mechanisms whereby infection may promote the development of RA, such as generation of neo-autoantigens, induction of loss of tolerance by molecular mimicry, and bystander activation of the immune system.

Autoantibodies From Single Circulating Plasmablasts React With Citrullinated Antigens and Porphyromonas gingivalis in Rheumatoid Arthritis

Anti–citrullinated protein antibodies (ACPAs) are highly specific for rheumatoid arthritis (RA). However, the molecular basis for ACPA production is still unclear. The purpose of this study was to determine if circulating plasmablasts from RA patients produce ACPAs and whether Porphyromonas gingivalis facilitates the generation of ACPAs.

Celastrol Inhibits Inflammatory Stimuli-Induced Neutrophil Extracellular Trap Formation

Neutrophil extracellular traps (NETs) are web-like structures released by activated neutrophils. Recent studies suggest that NETs play an active role in driving autoimmunity and tissue injury in diseases including rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). The purpose of this study was to investigate if celastrol, a triterpenoid compound, can inhibit NET formation induced by inflammatory stimuli associated with RA and SLE. We found that celastrol can completely inhibit neutrophil oxidative burst and NET formation induced by tumor necrosis factor alpha (TNFα) with an IC50 of 0.34 µM and by ovalbumin:anti-ovalbumin immune complexes (Ova IC) with an IC50 of 1.53 µM. Celastrol also completely inhibited neutrophil oxidative burst and NET formation induced by immunoglobulin G (IgG) purified from RA and SLE patient sera. Further investigating into the mechanisms, we found that celastrol treatment downregulated the activation of spleen tyrosine kinase (SYK) and the concomitant phosphorylation of mitogen-activated protein kinase kinase (MAPKK/MEK), extracellular-signal-regulated kinase (ERK), and NFκB inhibitor alpha (IκBα), as well as citrullination of histones. Our data reveals that celastrol potently inhibits neutrophil oxidative burst and NET formation induced by different inflammatory stimuli, possibly through downregulating the SYK-MEK-ERK-NFκB signaling cascade. These results suggest that celastrol may have therapeutic potentials for the treatment of inflammatory and autoimmune diseases involving neutrophils and NETs.

Galectins in the Pathogenesis of Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is a complex and common systemic autoimmune disease characterized by synovial inflammation and hyperplasia. Multiple proteins, cells, and pathways have been identified to contribute to the pathogenesis of RA. Galectins are a group of lectins that bind to β-galactoside carbohydrates on the cell surface and in the extracellular matrix. They are expressed in a wide variety of tissues and organs with the highest expression in the immune system. Galectins are potent immune regulators and modulate a range of pathological processes, such as inflammation, autoimmunity, and cancer. Accumulated evidence shows that several family members of galectins play positive or negative roles in the disease development of RA, through their effects on T and B lymphocytes, myeloid lineage cells, and fibroblast-like synoviocytes. In this review, we will summarize the function of different galectins in immune modulation and their distinct roles in RA pathogenesis.

Regulation of VH Replacement by B Cell Receptor–Mediated Signaling in Human Immature B Cells

VH replacement provides a unique RAG-mediated recombination mechanism to edit nonfunctional IgH genes or IgH genes encoding self-reactive BCRs and contributes to the diversification of Ab repertoire in the mouse and human. Currently, it is not clear how VH replacement is regulated during early B lineage cell development. In this article, we show that cross-linking BCRs induces VH replacement in human EU12 μHC+ cells and in the newly emigrated immature B cells purified from peripheral blood of healthy donors or tonsillar samples. BCR signaling–induced VH replacement is dependent on the activation of Syk and Src kinases but is inhibited by CD19 costimulation, presumably through activation of the PI3K pathway. These results show that VH replacement is regulated by BCR-mediated signaling in human immature B cells, which can be modulated by physiological and pharmacological treatments.

Effects of IL8 and immune cells on the regulation of luteal progesterone secretion

