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OBJECTIVE@#To investigate the effect of miR-125b on T cell activation in patients with aplastic anemia (AA) and its molecular mechanism.@*METHODS@#A total of 30 AA patients were enrolled in department of hematology, Binzhou Medical University Hospital from January 2018 to October 2021, as well as 15 healthy individuals as healthy control (HC) group. Peripheral blood mononuclear cells (PBMCs) were isolated, in which the levels of miR-125b and B7-H4 mRNA were detected by RT-qPCR. Immunomagnetic beads were used to separate naive T cells and non-naive T cells from AA patients and healthy people to detect the levels of miR-125b and B7-H4 mRNA. Lentivirus LV-NC inhibitor and LV-miR-125b inhibitor were transfected into cells, and T cell activation was detected by flow cytometry. The dual-luciferase reporter gene assay was used to detect the targetting relationship between miR-125b and B7-H4. RT-qPCR and Western blot were used to detect the levels of miR-125b, CD40L, ICOS, IL-10 mRNA and B7-H4 protein.@*RESULTS@#Compared with HC group, the expression of miR-125b was up-regulated but B7-H4 mRNA was down-regulated in PBMCs of AA patients (P <0.05), and the proportions of CD4+CD69+ T cells and CD8+CD69+ T cells in PBMCs of AA patients were higher (P <0.05). The expression of miR-125b was significantly up-regulated but B7-H4 mRNA was down-regulated in both naive T cells and non-naive T cells of AA patients (P <0.05), and non-naive T cells was more significant than naive T cells (P <0.05). Compared with NC inhibitor group, the expression of miR-125b was significantly decreased, the expression level of CD69 on CD4+ and CD8+ T cells in PBMCs was also significantly decreased, while the luciferase activity was significantly increased after co-transfection of miR-125b inhibitor and B7-H4-3'UTR-WT in the miR-125b inhibitor group (P <0.05). Compared with NC inhibitor group, the mRNA and protein levels of B7-H4 were significantly increased in the miR-125b inhibitor group (P <0.05). Compared with miR-125b inhibitor+shRNA group, the expression levels of CD69 on CD4+ and CD8+ T cells were significantly increased, and the levels of CD40L, ICOS and IL-10 mRNA were also significantly increased in the miR-125b inhibitor+sh-B7-H4 group (P <0.05).@*CONCLUSION@#MiR-125b may promote T cell activation by targetting B7-H4 in AA patients.
Subject(s)
Humans , Anemia, Aplastic/genetics , CD40 Ligand/metabolism , Interleukin-10 , Leukocytes, Mononuclear/metabolism , Luciferases , MicroRNAs/genetics , RNA, Messenger/metabolism , Lymphocyte Activation , T-Lymphocytes/metabolismABSTRACT
OBJECTIVE@#To investigate the effects of the B7-H4 gene rs10754339 and miR-125a gene rs12976445 on cancer susceptibility through a case-control study and meta-analysis.@*METHODS@#A total of 1,490 cancer patients (lung/gastric/liver/: 550/460/480) and 800 controls were recruited in this case-control study. The meta-analysis was performed by pooling the data from previous related studies and the present study.@*RESULTS@#The results of this study showed that in the Hubei Han Chinese population, the rs10754339 gene was significantly associated with the risk of lung and gastric cancer but not liver cancer, and the rs12976445 gene was significantly associated with the risk of lung cancer but not liver or gastric cancer. The meta-analysis results indicated that rs10754339 and rs12976445 contributed to cancer susceptibility in the Chinese population and also revealed a significant association between rs10754339 and breast cancer risk, as well as between rs12976445 and lung cancer risk.@*CONCLUSION@#The B7-H4 gene rs10754339 and miR-125a gene rs12976445 may be the potential genetic markers for cancer susceptibility in the Chinese population, which should be validated in future studies with larger sample sizes in other ethnic populations.
