AIM： To investigate the expression of B7-H1 in human colorectal carcinoma （CRC） to define its regulating ef- fects on T cells in tumor microenvironment.
Using a newly generated monoclonal antibody （2E6） against human B7-H3, we explored the expression of the molecule on dendritic cells derived from monocytes （Mo-DCs）. Its expression was examined by means of immunos...Using a newly generated monoclonal antibody （2E6） against human B7-H3, we explored the expression of the molecule on dendritic cells derived from monocytes （Mo-DCs）. Its expression was examined by means of immunostaining and flow cytometric （FCM） analysis. The results showed that B7-H3 was expressed in the course of Mo-DC maturation induced with interleukin 4 （IL-4） and granulocyte/macrophage colony-stimulating factor （GM-CSF）. The expression could be detected at all the stages of Mo-DC differentiation, and remained at a quite stable level. Interestingly, B7-H3 was not expressed by T cells and B cells, even these cells were activated respectively by PHA or PWM. A weak expression could be detected on resting monocytes. These data showed that constitutive expression of B7-H3 at a high level was found on imDCs and mDCs derived from monocytes. Due to no expression on T cells and B cells, we speculate that B7-H3 might be another valuable molecule marker for Mo-DCs.展开更多
AIM: To explore the probable pathway by which curcumin(Cur) regulates the function of Treg cells by observing the expression of costimulatory molecules of dendritic cells(DCs).METHODS: Experimental colitis was induced...AIM: To explore the probable pathway by which curcumin(Cur) regulates the function of Treg cells by observing the expression of costimulatory molecules of dendritic cells(DCs).METHODS: Experimental colitis was induced by administering 2, 4, 6-trinitrobenzene sulfonic acid(TNBS)/ethanol solution. Forty male C57BL/6 mice were randomly divided into four groups: normal, TNBS + Cur, TNBS + mesalazine(Mes) and TNBS groups. The mice in the TNBS + Cur and TNBS +Mes groups were treated with Cur and Mes, respectively, while those in the TNBS group were treated with physiological saline for 7 d. After treatment, the curative effect of Cur was evaluated by colonic weight, colonic length, weight index of the colon, and histological observation and score. The levels of CD4+CD25+Foxp3+ T cells(Treg cells) and costimulatory molecules of DCs were measured by flow cytometry. Also, related cytokines were analyzed by enzyme-linked immunosorbent assay. RESULTS: Cur alleviated inflammatory injury of the colonic mucosa, decreased colonic weigh and histological score, and restored colonic length. The number of Treg cells was increased, while the secretion of TNF-α, IL-2, IL-6, IL-12 p40, IL-17 and IL-21 and the expression of costimulatory molecules(CD205, CD54 [ICAM-1], TLR4, CD252[OX40 L], CD256 [RANK] and CD254 [RANK L]) of DCs were notably inhibited in colitis mice treated with Cur.CONCLUSION: Cur potentially modulates activation of DCs to enhance the suppressive functions of Treg cells and promote the recovery of damaged colonic mucosa in inflammatory bowel disease.展开更多
To investigate the effect of costimulatory factors in the pathogenesis of chronic idiopathic thrombocytopenic purpura(CITP), we examined the expression of CD80 on platelets and megakaryocytes in patients with CITP and...To investigate the effect of costimulatory factors in the pathogenesis of chronic idiopathic thrombocytopenic purpura(CITP), we examined the expression of CD80 on platelets and megakaryocytes in patients with CITP and the controls by FACS. By using CD80 monoclonal antibody (McAb) to inhibit interaction among cells which is mediated by costimulatory factors, we observed the effect of CD80 McAb on the growth and maturation of megakaryocytic progenitors of patients with CITP in vitro . The results showed the expression of CD80 on platelets and megakaryocytes in CITP group was significantly higher than that in controls ( P <0.01). There was a significantly positive correlation between the expression of CD80 on platelets and serum PAIgG in CITP (r=0.86, P <0.05). The mean of various clone numbers (CFU MK, BFU MK and mCFU MK) in CITP were all lower than those in controls ( P <0.05). In megakaryocytes co cultured with CD80 McAb, there was an increasing tendency of the number of CFU MK and big CFU MK (the number of megakaryocyte with GPⅢ a positive was more than 20) and mediate CFU MK (the number of megakaryocyte with GPⅢ a positive was 11-20). When the concentration of CD80 McAb was 10 μg/L, there was a significant difference in the number of megakaryocytic colony formation (CFU MK, BFU MK and mCFU MK) between the group with CD80 McAb and that without it ( P <0.05).These showed the abnormality of costimulatory factors had important effect in the pathogenesis of CITP.展开更多
B7-H1, a recently described member of the B7 family of costimulatory molecules, is thought to be involved in tumor immune escape by inducing T-cell apoptosis. In order to investigate the relationship between B7-H1 and...B7-H1, a recently described member of the B7 family of costimulatory molecules, is thought to be involved in tumor immune escape by inducing T-cell apoptosis. In order to investigate the relationship between B7-H1 and immune escape of bladder cancer, B7-H1 expression in 50 cases of bladder cancer was detected by using immunohistochemical method. Survival curves were con- structed using the Kaplan-Meier method and independent prognostic factors were evaluated using the Cox regression model. Our results showed that the positive rate of B7-H1 immunostaining in normal bladder tissue and bladder cancer was 0 and 72% respectively. The expression of B7-H1 was strongly associated with the pathological grade, clinical stage and recurrence （P〈0.05）. The survival rate was significantly lower in patients with B7-H1 positive group than in those with B7-H1 negative group and multi-variable analysis revealed that B7-H1 could be regarded as an independent factor in evaluating the prognosis of bladder cancer. It is concluded that the expression of B7-H1 is strongly associated with neoplastic progression and prognosis of bladder cancer. The manipulation of B7-H1 may become a beneficial target for immunotherapy in human bladder cancer.展开更多
