AIM： To observe biological characteristics of hepatocarcinoma cells before and after CD80 transfection and to compare the effect of CD80-transfected hepatocarcinoma cells on T lymphocyte activation. METHODS： Retro v...AIM： To observe biological characteristics of hepatocarcinoma cells before and after CD80 transfection and to compare the effect of CD80-transfected hepatocarcinoma cells on T lymphocyte activation. METHODS： Retro virus vector carrying CD80 gene was transfected into HepG2 cells to establish CD80transfected hepatocarcinoma cells （HepG2/hCD80）. Flow cytometry （FCM） was performed to detect CD80 expression in the transfected cells. RT-PCR was used to evaluate CD80 expression at mRNA level. In the presence of anti-CD3 mAb, the proliferation of T lymphocyte was observed by M＇n＇. Meanwhile, the expression of activated molecule marker CD25 was analyzed through FCM. RESULTS： A stable cell line HepG2/hCD80 expressing the human CD80 was established. Growth curve showed that the molecule CD80 could obviously decrease the growth of tumor cells. HepG2/hCD80 was evidenced to have a potency to enhance T cell proliferation and upregulate CD25 expression. CONCLUSION： CD80 transfection can lower malignant phenotype of hepatocarcinoma cells. CD80 transfection has a down-regulatory effect to activated T cells in vitro.展开更多
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 prolif...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.展开更多
Introduction:Hyperthyroidism is known to affect various physiological systems,including the immune system.Thyroid hormones(THs)play a crucial role in regulating immune function,and alterations in THs levels can lead t...Introduction:Hyperthyroidism is known to affect various physiological systems,including the immune system.Thyroid hormones(THs)play a crucial role in regulating immune function,and alterations in THs levels can lead to immune dysregulation.Objective:Currently,we aimed to elucidate the effects of hyperthyroidism on immune function in BALB/c mice,with a focus on anatomical and histological changes in lymphoid organs,the immune response to mitogenic stimulation,mitochondrial dynamics,and reactive oxygen species(ROS)production.Methods:Hyperthyroidism was induced in BALB/c mice by administering thyroxine(T4;14 mg/L)in their drinking water for 30 days.Thyroid function was assessed by measuring triiodothyronine(T3),T4,and Thyroid-Stimulating Hormone(TSH)levels.Lymphoid organ hyperplasia was evaluated through anatomical dissection.Lymphoid responses were analyzed by subcutaneous inoculation with lipopolysaccharide(LPS),followed by histological analysis of lymphoid follicles and evaluation of the morphometric parameters of lymphoid cells using flow cytometry.In vitro cell proliferation was quantified using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay.Mitochondrial morphology and density were assessed by Transmission Electron Microscopy(TEM).ROS and superoxide anion(O2-)production were measured using 2′,7′-dichlorodihydrofluorescein diacetate(DCFH-DA)and Nitroblue Tetrazolium(NBT)assays.Results:Hyperthyroid mice exhibited significantly increased T3 and T4 levels,with decreased TSH levels.Lymphoid organs,including the spleen and lymph nodes,were notably enlarged in hyperthyroid mice,with a corresponding increase in lymphoid cell number.LPS stimulation enhanced the number and size of lymphoid follicles,with hyperthyroid mice showing a greater proliferative response.TEM analysis revealed increased mitochondrial density and changes in mitochondrial structure in hyperthyroid lymphoid cells.ROS and O2-production were significantly higher in hyperthyroid mice,though no apoptotic activity was detected.Conclusion:Hyperthyroidism leads to significant alterations in immune responses,including enhanced lymphoid organ size,increased proliferation of immune cells,and elevated ROS production.These findings provide new insights into the immunomodulatory effects of thyroid dysfunction and its potential impact on immune system regulation,offering a deeper understanding of THs interactions with immune activation.展开更多
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 lym...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.展开更多
Objective：With the regular mixed lymphocytes culture （MLC） to detect the allograft rejection, the reactivity of the activated lymphocytes （primed lymphocytes） of a recipient shows sometimes increase and sometimes...Objective：With the regular mixed lymphocytes culture （MLC） to detect the allograft rejection, the reactivity of the activated lymphocytes （primed lymphocytes） of a recipient shows sometimes increase and sometimes decrease against the antigens from the donor, which is inconsistent with the clinical results. In order to establish a convenient method for testing the specificity of the activated lymphocytes in vitro, so as to know the rejection occurred or not by testing the existence of the specific activated lymphocytes against donor＇s HLA antigens in the recipient＇s peripheral blood. Methods： Anti-IL-2 neutralizing monoclonal antibody （anti-IL-2 N-mAb） and immunosuppressors were introduced in this test system in the presence of specific stimulators and activated lymphocytes. Results ： When the activated lymphocytes were chosen from the one-way MLC 4 d to undergo re-stimulation by specific stimulators, the activity of activated lymphocytes in the treatment group was suppressed significantly compared with that in the control group. The result of this test method is consistent with the biopsy in the clinical diagnosis of rejection. Conclusion：h suggests that the activated lymphocytes can be inactivated by specific antigens in certain conditions. This can be a useful tool to define the specificity of the activated lymphocytes.展开更多
