Long chikungunya?An overview to immunopathology of persistent arthralgia

Chikungunya fever(CF)is caused by an arbovirus whose manifestations are extremely diverse,and it has evolved with significant severity in recent years.The clinical signs triggered by the Chikungunya virus are similar to those of other arboviruses.Generally,fever starts abruptly and reaches high levels,followed by severe polyarthralgia and myalgia,as well as an erythematous or petechial maculopapular rash,varying in severity and extent.Around 40%to 60%of affected individuals report persistent arthralgia,which can last from months to years.The symptoms of CF mainly represent the tissue tropism of the virus rather than the immunopathogenesis triggered by the host's immune system.The main mechanisms associated with arthralgia have been linked to an increase in T helper type 17 cells and a consequent increase in receptor activator of nuclear factor kappa-Βligand and bone resorption.This review suggests that persistent arthralgia results from the presence of viral antigens post-infection and the constant activation of signaling lymphocytic activation molecule family member 7 in synovial macrophages,leading to local infiltration of CD4+T cells,which sustains the inflammatory process in the joints through the secretion of pro-inflammatory cytokines.The term"long chikungunya"was used in this review to refer to persistent arthralgia since,due to its manifestation over long periods after the end of the viral infection,this clinical condition seems to be characterized more as a sequel than as a symptom,given that there is no active infection involved.

Influence of REV and ALV-J Co-Infection on Immunologic Function of T Lymphocytes and Histopathology in Broiler Chickens

The aim of present investigation is to study the effect of single- and co-infection with REV and ALV-J on T lymphocytes bioactivities and histopathology in broiler chickens. The bioactivities of blood and spleen T lymphocytes including lymphoproliferation responses, cytotoxicitic responses, and histopathology of spleen were detected in broiler chickens singly- or co-infected with REV and ALV-J at different days post inoculation and the virus expressions in spleen of infected broiler chickens were detected with immunofluorescence assay （IFA）. The results indicated that blood and spleen T lymphocytes proliferation responses and cytotoxicity in broilers infected with REV or/and ALV-J were inhibited in the whole observed period compared with controls. In the co-infected chickens they were highly inhibited than in the single-infected. The histopathology of spleen in infected chickens at 17 and 37 d post inoculation （dpi） indicated that cell interium increased, the numbers of lymphocytes decreased, and the regrowth were destroyed or decreased, especially more significantly at 17 than at 37 dpi. The different numbers of virus were detected in spleen lymphocytes in REV- infected and/or ALV-J-infected chickens. In the spleen of co-infected chicken, both REV and ALV-J were detected and the total numbers of viruses were more than in chickens singly-infected with REV or ALV-J. Thus, the co-effect of REV and ALV-J caused more immunosuppression on T lymphocytes bioactivities in broiler chickens than single-effect of ALV-J or REV, which contributed to the sever histopathology and the product of tumor cells. This study will be helpful for understanding the effect of co-infection with many viruses and control them in poultry.

Correlation Analysis on Total Lymphocyte Count and CD4 Count in HIV-infected Patients: A Retrospective Evaluation