Background The B7/CD28 pathway provides critical costimulatory signals for complete T cell activation, and members of this pathway have served as useful targets for immunotherapeutic strategies. In this study, we inve...Background The B7/CD28 pathway provides critical costimulatory signals for complete T cell activation, and members of this pathway have served as useful targets for immunotherapeutic strategies. In this study, we investigated the RNA interference (RNAi) effect induced by small interfering RNA (siRNA) targeting CD28 mRNA on human lymphocytes and its specificity.Methods According to CD28 gene sequence, we designed and synthysized three different siRNAs ( siRNA-1,siRNA-2, siRNA-3 ) containing 21 bases using SilencerTM siRNA construction kit. These siRNAs were transfected into freshly isolated human lymphocytes with Lipofectamine 2000 reagent. At 24-hour, 48-hour and 72-hour post transfection, these cells were collected and analyzed. The changes of surface expression of CD28 gene were detected by flow cytometry, and the changes of CD28 mRNA levels were determined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). The cell viability of transfected lymphocytes was determined by methyl thiazolyl tetrazolium (MTT) assay and trypan blue dye exclusion assay.Results Three siRNAs (siRNA-1, siRNA-2, siRNA-3) specifically targeting CD28 mRNA were successfully designed and constructed. Flow cytometry analysis showed that a decrease in CD28 expression was detectable at 24-hour post transfection. Different siRNA showed different inhibition effects on CD28 expression. At 48-hour post transfection, the degrees of reduction with siRNA-1, siRNA-2 and siRNA-3 were 22. 10% ± 1.63% ,73.50% ± 1.02% and 42.90% ± 0.89% respectively compared with the control ( P < 0. 001 ). Neither of the groups transfected only with siRNA or lipo showed marked reduction in CD28 expression (3.15% ± 0.75% and 4. 55% ±0. 80% ) (P >0. 05). Moreover, lymphocytes treated with siRNA-co showed no marked reduction in CD28 expression (5.07% ± 0. 96% ) (P > 0. 05 ). The results of semi-quantitative RT-PCR assay indicated CD28 mRNA level was inhibited after transfection of specific siRNAs. At least 4-fold of reduction in siRNA-2 group 展开更多
The T cell costimulatory pathways are central to regulating immune responses,and targeting these pathways represents one of the most promising approaches for achieving immunotherapy.The molecular structures of costimu...The T cell costimulatory pathways are central to regulating immune responses,and targeting these pathways represents one of the most promising approaches for achieving immunotherapy.The molecular structures of costimulation revealed invaluable mechanistic insights underlying costimulatory receptor/ligand specificity,affinity,oligomeric state,and valency,which provided the bases for better manipulation of these signaling pathways.The incredible growth of this field led to identification of new members and unexpected interactions,revealing a complicated regulatory network of immune responses.The advances in structural biology of costimulation will promise unprecedented opportunities for furthering our understanding and therapeutic application of T cell costimulatory pathways.展开更多
PD-L1 is a member of the B7 protein family,most of whose members so far were identified as dimers in a solution and crystalline state,either complexed or uncomplexed with their ligand(s).The binding of PD-L1 with its ...PD-L1 is a member of the B7 protein family,most of whose members so far were identified as dimers in a solution and crystalline state,either complexed or uncomplexed with their ligand(s).The binding of PD-L1 with its receptor PD-1(CD279)delivers an inhibitory signal regulating the T cell function.Simultaneously with the Garboczi group,we successfully solved another structure of human PD-L1(hPD-L1).Our protein crystallized in the space group of C222_(1) with two hPD-L1 molecules per asymmetric unit.After comparison of reported B7 structures,we have found some intrinsic factors involved in the interaction of these two molecules.Based on these results,we tend to believe this uncomplexed hPD-L1 structure demonstrated its potential dimeric state in solution,althougt it could just be an evolutionary relic,too weak to be detected under present technology,or still a functional unit deserved our attentions.展开更多
Background This study was to evaluate whether anergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways can act as potent immunoregulatory cells in vitro and prolong cardiac allograft...Background This study was to evaluate whether anergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways can act as potent immunoregulatory cells in vitro and prolong cardiac allograft survival after adoptive transfer KH*2/5DMethods Anergic cells were induced in vitro by the addition of anti-CD154 and anti-CD80 monoclonal antibodies (mAbs) to primary MLR (mixed lymphocyte reaction) consisting of BALB/c as responder and C3H as stimulator Anergic cells were added to a newly formed MLR in assessing the regulatory capacity and antigen specificity of anergic cells The ability of anergic cells to respond to antigen and/or exogenous recombinant mouse interleukin-2 (rmIL-2) was tested For in vivo studies, anergic cells were intravenously injected into 3 0-Gy γ-irradiated BALB/c mice immediately after heterotopic abdominal cardiac transplantation To prolong allograft survival, recipient mice injected with anergic cells received rapamycin therapy (1 mg·day -1 ·kg -1 ) KH*2/5DResults Anergic cells strongly suppressed the proliferation of naǐve BALB/c splenocytes against the original (C3H) stimulator in a dose-dependent manner, but they failed to suppress the proliferation of naǐve BALB/c splenocytes against the third-party (C57BL/6J) stimulator The anergic state was reversed by both original (C3H) stimulator and additional exogenous IL-2 In in vivo studies, untreated irradiated BALB/c mice rejected C3H cardiac allografts with a mean survival time of (8 6±1 1) days, whereas those injected with the anergic cells rejected the allografts with a mean survival time of (11 8±1 9) days, which was slightly longer than that of the untreated mice The protocol based on anergic cells injection plus rapamycin therapy could prolong allograft survival significantly [(29 6±4 4) days] Conclusions Anergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways can act as potent immunoregulatory cells in vitro , and prolong ca展开更多
CD137,a costimulatory factor of TNFR family,is expressed on activated T cells and freshly isolated mouse dendritic cells (DCs).To date,there are only limited data dealing with the expression and the effect of CD137 on...CD137,a costimulatory factor of TNFR family,is expressed on activated T cells and freshly isolated mouse dendritic cells (DCs).To date,there are only limited data dealing with the expression and the effect of CD137 on human DCs.We report in this work that CD137 can coexpress with CD137L on immature peripheral blood-derived human DCs (9.77%).The mature DCs stimulated by LPS showed a much higher level of CD137 expression (36.06%).Ligation of CD137 on the surface of DCs with anti-CD137 monoclonal antibody (mAb) could enhance the direct anti-tumor effect mediated by human DCs in vitro.The agonistic anti-CD137 mAb was able to elevate by about 20% of the DC-mediated inhibition of tumor growth in five tumor cell lines.These results indicate that the appliance of anti-CD137 mAb might be used as a new strategy for tumor immunotherapy.Cellular & Molecular Immunology.2004;1(1):71-76.展开更多