Subject(s)
Humans , MicroRNAs/genetics , Stomach Neoplasms/genetics , Case-Control Studies , Lung Neoplasms/genetics , RiskABSTRACT
@#[摘 要] 目的：探讨干扰B7-H4表达对乳腺癌细胞增殖、凋亡、周期以及相关下游分子表达的影响。方法：利用脂质体转染技术分别将特异性靶向B7-H4的siRNA（siB7-H4）及其阴性对照（siNC）转染至对数生长期的乳腺癌T47D和MCF-7细胞，分别命名为T47D-siB7-H4、T47D-siNC、MCF-7-siB7-H4和MCF-7-siNC组。用qPCR法和WB法验证siRNA干扰效果及其对细胞周期分子cyclin D1表达的影响，CCK-8法和FCM分别检测干扰B7-H4表达对T47D和MCF-7细胞增殖、周期和凋亡的影响，qPCR法检测B7-H4干扰对E2F家族相关转录因子表达的影响。结果：成功构建干扰B7-H4表达的乳腺癌T47D和MCF-7细胞。与T47D-siNC和MCF-7-siNC组相比，T47D-siB7-H4和MCF-7-siB7-H4组细胞中B7-H4 mRNA和蛋白表达水平均显著降低、细胞增殖能力显著降低（均P<0.01），并伴有G1/S期细胞周期阻滞以及cyclin D1表达下调（均P<0.01），但细胞凋亡率差异无统计学意义（均P>0.05）。与T47D-siNC相比，干扰B7-H4后T47D细胞中E2F1、E2F2、E2F7和E2F8 mRNA水平有不同程度的降低（均P<0.01）；与MCF-7-siNC相比，干扰B7-H4后MCF-7细胞中E2F1、E2F2、E2F3、E2F7和E2F8 mRNA水平均有不同程度的降低（P<0.05或P<0.01）。结论：干扰乳腺癌细胞B7-H4表达可下调cyclin D1和E2F家族相关转录因子的表达，导致细胞周期阻滞并抑制细胞增殖。
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@#[摘 要] B7/CD28家族分子作为主要的共信号分子，在T细胞功能调控及免疫应答中发挥着至关重要的作用，关于其功能的研究及应用在世界范围内广泛开展。其中，CD28和CTLA-4都可以与B7-1/B7-2结合，但CD28能够促进T细胞增殖，维持Treg细胞稳态；而CTLA-4则可以抑制T细胞增殖，影响CD4+T细胞的分化；抗CTLA-4单抗ipilimumab与抗PD-1单抗联用能够用于治疗PD-L1+的非小细胞肺癌以及无症状的Ⅳ期黑色素瘤。PD-L1/PD-L2：PD-1途径则可以通过多种方式调节T细胞功能，参与CD8+T细胞“耗竭”状态的形成与维持。目前，针对PD-L1/PD-L2：PD-1途径的免疫治疗相关药物开发广泛且较为成熟，抗PD-1单抗主要通过诱导肿瘤浸润的部分耗竭的CD8+T细胞亚群的扩增发挥抗肿瘤作用。ICOS：ICOSL途径在T细胞分化、细胞因子分泌以及体液免疫应答中起着重要作用，抗ICOS抗体药物feladilimab与抗PD-1单抗联用能够有效治疗多发性或难治性骨髓瘤。B7-H3同时具有共刺激效应和共抑制效应，阻断B7-H3后能够有效增强TIL的抗肿瘤效应，其单抗enoblituzumab能够与抗PD-1单抗联用治疗非小细胞肺癌。B7-H4、人内源性逆转录病毒‑H长末端重复关联蛋白（human endogenous retrovirus‑H long terminal repeat‑associating protein，HHLA）以及含V结构域抑制T细胞活化的免疫球蛋白（V‑domain Ig‑cotaining suppressor of T cell activation，VISTA）均能够提供抑制信号，针对B7-H4靶点的CAR-T治疗以及VISTA的小分子抑制剂CA-170均有临床试验正在进行中。目前，对共信号分子的研究已经获得了长足进展，针对某些活化性分子或者抑制性分子开发了相关抗肿瘤药物并在临床中取得一定成效，但如何进一步提高其治疗有效率、减少不良反应等仍有待探索。
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Objective: To detect the levels of B7-H4 protein, adenosine deaminase (ADA) and carcinoembryonic antigen (CEA) in the malignant pleural effusion associated with lung cancer (LC-MPE) and tuberculous pleural effusion (TPK), and to evaluate the values of single and combined detection of B7-H4 protein, ADA and CEA in differential diagnosis of LC-MPE and TPK. Methods: A total of 202 samples of pleural effusion (PE) from 120 LC-MPE patients (LC-MPE group) and 82 TPE patients (TPE group) were collected. The levels of ADA and CEA and B7-H4 protein in PE were detected by the enzymatic, electrochemiluminescence and ELISA methods, respectively; and the receiver operating characteristic curve (ROC curve) was used to determine the diagnostic efficiencies of the above indexes alone or in combination, such as the sensitivity, the specificity, the Yoden index (YI)» and the area under ROC curve ( AUC). Results: The level of ADA in PE of the patients in TPE group was higher than that in LC-MPE group (P<0. 