CD4 count is the standard method for determining eligibility for highly active antiretroviral therapy （HAART） and monitoring HIV/AIDS disease progression, but it is not widely available in resource-limited settings. This study examined the correlation between total lymphocyte count （TLC） and CD4 count of HIV-infected patients before and after HAART, and assessed the thresholds of TLC for making decisions about the initiation and for monitoring HAART. A retrospective study was performed, and 665 HIV-infected patients with TLC and CD4 count from four counties （Shangcai, Queshan, Shenqiu and Weishi） were included in the study. Pearson correlation and receiver operating characteristic （ROC） were used. TLC and CD4 count after HAART was significantly increased as compared with pre-HAART （P〈0.01）. An overall positive correlation was noted between TLC and CD4 count （pre-HAART, r=0.73, P=0.0001; follow-up HAART, r=0.56, P=0.0001）. The ROC curve between TLC and CD4 count showed that TLC ≤ 1200 cells/mm3 could predict CD4 〈 200 cells/mm3 with a sensitivity of 71.12%, specificity of 66.35% at pre-HAART. After 12-month HAART, the optimum prediction for CD4 count 〈 200 cells/mm3 was a TLC ≤ 1300 cells/mm3, with a sensitivity of 63.27%, and a specificity of 74.84%. Further finding indicated that TLC change was positively correlated to CD4 change （r=0.77, P=0.0001） at the time point of 12-month treatment, and the best prediction point of TLC change for CD4 increasing was 135 cells/mm3. TLC and its change can be used as a surrogate marker for CD4 count and its change of HIV-infected individuals for making decisions about the initiation and for monitoring HAART in resource-limited settings.

IN VITRO ANTITUMOR ACTIVITY OF TUMOR-INFILTRATING LYMPHOCYTES FROM HUMAN GASTRIC CARCINOMA

Tumor-infiltrating lymphocytes (TIL) isolated from 11 gastric carcinoma were studied. TIL could grow for a long-term in medium containing recombi-nant interleukin-2(rlL-2). The mean expansion fold achieved in 6 long-term cultures of 11 specimens was 15.1. RIL-2 expanded gastric TIL exhibited significant cytotoxicity against K562, BGC823, MCF-7 and more effective antitumor cytotoxicity against fresh autologous tumor targets and human gastric cancer cell line. Peak cytotoxicity was shown in the third or fourth week after cultures. Cryopreservation of gastric TIL didn't influence their expansion capacity and antitumor activity. Phenotypic analysis was demonstrated in this study. The results of present study indicate that TIL from human gastric carcinoma could be expanded and reach high levels of antitumor effector function in long-term cultures with rIL-2. Their function may be of clinical importance.

Study on Lymphocyte Activation and Proliferation Induced by Anti-CD3 McAb

T cell activation and proliferation via CD3-TCR complex were investigated by lymphocyte DNA synthesis in vitro.Several interfering factors were also discussed.The result indicated that lymphocyte activation and proliferation are calciumdependent.A rise of cytoplasmic free Ca2+ quickly following activation with CD3 McAb is mainly due to intracellular mobilization of Ca2+,while lymphocyte proliferation needs both intracellular mobilization of Ca2+ as well as influx of extracellular Ca2+, It was confirmed that CTX sensitive G protein plays a role in regulating T cell proliferation by pretreatment with CTX suppressing lymphocyte H-TdR incorporation obviously.PLC and PKC inhibitor neomycin and P.S.S could also decrease T cell proliferation.

Activated cytotoxic lymphocytes promote tumor progression by increasing the ability of 3LL tumor cells to mediate MDSC chemoattraction via Fas signaling

The Fas/FasL system transmits intracellular apoptotic signaling, inducing cell apoptosis. However, Fas signaling also exerts non-apoptotic functions in addition to inducing tumor cell apoptosis. For example, Fas signaling induces lung cancer tumor cells to produce prostaglandin E2 （PGE2） and recruit myeloid-derived suppressor cells （MDSCs）. Activated cytotoxic T lymphocytes （CTLs） induce and express high levels of FasL, but the effects of Fas activation initiated by FasL in CTLs on apoptosis-resistant tumor cells remain largely unclear. We purified activated CD8^＋ T cells from OT-1 mice, evaluated the regulatory effects of Fas activation on tumor cell escape and investigated the relevant mechanisms. We found that CTLs induced tumor cells to secrete PGE2 and increase tumor cell-mediated chemoattraction of MDSCs via Fas signaling, which was favorable to tumor growth. Our results indicate that CTLs may participate in the tumor immune evasion process. To the best of our knowledge, this is a novel mechanism by which CTLs play a role in tumor escape. Our findings implicate a strategy to enhance the antitumor immune response via reduction of negative immune responses to tumors promoted by CTLs through Fas signaling.