AIM: To explore the effects of interferon-α(IFN-α) application on peripheral circulating CD1αdendritic cells (DCs) in patients with chronic hepatitis B, and the expression of HLA-DR, CD80, and ICAM-1 on CD1αDCs in...AIM: To explore the effects of interferon-α(IFN-α) application on peripheral circulating CD1αdendritic cells (DCs) in patients with chronic hepatitis B, and the expression of HLA-DR, CD80, and ICAM-1 on CD1αDCs in order to explore the mechanism of immune modulation of IFN-α. METHODS: By flow cytometry technique, changes of CD1αDCs were monitored in 22 patients with chronic hepatitis B treated with IFN-αand in 16 such patients not treated with IFN-αwithin three months. Meanwhile, the expression of HLA-DR, CD80, and ICAM-1 on CD1αDCs was detected. RESULTS: In the group of IFN-αtreatment, the percentage of CD1αDCs in peripheral blood mono-nuclear cells was increased after three months of therapy. In patients who became negative for HBV-DNA after IFN-αtreatment, the increase of DCs was more prominent, while in control, these changes were not observed. Increased expression of HLA-DR, CD80, and ICAM-1 on CD1αDCs was also observed. CONCLUSION: CD1αDCs can be induced by IFN-αin vivo, and the immune related molecules such as HLA-DR, CD80, and ICAM-1 are up-regulated to some degree. This might be an important immune related mechanism of IFN-αtreatment for chronic hepatitis B.展开更多
Background The hygiene hypothesis has been proposed to explain the pathogenesis of asthma.Allergen exposure was shown to inhibit asthma in an animal model.But the optimal timing of allergen exposure remains unclear.Th...Background The hygiene hypothesis has been proposed to explain the pathogenesis of asthma.Allergen exposure was shown to inhibit asthma in an animal model.But the optimal timing of allergen exposure remains unclear.This study aims to explore the time effcct of allergen exposure and the possible mechanisms.Methods Neonate Wistar rats were randomly divided into asthma group,control group and day 1,day 3,day 7,and day 14 groups.The day 1,day 3,day 7 and day 14 groups were injected with ovalbumin （OVA） subcutaneously on days 1,3,7 and 14 after birth,respectively.Six weeks later,all groups,except the control group,were sensitized and stimulated with OVA to make the asthma model.We observed the pulmonary pathologic changes,detected the regulatory T cells,and CD28 expression level in thymus and spleen by flow cytometry.Results The asthmatic inflammation in the day 1,day 3 and day 7 groups,but not the day 14 group,was alleviated.The asthma group and day 14 group had lower proportions of regulatory T cells in the thymus compared with the control group,day 1,day 3,and day 7 groups.There was no significant difference in the CD28 expression levels on the regulatory and conventional T cells among groups.But the control group and the day 1,day 3,and day 7 groups had relatively higher proportions of CD28 positive regulatory T cells in the thymus than the day 14 group and the asthma group.Conclusions There is a “time-window” for early allergen exposure.The impairment of regulatory T cells may promote the development of asthma.Allergen exposure in the “time-window” can make the thymus produce normal quantity of regulatory cells.The CD28 signal on regulatory T cells may participate in the production of regulatory T cells.展开更多
Experimental allergic encephalomyelitis is a mouse model of human multiple sclerosis with similar pathology and pathogenesis. Thl cells play an important role in the pathogenesis of experimental allergic encephalomyel...Experimental allergic encephalomyelitis is a mouse model of human multiple sclerosis with similar pathology and pathogenesis. Thl cells play an important role in the pathogenesis of experimental allergic encephalomyelitis. This study determined the potential effect of programmed cell death 1 ligand 1 in the pathogenesis of experimental allergic encephalomyelitis induced by injecting myelin oligodendrocyte glycoprotein, complete Freund＇s adjuvant and Bordetella pertussis toxin into C57BL/6J mice. Experimental allergic encephalomyelitis mice developed disease and showed in- flammatory changes in the central nervous system by hematoxylin-eosin staining of spinal cord pathological sections, demyelination by Luxol fast-blue staining and clinical manifestations. The expression of programmed cell death 1 ligand 1 in mice was detected by immunohistochemistry, flow cytometry and western blot anatysis. The expression of programmed cell death 1 ligand 1 in the spinal cord and splenocytes of mice was significantly increased compared with normal mice. Our findings suggest the involvement of programmed cell death 1 ligand 1 in the pathogenesis of ex- perimental allergic encephalomyelitis and suggest this should be studied in multiple sclerosis.展开更多
Many malignancies lack satisfactory treatment and new therapeutic options are urgently needed.Gene therapy is a new modality to treat both inherited and acquired diseases based on the transfer of genetic material to t...Many malignancies lack satisfactory treatment and new therapeutic options are urgently needed.Gene therapy is a new modality to treat both inherited and acquired diseases based on the transfer of genetic material to the tissues.Different gene therapy strategies against cancers have been developed.A considerable number of preclinical studies indicate that a great variety of cancers are amenable to gene therapy.Among these strategies, induction of anti-tumor immunity is the most promising approach.Gene therapy with cytokines has reached unprecedented success in preclinical models of cancer.Synergistic rather than additive effects have been demonstrated by combination of gene transfer of cytokines/chemokines,costimulatory molecules or adoptive cell therapy.Recent progress in vector technology and in imaging techniques allowing in vivo assessment of gene expression will facilitate the development of clinical applications of gene therapy,a procedure which may have a notorious impact in the management of cancers lacking effective treatment.Cellular & Molecular Immunology.2004;1(2):105-111.展开更多
Objective： Heat shock protein （HSP） has the promiscuous abilities to chaperone and present a broad repertoire of tumor antigens to antigen presenting cells including DCs. In this report, we analyzed the modulation ...Objective： Heat shock protein （HSP） has the promiscuous abilities to chaperone and present a broad repertoire of tumor antigens to antigen presenting cells including DCs. In this report, we analyzed the modulation of immature DC by lISP 96 （gp96）. Method： Murine bone marro6-derived DC was induced by GM-CSF plus IL-4, which aped the immunostimulatory effects of DC. Cocultured DC and gp96-peptide complexes （gp96-PC） or inactivated H22 cells, the expression of MHC class Ⅱ , CD40, CD80 was quantified by flow cytometry. The concentration of IL-12 and TNF- in culture supematants were determined by ELISA. C1 release assay was used to test specific cytotoxic T cell. Results： Our study demonstrated that the extent of DC maturation induced by gp96-PC, which was reflected in surface density of costimulatory and MHC Ⅱmolecules, was correlated with the secretion of IL-12 and with the T cellactivating potential in vitro. Conclusion： Heat shock protein 96 could be isolated and purified from H22 cells and could induce maturation of dendritic cell. Our findings might be relevance to the use of DC vaccine in therapy of human tumors.展开更多