05); the levels of CEA and B7-H4 protein in PE of the patients in LC-MPE group were higher than those in TPE group ( P'-CO. 05). The ROC curve analysis results showed that the sensitivities of single detection of ADA, CEA and B7-H4 protein were 93.30%, 83.33% and 79.90%, respectively; the specificities were 86. 59%, 96.34% and 72. 50%, respectively; the AUC were 0. 927, 0. 925 and 0. 836, respectively. The combined detection of the three indexes had the highest diagnostic value; the sensitivity was 93. 90% , the specificity was 97. 50% , the YI was 0. 971, and the AUC was 0. 998. Conclusion: The combined detection of levels of ADA, CEA and B7-H4 protein in PE is superior to the single evaluation of each index, which can greatly improve the differential diagnosis efficiencies of LC-MPE and TPE.
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Objective To investigate the influence of sorafenib on the expressions of B7-H3 and B7-H4 proteins in different human hepatocellular carcinoma cell lines,including HepG2,Hep3B,BEL-7402,BEL-7404,BEL-7405,QGY-7701,QGY-7703,SMMC-7721,MHCC97H,MHCC97L,HCCLM3 and HCCLM6.Methods Western blotting and MTT assay were used to check the influence of sorafenib on the expressions of B7-H3 and B7-H4 proteins in different human hepatocellular carcinoma cell lines and to test the inhibitory effect of sorafenib on different human hepatocellular carcinoma cell lines.Results Compared with normal human liver cells (HL-7702),the expressions of B7-H3 and B-H4 proteins in different human hepatocellular carcinoma cell lines were significantly up-regulated (P＜0.01).The cytotoxic activity IC50 values of sorafenib to Hep3B,BEL-7404,MHCC97H,HCCLM3 and HCCLM6 were 14.56,9.14,9.46,17.21 and 9.29 μmol/L respectively.After treating Hep3B,BEL-7404,MHCC97H,HCCLM3 and HCCLM6 with sorafenib at the doses of 5,10 and 20 μmol/L separately,the expressions of B7-H3 and B7-H4 proteins were strikingly down-regulated when compared with the control group (P＜0.01).Conclusion The overexpressions of B7-H3 and B7-H4 proteins in different human hepatocellular carcinoma cell lines are a common finding,which can influence tumor immune escape.It may be a new target for prevention and treatment of liver cancer in future.
