Hayakawa K, Salmeron M A, Kornbluth J, Bucana C, Itoh K
Department of Immunology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
J Immunol. 1991 Apr 1;146(7):2453-60.
The role of IL-4 in proliferation and differentiation of human NK cells was studied using newly established sublines of an IL-4-dependent NK cell clone (IL4d-NK cells) and an IL-2-dependent NK cell clone (IL2d-NK cells) derived from a parental conditioned medium-dependent NK cell clone (CM-NK cells). IL-4 induced the higher proliferation of CM-NK cells, but abolished their NK activity and decreased CD16 and CD56 Ag expression. In contrast, IL-2 induced the higher NK activity and increased CD16 and CD56 Ag expression. Addition of anti-IL-4 antibody to the culture of CM-NK cells with CM inhibited the proliferation, but slightly increased NK activity, and largely increased CD56 Ag expression. Addition of anti-IL-2 antibody to the culture of CM-NK cells with CM inhibited both proliferation and cytotoxicity. Proliferation of IL4d-NK cells, which is totally dependent on rIL-4, is greater than that of IL2d-NK cells, which was greater than parental CM-NK cells. Morphologically, IL4d-NK cells are small and round, whereas IL2d-NK cells are large and elongated. Anti-IL-4 antibody inhibited proliferation of IL4d-NK but not IL2d-NK cells, whereas anti-IL-2 antibody inhibited that of IL2d-NK but not IL4d-NK cells. IL-2 was not detected in the supernatant from IL4d-NK cells, nor was IL-2-mRNA expressed in IL4d-NK cells. In contrast, IFN-gamma production and protein expression in IL4d- and IL2d-NK cells were detected. NK cell activation markers (CD16 and CD56) were expressed on IL2d-NK cells but not IL4d-NK cells. IL4d-NK cells were not cytotoxic to any tumor cells tested, whereas IL2d-NK cells displayed potent NK activity and lymphokine-activated killer activity. IL4d-NK cells failed to bind K562 tumor cells, whereas one-third of the IL2d-NK cells did. IL4d-NK cells responded to rIL-2, proliferated, and differentiated into cytotoxic NK cells, whereas IL2d-NK cells failed to respond to rIL-4 and died. These results raise a possibility that IL4d-NK cells or IL2d-NK cells primarily represent the immunologic properties of immature or activated types of human NK cells, respectively. Our results provide the first evidence of the capability of IL-4 to support continuous proliferation of a lymphocyte clone with immature NK cell characteristics and to stimulate IFN-gamma production in the clone. IL-4 is suggested as a potential growth factor for certain types of human NK cell progenitors.
利用新建立的源自亲本条件培养基依赖性NK细胞克隆（CM - NK细胞）的IL - 4依赖性NK细胞克隆亚系（IL4d - NK细胞）和IL - 2依赖性NK细胞克隆亚系（IL2d - NK细胞），研究了IL - 4在人NK细胞增殖和分化中的作用。IL - 4诱导CM - NK细胞更高的增殖，但消除了它们的NK活性，并降低了CD16和CD56抗原表达。相反，IL - 2诱导更高的NK活性并增加了CD16和CD56抗原表达。向用CM培养的CM - NK细胞培养物中添加抗IL - 4抗体可抑制增殖，但略微增加NK活性，并大幅增加CD56抗原表达。向用CM培养的CM - NK细胞培养物中添加抗IL - 2抗体可同时抑制增殖和细胞毒性。完全依赖重组IL - 4的IL4d - NK细胞的增殖大于IL2d - NK细胞，而IL2d - NK细胞的增殖又大于亲本CM - NK细胞。形态学上，IL4d - NK细胞小而圆，而IL2d - NK细胞大且细长。抗IL - 4抗体抑制IL4d - NK细胞的增殖，但不抑制IL2d - NK细胞，而抗IL - 2抗体抑制IL2d - NK细胞的增殖，但不抑制IL4d - NK细胞。在IL4d - NK细胞的上清液中未检测到IL - 2，IL4d - NK细胞中也未表达IL - 2 - mRNA。相反，在IL4d - 和IL2d - NK细胞中检测到IFN - γ的产生和蛋白表达。NK细胞活化标志物（CD16和CD56）在IL2d - NK细胞上表达，但在IL4d - NK细胞上不表达。IL4d - NK细胞对任何测试的肿瘤细胞均无细胞毒性，而IL2d - NK细胞显示出强大的NK活性和淋巴因子激活的杀伤活性。IL4d - NK细胞不能结合K562肿瘤细胞，而三分之一的IL2d - NK细胞可以。IL4d - NK细胞对重组IL - 2有反应，增殖并分化为细胞毒性NK细胞，而IL2d - NK细胞对重组IL - 4无反应并死亡。这些结果提示IL4d - NK细胞或IL2d - NK细胞可能分别主要代表未成熟或活化型人NK细胞的免疫特性。我们的结果首次证明了IL - 4支持具有未成熟NK细胞特征的淋巴细胞克隆持续增殖并刺激该克隆中IFN - γ产生的能力。IL - 4被认为是某些类型人NK细胞祖细胞的潜在生长因子。