Summary: The expression of the costimulatory molecules B7/CD28 in peripheral blood mononuclear cells (PBMC) of the patients with systemic lupus erythematosus (SLE) and its relation to the pathogenesis of SLE were stud...Summary: The expression of the costimulatory molecules B7/CD28 in peripheral blood mononuclear cells (PBMC) of the patients with systemic lupus erythematosus (SLE) and its relation to the pathogenesis of SLE were studied. The expression of the costimulatory molecules in PBMC in 30 patients with active SLE and 20 cases of healthy controls was detected by using the techniques of immunofluorescence and flow cytometer. The result showed that the expression percentage of CD28+, CD4+CD28+ in T cells of PBMC from the patients with SLE decreased significantly as compared with that in healthy control group, while the expression percentage of CD80+, CD19+CD80+ in B cells was significantly increased than that in healthy control group (P<0.01). It suggested that the abnormal expression of costimulatory molecules B7/CD28 played a role in the pathogenesis of SLE.展开更多
Objective To detect expression of intercellular adhesion molecule 1 (ICAM 1), B7.1 and thyroid peroxidase (TPO) on thyrocyte and study the possible mechanism of interferon alpha (IFN α) in the pathogenesis ...Objective To detect expression of intercellular adhesion molecule 1 (ICAM 1), B7.1 and thyroid peroxidase (TPO) on thyrocyte and study the possible mechanism of interferon alpha (IFN α) in the pathogenesis of autoimmune thyroid disease (AITD). Methods Thyrocytes were cultured from 6 normal persons. Antigen expression on thyrocytes induced by cytokines was examined using immunofluorescence staining with flow cytometer. Results IFN α significantly stimulated the expression of ICAM 1, B7.1 and TPO, as compared with those of control group. IFN γ markedly enhanced the expression of HLA DR and ICAM 1, but not B7.1. Prolactin (PRL) resulted in increased expression of ICAM 1, B7.1, as well as overexpression of TPO, which is more significant than that stimulated by IFN α. Conclusions Thyroid autoimmunity induced by IFN α is associated with the expression of ICAM 1, B7.1 and TPO. IFN γ could not induce the expression of B7.1, therefore it is not an initiator in AITD. In addition, we should pay more attention to PRL which possibly plays an important role in the initiation and perpetuation of postpartum thyroiditis.展开更多
Alzheimer disease （AD） is characterized by the .presence of β-amyloid （Aβ） plaques in the brain.More evidence of inflammatory parameters, such as, complement factors, proinflammatory cytokines and lymphocytes ha...Alzheimer disease （AD） is characterized by the .presence of β-amyloid （Aβ） plaques in the brain.More evidence of inflammatory parameters, such as, complement factors, proinflammatory cytokines and lymphocytes has been found to be co-localized with Aβ plaques. The research in the past decades has demonstrated abnormalities of both the humoral and cellular immune responses, suggesting an association of immunological aberration and AD. The total percentage of lymphocytes was not found to be altered,展开更多
AIM： Although the pathogenic mechanism underlying autoimmune hepatitis （AIH） remains unclear, the immune system is thought to be critical for the progression of the disease. Cellular immune responses may be linked ...AIM： Although the pathogenic mechanism underlying autoimmune hepatitis （AIH） remains unclear, the immune system is thought to be critical for the progression of the disease. Cellular immune responses may be linked to the hepatocellular damage in AIH. Recently, much attention has been focused on the critical functions of costimulatory molecules expressed on mononuclear cells in the generation of effective T cell-mediated immune responses. Analysis of costimulatory molecule expressed on mononuclear cells from the patients with AIH may give us insight into the pathogenic mechanism of hepatocellular damage in AIH. METHODS： Peripheral blood mononuclear cells （PBMC） were taken from the patients with AIH （34 cases） and healthy controls （25 cases）. Uver infiltrating mononuclear cells （LIMCs） were taken from the patients with AIH （18 cases）, the patient with chronic hepatitis C （CH-C） （13 cases） and the patients with fatty liver （2 cases）. Using flow cytometry, the cells were analyzed for the expression of costimulatory molecules, such as CD80, CD86, and CD152 （CTLA-4）. The results were compared with clinical data such as the level of gammaglobulin, histological grade, presence or absence of corticosteroids administration and the response to corticosteroids. RESULTS： The levels of CD80＋, CD86＋ and CD152＋ PBMC were significantly reduced in the patients with AIH as compared with healthy controls. By contrast, those cells were significantly higher in LIMC than in PBMC of the patients with AIH. Especially, the level of CD86＋ LIMC showed a marked increase irrespective of the degree of disease activity in the patients with AIH,although CD86＋ cells were rarely present in PBMC. The levels of CD86＋ cells were present in significantly higher frequency in patients with AIH than in the patients with CH-C. Furthermore, the patients with AIH with high levels of CD86＋ LIMC showed good responses to corticosteroids, whereas 2 cases of AIH with low levels of CD86＋ LIMC did not r展开更多
Background OX40/OX40 ligand （OX40/OX40L） and programmed death-1/programmed death ligand-1 （PD-1/PD-L1） co- stimulator/signals play important roles in T cell-induced immune responses. The aim of this study was to i...Background OX40/OX40 ligand （OX40/OX40L） and programmed death-1/programmed death ligand-1 （PD-1/PD-L1） co- stimulator/signals play important roles in T cell-induced immune responses. The aim of this study was to investigate the roles of OX40/OX40L and PD-1/PD-L1 costimulatory pathways in mouse islet allograft rejection. Methods Lentiviral vectors containing OX40L siRNA sequences and an adenovirus vector containing the PD-L1 gene were constructed. The streptozotocin-induced model of diabetes was established in C57BL/6 （H-2b） mice. Diabetic C57BL/6 mice were randomly allocated into five groups： group 1, untreated control; group 2, Ad-EGFP treatment; group 3, Ad-PD-L1 treatment; group 4, OX40L-RNAi-LV treatment; group 5, OX40L-RNAi-LV combined with Ad-PD-L1 treatment. Lentiviral vector and the adenovirus vector were injected, singly or combined, into the caudal vein one day before islet transplantation. The islets of DBA/2 （H-2d） mice were transplanted into the renal subcapsular space of the diabetic recipients. Recipient blood glucose and the survival time of the allografts were monitored. Antigen-specific mixed lymphocyte reaction was also evaluated.展开更多
基金Supported by Grants from the Major State Basic Research Development Program of China 973 Program,No.2007CB512402National Natural Science Foundation of China,No.31100634+1 种基金Natural Science Foundation of Jiangsu Province,No.BK2010161"333" Project of Wuxi City,Jiangsu Province,No.CAE00901-09
文摘AIM： To investigate the expression of B7-H1 in human colorectal carcinoma （CRC） to define its regulating ef- fects on T cells in tumor microenvironment.
