Structural and Functional Analysis of Human Interleukin-12 Receptor

我们用信号陷阱方法建立了一个由单克隆抗体（mAb）组成的小组，称为人类介素-12受体β 1链（IL-12 R）。在这种情况下，用于Tac信号序列的cDNA被替换为IL-12 R信号+外链域序列。化学cDAN是被表达出来的，而反毒性抗体则被用来检测化学蛋白质。被移植的细胞被用来免疫性。杂交瘤是由传统细胞融合和确认的，阳性的支架是由植入物和阴性的野生细胞。我们可以获得两种中性化和非中性化抗体，作为通过抑制IL-12诱导的T细胞增殖试验和放射性配体结合研究来确定的。免疫吸收被披露的转染者和PHA刺激的人类淋巴细胞中的两种可移植性p110.我们可以用新开发的mAb绘制IL-12和IL-12 R的后续组合。T细胞对IL-12的响应要求存在单细胞和其他抗原存在细胞。为了更具体，CD 2在T细胞上和CD 58上的单核细胞之间的相互作用在IL-12的T细胞激活中的基本作用，而T细胞对IL-12的反应作用在IL-4抑制和gammaIFN增强中。T细胞上IL-12 R的表达水平如何，不受细胞因子的影响。我们可以与IL-12 R中的T细胞中的T细胞一起免疫蛋白85 kDa蛋白质。这一新发现的蛋白质在IL-12刺激中被酪氨酸磷酸化。这与其他已知的STAT和JAK蛋白质不同。这种分子可能是分析IL-12/IL-12 R信号转换系统的有趣线索。

We established a panel of monoclonal antibodies(mAb)to human Interleukin-12 receptor beta1chain(IL-12R)by Signal Trap method. In this case, the cDNA for Tac signal sequence was replaced with IL-12R signal+extracellular domain sequence. The chimeric cDAN was expressed and anti-Tac antibody was used to detect the chimeric protein. The transfected cells were used for immunization. Hybridomas were derived by conventional cell fusion and confirmed by positive staining with transfectants and negative with wild-type cells. We could obtain both neutralizing and non-neutralizing antibodies as determined by inhibition of IL-12 induced T cell proliferation assay and of radiolabelled ligand binding study. Immunoprecipitation disclosed putative p110 both from transfectants and PHA-stimulated human lymphocytes.We could draw the following conclusions on IL-12 and IL-12R with newly developed mAb.・T CELL responsiveness TO IL-12 required presence of monocytes and other antigen presenting cells. To be more specific, interaction between CD2 onT cells and CD58 on monocytes was essential in the T cell activation by IL-12・T cell responsiveness to IL-12 was inhibited by IL-4 and enhanced by gammaIFN.Expression level of IL-12R on T cells was, however, not affected by either of cytokines.・We could co-immunoprecipitate 85 kDa protein along with IL-12R from T cells with our mAb. This newly identified protein was tyrosine-phosphorylated upon IL-12 stimulation. This was different from other known STAT and JAK proteins. This molecule could be an interesting clue to analyze IL-12/IL-12R signal transduction system.