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-t...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.展开更多
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 ...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.展开更多
AIM： To construct the plasmid pcHEV23 containing fragments of HEV ORF2 and ORF3 chimeric gene and to assess its ability to elicit specific immunologic response in mice. METHODS： The gene encoding the structural prot...AIM： To construct the plasmid pcHEV23 containing fragments of HEV ORF2 and ORF3 chimeric gene and to assess its ability to elicit specific immunologic response in mice. METHODS： The gene encoding the structural protein of HEV ORF2 fragment and full-length ORF3 was amplified by PCR. The PCR products were cloned into an eucaryotic expression plasmid pcDNA3. The resulting plasmid pcHEV23 was used as a DNA vaccine to inoculate BALB/c mice intramuscularly thrice at a dose of 100 or 200 ug. Mice injected with empty pcDNA3 DNA or saline served as control and then specific immune responses in the mice were detected. RESULTS： After 2-3 times of inoculation, all mice injected with pcHEV23 had anti-HEV IgG seroconversion and specific T lymphocyte proliferation. The lymphocyte stimulation index in the group immunized with pcHEV23 （3.1＋0.49） was higher than that in the control group （0.787±0.12, P〈0.01）. None in the control group had a detectable level of anti-HEV IgG. CONCLUSION： DNA vaccine containing HEV ORF2 and ORF3 chimeric gene can successfully induce specific humoral and cellular immune response in mice.展开更多
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 dea...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.展开更多
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 prot...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.展开更多
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 supp...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.展开更多
It has been reported that host defense responses, such as phagocytic function of neutrophils and natural killer (NK) cell activity of lymphocytes, are impaired in cirrhotic patients. This review will concentrate on th...It has been reported that host defense responses, such as phagocytic function of neutrophils and natural killer (NK) cell activity of lymphocytes, are impaired in cirrhotic patients. This review will concentrate on the impairment of innate immune responses in decompensated cirrhotic patients and the effect of the treatment by branched-chain amino acids (BCAA) on innate immune responses. We already reported that phagocytic function of neutrophils was significantly improved by 3-mo BCAA supplementation. In addition, the changes of NK activity were also significant at 3 mo of supplementation compared with before supplementation. Also, Fisher’s ratios were reported to be significantly increased at 3 mo of BCAA supplementation compared with those before oral supplementation. Therefore, administration of BCAA could reduce the risk of bacterial and viral infection in patients with decompensated cirrhosis by restoring impaired innate immune responses of the host. In addition, it was also revealed that BCAA oral supplementation could reduce the risk of development of hepatocellular carcinoma in cirrhotic patients. The mechanisms of the effects will also be discussed in this review article.展开更多
Background:Lymphocyte activation gene 3(LAG-3)is a promising immune checkpoint for combination im-munotherapy.This study aims to elucidate the exact synergistic anti-tumor mechanism of programmed death 1(PD-1)and LAG-...Background:Lymphocyte activation gene 3(LAG-3)is a promising immune checkpoint for combination im-munotherapy.This study aims to elucidate the exact synergistic anti-tumor mechanism of programmed death 1(PD-1)and LAG-3 dual inhibition in lung cancer.Methods:Multiple patient-derived xenograft(PDX)models of lung cancer were constructed and analyzed by single-cell RNA sequencing(scRNA-seq).Clustering of all human-derived cells,identification of biomarker genes of three cell types,trajectory analysis,and calculation of tumor heterogeneity scores were performed.Differen-tially expressed genes(DEGs)were identified and functional enrichment analyses of cancer-associated genes were conducted.The functional significance of DEGs in the immune system was evaluated