Agah R, Shau H, Mazumder A
Cancer Immunol Immunother. 1987;24(3):247-52. doi: 10.1007/BF00205638.
Little is known regarding the effectors of lymphokine-activated killer activity. Lysosomotropic agents such as quinacrine can be used to positively sort for lysosome rich cells in natural killer (NK) cell populations. We therefore decided to use this agent to sort lymphokine-activated killer (LAK) cells to characterize their lysosomal content. We found that the positively sorted population contained all the LAK activity, i.e., lysis of NK-resistant tumor cells (B16 melanoma cell line), with the negatively sorted cells having no killing activity. Therefore separation of interleukin-2-incubated cells for LAK activity could be accomplished using sorting after quinacrine staining. The treatment of positively sorted LAK cell populations with L-leucine methyl ester, a lysosomotropic dye which inhibits killing by lysosome rich cells, caused abrogation of killing of the B16 tumor by the treated populations. Single cell conjugate assays were also done on these sorted cells, with positively sorted cells forming the highest and negatively sorted cells the lowest percent of conjugates. Our data therefore indicates the important role of lysosome rich cells in the LAK cell population in the murine system.
关于淋巴因子激活的杀伤活性的效应细胞,人们了解甚少。诸如奎纳克林等溶酶体亲和剂可用于对自然杀伤(NK)细胞群体中富含溶酶体的细胞进行阳性分选。因此,我们决定使用这种试剂对淋巴因子激活的杀伤(LAK)细胞进行分选,以表征其溶酶体含量。我们发现,阳性分选群体包含所有的LAK活性,即对NK抗性肿瘤细胞(B16黑色素瘤细胞系)的裂解,而阴性分选细胞则没有杀伤活性。因此,使用奎纳克林染色后进行分选,可实现对白细胞介素-2孵育细胞的LAK活性分离。用L-亮氨酸甲酯(一种抑制富含溶酶体细胞杀伤作用的溶酶体亲和染料)处理阳性分选的LAK细胞群体,导致处理后的群体对B16肿瘤的杀伤作用消失。还对这些分选细胞进行了单细胞结合试验,阳性分选细胞形成的结合物百分比最高,阴性分选细胞形成的结合物百分比最低。因此,我们的数据表明富含溶酶体的细胞在小鼠系统的LAK细胞群体中起着重要作用。