Sarneva M, Vujanovic N L, Van den Brink M R, Herberman R B, Hiserodt J C
Pittsburgh Cancer Institute, University of Pittsburgh, Pennsylvania 15213.
Cell Immunol. 1989 Feb;118(2):448-57. doi: 10.1016/0008-8749(89)90392-4.
The coculture of rat bone marrow cells with recombinant interleukin-2 induced the generation of cells mediating natural killer (NK) activity and subsequent lymphokine-activated killer (LAK) activity depending upon the dose of IL-2 and time of culture. NK activity was detected as early as 4 to 5 days after the addition of IL-2 and could be evoked with as little as 5 to 50 U/ml. The induced NK cells had large granular lymphocyte (LGL) morphology and expressed 0X8 and asialo GM1 surface markers but did not express 0X19 or W3/25 markers. LAK activity was detected only after 5 days of culture, and required above 100 U/ml IL-2. Cells mediating LAK activity also expressed 0X8 and asialo GM1 but not 0X19. The generation of detectable NK and subsequent LAK activity was due to induction of early progenitor cells and not contaminating mature LGL/NK cells within the bone marrow population since of removal of such mature NK cells with L-leucine methyl ester (L-LME) did not affect the subsequent generation of either activity. Moreover, the removal of actively dividing cells as well as mature NK cells from the bone marrow by treatment with 5-fluorouracil (5-FU) in vivo enriched the remaining bone marrow population for both NK and LAK progenitor cells. The phenotype of the L-LME- and 5-FU-resistant NK and LAK progenitor cells within populations of bone marrow was determined by antibody plus complement depletion analysis. Although treatment of normal bone marrow with anti-asialo GM1 + C reduced the induction of NK and LAK activity in 5-day cultures, treatment of 5-FU marrow with anti-asialo GM1 + C did not affect either activity. Treatment with a pan-T cell antibody + C did not affect the development of NK or LAK activity under any conditions. Thus, the 5-FU-resistant NK/LAK progenitors were asialo GM1 negative but became asialo GM1+ after induction by IL-2. Finally, evidence that bone marrow-derived LAK cells were generated directly from the IL-2-induced NK cells was obtained by treating the IL-2-induced LGL/NK cells with L-LME.(ABSTRACT TRUNCATED AT 400 WORDS)
大鼠骨髓细胞与重组白细胞介素-2共培养可诱导产生介导自然杀伤（NK）活性及随后的淋巴因子激活的杀伤（LAK）活性的细胞，这取决于白细胞介素-2的剂量和培养时间。早在添加白细胞介素-2后4至5天就能检测到NK活性，低至5至50 U/ml即可诱发。诱导产生的NK细胞具有大颗粒淋巴细胞（LGL）形态，表达OX8和脱唾液酸GM1表面标志物，但不表达OX19或W3/25标志物。LAK活性仅在培养5天后才能检测到，且需要高于100 U/ml的白细胞介素-2。介导LAK活性的细胞也表达OX8和脱唾液酸GM1，但不表达OX19。可检测到的NK及随后的LAK活性的产生是由于早期祖细胞的诱导，而非骨髓群体中污染的成熟LGL/NK细胞，因为用L-亮氨酸甲酯（L-LME）去除此类成熟NK细胞并不影响随后两种活性的产生。此外，通过体内用5-氟尿嘧啶（5-FU）处理从骨髓中去除活跃分裂细胞以及成熟NK细胞，可使剩余骨髓群体中NK和LAK祖细胞富集。通过抗体加补体耗竭分析确定骨髓群体中对L-LME和5-FU有抗性的NK和LAK祖细胞的表型。尽管用抗脱唾液酸GM1 + C处理正常骨髓会降低5天培养物中NK和LAK活性的诱导，但用抗脱唾液酸GM1 + C处理5-FU处理后的骨髓对两种活性均无影响。在任何条件下，用泛T细胞抗体 + C处理均不影响NK或LAK活性的发展。因此，对5-FU有抗性的NK/LAK祖细胞脱唾液酸GM1阴性，但在白细胞介素-2诱导后变为脱唾液酸GM1阳性。最后，通过用L-LME处理白细胞介素-2诱导的LGL/NK细胞，获得了骨髓来源的LAK细胞直接由白细胞介素-2诱导的NK细胞产生的证据。（摘要截短于400字）
