Apoptosis of sea bass (Dicentrarchus labrax L.) neutrophils and macrophages induced by experimental infection with Photobacterium damselae subsp piscicida
Apoptosis of sea bass (Dicentrarchus labrax L.) neutrophils and macrophages induced by experimental infection with Photobacterium damselae subsp piscicida
复制标题
DOI：
10.1016/s1050-4648(02)00144-4
复制
发表时间：
2003-08-01
影响因子：
4.7
通讯作者：
Silva, MT
中科院分区：
文献类型：
--
作者：
do Vale, A;Marques, F;Silva, MT
本文报道了花斑石斑鱼(Dicentarchus Labrax L.)经腹膜腔注射(Ip)鱼巴氏杆菌淡色杆菌亚种的注射制剂。鱼腥藻可引起腹膜中性粒细胞和巨噬细胞的凋亡。所有八个毒力菌株和两个非毒力菌株都没有表现出致细胞凋亡的活性。一种分泌的细菌蛋白(S)可能是导致这种活性的因素(S)，因为腹腔注射没有诱导细胞凋亡。注射紫外线灭活的强毒菌株和强毒培养上清液显示出与活细菌相同的热不稳定的促凋亡活性。通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)染色和DNA电泳法检测到DNA片段化，以及典型的超微结构改变，即细胞皱缩、染色质凝集、核碎裂和产生泡并脱落的凋亡小体，是细胞凋亡过程的特征。在致死剂量的强毒细菌或培养上清液诱导的细胞凋亡过程中，腹腔巨噬细胞和中性粒细胞均受到广泛影响。这些细胞中的大多数是凋亡的，在注射后24it时，每种细胞类型的每种腹膜约有10(7)个细胞。此外，静息腹膜腔内非凋亡性巨噬细胞的数量始终低于初始数量。由于巨噬细胞是消除细菌和凋亡性死亡细胞和凋亡体的关键细胞，因此丹参亚种的诱导作用。同时发生巨噬细胞和中性粒细胞凋亡的鱼丝，一方面会破坏限制细菌繁殖的两种吞噬细胞类型，另一方面会导致细胞凋亡过程失控地向继发性坏死发展，最终导致大量垂死的中性粒细胞和中性粒细胞凋亡小体的溶解，从而导致其高度细胞毒性成分的广泛释放。在实验性巴氏杆菌病死亡的鱼的头肾切片中也可以看到大量的凋亡细胞。反之，石斑鱼头肾巨噬细胞培养的毒力培养上清液或用毒力细菌或培养上清液处理腹膜渗出的白细胞，在体外均未见明显的细胞凋亡。这里描述的凋亡过程似乎是一种新的和非常强大的微生物致病策略。(C)2003爱思唯尔有限公司。保留所有权利。
The infection of sea bass (Dicentrarchus labrax L.) by intraperitoneal (i.p.) injection of the agent of fish pasteurellosis Photobacterium damselae subsp. piscicida resulted in the apoptosis of peritoneal neutrophils and macrophages. All the eight virulent and none of the two non-virulent strains tested exhibited apoptogenic activity. A secreted bacterial protein(s) is a likely candidate as the factor(s) responsible for this activity, since no apoptosis was induced by i.p. injected UV-killed virulent strains and the virulent culture supernatants exhibited a thermo-labile apoptogenic activity identical to that of live bacteria. The apoptotic process was characterized by the occurrence of DNA fragmentation detected by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) staining and DNA electrophoresis, and of typical ultrastructural alterations namely cell shrinkage, chromatin condensation, nuclear fragmentation and production of blebs with shedding of apoptotic bodies. In the apoptotic process induced by lethal doses of virulent bacteria or culture supernatants both peritoneal macrophages and neutrophils were extensively affected. the majority of these cells being apoptotic and reaching values around 10(7) per peritoneal cavity for each cell type at 24 It post-injection. Moreover, the number of non-apoptotic macrophages was always below the initial number in the resting peritoneal cavity. Since macrophages are key cells in the elimination of both bacteria and apoptotic moribund cells and apoptotic bodies, the induction by Ph. damselae subsp. piscicida of simultaneous macrophage and neutrophil apoptosis results, on the one hand, in the destruction of the two phagocytic cell types involved in the restriction of multiplication of the bacteria and, on the other hand, in the uncontrolled progression of the apoptotic process towards secondary necrosis and eventual lysis of high numbers of moribund neutrophils and of neutrophilic apoptotic bodies, with the consequent extensive release of their highly cytotoxic components. Abundant apoptotic cells were also seen in sections of head-kidney from fish dying from experimental pasteurellosis. In contrast, no apoptosis was seen in vitro after the treatment with virulent culture supernatants of sea bass head-kidney macrophage cultures or after the treatment ex vivo of peritoneal exudate leukocytes with virulent bacteria or culture supernatants. The apoptotic process described here appears as a novel and very powerful microbial pathogenic strategy. (C) 2003 Elsevier Ltd. All rights reserved.