Higher Expression Of CD86 Research Articles

Adenosine and the adenosineA2A receptor agonist, CGS21680, upregulate CD39 and CD73 expression through E2F-1 and CREB in regulatory Tcells isolated from septic mice.

The number of regulatory Tcells(Treg cells) and the expression of ectonucleoside triphosphate diphosphohydrolase1(ENTPD1; also known as CD39) and 5'-ectonucleotidase (NT5E; also known as CD73) on the Treg cell surface are increased during sepsis. In this study, to determine the factors leading to the high expression of CD39 and CD73, and the regulation of the CD39/CD73/adenosine pathway in Treg cells under septic conditions, we constructed a mouse model of sepsis and separated the Treg cells using a flow cytometer. The Treg cells isolated from the peritoneal lavage and splenocytes of the mice were treated with adenosine or the specific adenosineA2A receptor agonist, CGS21680, and were transfected with specific siRNA targeting E2F transcription factor1(E2F-1) or cyclic adenosine monophosphate(cAMP) response element-binding protein(CREB), which are predicted transcription regulatory factors of CD39 or CD73. The regulatory relationships among these factors were then determined by western blot analysis and dual-luciferase reporter assay. In addition, changes in adenosine metabolism were measured in the treated cells. The results revealed that adenosine and CGS21680 significantly upregulated CD39 and CD73 expression(P<0.01). E2F-1 and CREB induced CD39 and CD73 expression, and were upregulated by adenosine and CGS21680. Adenosine triphosphate(ATP) hydrolysis and adenosine generation were inhibited by the knockdown of E2F-1 or CREB, and were accelerated in the presence of CGS21680. Based on these results, it can be inferred that adenosine, the adenosineA2Areceptor agonist, E2F-1 and CREB are the possible factors contributing to the high expression of CD39 and CD73 on the Treg cell surface during sepsis. Adenosine and its A2Areceptor agonist served as the signal transducer factors of the CD39/CD73/adenosine pathway, accelerating adenosine generation. Our study may benefit further research on adenosine metabolism for the treatment of sepsis.

Enhanced Th1/Th17 Functions of CD161+ CD8+ T Cells in Mucosal Tissues of Rhesus Macaques.

Expression of the C-type lectin-like receptor CD161 by human T cells is associated with type-17 responses, which play critical regulatory roles in immunity and inflammation at mucosal sites. However, the functions of CD161-expressing T cells in macaques, the pre-clinical model of several human diseases, remain unknown. This study examined the phenotypic and functional characteristics of CD161+ T cells in peripheral blood, mucosal tissues and lymph nodes of rhesus macaques. Majority of CD161-expressing T cells in peripheral blood and lung/intestinal mucosal tissues of rhesus macaques were found to be CD8+CD4– in phenotype. There was a significant enrichment of CD161+CD8+ T cells in the lungs and colonic mucosa (16.1%±6.6 and 16.8%±5.7) in comparison to peripheral blood (4.2%±1.2) and mesenteric lymph nodes (1.3%±0.8). Regardless of the tissue compartment, CD161+CD8+ T cells mainly comprised of γδ T cells and TCR Vα7.2+ MAIT cells (up to 80%), and displayed Th1 and Th17 cytokine responses to mitogen stimulation. Mucosal CD161+CD8+ T cells were characterized by very high expression of CD69, a recent activation marker that is preferentially expressed on tissue resident cells. Furthermore, lung and colonic mucosal CD161+CD8+ T cells showed enhanced IFN-γ, IL-17, and Perforin production in comparison to those in blood. Thus, macaque CD161+CD8+ T cells represent mucosal tissue-homing innate-like CD8+ T-cell populations with Th1/Th17 type cytokine and cytotoxic effector functions that can potentially enhance the recruitment of adaptive immune cells and control initial pathogen burden/dissemination in tissues. Analysis of their role in early immune responses to mucosal pathogens will be valuable in the design of vaccines and therapeutics.

Regulation of CD40 signaling in colon cancer cells and its implications in clinical tissues.