THE IMMUNOREGULATORY EFFECT OF TLSF_(JM) ON THE EXPRESSION OF T CELL IL-2R AND PROTEIN TYROSINE PHOSPHORYLATION

The immunoregulatory effect of TLSFJM on the expression of T cell IL- 2R and protein tyrosine phosphorylation ( PTP ) was investigated by immunohistochemistry technique. The results showed that TLSFJMcan markedly suppress the expression of IL-2R and PTP on PHA or TPA-stimulated human PBMC and murine IL-2 dependent cell line CTLL-2. However, there was no effect of TLSFJMon the production of IL-1, IL-2 and IL-6 that play an important role in the course of T lymphocyte proliferation and differentiation.

A NEW EXPERIMENTAL AND CLINICAL APPROACH OF COMBINING USAGE OF HIGHLY ACTIVE TUMOR-INFILTRATING LYMPHOCYTES AND HIGHLY SENSITIVE ANTITUMOR DRUGS FOR THE ADVANCED MALIGNANT TUMOR

In recent years, tumor-nfiltrating lymphocytes (TILs) have been reported to be effective for tumors in experimental and clinical research. In order to increase the therapeutical effect, we modified some steps of Rosenberg's approach a. cold digestion with collagenase at 4C for 24 hours; b. sedimentation instead of centrifugation; c. elimination of tumor cells before the cultivation procedure. Compared with the original approach, the proliferation, activity and cytotoxicity of TILs obtained by the modified procedure were much improved. TILs' expansion-old was greater than that with the original approach. Cytotoxicity against rumor cells was more potent. Increased TILs' subsets were CD3 and CD8 cells. Meanwhile, we took tumor cells from tumor tissues to test their in vitro chemosensitivities to different drugs in order to select highly sensitive antitumor drugs for treatment of cases with advanced tumors. According to the design of using highly active TILs and highly sensitive drugs (H & H therapy), preliminary clinical results of 50 cases showed higher response rates than those in treatment with TIL / IL2, LAK / 1L2 and TIL+IL2+CTX. Less toxic side effects were observed in 14 patients.

Vitamin C promotes the proliferation and effector functions of human γδ T cells

γδT cells are of interest as effector cells for cellular immunotherapy due to their HLA-non-restricted lysis of many different tumor cell types.Potential applications include the adoptive transfer of in vitro-expandedγδT cells.Therefore,it is important to optimize the culture conditions to enable maximal proliferative and functional activity.Vitamin C(L-ascorbic acid)is an essential vitamin with multiple effects on immune cells.It is a cofactor for several enzymes,has antioxidant activity,and is an epigenetic modifier.Here,we investigated the effects of vitamin C(VC)and its more stable derivative,L-ascorbic acid 2-phosphate(pVC),on the proliferation and effector function of humanγδT cells stimulated with zoledronate(ZOL)or synthetic phosphoantigens(pAgs).VC and pVC did not increaseγδT-cell expansion within ZOL-or pAg-stimulated PBMCs,but increased the proliferation of purifiedγδT cells and 14-day-expandedγδT-cell lines in response toγδT-cell-specific pAgs.VC reduced the apoptosis ofγδT cells during primary stimulation.While pVC did not prevent activation-induced death of pAg-restimulatedγδT cells,it enhanced the cell cycle progression and cellular expansion.Furthermore,VC and pVC enhanced cytokine production during primary activation,as well as upon pAg restimulation of 14-day-expandedγδT cells.VC and pVC also increased the oxidative respiration and glycolysis ofγδT cells,but stimulus-dependent differences were observed.The modulatory activity of VC and pVC might help to increase the efficacy ofγδT-cell expansion for adoptive immunotherapy.

