卵巢上皮性癌肿瘤浸润淋巴细胞与腹水中肿瘤相关淋巴细胞生物学特性的比较

The Comparsion of Biological Characteristics between Tumor Infiltrating Lymphocytes and Tumor Associa ted Lymphocytes in Ascites of Epithelial Ovarian Carcinoma

目的比较卵巢上皮性癌肿瘤浸润淋巴细胞（ＴＩＬ）及腹水中肿瘤相关淋巴细胞（ＴＡＬ）的生物学特性。方法收集１２例上皮性卵巢癌组织及腹水，并将其分为实体瘤、血性腹水和非血性腹水３组。将３组标本分别在体外进行ＴＩＬ及ＴＡＬ分离，并在自体肿瘤细胞（ＡＴＣ）以及重组白细胞介素２（ｒＩＬ２）存在的条件下共同培养。其间应用流式细胞仪及４小时５１Ｃｒ释放实验分别测定每组标本的淋巴细胞表型及其抗肿瘤活性。结果３组淋巴细胞具有相似的ＣＤ３、ＣＤ４、ＣＤ８及ＣＤ２５４种表型。差异无显著性（Ｐ＞０．０５）。在全部培养过程中，每组标本ＣＤ４／ＣＤ８始终大于１。３组效应细胞均显示较高的ＮＫ杀伤活性（Ｐ＞０．０５）。实体瘤组ＴＩＬ和非血性腹水组ＴＡＬ在５０∶１的效靶比时具有明显的抗ＡＴＣ特异性，而血性腹水组ＴＡＬ未显示这种特异性，经５１Ｃｒ检测的杀伤活性分别为３０２％～５１０％和１０８％～１５８％，差异有极显著性（Ｐ＜０．０１）。结论卵巢上皮性癌ＴＩＬ及其非血性腹水组ＴＡＬ具有相似的生物学特性。

Objective To compare biological charicteristics between tumor infiltrating lymphocytes (TIL) and tumor associated lymphocytes (TAL) in epithelial ovarian carcinomas. Methods The surgical samples of four ovarian specimens and eight cancer ascites from epithelial ovarian carcinoma were collected, and ascites were divided into bloody and non bloody groups. The cancer tissue was dissected to yield single cells suspensions. TIL and TAL were grown in coculture with autologous tumor cells and 500 U/ml recombinant interleukin 2 (rIL 2). Phenotype was studied using flow cytometry for a varity of human immunocompetent cell surface markers (CD3, CD4, CD8, CD25). Cytotoxicity was investigated using 4 hr 51 Cr release assays with primary ovarian carcinoma cell cultures and the CAOV 3 cell line and the K 562 cell line as target cells in cultured period. Results (1) There was no significant difference in CD3, CD4, CD8 and CD25 expression among the three groups ( P >0.05). CD4 to CD8 (CD4/CD8) ratio was consistently more than 1 in cultured period. (2) In all three groups, Natural killer (NK) acitivity was high and no difference among them ( P >0.05). (3) In all twelve cases, TIL and TAL in the non bloody ascites possess higher potential to lyse the autologous tumor target than TAL in the bloody ascites ( P < 0.01) when effector cell to target cell (E∶T) was 50 to 1 (50∶1). Lytic activity was 30.2%～51.0% and 10.8% ～15.8% respectively. Conclusions TIL and TAL in non bloody ascites in epithelial ovarian cancer have the same biological characteristics.