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Objective To detect the expression of costimulatory molecules B7-H4 and programmed death ligand 1 (PD-L1) in bladder cancer,and to explore the correlation between them and clinicopathological features of bladder cancer.Methods Immunohistochemical staining was used to detect the expression of B7-H4 and PD-L1 in 98 cases of bladder urothelial carcinoma,which were confirmed by pathology from August 2014 to December 2015 in our hospital.There were 23 females,aged 45-82 years,with an average age of 67.8 years.Among them,42 cases of adjacent normal tissues were used as controls.The clinical stage,histological grade and recurrence of bladder cancer were collected,and the correlation between them was analyzed.Results The positive rates of B7-H4 and PD-L1 in bladder urothelial carcinoma were 54.1％ (53/98) and 59.2％ (58/98),respectively,and there was no expression in normal bladder tissues (P ＜ 0.05).The positive expression rate of B7-H4 in muscle invasive bladder cancer (MIBC) patients was higher than that in non-muscle invasive bladder cancer (NMIBC) [73.5％ (25/34) vs.43.8％ (28/64),P =0.005].The positive expression rate of B7-H4 in high-grade patients was higher than that of low-grade [70.0％ (21/30) vs.47.1％ (32/68),P =0.036].The expression rate of B7-H4 in high-risk group was higher than that of low-intermediate risk group [57.1％ (20/35) vs.27.6％ (8/29),P =0.018].The positive expression rate of PD-L1 in patients with MIBC was higher than that in NMIBC [79.4％ (27/34) vs.48.4％ (31/64),P =0.003].The PD-L1 expression rate of histological high-level group was higher than that of low-level group [73.3％ (22/30) vs.52.9％ (36/68)],but the difference was not statistically significant (P =0.058).The PD-L1 expression rate in high-risk group was 68.6％ (24/35),and also higher than low-middle group 24.1％ (7/29) (P ＜ 0.05).There was a positive correlation between the expression of B7-H4 and PD-L1 in bladder urothelial carcinoma (r =0.318,P=0.002).The combined recurrence rate of the two groups was significantly higher than that of the negative expression of the two groups [66.7％ (14/21) vs.30.8％ (8/26),P=0.014].Conclusions The expression of B7-H4 and PD-L1 is up-regulated in bladder urothelial carcinoma,which is closely related to the clinical stage,histological grade,risk classification and recurrence of NMIBC.
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Objective: To explore the expressions of B7-H1 and B7-H4 in colorectal cancer (CRC) tissue and their relationships with the Foxp3 regulated T-cell (Treg) infiltration in tumor tissue, and to provide the basis for judging the biological behaviours and prognosis of CRC. Methods: Immunohistochemistry was used to detect the expressions of B7-H1 and B7-H4 and the infiltration of Treg in 56 cases of CRC tissue samples and adjacent normal tissue samples, and their relationships with the clinicopathological features of CRC patients were analyzed. Results: The positive expression rate of B7-H1 in CRC tissue of the CRC patients was higher than that in adjacent normal tissue (χ2= 72. 34, P<0. 01). The differences of expression frequencies of B7-H1 in CRC tissues with different infiltration depths were significant (P<0. 01). The high expression frequency of B7-H1 in CRC tissue of the patients with positive lymphnode metastasis was higher than that in the patients with negative lymphnode metastasis (P<0. 01). The high expression frequency of B7-H1 in CRC tissue of the patients at Duke' s stage (C+D) was higher than that of the patients at Duke' s stage (A+B) (P<0. 01). The positive expression rate of B7-H4 in CRC tissue was higher than that in adjacent normal tissue (χ2=78. 92, P<0. 01). The differences of expression frequencies of B7-H4 in CRC tissues with different infiltration depths were significant (P<0. 05). The high expression frequency of B7-H4 in CRC tissue of the patients with positive lymphnode metastasis was higher than that of the patients with negative lymphnode metastasis (P<0. 05). The number of Treg positive expression cells in CRC tissue was significantly higher than that in adjacent normal tissue (P<0. 01), the number of Treg postive expression cells in CRC tissues with different differentiation degrees were different (P<0. 01), and the number of Treg positive expression cells in CRC tissue of the patients with positive lymphnode metastasis was higher than that of the patients with negative lymph node metastasis (P<0. 01). The expression of B7-H1 was correlated with Treg infiltration (P<0. 01, Cramer ' s = 0. 463); the expression of B7-H4 was correlated with Treg infiltration (P<0. 05, Cramer' s = 0. 320). Conclusion: B7-H1 and B7-H4 are highly expressed in human CRC tissue, and they are associated with the increasing of Treg infiltration; they can be used as the indicators to judge the prognosis of CRC.