using the Reactome online server.Major histocompatibility complex(MHC)pathways and angiogenesis-associated pathways were analyzed.The Cancer Genome Atlas(TCGA)was used for further verification.Results:PD-1 and LAG-3 dual inhibition achieved synergistic tumor inhibition in squamous cell carcinoma(SCC)PDX models,but not in adenocarcinoma and small cell lung cancer PDX models.A total of 8127 cells,including 2699 basal,4109 malignant,and 1319 epithelial cells,were identified by scRNA-seq.Malignant cells evolved from basal and epithelial cells in the trajectory analysis.The responders to the combination therapy of PD-1 and LAG-3 inhibitors had lower heterogeneity scores than non-responders.Compared with anti-PD-1 monotherapy,the combination group exhibited higher levels of neutrophil degranulation.The DEGs were correlated with disease,metabolism,and programmed cell death-associated pathways.The MHC classⅠ-associated pathways and pericyte pathways were upregulated,whereas the vascular endothelial growth factor pathway was downregulated in the combination group.Conclusion:We discovered the superior efficacy of PD-1 and LAG-3 dual inhibition in SCC PDX models,and showed that it may be associated with low tumor heterogeneity scores,upregulation of the MHC classⅠpathway,and normalization of tumor angiogenesis.展开更多
Background:The mechanisms underlying B-cell hyperactivation in patients with chronic hepatitis B virus(HBV)infection remain largely undefined.The present study assessed the clinical characteristics of the CD39/CD73/ad...Background:The mechanisms underlying B-cell hyperactivation in patients with chronic hepatitis B virus(HBV)infection remain largely undefined.The present study assessed the clinical characteristics of the CD39/CD73/adenosine pathway in patients with chronic hepatitis B(CHB).Methods:We examined CD39 and CD73 expression and adenosine production by B-cells from 202 HBV-infected patients.B-cell-activation phenotypes were assessed by flow cytometry after CpG+CD40 ligand stimulation with or without blockade and activation of the adenosine pathway.Results:CD39 and CD73 expression on circulating B-cells was decreased in CHB patients with high HBV DNA,HBeAg positivity,high HBsAg levels,and active liver inflammation,and was hierarchically restored in complete responders according to HBeAg seroconversion or HBsAg reduction.However,CD39 and CD73 expression on activated memory and tissue-like memory B-cell subsets in complete responders was not increased despite effective antiviral treatments.Furthermore,CD39 and CD73 expression on intra-hepatic B-cells was decreased in inflammatory livers.In vitro,B-cells from CHB patients showed a markedly reduced capacity to generate CD39/CD73-dependent extracellular adenosine and expressed increased levels of activation markers after adenosine-production blockade.Contrastingly,metformin significantly reduced activation-marker expression via regulating AMP-activated protein kinase.Conclusions:The skewed CD39 and CD73 expression on B-cells was associated with a high viral burden,liver inflammation,and antiviral efficacy in CHB patients,and the skewed CD39/CD73/adenosine pathway contributed to B-cell hyperactivation.Regulation of the CD39/CD73/adenosine pathway using metformin may represent a therapeutic option to reverse HBVinduced immune pathogenesis.展开更多
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 signa...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.展开更多
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 Rosen...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.展开更多
Hepatocellular carcinoma(HCC)stands as a primary malignant liver tumor characterized by chronic inflammation and complex alterations within the tumor microenvironment(TME).Lymphocyte activation gene 3(LAG-3),also know...Hepatocellular carcinoma(HCC)stands as a primary malignant liver tumor characterized by chronic inflammation and complex alterations within the tumor microenvironment(TME).Lymphocyte activation gene 3(LAG-3),also known as CD223,has gained prominence as a potential next-generation immune checkpoint,maintaining continuous expression in response to persistent antigen exposure within the TME,warranting our attention.In patients with HCC,LAG-3 expression on T cells,regulatory T cells(Tregs),and natural killer(NK)cells contributes to immune evasion,while high expression of LAG-3 leads to increased angiogenesis and poor prognosis.By interacting with major histocompatibility complex class II molecules,LAG-3 promotes T cell exhaustion and suppresses antitumor responses,often in collaboration with other immune checkpoints like programmed cell death protein 1(PD-1),while on Tregs and NK cells,LAG-3 modulates their suppressive functions,indirectly facilitating tumor immune escape.LAG-3 expression may offer prognostic insights,correlating with disease progression and outcomes in HCC patients,while various preclinical studies highlight the potential of LAG-3-targeted therapies in reinvigorating immune responses against HCC,with a few combination approaches targeting LAG-3 alongside other checkpoints demonstrating synergistic effects in restoring T cell function.Therefore,harnessing LAG-3 as a therapeutic target holds promise for enhancing antitumor immunity and potentially improving HCC treatment outcomes.Our narrative review aims to delve into the full spectrum of LAG-3 signaling in HCC,with the goal of a better understanding of the pathophysiological and immunological basis of its use to arrest HCC growth and development.展开更多
