Peng Bao-Gang, Liang Li-Jian, He Qiang, Li Jian, Lu Ming-De
Department of Hepatobiliary Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, P. R. China .
Ai Zheng. 2005 Oct;24(10):1287-9.
BACKGROUND & OBJECTIVE: Natural killer (NK) cells have strong potential of killing tumor cells, but it is difficult to expand sufficient NK cells to satisfy the treatment of tumor. This study was to explore the feasibility of efficiently expand NK cells through co-culturing peripheral blood mononuclear cells (PBMCs) and Wilms tumor cell line HFWT.
PBMCs and HFWT cells were co-cultured to expand NK cells. The cytotoxicity of NK cells was measured using 51Cr release assay and crystal violet (CV) staining. The proportions of CD3(+), CD4(+), CD8(+), CD16(+), and CD56(+) cells were detected using flow cytometry.
When PBMCs were co-cultured with irradiated HFWT cells for 10-21 days, NK cells were selectively expanded; the proportion of CD56(+)/CD16(+) cells was over 50%. The NK cells killed more than 80% of fresh HFWT cells at an effector/target ratio of 2. The expansion of NK cells required continuous stimulation of PBMCs by HFWT cells.
HFWT cells can effectively stimulating PBMCs to sensitively expand human NK cells for adoptive immunotherapy of human tumors.
自然杀伤(NK)细胞具有强大的杀伤肿瘤细胞的潜能,但难以扩增出足够数量的NK细胞以满足肿瘤治疗的需求。本研究旨在探讨通过外周血单个核细胞(PBMC)与肾母细胞瘤细胞系HFWT共培养来高效扩增NK细胞的可行性。
将PBMC与HFWT细胞共培养以扩增NK细胞。采用51Cr释放试验和结晶紫(CV)染色法检测NK细胞的细胞毒性。运用流式细胞术检测CD3(+)、CD4(+)、CD8(+)、CD16(+)和CD56(+)细胞的比例。
当PBMC与经辐照的HFWT细胞共培养10 - 21天时,NK细胞被选择性扩增;CD56(+)/CD16(+)细胞的比例超过50%。在效应细胞与靶细胞比例为2时,NK细胞对新鲜HFWT细胞的杀伤率超过80%。NK细胞的扩增需要HFWT细胞持续刺激PBMC。
HFWT细胞能够有效刺激PBMC,灵敏地扩增人NK细胞用于人类肿瘤的过继性免疫治疗。