Phosphorylation of src family lck tyrosine kinase following interleukin-12 activation of human natural killer cells.
Phosphorylation of src family lck tyrosine kinase following interleukin-12 activation of human natural killer cells.
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DOI：
10.1006/cimm.1995.1207
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发表时间：
1995-10
影响因子：
4.3
通讯作者：
Claudio Pignata;Kanteti V.S. Prasad;M. Hallek;B. Druker;C. E. Rudd;Michael J. Robertson;Jerome Ritz
中科院分区：
文献类型：
--
作者：
Claudio Pignata;Kanteti V.S. Prasad;M. Hallek;B. Druker;C. E. Rudd;Michael J. Robertson;Jerome Ritz
白细胞介素-12 （IL-12）是一种异二聚体细胞因子，可增强人NK细胞和T细胞的细胞溶解活性，但对静止淋巴细胞的有丝分裂活性很小。参与NK细胞被IL-12激活的细胞内信号通路尚未完全表征。在目前的研究中，我们发现IL-12诱导静止NK细胞中56-kDa蛋白的酪氨酸磷酸化，在Western blot实验中鉴定为p56lck。IL-12的活性范围为0.1 ~ 1000u /ml，最大活性范围为10 ~ 100u /ml （30 ~ 300pm）。刺激后5分钟达到最大效果，染料木素几乎完全抑制。IL-12在静止NK细胞和IL-2活化NK细胞中也有类似的作用。IL-12刺激静息NK细胞后，免疫沉淀的ck激酶在激活后5和10分钟的体外自磷酸化活性增加。另外几种底物在IL-12刺激下被体外磷酸化，包括70和110 kDa的蛋白。这些研究表明，缺乏酪氨酸激酶参与了人NK细胞IL-12信号通路。由于IL-12对静止NK细胞的主要功能作用是增强细胞溶解活性和分泌功能，而不是细胞增殖，这些发现表明缺乏酪氨酸激酶参与了人类NK细胞的这些信号通路。
Interleukin-12 (IL-12) is a heterodimeric cytokine that augments the cytolytic activity of human NK cells and T cells but has little mitogenic activity on resting lymphocytes. The intracellular signaling pathways involved in NK cell activation by IL-12 have not been fully characterized. In the present studies we found that IL-12 induces tyrosine phosphorylation of a 56-kDa protein, identified in Western blot experiments as p56lck, in resting NK cells. IL-12 was active in the range of 0.1 to 1000 U/ml, with maximal activity between 10 and 100 U/ml (30 and 300 pM). The maximal effect was noted 5 min after stimulation and was almost completely inhibited by genistein. IL-12 induced similar effects in resting and IL-2 activated NK cells. Following IL-12 stimulation of resting NK cells, immunoprecipitated lck kinase exhibited increased in vitro autophosphorylation activity 5 and 10 min after activation. Several additional substrates were phosphorylated in vitro following IL-12 stimulation, including proteins of 70 and 110 kDa. These studies indicate that lck tyrosine kinase is involved in the IL-12 signaling pathway in human NK cells. As the primary functional effect of IL-12 on resting NK cells is the enhancement of cytolytic activity and secretory function rather than cell proliferation, these findings suggest that lck tyrosine kinase is involved in these signaling pathways in human NK cells.