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Objective:To construct anti-B7-H4-scFv-PE38KDEL,a recombinant toxin based on anti-B7-H4 single chain antibody (scFv),to detect anti-tumor effect of toxin protein.Methods:The anti-B7-H4-scFv gene was ligated with the toxin PE38KDEL gene by overlapping extension PCR(SOE-PCR).The recombinant gene was cloned into prokaryotic expression vector pET28a(+),and the protein was renatured and purified by chromatography (Ni-NTA),and was identified by Western blot.Indirect ELISA and flow analysis technology were used for specific identification.The inhibitory effects of toxins on tumor cells were detected by MTT assay and subcutaneous xenograft model in vitro and in vivo.HE staining and immunohistochemical analysis were performed on tumor tissues.Results:The recombinant expression vector pET28a-anti-B7-H4-scFv-PE38KDEL was obtained by restriction endonuclease di-gestion.The purified toxin protein was inoculated on the tumor cells.The tumor growth was inhibited in the tumor model.Conclusion:The recombinant toxin expression system based on anti-B7-H4 single chain antibody was successfully constructed.The recombinant toxin protein had good biological activity and anti-tumor activity.
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The purified 3C8 was obtained by two step column purification,including Protein G affinity purification and DEAE anion exchange purification.The purity of purified 3C8 was about 93％ when analyzed by reverse column.SDS-PAGE showed that the purity of 3C8 was increased greatly by two step purification.By flowcytometry we found that 3C8 specifically binded with B7-H4/293T cells and did not bind with Mock/293T cells,moreover 3C8 did not bind with other B7 family members transgene cells.In confocal experiment 3C8 could specifically stained B7-H4/293T cells.In Western blot only B7-H4/293T cells showed positive band while Mock/293T cells showed negative result.The result of immunohistochemistry showed that B7-H4 was highly expressed in prostate cancer and renal cell carcinoma,while para-cancer tissues did not express B7-H4.The T cell proliferation experiment showed that B7-H4-Ig could bind to activate T cells and inhibit T cell proliferation,while 3C8 could block the binding of B7-H4-Ig and reverse the T cell proliferation inhibition effect of B7-H4-Ig by CFSE and CCK8 assay.The cytokine IFN-γ and IL-2 secreted by activating T cells was decreased by B7-H4-Ig and 3C8 could reverse the effect of B7-H4-Ig.
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Objective To investigate the intensive atorvastatin therapy on B7-H4 in peripheral blood monocytes of patients with unstable angina PCI undergoing percutaneous coronary intervention. Methods 80 patients with unstable angina were randomized to pretreatment with intensive dose (80 mg/day ,n = 40) and conventional dose(20 mg/day,n=40)of atorvastatin. Peripheral blood were subsequently obtained prior to PCI,and 18 ~ 24 h after PCI. Peripheral blood serum level of IL-4,IL-10 and IFN-γ were quantified using enzyme-linked immunosorbent assays. Fluorescence-based quantitative real-time PCR was used to measure levels of periph-eral blood monocytes B7-H4 mRNA. Results Levels of IL-10 ,sB7-H4 and B7-H4 mRNA increased in patients of both groups after PCI. The increase in intensive dose group is more significant (P < 0.05). IL-4 and IFN-γ decreased in patients of both groups after PCI. The decrease in intensive dose group is more significant (P <0.05). Conclusion Intensive dose atorvastatin treatment improve post-PCI immune inflammation in patients with unstable angina,possibly by promoting the expression of B7-H4 in peripheral blood monocytes.
