Improvement of deficient natural killer activity and delayed bactericidal activity by a thiol proteinase inhibitor, E-64-d, in leukocytes from Chediak-Higashi syndrome patients in vitro.
Improvement of deficient natural killer activity and delayed bactericidal activity by a thiol proteinase inhibitor, E-64-d, in leukocytes from Chediak-Higashi syndrome patients in vitro.
复制标题
DOI：
10.1016/j.intimp.2009.01.003
复制
发表时间：
2009-03
影响因子：
5.6
通讯作者：
F. Tanabe;Hirotake Kasai;Limin He;T. Kin;T. Fujikado;T. Kumamoto;T. Hara;T. Iwata;Masahiko Ito-Masahiko
中科院分区：
文献类型：
--
作者：
F. Tanabe;Hirotake Kasai;Limin He;T. Kin;T. Fujikado;T. Kumamoto;T. Hara;T. Iwata;Masahiko Ito-Masahiko
我们以前报道过，在Chediak-Higashi综合征（CHS）小鼠模型（米色小鼠）中，给予一种有效的钙蛋白酶抑制剂E-64-d，可保护蛋白激酶C（PKC）免受蛋白水解，降低其对金黄色葡萄球菌感染的易感性。在本研究中，我们研究了E-64-d对6例CHS患者白细胞的自然杀伤（NK）缺陷和延迟杀菌活性的体外作用。结果表明，E-64-d（1 μg/ml）预处理CHS患者外周血单个核细胞（PBMCs）后，能显著增强NK细胞对K562细胞的杀伤活性。研究了多形核白细胞（PMNs）对沙门氏菌的延迟杀菌活性。金黄色葡萄球菌也表现出明显的改善。用E-64-d（1 μg/ml）预处理后，PMN的凋亡率几乎恢复到正常水平。另一方面，相同浓度的E-64-d不影响正常对照的NK或杀菌活性。此外，我们证实，E-64-d治疗后，伴刀豆球蛋白A（Con A）刺激后PKC活性的异常下调被消除在从CHS患者获得的PBMC中。为了检查PKC是否参与NK细胞介导的中性粒细胞的细胞溶解和杀菌活性，使用了两种有效的PKC抑制剂，白屈菜红蛋白和GÖ 6976。我们发现白屈菜红蛋白以剂量依赖性方式抑制正常PBMC的NK活性，而GÖ 6976在抑制Ca 2+依赖性PKC同工酶的剂量下抑制NK活性。这些抑制剂也抑制了PMN对S.金黄色。综上所述，我们的研究结果表明，E-64-d通过逆转PKC活性的下调来改善CHS患者白细胞的NK和杀菌活性。
We previously reported that administration of a potent calpain inhibitor, E-64-d, which protects protein kinase C (PKC) from proteolysis, in a mouse model of Chediak–Higashi syndrome (CHS) (beige mice), decreases its susceptibility to Staphylococcus aureus infection. In the present study, we examined the in vitro effect of E-64-d on both deficient natural killer (NK) and delayed bactericidal activities of leukocytes from six CHS patients. Our results showed that pretreatment of peripheral blood mononuclear cells (PBMCs) obtained from CHS patients with E-64-d (1 μg/ml) significantly enhanced NK activity against K562 cells. The delayed bactericidal activity of polymorphonuclear cells (PMNs) against S. aureus also showed marked improvement. This was recovered to almost normal levels when PMNs were pretreated with E-64-d (1 μg/ml). On the other hand, the same concentration of E-64-d did not affect either the NK or bactericidal activity of normal controls. In addition, we confirmed that following E-64-d treatment, the abnormal down-regulation of PKC activity after concanavalin A (Con A) stimulation was eliminated in PBMCs obtained from CHS patients. To examine whether PKC is involved in the NK cell-mediated cytolysis and bactericidal activity of PMNs, two potent PKC inhibitors, chelerythrin and GÖ6976, were used. We found that chelerythrin inhibits NK activity of normal PBMCs in a dose-dependent manner, and GÖ6976 inhibits NK activity at doses that inhibit Ca2+-dependent PKC isozymes. These inhibitors also suppressed the bactericidal activity of PMNs against S. aureus. Taken together, our findings suggested that E-64-d improved the compromised NK and bactericidal activity of leukocytes from CHS patients by reversing the down-regulation of PKC activity.