基金supported by a grant from the National Natural Science Foundation of China(N0.30330540).
文摘Using a newly generated monoclonal antibody （2E6） against human B7-H3, we explored the expression of the molecule on dendritic cells derived from monocytes （Mo-DCs）. Its expression was examined by means of immunostaining and flow cytometric （FCM） analysis. The results showed that B7-H3 was expressed in the course of Mo-DC maturation induced with interleukin 4 （IL-4） and granulocyte/macrophage colony-stimulating factor （GM-CSF）. The expression could be detected at all the stages of Mo-DC differentiation, and remained at a quite stable level. Interestingly, B7-H3 was not expressed by T cells and B cells, even these cells were activated respectively by PHA or PWM. A weak expression could be detected on resting monocytes. These data showed that constitutive expression of B7-H3 at a high level was found on imDCs and mDCs derived from monocytes. Due to no expression on T cells and B cells, we speculate that B7-H3 might be another valuable molecule marker for Mo-DCs.
基金Supported by project of National Natural Science Foundation of ChinaNo.81260595 and No.81460679+3 种基金Chinese Scholarship Council and Jiangxi province as visiting scholarNo.201408360106 and No.201408360110project of Jiangxi University of Traditional Chinese MedicineNo.JZYC15S13
文摘AIM: To explore the probable pathway by which curcumin(Cur) regulates the function of Treg cells by observing the expression of costimulatory molecules of dendritic cells(DCs).METHODS: Experimental colitis was induced by administering 2, 4, 6-trinitrobenzene sulfonic acid(TNBS)/ethanol solution. Forty male C57BL/6 mice were randomly divided into four groups: normal, TNBS + Cur, TNBS + mesalazine(Mes) and TNBS groups. The mice in the TNBS + Cur and TNBS +Mes groups were treated with Cur and Mes, respectively, while those in the TNBS group were treated with physiological saline for 7 d. After treatment, the curative effect of Cur was evaluated by colonic weight, colonic length, weight index of the colon, and histological observation and score. The levels of CD4+CD25+Foxp3+ T cells(Treg cells) and costimulatory molecules of DCs were measured by flow cytometry. Also, related cytokines were analyzed by enzyme-linked immunosorbent assay. RESULTS: Cur alleviated inflammatory injury of the colonic mucosa, decreased colonic weigh and histological score, and restored colonic length. The number of Treg cells was increased, while the secretion of TNF-α, IL-2, IL-6, IL-12 p40, IL-17 and IL-21 and the expression of costimulatory molecules(CD205, CD54 [ICAM-1], TLR4, CD252[OX40 L], CD256 [RANK] and CD254 [RANK L]) of DCs were notably inhibited in colitis mice treated with Cur.CONCLUSION: Cur potentially modulates activation of DCs to enhance the suppressive functions of Treg cells and promote the recovery of damaged colonic mucosa in inflammatory bowel disease.
基金Theprojectwassupportedbya grantforreturnedscholarsfromtheMinistryofEducation (No .6 - 74 )
文摘To investigate the effect of costimulatory factors in the pathogenesis of chronic idiopathic thrombocytopenic purpura(CITP), we examined the expression of CD80 on platelets and megakaryocytes in patients with CITP and the controls by FACS. By using CD80 monoclonal antibody (McAb) to inhibit interaction among cells which is mediated by costimulatory factors, we observed the effect of CD80 McAb on the growth and maturation of megakaryocytic progenitors of patients with CITP in vitro . The results showed the expression of CD80 on platelets and megakaryocytes in CITP group was significantly higher than that in controls ( P <0.01). There was a significantly positive correlation between the expression of CD80 on platelets and serum PAIgG in CITP (r=0.86, P <0.05). The mean of various clone numbers (CFU MK, BFU MK and mCFU MK) in CITP were all lower than those in controls ( P <0.05). In megakaryocytes co cultured with CD80 McAb, there was an increasing tendency of the number of CFU MK and big CFU MK (the number of megakaryocyte with GPⅢ a positive was more than 20) and mediate CFU MK (the number of megakaryocyte with GPⅢ a positive was 11-20). When the concentration of CD80 McAb was 10 μg/L, there was a significant difference in the number of megakaryocytic colony formation (CFU MK, BFU MK and mCFU MK) between the group with CD80 McAb and that without it ( P <0.05).These showed the abnormality of costimulatory factors had important effect in the pathogenesis of CITP.
基金supported by a grant from Hubei Provin-cial Science and Technology Key Program Foundation of China (No. 2007AA402C60).