Uterine Natural Killer Cells:Their Choices,Their Missions

Uterine natural killer(uNK)cells,sharing many characters with peripheral blood natural killer(pNK)cells,are a major uterine lymphocyte population at early gestational stages during normal pregnancy in placental mammals. The functions of uNK cells include cytokine production and cytotoxcity that are regulated by signals through activating and inhibitory receptors.UNK cells differ from pNK cells however and contribute to the structural changes that accompany the differentiation of the maternal-fetal interface.Immunological mechanisms must provide a balanced environment for uNK cell proliferation,differentiation and activation through intricate signaling pathways.An improved knowledge of mechanisms regulating uNK cells development and the cytokine network at the maternal-fetal interface of mice and humans might be useful to harness the power of these cells for maintenance of pregnancy.Cellular & Molecular Immunology.2005;2(2):123-129.

Tumor immune checkpoints and their associated inhibitors

Immunological evasion is one of the defining characteristics of cancers,as the immune modification of an immune checkpoint(IC)confers immune evasion capabilities to tumor cells.Multiple ICs,such as programmed cell death protein-1(PD-1)and cytotoxic T-lymphocyte-associated antigen-4(CTLA-4),can bind to their respective receptors and reduce tumor immunity in a variety of ways,including blocking immune cell activation signals.IC blockade(ICB)therapies targeting these checkpoint molecules have demonstrated significant clinical benefits.This is because antibody-based IC inhibitors and a variety of specific small molecule inhibitors can inhibit key oncogenic signaling pathways and induce durable tumor remission in patients with a variety of cancers.Deciphering the roles and regulatory mechanisms of these IC molecules will provide crucial theoretical guidance for clinical treatment.In this review,we summarize the current knowledge on the functional and regulatory mechanisms of these IC molecules at multiple levels,including epigenetic regulation,transcriptional regulation,and post-translational modifications.In addition,we provide a summary of the medications targeting various nodes in the regulatory pathway,and highlight the potential of newly identified IC molecules,focusing on their potential implications for cancer diagnostics and immunotherapy.

Self-assembling, pH-responsive nanoflowers for inhibiting PAD4 and neutrophil extracellular trap formation and improving the tumor immune microenvironment

Self-assembling carrier-free nanodrugs are attractive agents because they accumulate at tumor by an enhanced permeability and retention(EPR) effect without introduction of inactive substances,and some nanodrugs can alter the immune environment. We synthesized a peptidyl arginine deiminase 4(PAD4) molecular inhibitor, ZD-E-1 M. It could self-assembled into nanodrug ZD-E-1. Using confocal laser scanning microscopy, we observed its cellular colocalization, PAD4 activity and neutrophil extracellular traps(NETs) formation. The populations of immune cells and expression of immune-related proteins were determined by single-cell mass cytometry. ZD-E-1 formed nanoflowers in an acidic environment, whereas it formed nanospheres at pH 7.4. Accumulation of ZD-E-1 at tumor was pHresponsive because of its pH-dependent differences in the size and shape. It could enter the nucleus and bind to PAD4 to prolong the intracellular retention time. In mice, ZD-E-1 inhibited tumor growth and metastasis by inhibiting PAD4 activity and NETs formation. Besides, ZD-E-1 could regulate the ratio of immune cells in LLC tumor-bearing mice. Immunosuppressive proteins like LAG3 were suppressed,while IFN-γ and TNF-a as stimulators of tumor immune response were upregulated. Overall, ZD-E-1 is a self-assembling carrier-free nanodrug that responds to pH, inhibits PAD4 activity, blocks neutrophil extracellular traps formation, and improves the tumor immune microenvironment.