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Objective@#To evaluate the value of combined detection of negative costimulatory molecule B7-H4 and carcinoembryonic antigen (CEA) in diagnosing malignant and benign pleural effusion.@*Methods@#Ninety-seven pleural effusion specimen were collected, 55 of which were diagnosed as malignant pleural effusion and 42 were benign pleural effusion. Enzyme-linked immunosorbent assay(ELISA) was used to examine the concentration of B7-H4 and CEA in pleural effusion. Electro-chemiluminescence immunoassay was used to detect the CEA level in pleural effusion. Receiver operating characteristic (ROC) curve was established to analyze and evaluate the single or combined detection of B7-H4 and CEA in diagnosing malignant and benign pleural effusion.@*Results@#The concentrations of B7-H4 and CEA in malignant pleural effusion (MPE) group were (60.08±35.04) ng/ml and (41.49±37.16) ng/ml, respectively, obviously higher than (27.26±9.55) ng/ml and (2.41±0.94) ng/ml of benign pleural effusion (BPE) group (both P<0.01). Area under curve (AUC) of B7-H4 was 0.884 in MPE groupand the diagnostic sensitivity and specificity were 81.8% and 90.5%, respectively, at the optimized cut off value of 37.25 ng/ml. Likewise, area under curve (AUC) of CEA was 0.954 and the sensitivity and specificity were 87.3% and 95.2%, respectively, at the cut off value of 4.18 ng/ml. When B7-H4 >37.25 ng/ml or CEA>4.18 ng/ml, the sensitivity of diagnosis as MPE was down-regulated to 90.9% and the specificity was elevated to 88.1%. When B7-H4 >37.25 ng/ml and CEA>4.18 ng/ml, the sensitivity of diagnosis as MPE was down-regulated to 78.2% and the specificity was elevated to 97.6%. The sensitivity and specificity of combined detection of B7-H4 and CEA to diagnose MPE were elevated to 90.9% and 97.6%, respectively. The level of B7-H4 in MPE and BPE were both positively correlated with CEA (r=0.670, P=0.001 in MPE and r=0.002, P=0.001 in BEP).@*Conclusions@#B7-H4 is a potential tumor marker in diagnosing the benign and malignant pleural effusion. Although the diagnostic value of B7-H4 may not precede to CEA, the combined detection of B7-H4 and CEA can improve the diagnostic sensitivity and specificity of MPE.
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Objective To investigate the role and mechanism of B7-H4, a negative costimulatory molecule, in mediating the immunomodulatory effects of mesenchymal stem cells C3H10T1/2 (C3H10) on T cell polarization. Methods The lentiviral vectors that carried the shRNA targeting mouse B7-H4 were transfected into mouse mesenchymal stem cells (C3H10-B7-H4). The cells were co-cultured with PHA-acti-vated mice spleen lymphocytes before and after the transfection. ELISA was performed to detect the concen-trations of cytokines in supernatants of cell culture in order to elucidate the effects of B7-H4 expressed by C3H10 on T cell polarization. A mouse model of experimental allergic encephalitis (EAE) was established. Fifty C57BL/6 mice were divided into five groups including control group, EAE group, C3H10 group (injec-ting EAE mice with C3H10 cells), C3H10-NC group ( injecting EAE mice with C3H10-NC cells) and C3H10-B7-H4 group (injecting EAE mice with C3H10-B7-H4 cells). ELISA was performed to detect the soluble form of IL-2, IL-17, IFN-γ and IL-4 in plasma samples. Results Knocking down the B7-H4 gene with shRNA significantly decreased the expression of B7-H4 on C3H10 cells, which weakened the inhibitory effects of C3H10 cells on the secretion of IL-2, IL-17 and IFN-γ by spleen lymphocytes. The therapeutic effects of C3H10-B7-H4 cells on mice with EAE were weakened after silencing the B7-H4 gene expression, which was manifested as higher nerve function score and earlier onset and bring forwarded peak time of EAE than those of the C3H10 group. Treating EAE mice with C3H10-B7-H4 cells was less efficient in inhibiting the expression of IL-2, IL-17 and IFN-γin plasma. However, knocking down the B7-H4 gene had no signif-icant effect on the expression of IL-4 in terms of treating EAE with C3H10 cells. Conclusion The co-inhib-itor molecule B7-H4 expressed on C3H10 cells mediated the treatment of EAE with C3H10 cells by regula-ting Th1 and Th17 effector T cells.