文摘B7-H1, a recently described member of the B7 family of costimulatory molecules, is thought to be involved in tumor immune escape by inducing T-cell apoptosis. In order to investigate the relationship between B7-H1 and immune escape of bladder cancer, B7-H1 expression in 50 cases of bladder cancer was detected by using immunohistochemical method. Survival curves were con- structed using the Kaplan-Meier method and independent prognostic factors were evaluated using the Cox regression model. Our results showed that the positive rate of B7-H1 immunostaining in normal bladder tissue and bladder cancer was 0 and 72% respectively. The expression of B7-H1 was strongly associated with the pathological grade, clinical stage and recurrence （P〈0.05）. The survival rate was significantly lower in patients with B7-H1 positive group than in those with B7-H1 negative group and multi-variable analysis revealed that B7-H1 could be regarded as an independent factor in evaluating the prognosis of bladder cancer. It is concluded that the expression of B7-H1 is strongly associated with neoplastic progression and prognosis of bladder cancer. The manipulation of B7-H1 may become a beneficial target for immunotherapy in human bladder cancer.
文摘Background The B7/CD28 pathway provides critical costimulatory signals for complete T cell activation, and members of this pathway have served as useful targets for immunotherapeutic strategies. In this study, we investigated the RNA interference (RNAi) effect induced by small interfering RNA (siRNA) targeting CD28 mRNA on human lymphocytes and its specificity.Methods According to CD28 gene sequence, we designed and synthysized three different siRNAs ( siRNA-1,siRNA-2, siRNA-3 ) containing 21 bases using SilencerTM siRNA construction kit. These siRNAs were transfected into freshly isolated human lymphocytes with Lipofectamine 2000 reagent. At 24-hour, 48-hour and 72-hour post transfection, these cells were collected and analyzed. The changes of surface expression of CD28 gene were detected by flow cytometry, and the changes of CD28 mRNA levels were determined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). The cell viability of transfected lymphocytes was determined by methyl thiazolyl tetrazolium (MTT) assay and trypan blue dye exclusion assay.Results Three siRNAs (siRNA-1, siRNA-2, siRNA-3) specifically targeting CD28 mRNA were successfully designed and constructed. Flow cytometry analysis showed that a decrease in CD28 expression was detectable at 24-hour post transfection. Different siRNA showed different inhibition effects on CD28 expression. At 48-hour post transfection, the degrees of reduction with siRNA-1, siRNA-2 and siRNA-3 were 22. 10% ± 1.63% ,73.50% ± 1.02% and 42.90% ± 0.89% respectively compared with the control ( P < 0. 001 ). Neither of the groups transfected only with siRNA or lipo showed marked reduction in CD28 expression (3.15% ± 0.75% and 4. 55% ±0. 80% ) (P >0. 05). Moreover, lymphocytes treated with siRNA-co showed no marked reduction in CD28 expression (5.07% ± 0. 96% ) (P > 0. 05 ). The results of semi-quantitative RT-PCR assay indicated CD28 mRNA level was inhibited after transfection of specific siRNAs. At least 4-fold of reduction in siRNA-2 group
文摘The T cell costimulatory pathways are central to regulating immune responses,and targeting these pathways represents one of the most promising approaches for achieving immunotherapy.The molecular structures of costimulation revealed invaluable mechanistic insights underlying costimulatory receptor/ligand specificity,affinity,oligomeric state,and valency,which provided the bases for better manipulation of these signaling pathways.The incredible growth of this field led to identification of new members and unexpected interactions,revealing a complicated regulatory network of immune responses.The advances in structural biology of costimulation will promise unprecedented opportunities for furthering our understanding and therapeutic application of T cell costimulatory pathways.
基金This work was supported by a grant from Ministry of Science and Technology(MOST)of China for the basic research program 973,Grant No.2006CB504204a grant from National Natural Science Foundation(NSFC)of China,Grant No.30671903+3 种基金a grant from Chinese Academy of Sciences(CAS)Knowledge Innovation Project,Grant No.KSCX2-SW-227G.F.G.is a distinguished young investigator of the NSFC(Grant No.30525010)Y.C.is a Ph.D.student supported by Science Innovation Project,Graduate University of Chinese Academy of Sciences(GUCAS),No.0729031EE1The China-Japan Joint Laboratory of Molecular Immunology and Molecular Microbiology is partlially supported by Japan MEXT(Ministry of Education,Culture,Sports,Science and Technology).
文摘PD-L1 is a member of the B7 protein family,most of whose members so far were identified as dimers in a solution and crystalline state,either complexed or uncomplexed with their ligand(s).The binding of PD-L1 with its receptor PD-1(CD279)delivers an inhibitory signal regulating the T cell function.Simultaneously with the Garboczi group,we successfully solved another structure of human PD-L1(hPD-L1).Our protein crystallized in the space group of C222_(1) with two hPD-L1 molecules per asymmetric unit.After comparison of reported B7 structures,we have found some intrinsic factors involved in the interaction of these two molecules.Based on these results,we tend to believe this uncomplexed hPD-L1 structure demonstrated its potential dimeric state in solution,althougt it could just be an evolutionary relic,too weak to be detected under present technology,or still a functional unit deserved our attentions.
文摘Background This study was to evaluate whether anergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways can act as potent immunoregulatory cells in vitro and prolong cardiac allograft survival after adoptive transfer KH*2/5DMethods Anergic cells were induced in vitro by the addition of anti-CD154 and anti-CD80 monoclonal antibodies (mAbs) to primary MLR (mixed lymphocyte reaction) consisting of BALB/c as responder and C3H as stimulator Anergic cells were added to a newly formed MLR in assessing the regulatory capacity and antigen specificity of anergic cells The ability of anergic cells to respond to antigen and/or exogenous recombinant mouse interleukin-2 (rmIL-2) was tested For in vivo studies, anergic cells were intravenously injected into 3 0-Gy γ-irradiated BALB/c mice immediately after heterotopic abdominal cardiac transplantation To prolong allograft survival, recipient mice injected with anergic cells received rapamycin therapy (1 mg·day -1 ·kg -1 ) KH*2/5DResults Anergic cells strongly suppressed the proliferation of naǐve BALB/c splenocytes against the original (C3H) stimulator in a dose-dependent manner, but they failed to suppress the proliferation of naǐve BALB/c splenocytes against the third-party (C57BL/6J) stimulator The anergic state was reversed by both original (C3H) stimulator and additional exogenous IL-2 In in vivo studies, untreated irradiated BALB/c mice rejected C3H cardiac allografts with a mean survival time of (8 6±1 1) days, whereas those injected with the anergic cells rejected the allografts with a mean survival time of (11 8±1 9) days, which was slightly longer than that of the untreated mice The protocol based on anergic cells injection plus rapamycin therapy could prolong allograft survival significantly [(29 6±4 4) days] Conclusions Anergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways can act as potent immunoregulatory cells in vitro , and prolong ca
基金supported by the National Natural Science Foundation of China (No.30271247)
文摘CD137,a costimulatory factor of TNFR family,is expressed on activated T cells and freshly isolated mouse dendritic cells (DCs).To date,there are only limited data dealing with the expression and the effect of CD137 on human DCs.We report in this work that CD137 can coexpress with CD137L on immature peripheral blood-derived human DCs (9.77%).The mature DCs stimulated by LPS showed a much higher level of CD137 expression (36.06%).Ligation of CD137 on the surface of DCs with anti-CD137 monoclonal antibody (mAb) could enhance the direct anti-tumor effect mediated by human DCs in vitro.The agonistic anti-CD137 mAb was able to elevate by about 20% of the DC-mediated inhibition of tumor growth in five tumor cell lines.These results indicate that the appliance of anti-CD137 mAb might be used as a new strategy for tumor immunotherapy.Cellular & Molecular Immunology.2004;1(1):71-76.