Recent studies have suggested that chemokines may mediate the luteolytic action of prostaglandin F2α (PGF). Our objective was to identify chemokines induced by PGF in vivo and to determine the effects of interleukin 8 (IL8) on specific luteal cell types in vitro. Mid-cycle cows were injected with saline or PGF, ovaries were removed after 0.5-4 h, and expression of chemokine was analyzed by qPCR. In vitro expression of IL8 was analyzed after PGF administration and with cell signaling inhibitors to determine the mechanism of PGF-induced chemokine expression. Purified neutrophils were analyzed for migration and activation in response to IL8 and PGF. Purified luteal cell types (steroidogenic, endothelial, and fibroblast cells) were used to identify which cells respond to chemokines. Neutrophils and peripheral blood mononuclear cells (PBMCs) were cocultured with steroidogenic cells to determine their effect on progesterone production. IL8, CXCL2, CCL2, and CCL8 transcripts were rapidly increased following PGF treatment in vivo. The stimulatory action of PGF on IL8 mRNA expression in vitro was prevented by inhibition of p38 and JNK signaling. IL8, but not PGF, TNF, or TGFB1, stimulated neutrophil migration. IL8 had no apparent action in purified luteal steroidogenic, endothelial, or fibroblast cells, but stimulated ERK phosphorylation in neutrophils. In coculture experiments neither IL8 nor activated neutrophils altered basal or LH-stimulated luteal cell progesterone synthesis. In contrast, activated PBMCs inhibited LH-stimulated progesterone synthesis from cultured luteal cells. These data implicate a complex cascade of events during luteolysis, involving chemokine signaling, neutrophil recruitment, and immune cell action within the corpus luteum.

Contribution of VH Replacement Products in Mouse Antibody Repertoire

VH replacement occurs through RAG-mediated recombination between the cryptic recombination signal sequence (cRSS) near the 3′ end of a rearranged VH gene and the 23-bp RSS from an upstream unrearranged VH gene. Due to the location of the cRSS, VH replacement leaves a short stretch of nucleotides from the previously rearranged VH gene at the newly formed V-D junction, which can be used as a marker to identify VH replacement products. To determine the contribution of VH replacement products to mouse antibody repertoire, we developed a Java-based VH Replacement Footprint Analyzer (VHRFA) program and analyzed 17,179 mouse IgH gene sequences from the NCBI database to identify VH replacement products. The overall frequency of VH replacement products in these IgH genes is 5.29% based on the identification of pentameric VH replacement footprints at their V-D junctions. The identified VH replacement products are distributed similarly in IgH genes using most families of VH genes, although different families of VH genes are used differentially. The frequencies of VH replacement products are significantly elevated in IgH genes derived from several strains of autoimmune prone mice and in IgH genes encoding autoantibodies. Moreover, the identified VH replacement footprints in IgH genes from autoimmune prone mice or IgH genes encoding autoantibodies preferentially encode positively charged amino acids. These results revealed a significant contribution of VH replacement products to the diversification of antibody repertoire and potentially, to the generation of autoantibodies in mice.

Accumulation of VH Replacement Products in IgH Genes Derived from Autoimmune Diseases and Anti-Viral Responses in Human

VH replacement refers to RAG-mediated secondary recombination of the IgH genes, which renews almost the entire VH gene coding region but retains a short stretch of nucleotides as a VH replacement footprint at the newly generated VH–DH junction. To explore the biological significance of VH replacement to the antibody repertoire, we developed a Java-based VH replacement footprint analyzer program and analyzed the distribution of VH replacement products in 61,851 human IgH gene sequences downloaded from the NCBI database. The initial assignment of the VH, DH, and JH gene segments provided a comprehensive view of the human IgH repertoire. To our interest, the overall frequency of VH replacement products is 12.1%; the frequencies of VH replacement products in IgH genes using different VH germline genes vary significantly. Importantly, the frequencies of VH replacement products are significantly elevated in IgH genes derived from different autoimmune diseases, including rheumatoid arthritis, systemic lupus erythematosus, and allergic rhinitis, and in IgH genes encoding various autoantibodies or anti-viral antibodies. The identified VH replacement footprints preferentially encoded charged amino acids to elongate IgH CDR3 regions, which may contribute to their autoreactivities or anti-viral functions. Analyses of the mutation status of the identified VH replacement products suggested that they had been actively involved in immune responses. These results provide a global view of the distribution of VH replacement products in human IgH genes, especially in IgH genes derived from autoimmune diseases and anti-viral immune responses.

Circulating plasmablasts from chronically human immunodeficiency virus-infected individuals predominantly produce polyreactive/autoreactive antibodies