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The new costimulatory molecule B7-H4,also called V-set domain containing T cell activa-tion inhibitor 1(VTCN-1),is a member of the B7 family.B7-H4 can negatively regulate the immune response of T lymphocytes through inhibiting their proliferation and cytokine production .The expression of B7-H4 plays an important role in initiation ,progression and regression of malignant tumors ,including female malignant tumor , such as breast cancer ,ovarian cancer and uterine neoplasm .The association between the female tumors and B 7-H4 causes great concern in recent years .In this review,the function of B7-H4 in female tumor research and the potential target of B7-H4 in the diagnosis and treatment of female tumor are summarized .
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AIM: To study the expressions and clinical significance of PDCD4 and B7-H4 in optic gliomas in children. METHODS:We used immunohistochemistry to evaluate the expressions of PDCD4 and B7-H4 in specimens from 52 optic gliomas in children cases, and analyzed the correlation of PDCD4 and B7 - H4 expression with clinicopathological factors children optic gliomas. RESULTS: PDCD4 expression reduced in optic glioma tissue:PDCD4 expression rate in 52 cases of optic glioma tissues decreasing was correlated with tumor malignancy increase (P CONCLUSION: Expressions of PDCD4 and B7-H4 may be clinically relevant to tumor malignancy of optic gliomas in children and prevention of metastasis and prediction of prognosis.
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Objective:To observe the expression of B7-H4 in experimental autoimmune myocarditis (EAM).Methods:BALB/c mice were randomly divided into 2 groups:the control group and the experimental group.The mice of experimental group were injected with myosin to establish EAM models , while the mice of control group were injected with complete Freund 's adjuvant and normal saline.All the mice were killed separately at the 14th,21st,30th and 45th day for lymphocyte proliferation assay ,hematoxylin-eosin staining,immunohistochemical staining and real-time PCR.Results:The inflammation infiltration of heart was most serious at the 14th and 21st day,then it was gradually relieved with time;the results of lymphocyte proliferation assay and real-time PCR were similar to that of the inflammation infiltration of heart ,which were in high level at the 14th and 21st day,and they were both higher than that of the control group ( P<0.05 );B7-H4 protein were only detected in the experimental group ,and it was constantly expressed during the whole experiment on the endothelium of heart with myocarditis.Conclusion:B7-H4 participates in the progress of EAM ,and it may be a new way of studying the mechanism of myocarditis.
