Caspase-independent cell death induced by anti-CD2 or staurosporine in activated human peripheral T lymphocytes.
Caspase-independent cell death induced by anti-CD2 or staurosporine in activated human peripheral T lymphocytes.
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在活化的人外周 T 淋巴细胞中，抗 CD2 或十字孢菌素诱导的不依赖于半胱天冬酶的细胞死亡。
作者：
O. Deas;C. Dumont;M. MacFarlane;M. Rouleau;C. Hebib;F. Harper;F. Hirsch;B. Charpentier;G. Cohen;A. Senik
我们研究了细胞通透性的广谱多肽半胱氨酸氨基转移酶抑制剂benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethyl酮(Z-VAD.fmk)和BOC-Asp(OMe)-氟甲基酮(BOC-D.fmk)对抗CD2、抗Fas和蛋白激酶抑制剂星形孢子素诱导活化的人外周T淋巴细胞凋亡的影响。我们监测了超微结构、流式细胞仪和生化的凋亡变化，包括磷脂酰丝氨酸外化，多聚ADP核糖聚合酶(PARP)和层粘连蛋白的切割，caspase-3和caspase-7的激活，线粒体膜电位的降低和DNA片段化。Z-VAD.fmk和BOC-D.fmk完全抑制抗Fas处理细胞的所有生化和超微结构变化。与之形成鲜明对比的是，Z-VAD.fmk和BOC-D.fmk都不能抑制CD2或星形孢菌素介导的细胞收缩、内质网扩张(在抗CD2处理的细胞中可见)、磷脂酰丝氨酸的外化以及伴随细胞死亡的线粒体膜电位的丧失。然而，这些抑制剂确实抑制了PARP和lamins的切割和亚二倍体细胞的形成，并部分抑制了染色质的凝聚。这些结果表明，在活化的T细胞中，抗CD2抗体和星形孢子素诱导的细胞死亡途径不依赖于caspase，表现出明显的细胞质特征的凋亡。然而，caspase的激活对于核底物如PARP和lamins的蛋白降解以及在凋亡细胞中发生的DNA断裂和极端的染色质凝聚是必需的。
We examined the effects of the cell-permeable, broad spectrum peptide caspase inhibitors, benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethyl ketone (Z-VAD.fmk), and BOC-Asp(OMe)-fluoromethyl ketone (BOC-D.fmk), on apoptosis induced by anti-CD2, anti-Fas, and the protein kinase inhibitor staurosporine in activated human peripheral T lymphocytes. We monitored ultrastructural, flow cytometric, and biochemical apoptotic changes, including externalization of phosphatidylserine, cleavage of poly(ADP-ribose) polymerase (PARP) and lamins, activation of caspase-3 and caspase-7, decrease in mitochondrial membrane potential, and DNA fragmentation. Z-VAD.fmk and BOC-D.fmk completely inhibited all the biochemical and ultrastructural changes of apoptosis in anti-Fas-treated cells. In marked contrast, neither Z-VAD.fmk nor BOC-D.fmk inhibited CD2- or staurosporine-mediated cell shrinkage, dilatation of the endoplasmic reticulum (seen in anti-CD2-treated cells), externalization of phosphatidylserine, and loss of mitochondrial membrane potential that accompanied cell death. However, these inhibitors did inhibit the cleavage of PARP and lamins and the formation of hypodiploid cells, and partially inhibited chromatin condensation. These results demonstrate that in activated T cells, anti-CD2 and staurosporine induce a caspase-independent cell death pathway that exhibits prominent cytoplasmic features of apoptosis. However, caspase activation is required for the proteolytic degradation of nuclear substrates such as PARP and lamins together with the DNA fragmentation and extreme chromatin condensation that occur in apoptotic cells.
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DOI：
10.1073/pnas.93.25.14486
发表时间：
1996-12-10
影响因子：
11.1
作者：
Srinivasula, SM;Ahmad, M;Alnemri, ES
通讯作者：
Alnemri, ES
DOI：
10.1073/pnas.93.16.8395
发表时间：
1996-08
影响因子：
11.1
作者：
A. Takahashi;A. Takahashi;E. Alnemri;Y. Lazebnik;Y. Lazebnik;T. Fernandes‐Alnemri;G. Litwack;R. Moir;R. Goldman;G. Poirier;S. Kaufmann;W. Earnshaw;W. Earnshaw
通讯作者：
A. Takahashi;A. Takahashi;E. Alnemri;Y. Lazebnik;Y. Lazebnik;T. Fernandes‐Alnemri;G. Litwack;R. Moir;R. Goldman;G. Poirier;S. Kaufmann;W. Earnshaw;W. Earnshaw
影响因子：
11.2
作者：
T. Fernandes‐Alnemri;Atsushi Takahashi;R. Armstrong;J. Krebs;L. Fritz;K. Tomaselli;Lijuan Wang;Zailin Yu;C. Croce;Guy Salveson;W. Earnshaw;G. Litwack;E. Alnemri
通讯作者：
T. Fernandes‐Alnemri;Atsushi Takahashi;R. Armstrong;J. Krebs;L. Fritz;K. Tomaselli;Lijuan Wang;Zailin Yu;C. Croce;Guy Salveson;W. Earnshaw;G. Litwack;E. Alnemri
DOI：
10.1073/pnas.93.15.7464
发表时间：
1996-07-23
影响因子：
11.1
作者：
FernandesAlnemri, T;Armstrong, RC;Alnemri, ES
通讯作者：
Alnemri, ES