Tilden A B, Itoh K, Balch C M
J Immunol. 1987 Feb 15;138(4):1068-73.
We analyzed the antigenic phenotype of lymphokine-activated killer (LAK) effector cells. Human blood lymphocytes were cultured for 3 days with 100 U/ml recombinant interleukin 2 (rIL 2), subpopulations isolated with monoclonal antibodies and a fluorescence-activated cell sorter (FACS) and assayed for cytotoxic activity against 51chromium labeled noncultured melanoma tumor cells. Initial experiments compared the LAK effector function of CD5+ T lymphocytes vs CD5- cells (predominantly CD16+ NK cells). The mean percent specific release at a 10:1 effector:target (E:T) ratio was 25% +/- 16 for CD5- cells, 10% +/- 6 for CD5+ cells, and 22% +/- 9 for unsorted cells. In contrast, when lymphocyte subpopulations were isolated before rIL 2 culture (LAK precursors), CD5- cells but not CD5+ cells developed LAK activity (28% +/- 12 vs 1% +/- 1, mean percent specific release, 10:1 E:T ratio), confirming our previous results showing that only CD16+ cells were LAK precursors. The discrepancy between LAK effector and precursor phenotypes suggested that LAK precursors acquired CD5 determinants during rIL 2 culture; however, double label immunofluorescence of rIL 2 cultured CD16+ cells showed that this was not the case. The data suggested that in the presence of other cell types, some T lymphocytes may develop LAK activity, but purified blood T lymphocytes do not develop LAK function when cultured with rIL 2 alone. We also analyzed LAK effector function in lymphocyte subpopulations defined by CD4 and CD8 antigens. The data showed that lymphocytes with a low density expression of CD8 and no expression of CD4 were enriched for LAK effector cells, whereas CD4+ and CD8- had less activity than unsorted cells. Lymphocytes with a high density expression of CD8 had activity similar to unsorted cells. We also assessed the contribution of Leu-7 (HNK-1) granular lymphocytes to LAK effector function. After culture with IL 2, lymphocytes were depleted of Leu-7+ cells by antibody and complement treatment and then were sorted into CD5+ and CD5- fractions. The cytotoxic activity of Leu-7-CD5+ cells was a mean 5% +/- 5 vs a mean 14% +/- 8 for the total CD5+ population (20:1 E:T ratio). The activity of Leu-7- CD5- was slightly less than the total CD5- fraction (21% +/- 9 vs 28% +/- 14, 10:1 E:T ratio). In conclusion, LAK effector function was highest in non-T cell (CD5- CD16+) populations and some activity was also present in T cell populations (CD5+ and predominantly Leu-7+).
我们分析了淋巴因子激活的杀伤（LAK）效应细胞的抗原表型。人血淋巴细胞用100 U/ml重组白细胞介素2（rIL-2）培养3天，用单克隆抗体和荧光激活细胞分选仪（FACS）分离亚群，并检测其对51铬标记的未培养黑色素瘤肿瘤细胞的细胞毒活性。初始实验比较了CD5+ T淋巴细胞与CD5-细胞（主要是CD16+ NK细胞）的LAK效应功能。在效应细胞与靶细胞比例为10:1（E:T）时，CD5-细胞的平均特异性释放百分比为25%±16，CD5+细胞为10%±6，未分选细胞为22%±9。相比之下，当在rIL-2培养前分离淋巴细胞亚群（LAK前体）时，CD5-细胞而非CD5+细胞产生了LAK活性（平均特异性释放百分比分别为28%±12和1%±1，E:T比例为10:1），证实了我们之前的结果，即只有CD16+细胞是LAK前体。LAK效应细胞和前体表型之间的差异表明LAK前体在rIL-2培养过程中获得了CD5决定簇；然而，对rIL-2培养的CD16+细胞进行双标记免疫荧光显示并非如此。数据表明，在存在其他细胞类型的情况下，一些T淋巴细胞可能产生LAK活性，但纯化的血T淋巴细胞单独用rIL-2培养时不会产生LAK功能。我们还分析了由CD4和CD8抗原定义的淋巴细胞亚群中的LAK效应功能。数据显示，CD8低密度表达且无CD4表达的淋巴细胞富含LAK效应细胞，而CD4+和CD8-细胞的活性低于未分选细胞。CD8高密度表达的淋巴细胞活性与未分选细胞相似。我们还评估了Leu-7（HNK-1）颗粒淋巴细胞对LAK效应功能的贡献。用IL-2培养后，通过抗体和补体处理去除Leu-7+细胞，然后将淋巴细胞分选到CD5+和CD5-组分中。Leu-7-CD5+细胞的细胞毒活性平均为5%±5，而总CD5+群体为14%±8（E:T比例为20:1）。Leu-7-CD5-细胞的活性略低于总CD5-组分（分别为21%±9和28%±14，E:T比例为10:1）。总之，LAK效应功能在非T细胞（CD5-CD16+）群体中最高，T细胞群体（CD5+且主要是Leu-7+）中也存在一些活性。