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Objective: To investigate the percentages of splenic CD4+T, CD8+T and CD4+CD2 5 ± forkhead-winged helix transcription factor 3 (foxp3)+ T regulatory cells and the expression of costimulatory molecule B7 homologue 4 (B7-H4) during different stages of progression of precancerous lesions in esophagus in mice. Methods: 4-Nitroquinoline-N-oxide (4NQO) in drinking water was used to induce esophageal precancerous conditions in mice. The hematoxylin-eosin (HE) staining was used to evaluate the pathological changes in different stages of precancerous conditions. The percentages of CD4+T, CD8+T and CD4+CD25+foxp3+ T regulatory cells in splenic lymphocytes and the positive rates of B7-H4 expression on CD4+T, CD8+T and CD4+CD25+foxp3+ T regulatory cells in mice in different stages of esophageal precancerous conditions were detected by flow cytometry (FCM). Results: At the time of the 12th week of giving 4-NQO in drinking water to induce esophageal cancer, the pathological abnormalities with low-grade intraepithelial neoplasia occurred in esophageal tissues in mice, and the high-grade intraepithelial neoplasia and carcinoma in situ were developed along with the time. The percentages of CD4+T and CD4+CD2 5+foxp3+ T regulatory cells in splenic lymphocytes in mice induced by drinking of 4-NQO were higher than those in the control mice (by drinking of distilled water) (both P +T cells in mice induced by drinking of 4-NQO was higher (P +T cells, T regulatory cells and B7-H4 may be involved in tumor immune escape mechanism.
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B7-H4 is a new member of costimulatory molecules,it negatively regulates T cell immunity by the inhibition of T cell proliferation, cytokine production, and cell cycle progression. Recent studies indicate that B7- H4 is associated with the generation and progression of tumor and is an important indication in the diag-nosis and treatment of tumor.
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Objective: To study the expression of negative costimulatroy molecule B7-H4 in non-small cell lung cancer (NSCLC) tissues and its relationship with the clinical features of NSCLC. Methods: Fifty-two NSCLC specimens from pa-tients who were pathologically diagnosed in our hospital during January 2008 to April 2009 were included in the present study. B7-H4 expression and infiltration of CD3~+ T cells in NSCLC tissues were detected by immunohistochemistry. The correlation between B7-H4 expression, CD3~+ T infiltration, and the clinical features of NSCLC was studied. Results: The positive rate of B7-H4 in 52 NSCLC tissues was 48.08% (25/52), and B7-H4 expression in normal lung tissues was neg-ative or low (P <0.05). B7-H4 expression was positively correlated with the clinical tumor stages and lymph node metas-tasis of NSCLC (P < 0.05), and negatively correlated with tumor infiltration of CD3~+ T cells (P < 0.05), but had no re-lationship with clinicopathologie parameters of NSCLC (P > 0.05). Conclusion: Negative costimulatroy molecule B7-H4 may play important roles in the development of NSCLC. Positive expression of BT-H4 is correlated with the clinical tumor stages and lymph node metastasis of NSCLC, which provides a foundation for diagnosis and therapy of NSCLC.
ABSTRACT
Objective To study the relationship between the expression of eostimulating factor B7-H4 in patients with primary biliary cirrhosis (PBC) and the pathogenesis of PBC. Methods The expression of B7-H4 mRNA on peripheral blood mononuclear cell (PBMC) of 65 patients with PBC was tested by real-time PCR. Serum levels of IL-2 were assayed by ELISA. CD4+, CD8+ T lymphocytes expression level and B7-H4 expression rate before and after activation were measured by three-color flow cytometry (FCM). Re-sults (1) Expression of B7-H4 mRNA and BT-H4 percentage in PBC group were significantly lower than that in none-PBC group and healthy controls(P<0.01);(2)After 72 h activation, the percentage of CD4+, CD8+, and CD4+ CD8+T lymphoeytes and serum levels of IL-2 decreased (P<0.05), and the percentage of CD4+, CD4+ CD8+T lymphocytes and serum levels of IL-2 were significantly higher than that of none-PBC group and healthy controls(P<0.01);(3)Levels of alanine aminotransferase(ALT), aspartate amin-otransferase(AST), alkaline phosphatase(ALP) and γ-glutamyl transpeptidase(GGT) of patients with posi-tive anti-mitochondrial antibody(AMA)-M2 rose. There was no significant difference of B7-H4 expressions on T cells between patients either with or without AMA-M2 antibody. Conclusion The costimulating factor B7-H4 can express on T lymphocytes which is activated by phytohemagglatinin(PHA) aad plays a negative role on T cells responses.