Deficiency of Src family kinases p59/61hck and p58c-fgr results in defective adhesion-dependent neutrophil functions.
Deficiency of Src family kinases p59/61hck and p58c-fgr results in defective adhesion-dependent neutrophil functions.
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DOI：
10.1083/jcb.133.4.895
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发表时间：
1996-05
期刊：
影响因子：
--
通讯作者：
Berton G
中科院分区：
文献类型：
--
作者：
Lowell CA;Fumagalli L;Berton G
通过细胞外基质（ECM）蛋白或单克隆抗体（mAb）交联嗜中性粒细胞-β 2-或β 3-相关的白细胞应答整联蛋白刺激细胞骨架重排，导致细胞扩散和呼吸爆发。近来，多形核白细胞（PMN）内多种蛋白质的酪氨酸磷酸化和Src家族激酶的活化与白细胞整联蛋白产生的细胞内信号传导途径有关（Yan，S.R.，L. Fumagalli，and G Berton. 1995. J.炎症。45：217 - 311.）为了直接测试这些功能反应是否依赖于Src家族激酶p59/61 hck和p58 c-fgr，我们检测了从hck-/-、fgr-/-和hck-/-fgr-/-敲除小鼠分离的PMN中的粘附依赖性呼吸爆发。在活化剂如肿瘤坏死因子（TNF）或甲酰甲硫氨酰亮氨酰苯丙氨酸存在下，野生型小鼠的纯化骨髓PMNS在粘附于纤维蛋白原、纤连蛋白或胶原包被的表面时释放大量O2-，如对人PMNS所述。然而，来自hck-/-fgr-/-双突变小鼠的PMNs没有反应。这种缺陷对于整合素信号传导是特异性的，因为在免疫复合物或PMA刺激的hck-/-fgr-/-PMN中呼吸爆发是正常的。在用鼠细胞内粘附分子-1（ICAM-1）包被的表面上接种的TNF-致敏的野生型PMN中观察到呼吸爆发的刺激，而hck-/-fgr-/-PMN没有反应。直接交联的β 2和β 2整合素的亚基的表面结合的单克隆抗体引起的O2-生产的野生型中性粒细胞，而双突变hck-/-fgr-/-细胞未能作出反应。光学显微镜和细胞粘附试验表明，受损的功能反应的hck-/-fgr-/-中性粒细胞的缺陷造成的扩展和紧密粘附ECM蛋白或单克隆抗体包被的表面。与此相反，hck-/-或fgr-/-单突变细胞产生的O2-水平相当于野生型细胞的ECM蛋白，鼠ICAM-1，抗整合素单克隆抗体包被的表面。因此，p59/61 hck和p58 c-fgr是通过白细胞β 2和β 3整联蛋白进行信号传导所必需的，从而导致PMN扩散和呼吸爆发。这是Src家族激酶在白细胞内整合素信号传导中重要性的第一个直接遗传证据，也是该基因家族成员在确定的信号转导途径内重叠功能的最佳实例。
Cross-linking of the neutrophil-beta 2- or beta 3-related leukocyte response integrins by extracellular matrix (ECM) proteins or monoclonal antibodies (mAb) stimulates cytoskeletal rearrangement leading to cell spreading and respiratory burst. Tyrosin phosphorylation of a variety of proteins and activation of the Src family kinases within polymorphonuclear leukocytes (PMN) have recently been implicated in the intracellular signaling pathways generated by leukocyte integrins (Yan, S.R., L. Fumagalli, and G Berton. 1995. J. Inflammation. 45:217-311.) To directly test whether these functional responses are dependent on the Src family kinases p59/61hck and p58c-fgr, we examined adhesion- dependent respiratory burst in PMNs isolated from hck -/-, fgr -/-, and hck -/- fgr -/- knockout mice. Purified bone marrow PMNS from wild-type mice released significant amounts of O2- when adherent to fibrinogen-, fibronectin-, or collagen-coated surfaces, in the presence of activating agents such as tumor necrosis factor (TNF) or formyl- methionyl-leucyl-phenylalanine, as described for human PMNs. PMNs from hck-/-fgr-/- double-mutant mic, however, failed to respond. This defect was specific for integrin signaling, since respiratory burst was normal in hck-/-fgr-/-PMNs stimulated by immune complexes or PMA. Stimulation of respiratory burst was observed in TNF-primed wild-type PMN plated on surfaces coated with murine intracellular adhesion molecule-1 (ICAM-1), while hck-/-fgr-/- PMNs, failed to respond. Direct cross-linking of the subunits of beta 2 and beta 2 integrins by surface-bound mAbs was elicited O2- production by wild-type PMNs, while the double-mutant hck- /-fgr-/- cells failed to respond. Photomicroscopy and cell adhesion assays revealed that the impaired functional responses of hck-/-fgr-/- PMNs were caused by defective spreading and tight adhesion on either ECM protein- or mAb-coated surfaces. In contrast, hck-/-or fgr-/-single mutant cells produced O2- at levels equivalent to wild-type cells on ECM protein, murine ICAM-1, and antiintegrin mAb-coated surfaces. Hence, either p59/61 hck and p 58c-fgr is required for signaling through leukocyte beta 2 and beta 3 integrins leading to PMN spreading and respiratory burst. This is the first direct genetic evidence of the importance of Src family kinases in integrin signaling within leukocytes, and it is also the best example of overlapping function between members of this gene family within a defined signal transduction pathway.
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DOI：
10.1083/jcb.126.6.1585
发表时间：
1994-09
期刊：
The Journal of cell biology
影响因子：
--
作者：
Lin TH;Yurochko A;Kornberg L;Morris J;Walker JJ;Haskill S;Juliano RL
通讯作者：
Juliano RL
影响因子：
7.8
作者：
Graham, I L;Anderson, D C;Holers, V M;Brown, E J
通讯作者：
Brown, E J
影响因子：
56.9
作者：
HIBBS, ML;XU, H;SPRINGER, TA
通讯作者：
SPRINGER, TA
影响因子：
5.5
作者：
BELLAVITE, P;CHIRUMBOLO, S;DRI, P
通讯作者：
DRI, P
影响因子：
7.8
作者：
Huang, M M;Lipfert, L;Cunningham, M;Brugge, J S;Ginsberg, M H;Shattil, S J
通讯作者：
Shattil, S J