Gorelik E, Jay G, Kwiatkowski B, Herberman R B
Pittsburgh Cancer Institute, Department of Pathology, University of Pittsburgh, PA 15213.
J Immunol. 1990 Sep 1;145(5):1621-32.
An H-2Kb- negative clone of BL6 melanoma (BL6-8) was transfected with neor, H-2Kb, or H-2IAk genes. In an 18-h cytotoxicity assay clones with high levels of H-2Kb Ag expression were found more sensitive to lysis by spleen cells of syngenic and allogeneic mice than H-2Kb low clones. NK cells were involved in the lysis of H-2Kb+ BL6 melanoma clones, with spleen cell cytotoxicity of mice increased after poly I:C stimulation or decreased after pretreatment with anti-asialo GM1 serum or NK1.1 mAb. Anti-TNF Ab were also able to reduce the cytotoxicity of normal spleen cells and completely abolished the cytotoxicity of the NK-depleted spleen cells suggesting involvement of NC cells in lysis of H-2Kb+ BL6 melanoma clones. Increase in sensitivity of H-2Kb+ BL6 cells to natural cell-mediated cytotoxicity was associated with the appearance of NK recognizable determinants as assessed by the cold target inhibition assay. All BL6 clones, irrespective of sensitivity to natural cell-mediated cytotoxicity, showed high sensitivity to lysis by LGL-derived granules. In contrast, all H-2Kb low BL6 clones were resistant and all H-2Kb highly positive clones were sensitive to lysis by TNF-alpha. When an H-2Kb highly positive clone was selected in vitro for resistance to TNF, it concomitantly showed increased resistance to cytotoxicity by spleen cells, confirming the importance of TNF in spleen cell cytotoxicity against H-2Kb+ melanoma cells. Taken together, the data indicate that class I H-2Kb but not class II H-2IAk gene product could increase the sensitivity of BL6 cells to lysis by NK and natural cytotoxic cells as well as TNF. We hypothesize that these effects could be due to pleiotropic effects of H-2Kb gene products on various biologic properties of BL6 melanoma cells some of which may be more directly involved in regulation of tumor cell sensitivity to lysis by NK and/or natural cytotoxic cells.
将neo、H-2Kb或H-2IAk基因转染至BL6黑色素瘤的H-2Kb阴性克隆（BL6-8）。在一项18小时细胞毒性试验中，发现H-2Kb Ag表达水平高的克隆比H-2Kb低的克隆对同基因和异基因小鼠的脾细胞裂解更敏感。NK细胞参与了H-2Kb+BL6黑色素瘤克隆的裂解，聚肌胞苷酸（poly I:C）刺激后小鼠脾细胞细胞毒性增加，或用抗唾液酸GM1血清或NK1.1单克隆抗体预处理后细胞毒性降低。抗TNF抗体也能够降低正常脾细胞的细胞毒性，并完全消除NK细胞耗竭的脾细胞的细胞毒性，提示NC细胞参与了H-2Kb+BL6黑色素瘤克隆的裂解。通过冷靶抑制试验评估，H-2Kb+BL6细胞对自然细胞介导的细胞毒性敏感性增加与NK可识别决定簇的出现有关。所有BL6克隆，无论对自然细胞介导的细胞毒性的敏感性如何，对LGL衍生颗粒的裂解均表现出高敏感性。相反，所有H-2Kb低的BL6克隆均具有抗性，所有H-2Kb高度阳性的克隆对TNF-α的裂解敏感。当在体外选择一个对TNF具有抗性 的H-2Kb高度阳性克隆时，它同时显示出对脾细胞细胞毒性的抗性增加，证实了TNF在脾细胞对H-2Kb+黑色素瘤细胞的细胞毒性中的重要性。综上所述，数据表明I类H-2Kb而非II类H-2IAk基因产物可增加BL6细胞对NK和自然细胞毒性细胞以及TNF裂解的敏感性。我们推测，这些效应可能是由于H-2Kb基因产物对BL6黑色素瘤细胞各种生物学特性的多效性作用，其中一些可能更直接参与调节肿瘤细胞对NK和/或自然细胞毒性细胞裂解的敏感性。
