Guo Jianwei, Cai Meiying, Wei Dapeng, Qin Liwei, Huang Jian, Wang Xia
Department of Immunology, School of Basic Medical Sciences, Sichuan University or West China University of Medical Science, Chengdu 610041, China.
Zhonghua Gan Zang Bing Za Zhi. 2002 Jun;10(3):178-80.
To study the immune responses of lymphocytes after activated by dendritic cells (DCs) loaded with cytotoxicity T lymphocyte (CTL) epitope based peptide of human alpha-fetoprotein (hAFP, 218-226 LLNQHACAV).
Get high purity DCs by cultured plastic-adherent monocytes isolated from healthy donor of HLA-A2(+) peripheral blood with granulocyte-monocyte colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) for 7 days. Stimulate self-lymphocytes with DCs that loaded with CTL epitope based peptide of hAFP under the culture medium contains interleukin-2 (IL-2) for 7 days. Analyse IL-12 and TNF in culture medium and also the specific lysis activity of lymphocytes against four strains of primary hepatocellular carcinoma cells.
After stimulated by DC loaded with CTL epitope based peptide derived from hAFP, lymphocytes appeared a good characteristics and the culture medium of activated lymphocytes contained a high level Th1 type cytokines of IL-12 and TNF. Activated lymphocytes not only specifically lysed HLA-A2(+) HepG2 line but also had the cytotoxicity against other three primary hepatocellular carcinoma cell lines and T2 target cell loaded with peptide of hAFP.
The results of this research supply the basic materials for the DC based vaccine with HLA-A2 restricted peptide epitope derived from hAFP against AFP positive primary hepatocellular carcinoma.
研究负载人甲胎蛋白(hAFP,218 - 226 LLNQHACAV)细胞毒性T淋巴细胞(CTL)表位肽的树突状细胞(DCs)激活淋巴细胞后的免疫反应。
用粒细胞 - 巨噬细胞集落刺激因子(GM - CSF)和白细胞介素 - 4(IL - 4)培养从HLA - A2(+)外周血健康供体分离的贴壁单核细胞7天,获得高纯度DCs。在含白细胞介素 - 2(IL - 2)的培养基中,用负载hAFP的CTL表位肽的DCs刺激自体淋巴细胞7天。分析培养基中的IL - 12和TNF以及淋巴细胞对四株原发性肝癌细胞的特异性杀伤活性。
用负载源自hAFP的CTL表位肽的DCs刺激后,淋巴细胞表现出良好特性,活化淋巴细胞的培养基中含有高水平的Th1型细胞因子IL - 12和TNF。活化的淋巴细胞不仅能特异性杀伤HLA - A2(+) HepG2细胞系,还对其他三株原发性肝癌细胞系以及负载hAFP肽的T2靶细胞具有细胞毒性。
本研究结果为基于DCs的、含源自hAFP的HLA - A2限制性肽表位的疫苗用于抗AFP阳性原发性肝癌提供了基础材料。