Understanding the B-cell response during chronic human immunodeficiency virus (HIV) infection is essential for eliciting broad and potent neutralizing antibodies (Abs). In this study, we analyzed the plasmablast repertoire of chronically HIV-infected individuals in combination with antiretroviral therapy (ART). Among the obtained 72 recombinant monoclonal antibodies (mAbs), 27.8% weakly bound to HIV gp140 and were non-neutralizing. Remarkably, 56.9% were polyreactive and 55.6% were autoreactive. The prominent feature of being polyreactive/autoreactive is not limited to anti-gp140 Abs. Furthermore, these polyreactive/autoreactive Abs displayed striking cross-reactivity with DWEYS in the N-methyl-d-aspartate receptor (NMDAR), and this binding induced SH-SY5Y cell apoptosis. We also found higher frequencies of VH4-34 utilization and VH replacement in the plasmablast repertoire of chronically HIV-infected individuals, which may contribute to the generation of poly/autoreactive Abs. Taken together, these data demonstrate that circulating plasmablasts in chronically HIV-infected individuals experienced with ART predominantly produce poly/autoreactive Abs with minimal anti-HIV neutralizing capacity and potential cross-reactivity with autoantigens. This may represent another dysfunction of B cells during chronic HIV infection.

The accumulated VH replacement products contribute to the generation of auto/polyreactive antibodies in systemic lupus erythematosus (P1721)

Autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis and others affect millions of americans. The polymorphism rs2004640, found in the Interferon regulatory factor 5 (IRF5) promoter has been found to be a risk factor for several autoimmune diseases. We sought to define functional molecular effects due to this risk factor in cells from healthy individuals with the risk factor compared to those without. In cells with this polymorphism, we found >2-fold higher levels of IRF5 mRNA and protein (p<0.05). IRF5 is proapoptotic, so we expected higher levels of IRF5 would lead to increased apoptosis. Apoptosis levels were found to be 2.1 fold higher in risk cells (p=0.012). Since the polymorphism allows use of the IRF5 alternate first exon 1B, we analyzed four potential IRF5 promoters using ChIP-Seq data (ENCODE database) and the FactorBook database to define transcription factor binding sites. From this analysis, a p53 binding site was found only on the promoter for exon 1B. Further analysis measured the proportion of usage of each exon using qPCR. Usage of exon 1C was 2.3-fold lower (p=0.013) and 1D was 3.6-fold lower (p=0.017) in risk cells. We discovered 5 new splice variants of IRF5 mRNA, three of which used the exon 1B promoter. Each new variant was missing part of an instability domain, which may affect protein stability. These changes in expression, apoptosis and splicing help to explain how this polymorphism affects cellular and molecular processes and contributes to the development of autoimmune disease.T cells play a critical role in the pathogenesis of autoimmune diseases. Differentiation of Th17 cells requires the nuclear receptor RORγt, and inhibition of RORγt by small molecule inhibitors prevents Th17 cell differentiation in vitro. RORγt inhibitors also modulate RORγt activity in vivo as demonstrated by a significant reduction in the severity of experimental autoimmune encephalomyelitis. However, the effect of RORγt small molecule inhibitors on the in vivo generation of Th17 cells has not yet been clearly demonstrated. Here we report a novel, potent, and selective RORγt inverse agonist TMP778 and demonstrate its role in the in vivo differentiation of Th17 cells. TMP778 blocks both human and mouse Th17 cell differentiation in vitro. In vivo, administration of TMP778 to C57BL/6 mice immunized with myelin oligodendrocyte peptide 35-55 (MOG3555) blocked MOG-specific IL-17A expression. In addition, expression of Th17 signature genes (Il17a, Il17f, Il22, and CCL20) as well as Ifnγ was inhibited. Further, IL-17-producing CD4+ T cells were significantly reduced in IL-17-IRES-GFP transgenic mice administrated TMP778 after immunization with MOG35-55. Thus, our studies demonstrate that RORγt inverse agonist TMP778 suppress Th17 differentiation in vivo. Jianfei Yang, J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013H contacts (HHCs) of patients with tuberculosis (TB) are at higher risk of infection as well as the development of active disease. Longitudinal tracking of antigen-specific cytokines with the exposure may significantly help in understanding the dynamic changes in cytokine patterns associated with disease establishment. The aim of the present study is to investigate the role of candidate cytokines stimulated with r32-kDa antigen of M.bovis BCG (Ag85A-BCG) in Active Pulmonary Tuberculosis (APTB) patients, their HHC and Healthy Controls (HC) were assessed at 0, 4, 6, and 12 months after exposure. The in vitro T cell assays, ELISA of (Interferon gamma -IFN-γ), (Tumour Necrosis factor alpha-TNF-α) and (InterleukinIL-4) cytokines of culture supernatants were studied in APTB (n=50), HHC who were tuberculin-Purified Protein Derivative (PPD) skin test positive (n=50) and Healthy Controls (HC) n=50, were studied. The mean proliferative responses of stimulation cells was found to be significant between APTB patients at 4M, 6M, HHC at 4M compared to HC (2±0.55,1.061±0.508,0.99±0.3381.518±0.909 at p<0.02,0.01&0.03) respectively. The mean IFN-γ levels were significantly low in Pts/HHC compared to HC (24.5±19.8/33.55±25.44&107.5±59.0)at p<0.0001. APTB Pts at 0M compared to 4M &6M (24.5±19.8,64.13±43.88&81.97±56.22) at p<0.013&0.0008 and HHC at 0M compared to 4M (33.55±25.44&71.6±56.46) at p<0.031.TNF-α levels were significantly low in Pts at 0M compared to 12M(22.82±20.71&40.83±19.09) at p<0.02. IL-4 levels were significantly high at 0M in Pts and HHC compared to HC (7.46±5.826, 7.495±5.568&5.405±4.639) at p<0.01, 0.007. The ratios for IFN-γ/IL-4 and TNFα/IL-4 were significantly increasing from 0-6 months. Five APTB pts were turned into relapse during the follow up, 2 at 4th month and 3 at 6th month Whereas in HHC IFN-γ & IL-4 levels were decreasing during the follow up from 0-6 months. One of HHC developed the disease at 4th month of follow up. r32Kda M.bovis BCG antigen seems to be immunogenic, stimulating protective immune responses which may help us to assess the treatment outcome. In conclusion, it is important that there is a need to elucidate the mechanism of Th1 and Th2 responses in understanding recurrent TB or susceptibility to disease and also help in identifying the HHC by monitoring their immunological status who are at high risk. Suman Latha G et al., J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013I immune system acts as the first-line of host defense against infection by microbes to activate anti-microbial responses for elimination of invading pathogens. Recent rapid progress in studies on innate immunity has facilitated the identification of various pattern recognition receptors (PRRs), which sense pathogen-derived molecular patterns (PAMPs) and evoke robust innate immune responses through the gene induction of proinflammatory cytokines and type I interferons (IFNs). Particularly, during viral infection, virus-derived nucleic acids are mainly recognized as viral PAMPs by nucleic acid sensors. Among these, a DExD/H-box RNA helicase