文摘AIM: To explore the effects of interferon-α(IFN-α) application on peripheral circulating CD1αdendritic cells (DCs) in patients with chronic hepatitis B, and the expression of HLA-DR, CD80, and ICAM-1 on CD1αDCs in order to explore the mechanism of immune modulation of IFN-α. METHODS: By flow cytometry technique, changes of CD1αDCs were monitored in 22 patients with chronic hepatitis B treated with IFN-αand in 16 such patients not treated with IFN-αwithin three months. Meanwhile, the expression of HLA-DR, CD80, and ICAM-1 on CD1αDCs was detected. RESULTS: In the group of IFN-αtreatment, the percentage of CD1αDCs in peripheral blood mono-nuclear cells was increased after three months of therapy. In patients who became negative for HBV-DNA after IFN-αtreatment, the increase of DCs was more prominent, while in control, these changes were not observed. Increased expression of HLA-DR, CD80, and ICAM-1 on CD1αDCs was also observed. CONCLUSION: CD1αDCs can be induced by IFN-αin vivo, and the immune related molecules such as HLA-DR, CD80, and ICAM-1 are up-regulated to some degree. This might be an important immune related mechanism of IFN-αtreatment for chronic hepatitis B.
文摘Background The hygiene hypothesis has been proposed to explain the pathogenesis of asthma.Allergen exposure was shown to inhibit asthma in an animal model.But the optimal timing of allergen exposure remains unclear.This study aims to explore the time effcct of allergen exposure and the possible mechanisms.Methods Neonate Wistar rats were randomly divided into asthma group,control group and day 1,day 3,day 7,and day 14 groups.The day 1,day 3,day 7 and day 14 groups were injected with ovalbumin （OVA） subcutaneously on days 1,3,7 and 14 after birth,respectively.Six weeks later,all groups,except the control group,were sensitized and stimulated with OVA to make the asthma model.We observed the pulmonary pathologic changes,detected the regulatory T cells,and CD28 expression level in thymus and spleen by flow cytometry.Results The asthmatic inflammation in the day 1,day 3 and day 7 groups,but not the day 14 group,was alleviated.The asthma group and day 14 group had lower proportions of regulatory T cells in the thymus compared with the control group,day 1,day 3,and day 7 groups.There was no significant difference in the CD28 expression levels on the regulatory and conventional T cells among groups.But the control group and the day 1,day 3,and day 7 groups had relatively higher proportions of CD28 positive regulatory T cells in the thymus than the day 14 group and the asthma group.Conclusions There is a “time-window” for early allergen exposure.The impairment of regulatory T cells may promote the development of asthma.Allergen exposure in the “time-window” can make the thymus produce normal quantity of regulatory cells.The CD28 signal on regulatory T cells may participate in the production of regulatory T cells.
基金financially sponsored by the Natural Science Foundation of Jiangsu Province in China,(General Program),No.BK2011267
文摘Experimental allergic encephalomyelitis is a mouse model of human multiple sclerosis with similar pathology and pathogenesis. Thl cells play an important role in the pathogenesis of experimental allergic encephalomyelitis. This study determined the potential effect of programmed cell death 1 ligand 1 in the pathogenesis of experimental allergic encephalomyelitis induced by injecting myelin oligodendrocyte glycoprotein, complete Freund＇s adjuvant and Bordetella pertussis toxin into C57BL/6J mice. Experimental allergic encephalomyelitis mice developed disease and showed in- flammatory changes in the central nervous system by hematoxylin-eosin staining of spinal cord pathological sections, demyelination by Luxol fast-blue staining and clinical manifestations. The expression of programmed cell death 1 ligand 1 in mice was detected by immunohistochemistry, flow cytometry and western blot anatysis. The expression of programmed cell death 1 ligand 1 in the spinal cord and splenocytes of mice was significantly increased compared with normal mice. Our findings suggest the involvement of programmed cell death 1 ligand 1 in the pathogenesis of ex- perimental allergic encephalomyelitis and suggest this should be studied in multiple sclerosis.
文摘Many malignancies lack satisfactory treatment and new therapeutic options are urgently needed.Gene therapy is a new modality to treat both inherited and acquired diseases based on the transfer of genetic material to the tissues.Different gene therapy strategies against cancers have been developed.A considerable number of preclinical studies indicate that a great variety of cancers are amenable to gene therapy.Among these strategies, induction of anti-tumor immunity is the most promising approach.Gene therapy with cytokines has reached unprecedented success in preclinical models of cancer.Synergistic rather than additive effects have been demonstrated by combination of gene transfer of cytokines/chemokines,costimulatory molecules or adoptive cell therapy.Recent progress in vector technology and in imaging techniques allowing in vivo assessment of gene expression will facilitate the development of clinical applications of gene therapy,a procedure which may have a notorious impact in the management of cancers lacking effective treatment.Cellular & Molecular Immunology.2004;1(2):105-111.