OBJECTIVE: To investigate which of the two immunoglobulin (Ig)-like domains, the immunoglobulin variable region homologous domain IgV (hB7.2 IgV) and the immunoglobulin constant region homologous domain IgC (hB7.2 IgC...OBJECTIVE: To investigate which of the two immunoglobulin (Ig)-like domains, the immunoglobulin variable region homologous domain IgV (hB7.2 IgV) and the immunoglobulin constant region homologous domain IgC (hB7.2 IgC) on the human B7.2 molecule contains receptor binding sites, and to evaluate whether the B7.2 protein expressed in bacteria has biological activity in vitro. METHODS: Three fragments of hB7.2 IgV,hB7.2 IgC and the complete extracellular region of human B7.2 containing both the IgV and IgC domains,hB7.2 Ig (V+C), were amplified by PCR and subcloned into pGEM-Teasy. Three recombinants,pGEX-4T-3-hB7.2 IgV,pGEX-4T-3-hB7.2 IgC and pGEX-4T-3-hB7.2 Ig (V+C), were generated by cloning the fragments into a prokaryote expression plasmid (pGEX-4T-3) and transformed into the host strain E. coli DH5alpha. The relevant target fusion proteins consisting of GST and hB7.2 IgV,hB7.2 IgC and hB7.2 Ig (V+C), were identified by SDS-PAGE and Western blotting. With the presence of the first signal imitated by anti-CD3 antibody, T cell activation was observed by exposing purified T lymphocytes to each soluble form of the three bacterially-produced human B7.2 fusion proteins by [(3)H]-TdR incorporation. RESULTS: Three recombinant fusion proteins of human B7.2, GST-hB7.2 IgV, GST-hB7.2 IgC and GST-hB7.2 Ig (V+C) were produced and detected in inclusion body form from engineered bacteria. With the first signal present,T lymphocytes proliferated when co-stimulated by bacterially-produced either GST-hB7.2 Ig (V+C) or GST-hB7.2 IgV fusion proteins, but not by GST-hB7.2 IgC. CONCLUSIONS: Functional human B7.2 fusion protein can be produced in bacteria. The IgV-like domain of human B7.2 is sufficient for B7.2 to interact with its counter-receptors and co-stimulate T lymphocytes.展开更多
OBJECTIVE: To investigate the specific cell-mediated immune efficacy of the an HPV16 prophylactic vaccine. METHODS: C57BL/6 mice were randomly divided into 3 groups: experimental group I (treated with pcDNA L1), contr...OBJECTIVE: To investigate the specific cell-mediated immune efficacy of the an HPV16 prophylactic vaccine. METHODS: C57BL/6 mice were randomly divided into 3 groups: experimental group I (treated with pcDNA L1), control group II (treated with pcDNA3.1) and control group III (treated with PBS buffer). The mice were immunized three times during a three-week interval. Ten to fourteen days after the third inoculation, a footpad swelling test was used to detect delayed-type hypersensitivity (DTH) responses. Antigen-specific splenocyte proliferation assay and quantitation of IFN-gamma cells in splenocytes were performed by FACS assay. RESULTS: In the experimental group, splenocytes actively proliferated after stimulation with HPV16 VLP, and had developed a markedly larger amount of CD8(+) IFN-gamma(+) cells, which is an index for special CTL. Also, the footpad was significantly thickened upon inoculation with HPV16 VLP. CONCLUSION: Naked DNA vaccine of HPV16 L1 can induce specific cell-mediated immune responses in mice, which should be considered for evaluation of HPV16 DNA vaccine feasibility.展开更多
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-expan...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.展开更多
文摘AIM： To observe biological characteristics of hepatocarcinoma cells before and after CD80 transfection and to compare the effect of CD80-transfected hepatocarcinoma cells on T lymphocyte activation. METHODS： Retro virus vector carrying CD80 gene was transfected into HepG2 cells to establish CD80transfected hepatocarcinoma cells （HepG2/hCD80）. Flow cytometry （FCM） was performed to detect CD80 expression in the transfected cells. RT-PCR was used to evaluate CD80 expression at mRNA level. In the presence of anti-CD3 mAb, the proliferation of T lymphocyte was observed by M＇n＇. Meanwhile, the expression of activated molecule marker CD25 was analyzed through FCM. RESULTS： A stable cell line HepG2/hCD80 expressing the human CD80 was established. Growth curve showed that the molecule CD80 could obviously decrease the growth of tumor cells. HepG2/hCD80 was evidenced to have a potency to enhance T cell proliferation and upregulate CD25 expression. CONCLUSION： CD80 transfection can lower malignant phenotype of hepatocarcinoma cells. CD80 transfection has a down-regulatory effect to activated T cells in vitro.