CD40 is a member of the tumor necrosis factor receptor family. We reveal here a correlation between CD40 expression and colon cancer differentiation. Upon CD40 ligand (CD40L) binding, CD40/CD40L signaling inhibited colon cancer proliferation, induced apoptosis, stalled cells at G0/G1, and influenced cell adhesion and metastasis. Clustering analysis identified the elevation of aryl hydrocarbon receptor repressor (AHRR) expression along with activation of CD40/CD40L signaling. Examination of clinical specimens revealed that both AHR and AHRR levels correlated with colon cancer histological grade. In addition, high expression of AHRR was associated with high expression of CD40 in tumor cells, with CD40L expression being particularly high in the tumor interstitium. Real-time PCR and western blotting analysis showed that AHRR expression in colon cancer cells was up-regulated by CD40L binding. The likely mediating signaling pathways for the effects of CD40 are described herein.

Transcription Factor SP2 Enhanced the Expression of Cd14 in Colitis-Susceptible C3H/HeJBir.

Genetic analysis in the IL10-deficient mouse model revealed a modifier locus of experimental inflammatory bowel disease (IBD) on chromosome 18, with the allele of the strain C3H/HeJBir (C3Bir) conferring resistance and the allele of C57BL/6J (B6) conferring susceptibility. Differential Cd14 expression was associated with this background specific susceptibility to intestinal inflammation. Polymorphisms of the Cd14 promoter were found to be likely causative for strain specific expression, and Cd14-knockout mice revealed a protective role of this gene-product in experimental IBD. In this study, luciferase reporter assays confirmed an increased activity of the C3Bir derived Cd14 promoter compared to the one of B6. Promoter truncation experiments and site-directed mutagenesis in both strains resulted in reduced Cd14 promoter activity and confirmed that a central AP1 and the proximal SP1 transcription factor binding sites mediated the basal activity of the Cd14 promoter in the mouse. Moreover, a T to C exchange at position -259 replaced putative STAT1 and CDX1 sites in the Cd14 promoter from B6 by a SP2 site in C3Bir. Ablation of the Sp2 site through truncation was associated with a decreased promoter activity. Site-directed mutagenesis also demonstrated that the inactivation of SP2 led to a substantial loss of promoter activity in C3Bir. Performing electrophoretic mobility shift and supershift assays demonstrated interaction of SP2 with its potential binding site. In addition, retroviral—mediated overexpression of the SP2 transcription factor in primary bone marrow macrophages derived from C3Bir mice caused a significant increase in Cd14 transcription. These data characterized SP2 as important factor responsible for higher Cd14 expression and reduced IBD susceptibility mediated by the C3Bir allele.

Increased Treg cells and expression of CD39/CD73 during listeriosis in aging mice

Abstract Foodborne Listeria monocytogenes (LM) is a cause of serious illness &amp; death in the elderly. Aging is associated with increased susceptibility to pathogens. We investigated the cause of this increased susceptibility of aging mice to LM infection using an oral model. Young &amp; old C57BL/6 mice were gavaged with murinized LM and T-helper (Th) cell responses were evaluated. After infection, tissues were examined for bacterial burden &amp; histology. Isolated splenocytes were analyzed for intracellular Th1/Th2 cytokine expression (TNF-α, IL-10 and IFN-γ) &amp; distribution of Treg (CD4+CD25h) &amp; Th17 cells before &amp; after infection. The expression of adenosine producing enzymes, CD39/CD73, in Th cells was analyzed by FACS &amp; qRT-PCR. Old LM-infected mice lost more body-weight &amp; had a higher bacterial burden in the liver/spleen than young-infected mice. Uninfected old mice showed a higher baseline proinflammatory response when compared to young control mice. After infection, the proinflammatory cytokine, IFN-g, was higher in young mice as compared to old mice. An increase in Th-17 cells was observed in the older mice before &amp; after infection. Increased levels of IL-10 &amp; Treg cells were observed in older mice after infection. Higher expression of CD39/CD73 was observed in the Th cells of the older mice &amp; the Treg expression of CD39 doubled after infection, suggesting increased production of adenosine, an anti-inflammatory mediator. Increased LM infection in aging mice is associated with reduced IFN-g response &amp; increased anti-inflammatory responses from Treg cells with high expression of CD39 in splenocytes. Increased accumulation of splenic Th17 cells in LM-infected old mice can further contribute to increased inflammation leading to LM pathogenesis.