RIG-I, is known to be a key cytosolic RNA sensor that has an important role in triggering responses to many viruses, such as influenza A virus, measles virus, hepatitis C virus, most of which are causative agents for infectious diseases in human. Recently, our group has identified the poly(ADP-ribose) polymerase-13 (PARP-13) shorter isoform “ZAPS” as a potent stimulator of the RIG-I-mediated signaling. ZAPS interacts with RIG-I to promote its activity, leading to robust activation of IRF-3 and NF-kappaB transcription factors. We also found that ZAPS contributes to RIG-I-mediated antiviral activity during influenza virus infection. On the other hand, such innate immune signals are often targeted by viral proteins to evade host immune system. In our current study, we demonstrate that ZAPS is involved in innate immune evasion of influenza virus. Our data indicate that influenza virus expresses a viral protein to competitively inhibit the interaction of ZAPS with RIG-I. In addition, a study with ZAPS transgenic mice also revealed an important role of ZAPS in antiviral defense against influenza virus in vivo. These results may provide a therapeutic insight for the control of viral infection. Akinori Takaoka, J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013T forkhead box P3 (FOX P3) is a major transcriptional regulatory factor required for the development and functioning of T-regulatory cells (CD4+, CD25+, T-cells). Through interaction with the nuclear factor of the activated T-cells (NFAT), the FOX P3 molecule controls the functioning of the regulatory T-cells. In this presentation we analyze the association of two Single Nucleotide Polymorphisms (SNPs) in the promoter region of the FOX P3 gene -3279C/A and -2383C/T with Graves disease and Hashimoto’s thyroiditis respectively. An important feature of these two autoimmune diseases is that a predominant Th2 type of immune response to thyroid antigens (eg. Anti-TSHR antibodies) is reported in Graves disease contrary to a predominant Th1 response in Hashimoto’s thyroiditis. This indicates a profound influence of the genotypes of -3279C/A and -2383C/T SNPs of FOX P3 gene in determining the pattern of immune response (Th1 / Th2). The role of the above mentioned polymorphisms in affecting clonal expansion/functional efficiency of T regulatory cells and thus resulting in diverse patterns of immune response in autoimmune thyroid disorders will be discussed in combination with genotypes of certain proand anti-inflammatory cytokines. Ethnic differences in the association of genotypes of these SNPs and variations in the frequencies of their genotypes / alleles will also be discussed. Mohammed Ishaq, J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013A of sunshine a day keeps gestational diabetes away? It is established that vitamin D the sunshine vitamin, specifically the active metabolite 1,25(oH)(2)D is involved in homeostasis of bone structure-its mineralization and formation and also the metabolism of calcium and phosphorus. However it is emerging that this metabolite also has effect on other important physiological processes in the body at a cellular level including autoimmune, and also, appear to play a role on the sensitivity of insulin. This report will attempt to evaluate this assumption as well as the notion or possibility of incorporating as part of routine antenatal care in pregnant women in particular those at risk of gestational diabetes. The report will discuss current practices research not only discuss current practices , relevant research as well as metabolic, physiological, biochemical basis Susan Okafor, J Diabetes Metab 2013, 4:6 http://dx.doi.org/10.4172/2155-6156.S1.022Purpose: The nature of heightened endotoxin sensitivity state observed in Familial Mediterranean fever (FMF) at present remains unknown. To assess the possibility that IL-10 plays a role in setting inflammatory threshold we studied the IL-10 production by monocytes and dendritic cells and endotoxin tolerance induction in FMF patients. Methods: 46 attack free FMF patients included in this study. The production of IL-10 by NLRor TLR-agonists stimulated monocytes and dendritic cells assayed either by conventional ELISA and flow cytometry. Versatility of monocytes studied by measuring the production of IL-10 and IL-1β after stimulation by proand anti-inflammatory agents, and after stimulation arrest or a further counter stimulation. Monocyte endotoxin tolerance and cross-tolerance induction assayed by measuring the production of IL-1β, IL-10, TNF-α and IFN-γ after pre-stimulation by NLRor TLR-ligands and after re-stimulation with LPS. Results: In FMF patients we observed down-regulation of circulating CD36+ peripheral blood lymphoid cells but not monocytes, constitutively producing IL-10. The production of IL-10 by TLRand NLR-agonists stimulated monocytes and dendritic cells is declined in FMF patients. Monocytes isolated from FMF patients failed to switch from a pro-inflammatory activated state to antiinflammatory phenotype and still produce IL-1β but not IL-10, which cause impaired endotoxin tolerance and cross-tolerance induction. The IL-10 production and endotoxin tolerance induction by monocytes and dendritic cells restored by NOD2ligand MDP and colchicine treatment. Conclusion: Reduced IL-10 production associated with impaired setting of feedback inhibition of inflammatory response and caused impaired resolution of inflammation and endotoxin tolerance induction. Tigran K. Davtyan, J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013I has been long known that cytokines play important role in cancer initiation, development and progression. Until recently, the lack of suitable high-content and high-throughput approaches has hindered progress in understanding the role of complex cytokine networks in cancer. Since their introduction in 2001, cytokine antibody arrays, which have the ability to detect hundreds of cytokines