异常的CD39/CD73/腺苷通路导致慢性乙型肝炎患者B细胞超活化和疾病进展

背景:乙肝病毒(HBV)感染患者B细胞超活化的机制尚不明确。本研究旨在评估CD39/CD37/腺苷通路在慢性乙型肝炎(CHB)患者中的临床特点。方法:我们检测了202例HBV感染患者B细胞CD39和CD73表达及腺苷产生情况。CpG+CD40配体刺激B细胞后,阻断或激活CD39/CD73/腺苷通路,然后通过流式细胞术检测B细胞活化表型结果:在具有高HBV DNA载量,HBeAg阳性,高HBsAg水平以及肝脏炎症活跃的CHB患者中,外周血B细胞的CD39和CD73表达降低,并且在HBeAg血清转化或HBsAg降低的完全应答者中逐步恢复。尽管进行了有效的抗病毒治疗,但完全应答者中活化记忆B细胞亚群和组织样记忆B细胞亚群的CD39和CD73表达并未增加。此外,在炎性肝脏中,肝内B细胞的CD39和CD73表达降低。在体外,CHB患者的B细胞显示产生CD39/CD73依赖性细胞外腺苷的能力明显降低。阻断腺苷生成后B细胞活化标志物表达增加。相反,二甲双胍通过调节腺苷酸活化蛋白激酶,激活CD39/CD73/腺苷通路,从而使B细胞活化标志物表达显著降低。结论:B细胞中CD39和CD73表达下调可导致CHB患者的高病毒负荷,加重肝脏炎性反应,并影响抗病毒疗效。异常的CD39/CD73/腺苷通路导致B细胞超活化。使用二甲双胍调节CD39/CD73/腺苷通路可能作为一种能够逆转HBV诱导的免疫发病机制的治疗选择。

Cutaneous Dengue Virus Inoculation Triggers Strong B Cell Reactions but Contrastingly Poor T Cell Responses

Dengue is a global health problem without current specific treatment nor safe vaccines available.While severe dengue is related to pre-existing non-neutralizing dengue virus(DENV)antibodies,the role of T cells in protection or pathology is unclear.Using cutaneous DENV infection in immunocompetent mice we previously showed the generation of PNA+germinal centers(GCs),now we assessed the activation and proliferation of B and T cells in draining lymph nodes(DLNs).We found a drastic remodelling of DLN compartments from 7 to 14 days post-infection(dpi)with greatly enlarged B cell follicles,occupying almost half of the DLN area compared to*24%in na?ve conditions.Enormous clusters of proliferating(Ki-67+)cells inside B follicles were found 14 dpi,representing*33%of B cells in DLNs but only*2%in noninfected mice.Inside GCs,we noticed an important recruitment of tingle body macrophages removing apoptotic cells.In contrast,the percentage of paracortex area and total T cells decreased by 14–16 dpi,compared to controls.Scattered randomly distributed Ki-67+T cells were found,similar to non-infected mice.CD69 expression by CD4+and CD8+T cells was minor,while it was remarkable in B cells,representing 1764.7%of change from basal levels 3 dpi.The apparent lack of T cell responses cannot be attributed to apoptosis since no significant differences were observed compared to noninfected mice.This study shows massive B cell activation and proliferation in DLNs upon DENV infection.In contrast,we found very poor,almost absent CD4+and CD8+T cell responses.

Expression and significance of myeloid differentiation factor 88 in marrow dendritic cells in asthmatic rats with cigarette smoke exposure