Defining the phenotype and transcriptome of IgG3+ B cells

Abstract Recent studies suggest that highly polyfunctional non-neutralizing HIV-specific IgG3 antibodies are associated with protection from HIV infection in the RV144 HIV vaccine trial. While it is well established that IgG3 Abs can drive a wide range of Fc-mediated innate effector functions, it remains unclear whether intrinsic factors are important for selection and maintenance of IgG3-secreting B cells and what extrinsic signals preferentially induce IgG3 class-switch. We hypothesized that (1) IgG3+ B cells will generally express enhanced levels of activation-associated surface proteins and markers important for interactions with T cells as compared to IgG1+ B cells and (2) IgG3+ B cells will have a unique transcriptome that helps prevent downstream class switch recombination. Our initial flow cytometry and RNA-sequencing (RNA-seq) analyses have focused on comparing IgG3+ memory B cells to IgM+ and IgG1+ memory B cells in HIV negative donors. We have found that IgG3+ B cells have higher expression of CD80 and CD95, but lower expression of CD40, than IgG1+ B cells. Preliminary bulk RNA-seq analyses similarly suggest that IgG3+ B cells have a unique transcriptome, with distinct expression of genes involved in transcriptional regulation, nucleosome assembly and chromatin organization. Interestingly, IgM+ and IgG1+ memory B cells share an overlapping transcriptional regulation cluster that may exist to prevent class-switch to IgG3 (from IgM) and induce class-switch to IgG1 (from IgG3). We are currently investigating the heterogeneity of resting and stimulated IgG3+ memory B cells at the single-cell level to better understand why some, but not all, IgG3+ memory B cells retain expression of IgG3.

Tumor-derived exosomes regulate expression of immune function-related genes in human T cell subsets.

Tumor cell-derived exosomes (TEX) suppress functions of immune cells. Here, changes in the gene profiles of primary human T lymphocytes exposed in vitro to exosomes were evaluated. CD4+ Tconv, CD8+ T or CD4+ CD39+ Treg were isolated from normal donors’ peripheral blood and co-incubated with TEX or exosomes isolated from supernatants of cultured dendritic cells (DEX). Expression levels of 24–27 immune response-related genes in these T cells were quantified by qRT-PCR. In activated T cells, TEX and DEX up-regulated mRNA expression levels of multiple genes. Multifactorial data analysis of ΔCt values identified T cell activation and the immune cell type, but not exosome source, as factors regulating gene expression by exosomes. Treg were more sensitive to TEX-mediated effects than other T cell subsets. In Treg, TEX-mediated down-regulation of genes regulating the adenosine pathway translated into high expression of CD39 and increased adenosine production. TEX also induced up-regulation of inhibitory genes in CD4+ Tconv, which translated into a loss of CD69 on their surface and a functional decline. Exosomes are not internalized by T cells, but signals they carry and deliver to cell surface receptors modulate gene expression and functions of human T lymphocytes.

Methylated SFRP1,2 and CD25 Expression in Acute Myeloid Leukemia Play an Important Role in the Pathogenesis of the Disease and in Turn in its Treatment

Objectives: Recently, hyperactivation of the Wnt signaling pathway has been implicated in leukomogenesis, so we studied the epigenetic dysfunction of SFRP1,2 and expression of interleukin2 receptor α chain (IL2Rα, also known as CD25) and its prognostic impact in acute myeloblastic leukemia (AML). Methods: We studied the methylation profile of SFRP1,2 in AML cells by methylation-specific polymerase chain reaction (MSP) and the hyper expression of IL2Rα (CD25) by flowcytometry. Results: We analyzed the methylation profile of SFRP1,2 in 40 de novo AML patients. The percentage of hypermethylation in the patient samples were 37.5% for SFRP1, 12.5% for SFRP2. CD25 was positive in 12(30%) of 40 patients AML. We found that in patients whom 60 years and younger with intermediate risk cytogenetics in de novo AML, hypermethylation of SFRP1 and CD25 were accompanied with relapse (P=0.024). Conclusion: Our data indicates that in a subgroup of AML patients, hypermethylation of SFRP1 and high expression of CD25 predict relapse

Prognostic Impact of Hypermethylated SFRP1, SFRP2 and Over Expression of CD25 in Acutemyeloid Leukemia