simultaneously in highly sensitive, high-throughput format, significant advances have been made in our understanding of the role of tumor-stromal interactions and chronic inflammation in tumorigenesis, angiogenesis and metastasis, as well as the elucidation of several cytokine-driven mechanisms of cancer drug resistance in tumor cells. This presentation will discuss cytokine antibody array technologies and their recent contributions toward a better understanding of the mechanisms of cancer hallmarks. Ruo-Pan Huang, J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013T rodent T-cell-dependent antibody response (TDAR) assay is used to assess the effect of candidate therapeutic agents on the immune system by measuring primary and secondary IgM and IgG antibody responses to exogenous antigen challenge. TDAR responses require intact function of multiple immune cells including antigen presenting cells and T and B lymphocytes, as well as a cytokine-dependent isotype class switch from IgM to IgG, resulting in production of an antigen-specific antibody response. Alterations in the amount of antibody produced therefore can reflect effects on any or all of the cell populations involved in TDAR. TDAR is commonly used in preclinical drug development especially where increased cause for concern exists (ICH guideline S8). Development of combination therapy that engages multiple targets impacting the immune system poses unique opportunity for increased efficacy but also unique risk for increased immunotoxicity. For this work, a mouse TDAR evaluating primary and secondary KLH antibody responses in a KLH-Specific IgM and IgG sandwich enzyme-linked immunosorbent assay (ELISA) was used to assess immunotoxicologic potential of multiple single and combination therapies. Combination therapy did not result in enhanced TDAR immunotoxicity compared to single therapy alone for the molecules evaluated. Tier 1 immunotoxicology assessments similar to those in standard toxicity studies were added to the TDAR assessment. Together, our data support the use of the TDAR assay for early safety assessment of potential combination therapies. Amy L. Volk, J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013P is a common mechanism by which the mediators of innate immunity can fight off microbial infections. Our ongoing research demonstrates that human herpesviruses may have evolved a common mechanism to avoid phagocytic degradation by exploiting a phagocytosislike entrapment to gain entry into the cells of the eye. The three representative herpesviruses included in our study are herpes simplex virus-1 (HSV-1) that causes corneal keratitis, cytomegalovirus (CMV), which is associated with retinitis in immunocompromised individuals and the third herpesvirus, human herpesvirus-8 (HHV-8), is related to the pathogenesis of Kaposis sarcoma, a common AIDS-related tumor of eyelid and conjunctiva. Using laser scanning confocal microscopy imaging, we have found that successful infection of ocular cell types by all the three viruses, belonging to three divergent subfamilies of herpesviruses, is facilitated by induction of F-actin rich membrane pseudopods. Inhibitors of F-actin polymerization and pseudopod formation, cytochalasin D and latrunculin B, show strong efficacy in stopping the infection by all three viruses. We also found that an identical inhibition was seen by preventing phosphoinositide 3 kinase signaling, which is required for the phagocytosis of microbes. Transmission electron microscopy data using human corneal fibroblasts for HSV-1, human retinal pigment epithelial cells for CMV, and human conjunctival epithelial cells for HHV-8 provide further support to our argument that pseudopod-like membrane protrusions facilitate virus uptake by the ocular cells. Our findings suggest a novel mechanism by which the nonprofessional mediators of phagocytosis can be infected by human herpesviruses. Deepak Shukla, J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013I regulatory factor 5 (IRF5)is proapoptotic and has polymorphisms that confer risk for autoimmune disease. IRF5 has four first exon choices: 1D, 1A, 1B, and 1C respectively and the promoter region of exon 1A contains a CGGGG indel which is a risk factor for: Systemic lupus erythematosus, Sjogrens disease, Multiple Sclerosis, Crohns disease, and ulcerative colitis as determined by genome wide association studies (GWAS). The presence of this indel changes transcription factor binding in the promoter and correlates with higher levels of IRF5 as transcription levels are dependent on a gene’s first exon. Additionally, based on mRNA testing the 1A exon is also translated most efficiently. Thus the 1A risk indel may contribute to an increase in apoptosis through upregulation of IRF5 by increased transcription and efficient translation. To determine which transcription factors would bind to IRF5’s promoter, we used ChIP-sequencing data (ENCODE database).This data was used in conjunction with the FactorBook database to define where conserved genomic sequences where transcription factors are likely to bind in the promoter regions of IRF5. Exons 1A and 1D were found to contain putative PU.1 and NFkB binding sites. IRF5’s four promoters were cloned into luciferase plasmids to determine promoter activity for each exon using immune and epithelial cells. Imiquimod, a Toll-like receptor 7 ligand, and etoposide were used to activate the promoters. As a result of imiquimod treatment IRF5 levels were doubled (p<0.001), due to increased expression of exon 1A (2.2 fold, p=0.03) and 1D (2.8 fold, p=0.03), although when untreated there is less exon 1D usage in cells with the risk indel. IRF5 itself is a transcription factor for CCR7, one of its downstream targets, yet the risk indel is more likely to turn down CCR7 which suggests that it may be acting as a repressor. Understanding polymorphisms in the promoter regions of IRF5 and the effect that transcription