Triozzi P L, Aldrich W A, Rinehart J J
Department of Medicine, Ohio State University, Columbus 43210.
Mol Biother. 1991 Mar;3(1):22-5.
Depleting monocytes from human peripheral blood mononuclear cells (PBMC) enhances the in vitro activation of lymphokine-activated killer (LAK) cells. To determine if monocytes also altered LAK-cell expansion, we evaluated two methods of depleting monocytes from PBMC: nylon wool adherence (NWA) and phenylalanine methyl ester (PME) treatment. Both methods of depleting monocytes enhanced interleukin-2 (IL-2) driven, LAK-cell expansion; LAK expansion, however, was significantly greater after depletion with NWA than after PME. LAK cytotoxicity after NWA and PME depletion was equivalent. The degree of monocyte depletion, determined by evaluating morphology and the number of Leu-M3 (CD14) positive cells, and the proliferation of Leu 19 (CD56), OKT-3 (CD3), Leu2 (CD8), and Leu 3a (CD4) positive cells was also equivalent. Exposure of IL-2 activated cells to PME did not alter their cytotoxic activity. However, sequential treatment of PBMC with NWA, then PME, or with PME and then NWA, resulted in reduced expansion. This reduction in expansion was similar to PBMC treated with PME alone. Exposure of PME-depleted cells to nylon wool or to supernatants obtained from cells adherent to nylon wool further decreased LAK expansion relative to cells treated with NWA alone. We conclude that even at relatively low cell density, human monocytes markedly inhibit LAK-cell expansion in IL-2 driven PBMC cultures. Further, depletion of monocytes by NWA adherence is more effective than by treatment with PME, possibly due to subtle cellular damage induced by this latter treatment. These findings have implication for the in vitro and in vivo generation of LAK-cells by IL-2.
从人外周血单个核细胞（PBMC）中去除单核细胞可增强淋巴因子激活的杀伤细胞（LAK细胞）的体外激活。为了确定单核细胞是否也会改变LAK细胞的扩增，我们评估了两种从PBMC中去除单核细胞的方法：尼龙毛黏附法（NWA）和苯丙氨酸甲酯（PME）处理法。两种去除单核细胞的方法均增强了白细胞介素-2（IL-2）驱动的LAK细胞扩增；然而，用NWA去除单核细胞后的LAK细胞扩增明显大于用PME处理后的扩增。NWA和PME去除单核细胞后的LAK细胞细胞毒性相当。通过评估形态学和Leu-M3（CD14）阳性细胞数量确定的单核细胞去除程度，以及Leu 19（CD56）、OKT-3（CD3）、Leu2（CD8）和Leu 3a（CD4）阳性细胞的增殖情况也相当。将IL-2激活的细胞暴露于PME不会改变其细胞毒性活性。然而，先用NWA然后用PME，或先用PME然后用NWA顺序处理PBMC，会导致扩增减少。这种扩增减少类似于单独用PME处理的PBMC。相对于单独用NWA处理的细胞，将PME去除的细胞暴露于尼龙毛或从黏附于尼龙毛的细胞获得的上清液中会进一步降低LAK细胞扩增。我们得出结论，即使在相对较低的细胞密度下，人单核细胞也会在IL-2驱动的PBMC培养物中显著抑制LAK细胞扩增。此外，通过NWA黏附去除单核细胞比用PME处理更有效，这可能是由于后一种处理诱导的细微细胞损伤。这些发现对IL-2在体外和体内产生LAK细胞具有启示意义。