文摘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.
文摘Introduction:Hyperthyroidism is known to affect various physiological systems,including the immune system.Thyroid hormones(THs)play a crucial role in regulating immune function,and alterations in THs levels can lead to immune dysregulation.Objective:Currently,we aimed to elucidate the effects of hyperthyroidism on immune function in BALB/c mice,with a focus on anatomical and histological changes in lymphoid organs,the immune response to mitogenic stimulation,mitochondrial dynamics,and reactive oxygen species(ROS)production.Methods:Hyperthyroidism was induced in BALB/c mice by administering thyroxine(T4;14 mg/L)in their drinking water for 30 days.Thyroid function was assessed by measuring triiodothyronine(T3),T4,and Thyroid-Stimulating Hormone(TSH)levels.Lymphoid organ hyperplasia was evaluated through anatomical dissection.Lymphoid responses were analyzed by subcutaneous inoculation with lipopolysaccharide(LPS),followed by histological analysis of lymphoid follicles and evaluation of the morphometric parameters of lymphoid cells using flow cytometry.In vitro cell proliferation was quantified using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay.Mitochondrial morphology and density were assessed by Transmission Electron Microscopy(TEM).ROS and superoxide anion(O2-)production were measured using 2′,7′-dichlorodihydrofluorescein diacetate(DCFH-DA)and Nitroblue Tetrazolium(NBT)assays.Results:Hyperthyroid mice exhibited significantly increased T3 and T4 levels,with decreased TSH levels.Lymphoid organs,including the spleen and lymph nodes,were notably enlarged in hyperthyroid mice,with a corresponding increase in lymphoid cell number.LPS stimulation enhanced the number and size of lymphoid follicles,with hyperthyroid mice showing a greater proliferative response.TEM analysis revealed increased mitochondrial density and changes in mitochondrial structure in hyperthyroid lymphoid cells.ROS and O2-production were significantly higher in hyperthyroid mice,though no apoptotic activity was detected.Conclusion:Hyperthyroidism leads to significant alterations in immune responses,including enhanced lymphoid organ size,increased proliferation of immune cells,and elevated ROS production.These findings provide new insights into the immunomodulatory effects of thyroid dysfunction and its potential impact on immune system regulation,offering a deeper understanding of THs interactions with immune activation.
基金supported by the Special Fund for Agroscientific Research in the Public Interest, China(200803019)the Youth Innovation Foudation of Shandong Agricultural University, China (23477)
文摘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.
文摘Objective：With the regular mixed lymphocytes culture （MLC） to detect the allograft rejection, the reactivity of the activated lymphocytes （primed lymphocytes） of a recipient shows sometimes increase and sometimes decrease against the antigens from the donor, which is inconsistent with the clinical results. In order to establish a convenient method for testing the specificity of the activated lymphocytes in vitro, so as to know the rejection occurred or not by testing the existence of the specific activated lymphocytes against donor＇s HLA antigens in the recipient＇s peripheral blood. Methods： Anti-IL-2 neutralizing monoclonal antibody （anti-IL-2 N-mAb） and immunosuppressors were introduced in this test system in the presence of specific stimulators and activated lymphocytes. Results ： When the activated lymphocytes were chosen from the one-way MLC 4 d to undergo re-stimulation by specific stimulators, the activity of activated lymphocytes in the treatment group was suppressed significantly compared with that in the control group. The result of this test method is consistent with the biopsy in the clinical diagnosis of rejection. Conclusion：h suggests that the activated lymphocytes can be inactivated by specific antigens in certain conditions. This can be a useful tool to define the specificity of the activated lymphocytes.
文摘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.
文摘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.