Background Smoking causes frequent asthma attacks, leading to a rapid decline in lung function in patients with asthma, and it can also reduce the therapeutic effect of glucocorticoids in patients with asthma. Therefore, the present study aimed to investigate the effect of cigarette smoke on the expression of myeloid differentiation factor 88 （MyD88） in marrow dendritic cells （DCs） in asthmatic rats, and to explore the molecular mechanism of cigarette smoke exposure on asthma by DCs. Methods Forty Wistar rats were randomly divided into the following groups： control, smoke exposure, asthma, and asthma combined with smoke exposure. The animal model was established, and then rat bone marrow-derived DCs were collected. Additionally, rat spleen lymphocytes and bone marrow-derived DCs were cultured together for mixed lymphocyte responses. Interferon （IFN）-gamma and interleukin （IL）-4, IL-10, and IL-12 expressions were determined by enzyme-linked immunosorbent assay （ELISA）. MyD88 expression was determined by Western blotting. The proliferation of lymphocytes was examined with methyl thiazolyl tetrazolium （MTT） colorimetric assay. Results MyD88 expression was decreased in the asthma combined with smoke exposure group compared to the asthma group （P 〈0.01）, and IL-10 and IL-12 expressions were decreased in the asthma combined with smoke exposure group compared to control group （P 〈0.01）. In addition, DCs stimulating activity on allogeneic lymphocytes were significantly decreased in the smoke exposure combined with asthma group compared to the control and asthma groups （P 〈0.01）. After allogeneic mixed lymphocyte responses, IL-4 expression was increased and IFN-gamma was decreased in the asthma group and the asthma combined with smoke exposure group compared to control group （P 〈0.01）. IL-4 expression was increased and IFN-gamma was decreased in the asthma combined with smoke exposure group compared to the asthma group （P 〈0.01）. The study also showed that MyD88 expression was positively correlated with IL-12 and IFN-gamma expressions and the activity of lymphocytes （P 〈0.01）, and negatively correlated with IL-4 expression （P 〈0.01）. Conclusions Smoking aggravates asthma by weankening immunological mechanism. MyD88-dependent pathways may play a role in the immunological balance and activation of lymphocytes.

4-1BB gene expression in peripheral blood mononuclear cells from orthotopic liver transplant recipients with graft acceptance

Objective To investigate the gene expression of 4-1BB in peripheral blood mononuclear cells (PBMCs) and the possible significance of the 4-1BB pathway after clinical orthotopic liver transplantation (OLT). Methods 4-1BB mRNA levels in PBMCs from 22 OLT patients were analyzed by RT-PCR. 4-1BB protein expressed on the surface of CD 4 + and CD 8 + T cells were detected by flow cytometry, and visualized with direct immunofluorescence and confocal fluorescence microscopy. Patients with primary liver cancer (PLC) and healthy volunteers served as controls. Six cases of recently performed liver transplantation were also observed in this study. Results 4-1BB mRNA was detected in PBMCs from both liver transplant patients with long-term graft acceptance (22 cases) and from transplant patients on day 1 to day 3 post-transplantation (6 cases), but was not found in PBMCs from transplant patients on day 7 to day 30 post-transplantation (6 cases). 4-1BB mRNA was also not found in samples from 8 of the healthy controls and 7 of the PLC patients, though very low expression was detected in the other 4 healthy volunteers and 6 PLC patients. Simultaneously, 4-1BB protein was expressed at nearly undetectable levels on CD 4 + and CD 8 + T cells from healthy controls, PLC patients, as well as OLT patients within the first month post-transplantation (6 cases). However, 4-1BB expression was found on the surface of CD 4 + and CD 8 + T cells from liver transplant patients with long-term graft acceptance. Direct immunofluorescent staining and confocal fluorescence microscopy clearly revealed evidence of 4-1BB protein on cell membranes of CD 4 + and CD 8 + T cells from liver transplant patients with long-term graft acceptance. Simultaneously, a significantly higher percentage of CD 3 + CD 25 + T cells were found in liver transplant patients with long-term graft acceptance group as compared with the healthy control group (P<0.05). The expression of 4-1BB protein on T cells did not correlate with the survival time of OLT patients postoperation. Conclusions This study demonstrates that although patients remain in stable condition after liver transplantation under the treatment of immunosuppressants, activated T cells are present to some extent and 4-1BB protein may be involved in this process. Effector T-cells can exert permanent immunoresponses against grafts under these circumstances. Therefore, we conclude that a new immune response balance is established under the combination of both treatment with immunosuppressants and natural immune responses against alloantigens. Manipulation of the 4-1BB/4-1BBL pathway may provide a therapeutic technique for prolonging graft survival.