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温度对豚鼠吞噬细胞杀灭嗜肺军团菌及嗜肺军团菌在吞噬细胞中增殖的影响。

Temperature effects on Legionella pneumophila killing by and multiplication in phagocytes of guinea pigs.

作者信息

Miyamoto H, Ogawa M, Maruta K, Nikaido Y, Yamamoto C, Taniguchi H, Yoshida S

机构信息

Department of Microbiology, School of Medicine, University of Occupational and Environmental Health, Fukuoka, Japan.

出版信息

Microbiol Immunol. 1995;39(9):647-54. doi: 10.1111/j.1348-0421.1995.tb03252.x.

Abstract

We examined the effects of temperature on the interaction between Legionella pneumophila and phagocytes of guinea pigs. The body temperatures of guinea pigs infected with a sublethal dose (1.2 x 10(4) CFU) or a lethal dose (1.0 x 10(5) CFU) of L. pneumophila elevated from 38.4 +/- 0.15 C to 40.2 +/- 0.42 C or 40.3 +/- 0.62 C, respectively. The intracellular bacterial killing by and bacterial proliferation in the phagocytes were examined at 33, 37, 40, and 42 C, using in vitro culture systems of peritoneal macrophages or polymorphonuclear leukocytes (PMN) of guinea pigs. In all the macrophages incubated at different temperatures, significant intracellular bacterial killings were observed at 4 hr after in vitro phagocytosis. After 24 hr of incubation, there was about a 100-fold increase of CFU and the number reached a maximum after 48 hr of incubation in the macrophages incubated at 42 C as well as 37 and 40 C, suggesting that macrophages support the intracellular bacterial growth in hyperthermia. In the PMN, L. pneumophila CFU 4 hr or 12 hr after the infection were significantly lower at 42 C than those at 37 C (P < 0.05), indicating that the bactericidal capacity of PMN was enhanced at 42 C compared to 37 C. However, in all the PMN incubated at different temperatures, there were about 10-fold increases of CFU 24 hr after the infection, suggesting that PMN as well as macrophages support intracellular bacterial growth in hyperthermia. The extracellular bacterial growth was examined at 33, 37, 40, and 42 C in buffered yeast extract (BYE) broth or RPMI 1640 medium containing 50% guinea pig serum as a permissive or non-permissive liquid medium for the bacterial growth, respectively. Inhibition of bacterial growth in BYE broth at 42 C, and a decrease of CFU in RPMI 1640 medium containing 50% guinea pig serum at 42 C were observed. In conclusion, hyperthermia may be beneficial by restricting extracellular bacterial survival, but it exerts no beneficial effect on the restriction of intracellular bacterial growth in phagocytes, though PMN showed enhanced initial killing at 42 C. These results suggest that fever, or hyperthermia itself, may not largely contribute as a nonspecific host defense early in the course of legionellosis.

摘要

我们研究了温度对嗜肺军团菌与豚鼠吞噬细胞之间相互作用的影响。感染亚致死剂量(1.2×10⁴CFU)或致死剂量(1.0×10⁵CFU)嗜肺军团菌的豚鼠体温分别从38.4±0.15℃升高到40.2±0.42℃或40.3±0.62℃。利用豚鼠腹腔巨噬细胞或多形核白细胞(PMN)的体外培养系统,在33、37、40和42℃下检测吞噬细胞内细菌的杀伤情况以及细菌的增殖情况。在不同温度下孵育的所有巨噬细胞中,体外吞噬4小时后均观察到明显的细胞内细菌杀伤。孵育24小时后,在42℃以及37℃和40℃下孵育的巨噬细胞中,CFU增加了约100倍,且在孵育48小时后数量达到最大值,这表明巨噬细胞在高热状态下支持细胞内细菌生长。在PMN中,感染后4小时或12小时时,42℃下的嗜肺军团菌CFU显著低于37℃时(P<0.05),这表明与37℃相比,42℃时PMN的杀菌能力增强。然而,在不同温度下孵育的所有PMN中,感染后24小时CFU均增加了约10倍,这表明PMN以及巨噬细胞在高热状态下均支持细胞内细菌生长。分别在缓冲酵母提取物(BYE)肉汤或含有50%豚鼠血清的RPMI 1640培养基中,于33、37、40和42℃下检测细胞外细菌的生长情况,其中BYE肉汤或RPMI 1640培养基分别作为细菌生长的允许或非允许液体培养基。观察到42℃时BYE肉汤中细菌生长受到抑制,以及42℃时含有50%豚鼠血清的RPMI 1640培养基中CFU减少。总之,高热可能通过限制细胞外细菌存活而有益,但对限制吞噬细胞内细菌生长没有有益作用,尽管PMN在42℃时初始杀伤能力增强。这些结果表明,发热或高热本身在军团菌病病程早期作为一种非特异性宿主防御可能作用不大。

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