factors play in the activity of these promoters is critical to determining how IRF5 is regulated in those with risk factors for autoimmune disease. Rodney Till, J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013A of innate immune components such as Toll-like receptor (TLR)s during diseases such as infectious diseases, cancer, metabolic diseases and liver disorders are associated with disruptions in metabolism and clearance of drugs. Impairment of drug metabolism/clearance can cause adverse effects of drugs. Thus, understanding the mechanism by which TLRs disrupt drug metabolism is important to develop effective strategies to prevent undesirable effects of drugs in patients with activated immune responses. Drug disposition is regulated by drug metabolizing enzymes (DMEs) and transporters primarily present in the liver. We have previously shown that activation of TLRs reduce the expression of DME/transporter genes. Expression of these genes is regulated by basal transcription factors as well as by nuclear receptors which bind to the promoters of these genes. We hypothesize that activation of TLRs recruit downstream adaptor proteins, leading to induction of cell-signaling components (c-Jun N-terminal kinase (JNK) and NF-κΒ) to reduce the expression of nuclear receptors leading to reduced expression of DMEs/transporter genes. We determined the role of the bacterial receptors, TLR2 (for gram-positive bacteria) and TLR4 (for gram-negative bacteria), their downstream adaptor proteins (TIRAP and TRIF), cell-signaling components and nuclear receptors in the regulation of DMEs/transporters. Pharmacokinetc and pharmacodynamic studies with clinically-relevant medications were also performed. These studies establish the role of innate immune components as novel regulators of drug metabolism/ clearance. These regulators can be targeted to prevent or reduce the undesirable effects of drugs in patients. Romi Ghose, J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013Identifying the immune correlates of protection is a key aspect of developing vaccines against infectious pathogens. We designed a universally applicable strategy to profile the antibody (Ab) repertoire of protected vaccine recipients, using recombinant phages encoding random peptide libraries. The novel approach, termed “protection-linked (PL) biopanning,” uses subtractive biopanning and probes Ab paratopes of protected vaccinees (“winners”) versus those with vaccine failure (“losers”). As proof-of-concept, we screened plasma samples from vaccinated macaques that had completely resisted multiple mucosal challenges with CCR5-tropic simian-human immunodeficiency viruses (SHIVs). The animals had been immunized with multimeric HIV-1 gp160 (Env), HIV-1 Tat, and SIV Gag-Pol particles. PL biopanning yielded a panel of approximately 100 recombinant phages that specifically reacted with Abs from “winners” and partially protected macaques but not Abs from “losers” or non-vaccinated, virus-challenged controls. Only about half of the phagotopes selected revealed amino acid homologies with HIV-1 Env (Env mimotopes). Expanding the sequence analysis to include non-Env immunogens revealed an unexpected additional humoral correlate of protection: the neutralizing epitope of HIV-1 Tat. Functional assays confirmed the induction of neutralizing anti-Tat Abs in macaques with full or partial protection but not in those with vaccine failure. Our data suggest that Tat should be included in multicomponent HIV-1 vaccines – and highlight the power of the new PL-biopanning strategy to identify Ab responses with significant association to vaccine protection, regardless of the mechanism(s) or targets of the protective Abs. PL biopanning is unbiased with regard to pathogens or disease model, making it a universal tool. Biography: Ruprecht obtained her Ph.D. in Human Genetics from Columbia University, NY, and her M.D. from the University of Miami, FL. After training in internal medicine at UCLA and hematology-oncology at the Memorial Sloan-Kettering Cancer Center, NY, she joined the Dana-Farber Cancer Institute and Harvard Medical School, where she became a tenured professor of medicine. She recently joined the Texas Biomedical Research Institute/Southwest National Primate Research Center, where she plans to expand the AIDS Research Program. She is internationally known for her studies on AIDS vaccine development and lentiviral pathogenesis in primate models, especially mucosal virus transmission and its prevention.C of ligand-engaged cytotoxic T lymphocyte antigen-4 (CTLA-4) to the T cell receptor (TCR) during the early phase of T cell activation attenuates TCR signaling, leading to T cell inhibition. To promote this event, a bispecific fusion protein comprising a mutant mouse CD80 (CD80w88a) and lymphocyte activation antigen-3 was engineered to concurrently engage CTLA-4 and crosslink it to the TCR. Crosslinking is expected to be attained via ligation of CTLA-4 first to MHCII and then indirectly to the TCR, generating a CTLA-4-MHCII-TCR tri-molecular complex that forms between T cells and antigenpresenting cells during T cell activation. Treating T cells with this bispecific fusion protein in vitro inhibited T cell activation, induced production of IL-10 and TGF-β, and attenuated AKT and mTOR signaling. Intriguingly, the bispecific fusion protein also directed early T cell differentiation into Foxp3 positive regulatory T cells (Tregs) in a process that was dependent on the endogenous production of TGF-β. Treatment of non-obese diabetic (NOD) female mice between 4-13 weeks of age with the bispecific protein significantly delayed the onset of disease or protected animals from developing autoimmune diabetes. Thus, bispecific fusion proteins that engage CTLA-4 and co-ligate it to the TCR during the early phase of T cell activation can negatively regulate the T cell response at multiple levels. Bispecific biologics with such dual