B7-1分子诱导体外抗肝癌免疫反应

Study on the role of B7-1 molecules in antitumor immunity in vitro against hepatocellular carcinoma

目的：了解Ｂ７分子在体外抗肝癌免疫中的作用。方法：健康人外周血单个核细胞（ＰＢＭＣ）与ＨｅｐＧ２／ｈＢ７－１。ＨｅｐＧ２／ｎｅｏ及亲代ＨｅｐＧ２瘤苗混合培养（ＭＬＴＣ），检测淋巴细胞活化增殖能力，淋巴细胞ＨＬＡ－１类抗原的表达，培养上清ＩＦＮ－γ水平、ＴＮＦ活性及ＬＡＫ、ＣＴＬ细胞活性。结果：ＨｅｐＧ２／ｈＢ７－１瘤苗促淋巴细胞增殖最高达１４．６倍，明显高于ＨｅｐＧ２／ｎｅｏ和ＨｅｐＧ２瘤苗的作用（Ｐ＜０．０５）。ＨｅｐＧ２／ｈＢ７－１瘤苗刺激后淋巴细胞ＨＬＡ－Ｉ类抗原表达、ＩＦＮ－γ及ＴＮＦ分泌均高于ＨｅｐＧ２／ｎｅｏ、ＨｅｐＧ２瘤苗刺激组。Ｅ／Ｔ＝４０：１时ＬＡＫ细胞对ＨｅｐＧ２／ｈＢ７－１细胞的杀伤率为杀伤ＨｅｐＧ２／ｎｅｏ和ＨｅｏｐＧ２胞的２．０，２．４倍；ＣＴＬ细胞对ＨｅｐＧ２／ｈＢ７－１细胞杀伤为对ＨｅｐＧ２／ｎｅｏ及ＨｅｐＧ２杀伤的２．７倍，具有明显差异（Ｐ＜０．００１）。结论：抗肝癌免疫中Ｔ细胞活化也需双信号共刺激。Ｂ７－１分子能诱导体外抗肝癌免疫反应。

Objective: To study the effect of R7-1 molecules on antitummor immunity against hepatocellular carcinoma (HCC) in vitro.Methods: PBMC of healthy subjects were co-cultured with HepG2, HepG2/neo and HepG2/hB7-1 vaccines . Lymphocyte proliferation, TNF level in the supernatant and Cytotoxic activity of LAK and CTL cells were evaluated by MTF assay; HLA-I molecule expression of lymphocytes wasdetected by flow cytometry. IFN level was detected by ELISA assay. Results: HepG2/hB7-1 cells stimulated vigorous proliferation of PBMC(at a maximum of 14. 6 times), much higher than that of HepG2/neo and HepG2 vaccines. HLA-I expression on lymphocytes, IFN and TNF secretion in the MLTC with HepG2/hB7-1 group were higher for that of stimulated by HepG2 and HeoG2/neo vaccines. Cytotoxic activity ofLAK against HegG2/hB7-1 cells were 2.0,2.4 times that much of HepG2 and HepG2/neo groups,while or activity 2.7 times that much ofHepG2 and HepG2/neo groups at E/T4O: 1 (p < 0.001 ). Conclusion: B7-1 co-stimulation is able to enhance immunogenicity of HCC cells .Both CD4+ and CD8+ T cells participate in anti-tumor immunity against HCC induced by B7-1, in which enhanced expression of HLA-I molecules on lymphocytes may be involved.