基金Supported by the Grants from the Natural Science Foundation of Zhejiang Province, No. RC01054, Science Technology Department of Zhejiang Province, No. F11023 and Key Project of Health Bureau of Zhejiang Province
文摘AIM： To construct the plasmid pcHEV23 containing fragments of HEV ORF2 and ORF3 chimeric gene and to assess its ability to elicit specific immunologic response in mice. METHODS： The gene encoding the structural protein of HEV ORF2 fragment and full-length ORF3 was amplified by PCR. The PCR products were cloned into an eucaryotic expression plasmid pcDNA3. The resulting plasmid pcHEV23 was used as a DNA vaccine to inoculate BALB/c mice intramuscularly thrice at a dose of 100 or 200 ug. Mice injected with empty pcDNA3 DNA or saline served as control and then specific immune responses in the mice were detected. RESULTS： After 2-3 times of inoculation, all mice injected with pcHEV23 had anti-HEV IgG seroconversion and specific T lymphocyte proliferation. The lymphocyte stimulation index in the group immunized with pcHEV23 （3.1＋0.49） was higher than that in the control group （0.787±0.12, P〈0.01）. None in the control group had a detectable level of anti-HEV IgG. CONCLUSION： DNA vaccine containing HEV ORF2 and ORF3 chimeric gene can successfully induce specific humoral and cellular immune response in mice.
基金supported by the National Key Research and Development Program of China(No.2021YFC2700903)the National Natural Science Foundation of China(Nos.81672791 and 81872300)+2 种基金the Zhejiang Provincial Natural Science Fund for Distinguished Young Scholars of China(No.LR18C060002)the Huadong Medicine Joint Funds of the Zhejiang Provincial Natural Science Foundation of China(No.LHDMY22H160006)the ZJU-QILU Joint Research Institute and Qilu Group.
文摘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.
基金supported by a grant from CONACYT-Mexico(221102)to Leopoldo Flores-Romoby a CINVESTAV grant to Leticia Cedillo-Barrón
文摘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.
文摘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.
基金Supported by Grants(in part)from Ministry of Education,Culture,Sports,Science and Technology of Japan and the Ministry of Health,Labor and Welfare of Japan
文摘It has been reported that host defense responses, such as phagocytic function of neutrophils and natural killer (NK) cell activity of lymphocytes, are impaired in cirrhotic patients. This review will concentrate on the impairment of innate immune responses in decompensated cirrhotic patients and the effect of the treatment by branched-chain amino acids (BCAA) on innate immune responses. We already reported that phagocytic function of neutrophils was significantly improved by 3-mo BCAA supplementation. In addition, the changes of NK activity were also significant at 3 mo of supplementation compared with before supplementation. Also, Fisher’s ratios were reported to be significantly increased at 3 mo of BCAA supplementation compared with those before oral supplementation. Therefore, administration of BCAA could reduce the risk of bacterial and viral infection in patients with decompensated cirrhosis by restoring impaired innate immune responses of the host. In addition, it was also revealed that BCAA oral supplementation could reduce the risk of development of hepatocellular carcinoma in cirrhotic patients. The mechanisms of the effects will also be discussed in this review article.
基金supported in part by a grant from the National Key Research and Development Program of China(No.2022YFF0705300)the National Natural Science Foundation of China(No.52272281)+3 种基金the Shanghai Municipal Science and Technology Major Project(No.2021SHZDZX0100)the Fundamental Research Funds for the Central Universities,Shanghai Municipal Health Commission Health Industry Clinical Research Project,Clinical Research Project of Shanghai Pul-monary Hospital(No.FKLY20010)Young Talents in Shanghai(No.2019 QNBJ),Shanghai Shuguang Scholar,2021 Science and Technology Think Tank Youth Talent Plan of China Association for Science and Technology,“Dream Tutor”Outstanding Young Talents Program(No.fkyq1901)the National Key Research and Development Program of China(Nos.2021YFF1201200 and 2021YFF1200900).
文摘Background:Lymphocyte activation gene 3(LAG-3)is a promising immune checkpoint for combination im-munotherapy.This study aims to elucidate the exact synergistic anti-tumor mechanism of programmed death 1(PD-1)and LAG-3 dual inhibition in lung cancer.Methods:Multiple patient-derived xenograft(PDX)models of lung cancer were constructed and analyzed by single-cell RNA sequencing(scRNA-seq).Clustering of all human-derived cells,identification of biomarker genes of three cell types,trajectory analysis,and calculation of tumor heterogeneity scores were performed.Differen-tially expressed genes(DEGs)were identified and functional enrichment analyses of cancer-associated genes were conducted.The functional significance of DEGs in the immune system was evaluated using the Reactome online server.Major histocompatibility complex(MHC)pathways and angiogenesis-associated pathways were analyzed.The Cancer Genome Atlas(TCGA)was used for further verification.Results:PD-1 and LAG-3 dual inhibition achieved synergistic tumor inhibition in squamous cell carcinoma(SCC)PDX models,but not in adenocarcinoma and small cell lung cancer PDX models.A total of 8127 cells,including 2699 basal,4109 malignant,and 1319 epithelial cells,were identified by scRNA-seq.Malignant cells evolved from basal and epithelial cells in the trajectory analysis.The responders to the combination therapy of PD-1 and LAG-3 inhibitors had lower heterogeneity scores than non-responders.Compared with anti-PD-1 monotherapy,the combination group exhibited higher levels of neutrophil degranulation.The DEGs were correlated with disease,metabolism,and programmed cell death-associated pathways.The MHC classⅠ-associated pathways and pericyte pathways were upregulated,whereas the vascular endothelial growth factor pathway was downregulated in the combination group.Conclusion:We discovered the superior efficacy of PD-1 and LAG-3 dual inhibition in SCC PDX models,and showed that it may be associated with low tumor heterogeneity scores,upregulation of the MHC classⅠpathway,and normalization of tumor angiogenesis.