Objective: To study the epigenetic dysfunction of SFRP1, SFRP2 and expression of interleukin-2 receptor α chain (IL2Rα, also known as CD25) in acute myeloblastic leukaemia (AML). Methods: We study the methylation profile of SFRP1, SFRP2 in AML cells by methylation-specii¬c polymerase chain reaction (MSP) and the hyperexpression of IL2Rα (CD25) by flow-cytometry. Results: We analysed the methylation profile of SFRP1, SFRP 2 in 40 de novo AML patients. The percentage of hypermethylation in the patient samples were 37.5% for SFRP1 and 12.5% for SFRP2. CD25 was positive in 12 (30%) of 40 patients AML. In patients whom 60 years and younger with intermediate risk cytogenetics in de novo AML, hypermethylation of SFRP1 and CD25 were accompanied with relapse (P=0.024). Conclusion: Our data indicates that in a subgroup of AML patients, hypermethylation of SFRP1 and high expression of CD25 predict relapse.

CD28-mediated T cell response is upregulated by exogenous application of autologous Hsp70-peptide complex in a tumor-bearing host.

Hsp70, a highly conserved protein, has gained plenty of attention by virtue of its adjuvant capability to induce peptide-specific cytotoxic T lymphocyte responses. In this study, we have investigated the effect of autologous Hsp70-peptide complex (or simply autologous Hsp70) on the expression of CD28 on T cells and its effector functions through macrophage activation. Further, we investigated the effect of Hsp70 on the expression of CD80 and CD86 on macrophages isolated from normal and tumor-bearing host to provide costimulatory signal for T cell activation and secretion of IL-2 and IFN-γ during interaction. We found that treatment of autologous Hsp70 effectively activated TAMs to induce higher expression of CD28 on T cells through T cells-macrophage interaction. Treatment of autologous Hsp70 induces higher expression of CD80 and CD86 on TAMs, as a result, increases B7/CD28 interaction, which in turns activates T cells and induces higher production of IL-2 and IFN-γ, thereby increasing antigen-specific T cell proliferation. With our novel study, we have provided the strong insights into the role of extracellular Hsp70 on the expression of CD28 costimulatory molecule on T cells, which helps in the activation and generation of antigen-specific T cell effector functions in a tumor-bearing host to curb malignancy.

Heightened Expression of CD39 by Regulatory T Lymphocytes Is Associated with Therapeutic Remission in Inflammatory Bowel Disease

To evaluate whether changes in expression of CD39 by regulatory T lymphocytes (Treg) impact treatment response in inflammatory bowel disease. To then define the biological role of expression of CD39 on Treg in an animal model of colitis. A prospective study of consecutive patients commencing anti-tumor necrosis factor therapy with infliximab (IFX) or adalimumab (ADA), who were then followed for 12 months. Treatment responses were defined both symptomatically and by endoscopy showing mucosal healing. Peripheral blood Tregs were quantified by flow cytometry. Functional importance of CD39 expression by Treg was determined in an adoptive T-cell transfer model of colitis. Forty-seven patients (ulcerative colitis, n = 22; Crohn's disease, n = 25) were recruited; 16 patients were complete responders and 13 nonresponders to anti-tumor necrosis factor. CD39 expression by Treg was lower in active inflammatory bowel disease and increased significantly after treatment in responders (CD39Treg/total Treg; 8% at baseline to 22.5% at late time point, P < 0.001). Responders were more likely to have therapeutic drug levels and in multivariate analysis therapeutic drug levels were associated with higher expression of CD39 by FoxP3 Treg and lower frequencies of interleukin 17A expressing cells. Tregs with genetic deletion of CD39 exhibit decrements in potential to suppress intestinal inflammation in a murine (CD45RB) T-cell transfer model of colitis in vivo, when compared with wild-type Treg. Increased expression of CD39 by peripheral blood Treg is observed in the setting of clinical and endoscopic remission in inflammatory bowel disease. Deficiency of CD39 expression by Treg can be linked to inability to suppress experimental colitis.

