MGBA抗原特异性CD8+CTL细胞与CIK细胞杀瘤活性的对比研究
Cytotoxic activity of mammaglobin A-specific CD8+ cytotoxic T lymphocytes and cytokine-induced killers against breast cancer cells in vitro

摘要：目的 比较乳腺珠蛋白A(MGBA)抗原特异性CD8+CTL细胞与CIK细胞对乳腺癌细胞的杀伤效果。方法 从健康志愿者外周血中提纯单个核细胞，体外分离、诱导培养成树突状细胞(DC)、CTL和CIK细胞，用免疫磁珠法从CTL细胞中分选CD8+CTL细胞；用表达乳腺珠蛋白的重组腺病毒(Ad-MGBA)转染DC细胞后，经刺激制备MGBA抗原特异性CD8+CTL与CIK细胞，用流式细胞术检测两种杀伤细胞对乳腺癌细胞的杀伤效果。结果 CD8+CTL和CIK对表达 MGBA抗原的乳腺癌MDA-MB-415细胞杀伤率分别是63.07%和48.35%(P<0.05)；对不表达MGBA抗原的乳腺癌MDA-MB-231细胞的杀伤率是14.62%和29.29%(P<0.05)。结论 抗原特异性CD8+CTL对表达该抗原的乳腺癌杀伤效果高于CIK细胞，而对不表达该抗原的乳腺癌细胞的杀伤效果低于CIK细胞。
ABSTRACT: Objective To compare the cytotoxic activity of mammaglobin A-specific CD8+ cytotoxic T lymphocytes (CTLs) and cytokine-induced killers (CIKs) against breast cancer cells in vitro. Methods PBMCs were isolated in vitro from the peripheral blood of healthy volunteers. Then DCs, CTLs and CIKs were isolated, induced and cultured from PBMCs in vitro. CD8+ CTLs were purified with immunomagnetic beads from CTLs. DCs were infected with recombinant adenovirus encoding mammaglobin A(Ad-MGBA). CTLs and CIKs were co-cultured with DCs being infected with Ad-MGBA. The cytotoxic activity of mammaglobin A-specific CD8+ CTLs and CIKs against breast cancer cells was compared by flow cytometry. Results The apoptosis rate of breast cancer cell MDA-MB-415, which expressed MGBA was 63.07% by CD8+ CTL killing and 48.35% by CIK killing (P<0.05). The apoptosis rate of breast cancer cell MDA-MB-231, which could express MGBA was 14.62% by CD8+CTL killing and 29.29% by CIK killing (P<0.05). Conclusion The cytotoxic activity of antigen-specific CD8+CTLs for the same antigen-expressing oncology cells is higher than CIKs. However, for different antigen-expressing oncology cells, the cytotoxic activity of CD8+CTLs is lower than that of CIKs