functions may therefore represent a novel class of therapeutics for immune modulation. These findings presented here also reveal a potential new role for CTLA-4 in Treg differentiation. Yunxiang Zhu et al., J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013Objective: To analyze the risk factors affecting postoperative infectious complications in HIV-infected patients and explore the rational use of perioperative antibiotics. Methods: Retrospective analysis of 308 HIV-infected patients (male 272, female 36) who were operated at the Shanghai Public Health Clinical Center from Nov 2008 to Apr 2012. The patients were divided into postoperative infection and non-infection groups. Age and clinical variables were compared. The correlation between the method of surgical incision, surgical site infection (SSI) and postoperative sepsis were analyzed. Prophylactic antibiotics were used for patients with type I and II surgical incisions for no longer than 2 days. Patients with type III surgical incisions were administered antibiotics until infection was controlled. Antiretroviral therapy (ART) were used before operation for patients whose preoperative CD4 counts were <350 cells/μL,For those patients whose preoperative CD4 counts were <200 cells/μL, Sulfamethoxazole and fluconazole were administered preoperatively as a prophylaxis against Pneumocystis carinii pneumonia and fungal infection. Results: 196 patients developed postoperative infectious complications with 7 mortalities. Preoperative CD4 counts, ratio of CD4/ CD8 cells, hemoglobin level, and postoperative CD4 counts, hemoglobin and albumin levels were risk factors of perioperative infection in HIV-infected patients. Patients with a preoperative CD4 count < 200cell/μl, anemia, or a postoperative CD4 count < 200cell/μl and albumin levels < 35g/L correlated with a higher rate of postoperative infection. There was a significant correlation between SSI and the type of surgical incision. The rate of SSI in patients with type I surgical incision was 2% and the patients with type II surgical incision was 38%. All the patients who received type III surgical incisions developed SSI. Patients with SSI were more likely to develop postoperative sepsis. Conclusions: HIV-infected patients are more likely to develop postoperative infectious complications. Rational use of antibiotics in HIV-infected patients could help to reduce the rate of postoperative infectious complications in these patients. Baochi Liu et al., J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013N Diabetes after Transplantation (NODAT) is a serious metabolic complication after kidney transplantation, associated with poorer patient and graft survival. This study aimed to determine the incidence of NODAT and to identify risk factors for development of NODAT among kidney transplant recipients. Materials and Methods: This is an observational study of 401 patients who were transplanted between Jan 2000Jan 2008 at Mahavir hospital. NODAT was determined using criteria as per American Diabetes Association/ WHO guidelines. Logistic regression analyses were performed to identify predictors of NODAT. Results: Among 401 patients included in the study NODAT was observed in 59 patients (14.7%) after a mean follow-up time of 58.7 months. The mean age of the patients was 39.7±9.1 years and 69.1% were men and 30.1% were women; 26.9% received cyclosporine (CsA) and 73.1% tacrolimus (Tac). The incidence of NODAT was 2.74%, 4.98% and 6.98% at 1, 3, and 5 years following transplantation. In CsA-treated patients, the NODAT was 5.5% and 8.4% at 1and 5year post-transplantation, while in Tac-treated patients, it was 15.6% and 19.8%. Risk factors of NODAT were older age, use of CNIs, acute rejections and viral infections like HCV and CMV. Conclusion: We conclude that high incidences of NODAT are associated with the type of initial maintenance immunosuppression, acute rejections, obesity, hepatitis C infection and family history of diabetes. It is one of factor of graft failure and mortality. Sailaja kesiraju et al., J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013C molecules are targeted for immune intervention in inflammatory and autoimmune diseases. Among the costimulatory molecules, CD2-CD58 molecular pairs provide the adhesion between T cell and antigen presenting cell for signaling in the very early stage of immune response. The protein-protein interaction (PPI) between CD2 and CD58 (CD48 in rodents) helps enhance T cell-antigen-presenting cells (APC) adhesion and thus promotes T-cell activation. Studies related to structure of proteins indicated that PPI surfaces have “hot-spots” that are hydrophobic in nature and PPI can be modulated by targeting small molecules or peptides. Our strategy is to use conformationally constrained peptides to inhibit proteinprotein interactions in the key regions of the CD2-CD58 interactions. The result of blocking CD2-CD58 interaction will lead to suppression of T-cell activation and is clinically important for the treatment of autoimmune diseases. We have designed cyclic peptides and peptidomimetics from the surface epitopes of CD2 protein to inhibit CD2-CD58/CD48 interaction ultimately resulting in immunomodulation. The cell adhesion inhibition activity of the designed peptides (compound 6 and 7) were studied by cell adhesion assay. Compounds 6 and 7 inhibit the cell adhesion with an IC50 value of 6 and 11 nM respectively. Antibody binding inhibition assay indicated that these compounds bind to cells expressing CD58/CD48 proteins. Compounds were evaluated for their ability to modulate the immune response in collagen induced arthritis model (CIA). Results suggested that both the compounds were able to suppress the progression of arthritis in CIA model and were able to suppress T cell response in transgenic animal model. Seetharama Satyanarayanajois et al., J Clin Cell Immunol 2013, 4:5 http://dx.doi.org/10.4172/2155-9899.S1.013