IMPS-05CD40/CD40L EXPRESSION CORRELATES WITH THE SURVIVAL OF PATIENTS WITH GLIOBLASTOMAS AND AN AUGMENTATION IN CD40 SIGNALING PROLONGS SURVIVAL IN INTRACRANIAL GLIOMA- AND GLIOMA-INITIATING CELL-ISOGRAFTED TUMOR MODELS

BACKGROUND: The prognosis of glioblastomas remains poor; therefore, effective therapeutic strategies need to be developed. CD40 is a costimulatory molecule and its agonistic antibody have been shown to activate anti-tumor effects. Recently, CD40 has been extensively targeted for immunotherapeutic purposes. METHODS: The expression of CD40 and CD40L mRNAs were examined in 86 cases of grade IV glioblastomas and 36 cases of grade III gliomas and correlated with outcomes. CD40 signaling was employed to augment the efficacy of immunotherapy against gliomas. The efficacy of FGK45, dendritic cells, and OX86; an agonistic antibody for OX40 were examined by adding it to a tumor lysate-based subcutaneous vaccination against GL261 glioma model or NSCL61 glioma-initiating cell-like cell tumor model. RESULTS: We demonstrated for the first time using quantitative PCR that grade III gliomas expressed higher levels of CD40 and CD40L than grade IV glioblastomas. The higher expression of CD40 and CD40L was associated with good prognoses in patients with glioblastomas. Addition of FGK45 to the subcutaneous tumor cell lysate-based vaccination significantly prolonged survival in both tumor models. However, the efficacy was modest in NSCL61 model mice. Therefore, we established combination immunotherapeutic strategies using FGK45, dendritic cells, and OX86. We initially used a FGK45 and dendritic cells-based vaccination. The effects of the combination immunotherapy were not significantly different from those of the FGK45 immunotherapy. We then used a combination of FGK45 and OX86. Combination immunotherapy significantly prolonged survival with synergistic effects. Apoptosis increased and proliferation decreased in tumors treated with combination immunotherapy, especially when CD40 was stimulated. CONCLUSIONS: The high expression of CD40 and CD40L can be used as a biomarker for better prognoses in patients with gliomas. Immunotherapy using FGK45, dendritic cells, and OX86 significantly prolonged survival and represents a potential therapeutic strategy for gliomas including glioma-initiating cells.

IMPS-17BRANCHED-MULTIPEPTIDE IMMUNOTHERAPY FOR GLIOBLASTOMA USING HLA-A*0201-RESTRICTED CYTOTOXIC T-LYMPHOCYTE EPITOPE FROM BIRC5, CD99 AND ERBB2

BACKGROUND: Branched-multipeptide has the increased molecular weight of the peptide which can elevate stable immunogenicity in the body. We identified and synthesized BIRC5, CD99 and ERBB2 peptides and their corresponding branched-multipeptide for HLA-A*0201 molecules. We investigated the possibility of immunotherapy for glioblastoma using cytotoxic T lymphocytes (CTLs) generated by branched-multipeptides pulsed dendritic cells (DCs). MATERIALS AND METHODS: We selected three peptides (BIRC5, CD99 and ERBB2), on the basis of their binding affinity, as determined by a peptide-T2 binding assay and their corresponding branched-multipeptide was synthesized using mini-polyethylene glycol (mini-PEGs) spacer. The function of DCs pulsed with multipeptide- cocktail was compared with DCs pulsed with branched-multipeptide by immunephenotype and cytokine secretion. We measured the functional activity for CTLs by using IFN-γ-enzyme linked immune absorbent spot (ELISPOT) assay. RESULTS: The stability of branched-multipeptide-T2 cells binding was relatively well maintained up to 24 hours compared to multipeptide cocktail. Based on FACS analysis which compared the phenotypes of multipeptide-cocktail and branched-multipeptide pulsed DCs, both DCs exhibited high expression of CD80, CD83 and CD86, which meant that both DCs resulted in efficient generation of DCs without altering the phenotype of the mature DCs. The both multipeptides pulsed DCs showed similarly increased IL-12p70, IL-12p40and IL-10 production. The CTLs that were stimulated by both multipeptides pulsed DCs displayed a greater number of IFN-γ-secreting cells against T2 cells loaded with both multipeptides and glioblastoma cell lines compared to those stimulated by un-pulsed DCs. CONCLUSION: Branched-multipeptide of BIRC5, CD99 and ERBB2 showed the stable binding with T cells and similar cytotoxic activity to target cells compared with multipeptide-cocktail. Branched-multipeptide can be the candidate of useful immunotherapeutic modalities for glioblastoma. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (2014R1A1A1004469).