基金supported by grants from the Innovative Research Team in the National Natural Science Foundation of China[81721002]the National Science and Technology Major Project[2018ZX10301202].
文摘Background:The mechanisms underlying B-cell hyperactivation in patients with chronic hepatitis B virus(HBV)infection remain largely undefined.The present study assessed the clinical characteristics of the CD39/CD73/adenosine pathway in patients with chronic hepatitis B(CHB).Methods:We examined CD39 and CD73 expression and adenosine production by B-cells from 202 HBV-infected patients.B-cell-activation phenotypes were assessed by flow cytometry after CpG+CD40 ligand stimulation with or without blockade and activation of the adenosine pathway.Results:CD39 and CD73 expression on circulating B-cells was decreased in CHB patients with high HBV DNA,HBeAg positivity,high HBsAg levels,and active liver inflammation,and was hierarchically restored in complete responders according to HBeAg seroconversion or HBsAg reduction.However,CD39 and CD73 expression on activated memory and tissue-like memory B-cell subsets in complete responders was not increased despite effective antiviral treatments.Furthermore,CD39 and CD73 expression on intra-hepatic B-cells was decreased in inflammatory livers.In vitro,B-cells from CHB patients showed a markedly reduced capacity to generate CD39/CD73-dependent extracellular adenosine and expressed increased levels of activation markers after adenosine-production blockade.Contrastingly,metformin significantly reduced activation-marker expression via regulating AMP-activated protein kinase.Conclusions:The skewed CD39 and CD73 expression on B-cells was associated with a high viral burden,liver inflammation,and antiviral efficacy in CHB patients,and the skewed CD39/CD73/adenosine pathway contributed to B-cell hyperactivation.Regulation of the CD39/CD73/adenosine pathway using metformin may represent a therapeutic option to reverse HBVinduced immune pathogenesis.
基金The work was supported by the Specialized Research Fund for the Chinese National 973 Project （2013CB530502）, the Doctoral Program of Higher Education of China （20110101110105）, the Project of the Chinese National Nature Science Foundation （31370902, 31070795, 31270944）, the Projects in Science and Technology Plan of Zhejiang Province （013C33G2010434） of China, the National Key Science and Technology Specific Proiect of China （2012ZX10002006）, the National High Technology Research and Development Program （2012AA020900）, and the Project of the Chinese National Natural Science Fund Committee for Talent Cultivation （J1103603）.
文摘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.
文摘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.
文摘Hepatocellular carcinoma(HCC)stands as a primary malignant liver tumor characterized by chronic inflammation and complex alterations within the tumor microenvironment(TME).Lymphocyte activation gene 3(LAG-3),also known as CD223,has gained prominence as a potential next-generation immune checkpoint,maintaining continuous expression in response to persistent antigen exposure within the TME,warranting our attention.In patients with HCC,LAG-3 expression on T cells,regulatory T cells(Tregs),and natural killer(NK)cells contributes to immune evasion,while high expression of LAG-3 leads to increased angiogenesis and poor prognosis.By interacting with major histocompatibility complex class II molecules,LAG-3 promotes T cell exhaustion and suppresses antitumor responses,often in collaboration with other immune checkpoints like programmed cell death protein 1(PD-1),while on Tregs and NK cells,LAG-3 modulates their suppressive functions,indirectly facilitating tumor immune escape.LAG-3 expression may offer prognostic insights,correlating with disease progression and outcomes in HCC patients,while various preclinical studies highlight the potential of LAG-3-targeted therapies in reinvigorating immune responses against HCC,with a few combination approaches targeting LAG-3 alongside other checkpoints demonstrating synergistic effects in restoring T cell function.Therefore,harnessing LAG-3 as a therapeutic target holds promise for enhancing antitumor immunity and potentially improving HCC treatment outcomes.Our narrative review aims to delve into the full spectrum of LAG-3 signaling in HCC,with the goal of a better understanding of the pathophysiological and immunological basis of its use to arrest HCC growth and development.
