| Objective:1.B7-H4 is a recently described member of costimulation factor B7 family ligand which is known to be critically involved in the down regulation of antigen-specific T cell-mediated immunity.This study was designed to evaluate the expression of B7-H4 in epithelial ovarian tumor and to investigate the relationship between B7-H4 expression and the clinicopathological variables.We also tested the expression of B7-H4 in ovarian carcinioma cell lines including SKOV3,3AO,A2780,which might provide the cell carrier to investigate the bionomics of B7-H4 in vitro.2.CD4~+CD25~+T cells(Regulatory T cells,Tregs)are detectable in tumor effusion and blood,which are important in suppressing tumor-associated antigens specific immunity.CD8~+ T cell is one of the tumor infiltrating lymphocytes(TILs),which to be a high and specific performance and low side effect anti-tumor immunologically competent cell.In this study we evaluated the two T cells and associated them with the B7-H4 expression,which might reveal the contribution of B7-H4 in tumor immunity of ovarian carcinoma.We also tested the relationship between the two T cells infiltrations and the survival rate in ovarian serous carcinoma.Methods:1.Using RT-PCR and immunohistochemistry to evaluate the expression of B7-H4 mRNA and protein in ovarian epithelial tumor tissues respectively.Specimens were obtained from the 65 patients who were operated for primary ovarian cancer (unradiotherapy or unchemotherapy before operation),18 cases of bordline tumor,12 cases of benign ovarian tumor in the Department of Obstetrics and Gynecology of Qilu Hospital,Shandong University from January,2003 to July,2007.All the Specimens were diagnosed by HE pathology examination.The study of association B7-H4 with follow-up was done and statistical analysis was done using SPSS13.0 software.2.Double immunofluorescence was used to evaluate the number of CD4~+CD25~+ T lymphocytes and immunohistochemistry was used for CD8~+ T lymphocytes evaluation in the serous carcinoma tissues.We compared the relationship between the expression of B7-H4 and the number of CD4~+CD25~+ T lymphocytes and CD8~+ T lymphocytes to discover the contribution of B7-H4 to the T cell-immune response.Also the analysis of the relationship between the two T cells infiltrations and the survival rate in ovarian serous carcinoma was done.Statistical analysis was done using SPSS13.0 software.Results:1.The B7-H4 mRNA by RT-PCR amplification was 184bp.All the 95 ovarian epithelial tumor tissues were positive of B7-H4 mRNA.The positive proportions of B7-H4 protein in benign,borderline and malignant epithelial ovarian tumors were 4/12,11/18 and 59/65,which showed malignant cases were significantly higher than nonmalignant ones(P＜0.05).The positive proportion of B7-H4 protein in serous,endometrioid,clear cell and mucinous ovarian carcinomas were 41/41,5/5,2/2 and 4/10, which showed the first three were significantly higher than the last one(P＜0.05).There was no significant difference in the distribution of the proportion of B7-H4 positive cells in serous carcinoma in the four groups including＜10%,＞10%-50%,＞50-80%,＞80-100%(P＞0.05).The expression of B7-H4 protein in 41 ovarian serous carcinomas revealed that there was no correlation between B7-H4 expression and age,ascites cytology or lymph node metastasis(P＞0.05)except for stage and grade(P＜0.05).The three ovarian carcinioma cell lines were negative of B7-H4 mRAN and protein.2.The results showed CD4~+CD25~+ T lymphocytes was 19.95±11.50/10 High power Field(HPF)and CD8~+ T lymphocytes was 43.46±16.69 /10HPF.Mann-Whitney test showed that the higher expression of B7-H4 the larger number of Tregs was detected(P＜0.05),while the lower number of CD8~+ T lymphocytes was detected(P＜0.05). Kaplan-Meier survival analysis showed that high CD4~+CD25~+ T lymphocyte group was characterized by a significantly worse prognosis in term of overall survival(P＜0.05),while CD8~+ T lymphocyte group was characterized reversely(P＜0.05).However, no significant differences were found between the number of CD4~+CD25~+ / CD8~+ T lymphocytes and age,pathology grade,clinical stage,lympha metastasis or ascites cytology(P＞0.05).Conclusions:1.The B7-H4 overexpressed in ovarian carcinoma.The up-regulated expression of B7-H4 in ovarian serous carcinoma which has positive relationship with later stage and lower grade and the above results revealed that B7-H4 plays an important role in cancer generation and development,which provided a potential target for the therapy and diagnosis of ovarian carcinoma.2.The negative expression of B7-H4 in the three ovarian carcinoma cell lines provide the cell carriers for the study of B7-H4 in vitro.3.B7-H4 played a suppressive role in ovarian carcinoma specific immunity.4.CD4~+CD25~+ and CD8~+T cells play significant roles in the survival of ovarian serous carcinioma. |
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