Antigenic Stimulation of Kv1.3-Deficient Th Cells Gives Rise to a Population of Foxp3-Independent T Cells with Suppressive Properties.

Multiple sclerosis (MS) is an immune-mediated demyelinating disease of the CNS that has been linked with defects in regulatory T cell function. Therefore, strategies to selectively target pathogenic cells via enhanced regulatory T cell activity may provide therapeutic benefit. Kv1.3 is a voltage-gated potassium channel expressed on myelin-reactive T cells from MS patients. Kv1.3-knockout (KO) mice are protected from experimental autoimmune encephalomyelitis, an animal model of MS, and Kv1.3-KO Th cells display suppressive capacity associated with increased IL-10. In this article, we demonstrate that myelin oligodendrocyte glycoprotein-specific Kv1.3-KO Th cells exhibit a unique regulatory phenotype characterized by high CD25, CTLA4, pSTAT5, FoxO1, and GATA1 expression without a corresponding increase in Foxp3. These phenotypic changes result from increased signaling through IL-2R. Moreover, myelin oligodendrocyte glycoprotein-specific Kv1.3-KO Th cells can ameliorate experimental autoimmune encephalomyelitis following transfer to wild-type recipients in a manner that is partially dependent on IL-2R and STAT5 signaling. The present study identifies a population of Foxp3(-) T cells with suppressive properties that arises in the absence of Kv1.3 and enhances the understanding of the molecular mechanism by which these cells are generated. This increased understanding could contribute to the development of novel therapies for MS patients that promote heightened immune regulation.

Crossing the placental barrier: Preferential migration of microchimeric cells from maternal blood into fetal organs in mice

as in the 1st trimester trophoblast cell line HTR-8/SVneo. Isolated blood monocytes where cultured and differentiated in the presence of IL-34 for six days. When analyzed with flow cytometry, they showed high expression of CD14, CD163 (scavenger receptor), CD206 (mannose receptor) and CD209 (DC-SIGN), markers expressed on decidual macrophages. Taken together, our results show that IL-34 is produced both by placental and decidual cells and is able to induce macrophages with a phenotype resembling that of decidual macrophages. The findings suggest a role for IL-34 in the regulation of decidual macrophages during early pregnancy.

Stress exposure during pregnancy affects placental 11β-hydroxysteroid dehydrogenase type 2 and maternal immune repertoire

as in the 1st trimester trophoblast cell line HTR-8/SVneo. Isolated blood monocytes where cultured and differentiated in the presence of IL-34 for six days. When analyzed with flow cytometry, they showed high expression of CD14, CD163 (scavenger receptor), CD206 (mannose receptor) and CD209 (DC-SIGN), markers expressed on decidual macrophages. Taken together, our results show that IL-34 is produced both by placental and decidual cells and is able to induce macrophages with a phenotype resembling that of decidual macrophages. The findings suggest a role for IL-34 in the regulation of decidual macrophages during early pregnancy.

Paternal lymphocyte immunization in women with recurrent embryo implantation failures and/or recurrent miscarriages after IVF

as in the 1st trimester trophoblast cell line HTR-8/SVneo. Isolated blood monocytes where cultured and differentiated in the presence of IL-34 for six days. When analyzed with flow cytometry, they showed high expression of CD14, CD163 (scavenger receptor), CD206 (mannose receptor) and CD209 (DC-SIGN), markers expressed on decidual macrophages. Taken together, our results show that IL-34 is produced both by placental and decidual cells and is able to induce macrophages with a phenotype resembling that of decidual macrophages. The findings suggest a role for IL-34 in the regulation of decidual macrophages during early pregnancy.