基金National Natural Science Foundation of China (No.30200369)
文摘Objective： Heat shock protein （HSP） has the promiscuous abilities to chaperone and present a broad repertoire of tumor antigens to antigen presenting cells including DCs. In this report, we analyzed the modulation of immature DC by lISP 96 （gp96）. Method： Murine bone marro6-derived DC was induced by GM-CSF plus IL-4, which aped the immunostimulatory effects of DC. Cocultured DC and gp96-peptide complexes （gp96-PC） or inactivated H22 cells, the expression of MHC class Ⅱ , CD40, CD80 was quantified by flow cytometry. The concentration of IL-12 and TNF- in culture supematants were determined by ELISA. C1 release assay was used to test specific cytotoxic T cell. Results： Our study demonstrated that the extent of DC maturation induced by gp96-PC, which was reflected in surface density of costimulatory and MHC Ⅱmolecules, was correlated with the secretion of IL-12 and with the T cellactivating potential in vitro. Conclusion： Heat shock protein 96 could be isolated and purified from H22 cells and could induce maturation of dendritic cell. Our findings might be relevance to the use of DC vaccine in therapy of human tumors.
基金ThisprojectwassupportedbyagrantfromHubeiProvincialNaturalSciencesFoundationofChina (No .2 0 0 0 j0 5 3)
文摘Summary: The expression of the costimulatory molecules B7/CD28 in peripheral blood mononuclear cells (PBMC) of the patients with systemic lupus erythematosus (SLE) and its relation to the pathogenesis of SLE were studied. The expression of the costimulatory molecules in PBMC in 30 patients with active SLE and 20 cases of healthy controls was detected by using the techniques of immunofluorescence and flow cytometer. The result showed that the expression percentage of CD28+, CD4+CD28+ in T cells of PBMC from the patients with SLE decreased significantly as compared with that in healthy control group, while the expression percentage of CD80+, CD19+CD80+ in B cells was significantly increased than that in healthy control group (P<0.01). It suggested that the abnormal expression of costimulatory molecules B7/CD28 played a role in the pathogenesis of SLE.
文摘Objective To detect expression of intercellular adhesion molecule 1 (ICAM 1), B7.1 and thyroid peroxidase (TPO) on thyrocyte and study the possible mechanism of interferon alpha (IFN α) in the pathogenesis of autoimmune thyroid disease (AITD). Methods Thyrocytes were cultured from 6 normal persons. Antigen expression on thyrocytes induced by cytokines was examined using immunofluorescence staining with flow cytometer. Results IFN α significantly stimulated the expression of ICAM 1, B7.1 and TPO, as compared with those of control group. IFN γ markedly enhanced the expression of HLA DR and ICAM 1, but not B7.1. Prolactin (PRL) resulted in increased expression of ICAM 1, B7.1, as well as overexpression of TPO, which is more significant than that stimulated by IFN α. Conclusions Thyroid autoimmunity induced by IFN α is associated with the expression of ICAM 1, B7.1 and TPO. IFN γ could not induce the expression of B7.1, therefore it is not an initiator in AITD. In addition, we should pay more attention to PRL which possibly plays an important role in the initiation and perpetuation of postpartum thyroiditis.
文摘Alzheimer disease （AD） is characterized by the .presence of β-amyloid （Aβ） plaques in the brain.More evidence of inflammatory parameters, such as, complement factors, proinflammatory cytokines and lymphocytes has been found to be co-localized with Aβ plaques. The research in the past decades has demonstrated abnormalities of both the humoral and cellular immune responses, suggesting an association of immunological aberration and AD. The total percentage of lymphocytes was not found to be altered,
文摘AIM： Although the pathogenic mechanism underlying autoimmune hepatitis （AIH） remains unclear, the immune system is thought to be critical for the progression of the disease. Cellular immune responses may be linked to the hepatocellular damage in AIH. Recently, much attention has been focused on the critical functions of costimulatory molecules expressed on mononuclear cells in the generation of effective T cell-mediated immune responses. Analysis of costimulatory molecule expressed on mononuclear cells from the patients with AIH may give us insight into the pathogenic mechanism of hepatocellular damage in AIH. METHODS： Peripheral blood mononuclear cells （PBMC） were taken from the patients with AIH （34 cases） and healthy controls （25 cases）. Uver infiltrating mononuclear cells （LIMCs） were taken from the patients with AIH （18 cases）, the patient with chronic hepatitis C （CH-C） （13 cases） and the patients with fatty liver （2 cases）. Using flow cytometry, the cells were analyzed for the expression of costimulatory molecules, such as CD80, CD86, and CD152 （CTLA-4）. The results were compared with clinical data such as the level of gammaglobulin, histological grade, presence or absence of corticosteroids administration and the response to corticosteroids. RESULTS： The levels of CD80＋, CD86＋ and CD152＋ PBMC were significantly reduced in the patients with AIH as compared with healthy controls. By contrast, those cells were significantly higher in LIMC than in PBMC of the patients with AIH. Especially, the level of CD86＋ LIMC showed a marked increase irrespective of the degree of disease activity in the patients with AIH,although CD86＋ cells were rarely present in PBMC. The levels of CD86＋ cells were present in significantly higher frequency in patients with AIH than in the patients with CH-C. Furthermore, the patients with AIH with high levels of CD86＋ LIMC showed good responses to corticosteroids, whereas 2 cases of AIH with low levels of CD86＋ LIMC did not r
文摘Background OX40/OX40 ligand （OX40/OX40L） and programmed death-1/programmed death ligand-1 （PD-1/PD-L1） co- stimulator/signals play important roles in T cell-induced immune responses. The aim of this study was to investigate the roles of OX40/OX40L and PD-1/PD-L1 costimulatory pathways in mouse islet allograft rejection. Methods Lentiviral vectors containing OX40L siRNA sequences and an adenovirus vector containing the PD-L1 gene were constructed. The streptozotocin-induced model of diabetes was established in C57BL/6 （H-2b） mice. Diabetic C57BL/6 mice were randomly allocated into five groups： group 1, untreated control; group 2, Ad-EGFP treatment; group 3, Ad-PD-L1 treatment; group 4, OX40L-RNAi-LV treatment; group 5, OX40L-RNAi-LV combined with Ad-PD-L1 treatment. Lentiviral vector and the adenovirus vector were injected, singly or combined, into the caudal vein one day before islet transplantation. The islets of DBA/2 （H-2d） mice were transplanted into the renal subcapsular space of the diabetic recipients. Recipient blood glucose and the survival time of the allografts were monitored. Antigen-specific mixed lymphocyte reaction was also evaluated.