基金ThisworkwassupportedbyagrantfromtheNationalNaturalScienceFoundationofChina (No 3 9470 2 93 )
文摘OBJECTIVE: To investigate which of the two immunoglobulin (Ig)-like domains, the immunoglobulin variable region homologous domain IgV (hB7.2 IgV) and the immunoglobulin constant region homologous domain IgC (hB7.2 IgC) on the human B7.2 molecule contains receptor binding sites, and to evaluate whether the B7.2 protein expressed in bacteria has biological activity in vitro. METHODS: Three fragments of hB7.2 IgV,hB7.2 IgC and the complete extracellular region of human B7.2 containing both the IgV and IgC domains,hB7.2 Ig (V+C), were amplified by PCR and subcloned into pGEM-Teasy. Three recombinants,pGEX-4T-3-hB7.2 IgV,pGEX-4T-3-hB7.2 IgC and pGEX-4T-3-hB7.2 Ig (V+C), were generated by cloning the fragments into a prokaryote expression plasmid (pGEX-4T-3) and transformed into the host strain E. coli DH5alpha. The relevant target fusion proteins consisting of GST and hB7.2 IgV,hB7.2 IgC and hB7.2 Ig (V+C), were identified by SDS-PAGE and Western blotting. With the presence of the first signal imitated by anti-CD3 antibody, T cell activation was observed by exposing purified T lymphocytes to each soluble form of the three bacterially-produced human B7.2 fusion proteins by [(3)H]-TdR incorporation. RESULTS: Three recombinant fusion proteins of human B7.2, GST-hB7.2 IgV, GST-hB7.2 IgC and GST-hB7.2 Ig (V+C) were produced and detected in inclusion body form from engineered bacteria. With the first signal present,T lymphocytes proliferated when co-stimulated by bacterially-produced either GST-hB7.2 Ig (V+C) or GST-hB7.2 IgV fusion proteins, but not by GST-hB7.2 IgC. CONCLUSIONS: Functional human B7.2 fusion protein can be produced in bacteria. The IgV-like domain of human B7.2 is sufficient for B7.2 to interact with its counter-receptors and co-stimulate T lymphocytes.
基金ThisstudywassupportedbytheNationalNaturalScienceFoundationofChina (No 3 970 0 172 )
文摘OBJECTIVE: To investigate the specific cell-mediated immune efficacy of the an HPV16 prophylactic vaccine. METHODS: C57BL/6 mice were randomly divided into 3 groups: experimental group I (treated with pcDNA L1), control group II (treated with pcDNA3.1) and control group III (treated with PBS buffer). The mice were immunized three times during a three-week interval. Ten to fourteen days after the third inoculation, a footpad swelling test was used to detect delayed-type hypersensitivity (DTH) responses. Antigen-specific splenocyte proliferation assay and quantitation of IFN-gamma cells in splenocytes were performed by FACS assay. RESULTS: In the experimental group, splenocytes actively proliferated after stimulation with HPV16 VLP, and had developed a markedly larger amount of CD8(+) IFN-gamma(+) cells, which is an index for special CTL. Also, the footpad was significantly thickened upon inoculation with HPV16 VLP. CONCLUSION: Naked DNA vaccine of HPV16 L1 can induce specific cell-mediated immune responses in mice, which should be considered for evaluation of HPV16 DNA vaccine feasibility.
基金This work was supported by grant Ka 502/19-1 fromthe German Research Council(Deutsche Forschungsgemeinschaft)to D.K.the Cluster of Excellence ExC 306"Inflammation-at-Interfaces"(Deutsche Forschungs-gemeinschaft)to D.K+6 种基金L.K.was supported by a long-term fellowship from the German Academic Exchange Service(DAAD)C.P.is the recipient of a grant fromthe Erich und Gertrud Roggenbruck FoundationThis work was also supported by the Major International Joint Research Program of China(Grant 31420103901)the Key Program of the National Natural Science Foundation of China(Grant31830021)the"111"project(816021)the Incubating Program from the Science and Technology Department of Guangdong Province of China(Grant2014A030308003)Guangzhou Science and Technology Key Project(201604020006)to Z.Y.
文摘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.