Regulatory T cells in ankylosing spondylitis and the response after adalimumab treatment

ObjectivesThe aim of this study was to investigate the role of regulatory T cells (Tregs) in ankylosing spondylitis (AS). MethodsWe included 69 AS patients (15 of them received anti-tumor necrosis factor-apha agent-adalimumab) in the study and used a questionnaire to record the demographic data, disease activity index, functional index, human leukocyte antigen-B27, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). Thirty healthy subjects were used as controls. The peripheral blood mononuclear cells (PMBCs) were stained with anti-CD4, anti-CD25 and anti-Forkhead/winged helix transcription factor P3 (anti-FoxP3) antibodies and flow-cytometry was used to determine cell populations. ResultsThe percentages of Tregs in PMBCs were significantly higher in AS patients than in healthy controls. In AS patients who had poor disease functional index with higher levels of ESR and CRP were positively and significantly correlated with Tregs percentages in PMBCs. After adalimumab treatment in 15 patients, the percentages of Tregs, the ESR/CRP levels and the Bath Ankylosing Spondylitis Disease Activity Index/Bath Ankylosing Spondylitis Functional Index were significantly and gradually decreased over time. ConclusionsThe high expression of FoxP3 and CD25 on CD4+ T cells in PBMCs in AS patients was noted, and could be reversed by adalimumab therapy. These findings suggest that Tregs may play a role in modulating the inflammatory process in AS. Whether Tregs can be taken as a predictor for disease activity or treatment outcome is unclear and requires further study.

Monocyte-derived dendritic cells from late gestation cows have an impaired ability to mature in response to E. coli stimulation in a receptor and cytokine-mediated fashion

During late gestation the bovine immune system is less capable of eliciting inflammatory responses and eliminating invading pathogens. The maternal immune system is directed toward tolerance in order to prevent fetal rejection due to recognition of paternal antigens. In humans and mice, dendritic cell (DC) populations maintain a tolerogenic phenotype essential in the generation and preservation of maternal immune tolerance throughout pregnancy. However, the primary mechanisms which facilitate maternal immune tolerance involved in bovine gestation remain poorly understood. In order to determine if DC phenotype and function were regulated toward tolerance during bovine gestation, we compared in vitro generated monocyte-derived DC (mo-DC) from monocytes isolated from cows in late gestation (LG) to those from non-pregnant (NP) cows in their ability to mature following stimulation with UV irradiated Escherichia coli. Our results show mo-DC from LG cows have an impaired ability to mature in response to E. coli stimulation in a receptor and cytokine-mediated fashion in comparison to those from NP cows. Specifically, mo-DC from LG cows were unable to upregulate MHC II and maintained high expression of CD14, both indicative of an immature phenotype following E. coli-stimulation. Only mo-DC from LG showed significant increase in IL-10 production and had a significantly lower ratio of production of the Th1-polarizing cytokine IL-12 to regulatory cytokine IL-10 following E. coli stimulation compared to mo-DC from NP cows. Our findings demonstrate mo-DC from LG cows have a stifled capacity to develop a mature phenotype and drive pro-inflammatory Th1-type responses to E. coli stimulation. Results from this study provide insight into DC immune modulation in bovine pregnancy and elucidate host factors which may contribute to the heightened susceptibility to infection in late gestation.

High expression of CD39/ENTPD1 in malignant epithelial cells of human rectal adenocarcinoma.

The ectonucleotidase CD39 is pivotal in the conversion of immunostimulatory adenosine triphosphate (ATP) into immunosuppressive adenosine which potently inhibits host immune responses against cancer. This study investigated the expression level and prognostic significance of CD39 in human rectal adenocarcinoma. Our data demonstrated that CD39 staining strongly marked malignant epithelial cells where the protein and messenger RNA (mRNA) expression levels of CD39 were significantly increased compared with paracancerous controls. In addition to primary tumors, CD39 was also abundantly expressed in liver metastases and tumor-draining lymph nodes from metastatic rectal adenocarcinoma. Although patients with higher CD39 density in tumor cells were more likely to have favorable characteristics (early TNM and N stages) and overall survival, the singular parameter cannot be used as an independent factor for predicting patients' prognosis. Intriguingly, combined analysis of CD39 and CD73 expression was more efficient to foretell patient's outcome where patients with increased CD73 but decreased CD39 levels displayed a worst prognosis. Taken together, the current study revealed that malignant epithelial cells of human rectal adenocarcinoma strongly express CD39 that may play a potential role in the tumor invasion and metastasis. Although high expression of CD39 in tumor cells is correlated with favorable clinical outcome, the combination of CD39 and CD73 expression may have a better prognostic value.