| BackgroundEsophageal carcinoma is one of the most malignant tumor in China. In spite of the combined treatment of surgery, radiotherapy and chemotherapy, the 5-year survival rate remains low, so esophageal carcinoma terribly endangers the human life. It has been discovered in many clinical practice that several patients of carcinomas, such as esophageal carcinoma, colon carcinoma and carcinoma of gastric cardia were founded to recurrence and metastasis shortly after resection, while others either in progression staging or received R1 or R2 resection had long term survival. It is proved that many factors influence the therapeutic efficacy except TNM stage.Tumor-infiltrating lymphocytes (TILs) are mainly consisted of T lymphocytes infiltrated in the stroma ,as well as many different kinds of cells. The type, density and location of TILs are considered manifestations of host immune reactions against cancers. Recently, it was reported that the number of TILs and subsets was closely associated with survival of the patients, especially the balance of immune was associated with recurrence and survival following resection.Cytotoxic CD8~+T lymphocytes(CTLs) are potential effector cells in the control of tumor growth, Continued tumor growth despite the presence of a lymphocytic infiltration including tumor-specific T cells within and surrounding tumors suggests a failure of immune control. T regulatory lymphocytes (Treg cells)are a subgroup of CD4~+T cells with suppressor function characterized by expression of the interleukin-2 receptor a chain CD25 in the resting state. Treg cells suppress activity and proliferation of both CD4~+TCD25~-T and CD8~+T cells in a contact-dependent manner or through secreted inhibition cytokines. Foxp3, a critical regulator of the development and function of Tregs, fulfills the criteria of a Treg-specific marker. It was reported that high Foxp3 expression was associated not only with dismal prognosis in ovarian cancer but also represented an independent predictor for overall survival (OS) and progression-free survival. However, discrepancies remain among different cancer types. The increased number Treg cells are present in non-small cell lung, ovarian, breast, and pancreatic cancers. Murine studies had demonstrated that the rejection of early tumors is induced by the deletion of Treg cells. However, the prognostic role and distribution of TILs in patients with esophageal carcinoma is unclear. In this study, the significance of various subtypes of TILs and their relation to clinical pathological characteristics were examined, hopefully providing a independent predictor for recurrence and survival.ObjectiveTo investigate the number of CD3~+T cells, CD8~+T cells, Foxp3~+Treg cells and GrB~+T cells infiltrated in the stroma and within cancer cell nest and their correlations with clinicopathologic features in esophageal carcinoma. In the meantime, to determine the relationship between TILs subsets and prognosis.Materials and Methods1. Patients90 specimens were obtained from patients who underwent curative resection for esophageal carcinoma. All of the patients without any prior-anticancer treatment and complete clinicopathologic and follow-up data.2. Mathods and Evalution of ImmunostainingThe expression of CD3, CD8, Foxp3 and GrB in the stroma and within cancer cell nest was detected by SP immunohistochemistry. The number of these cells was counted .CD3, CD8 positive cells exhibited distinct cell membrane brown staining. Granzyme B positive lymphocytes display a granular cytoplasmic brown staining pattern, which reflects the granular localization of granzyme B. Foxp3 positive cells exhibited distinct nuclear brown staining.3. Statistical AnalysisAll the data was analyzed by SPSS13.0 statistics software. Mann-Whitney test, x~2 test, Log-rank test and Cox proportional hazards regression model were used to analyze the date. Size of test wasα=0.05.Results1 Most CD3~+T cells were distributed in the stroma, others were within cancer cell nest. The number of CD3~+T cells in the stroma and within cancer cell nest were 153.2 (10.8-369.8), 22.0 (0-146.6), respectively, with significant difference (P <0.01). The expression of CD3~+T cells in tumor invasion to plasma membrane were obviously lower than those in cases of no-invasion to muscle (P < 0.05). Univariate survival analysis suggested that better prognosis was found to correlate with the increased number of CD3~+T in the stroma and within the cancer cell nest (P<0.05).2 Most CD8 positive T cells were distributed in the stroma, a few were within cancer cell nest. The number of CD8~+T cells in the stroma and in cancer cell nest were 44.4 (0-172.8), 4.9(0-43.2), respectively, there was significant difference (P <0.01). CD8~+T cells counts in tumor invasion to plasma membrane were obviously lower than those in cases of no-invasion to muscle (P < 0.05). The quantity of CD8~+T cells in the stroma was associated negatively with differentiate degrees (P < 0.05). Univariate survival analysis suggested that better prognosis was found to correlate with the increased number of CD8~+T in the stroma and within the cancer cell nest (P < 0.05).3 Most Foxp3~+Treg were distributed in the stroma, a few were located in cancer cell nest. The number of Foxp3~+Treg cells in the stroma and in cancer cell nest werel2.5 (0-72.4), 2.9 (0-19.0), respectively,with significant difference (P<0.01). The quantity of Foxp3~+Treg in lymph node metastasis and tumor length longer than 5 centimeters were obviously higher than those in cases of without lymph node metastasis and tumor length shorter than 5 centimeters(P < 0.05). Better prognosis was found to correlate with the decreased number of Foxp3~+Treg cells in the stroma and within the cancer cell nest (P < 0.05).4 GrB~+T cells were diffused distributed in the stroma and within cancer cell nest. The number of GrB~+T cells in the stroma and in cancer cell nest were 12.5 (0-38.6), 2.9 (0-16.4), respectively, there was significant difference (P < 0.01). The quantity of GrB~+T in lymph node metastasis lower than those in cases of without lymph node metastasis (P < 0.05). There was no difference in survival rate between high and low-density of GrB~+T cells in esophageal cancer patients (P > 0.05).5 Cox multivariate analysis indicated that TNM stage, tumor length and the number of CD3~+T, CD8~+T and Foxp3~+Treg cells infiltrated in the stroma were the independent prognostic factors (P < 0.05).ConclusionThe number of CD3~+T, CD8~+T and Foxp3~+Treg cells infiltrated in the stroma are the independent factors for the prognosis of esophageal carcinoma. The increased number of Foxp3~+Treg cells is related to poor prognosis, while the increased number CD3~+T,CD8~+T cells are related to good prognosis. |
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