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[检测不同基因型簇的结核分枝杆菌在巨噬细胞中的体外生长情况]

[Examining the ex vivo growth in the macrophages of Mycobacterium tuberculosis of various genotypic clusters].

作者信息

Andreevskaia S N, Chernousova L N, Smirnova T G, Larionova E E, Kuz'min A V

出版信息

Probl Tuberk Bolezn Legk. 2006(12):43-8.

Abstract

To study the specific features of replication of Mycobacterium tuberculosis (MBT) strains of the W cluster in the macrophages (MP) on the ex vivo model of peritoneal MP of MBT-infected C57B1/6 mice, the authors estimated the viability of 6 antituberculous drug-sensitive MBT strains of the W-cluster from the incorporation of 5,6-[3H]-uracil into the mycobacterial cells and their induced specific MP from the level of LDH. Eight sensitive MBT strains of other genotypes clustered by the restriction fragment length polymorphism (PDRF) IS6110 and 2 laboratory strains M. tuberculosis H37Rv and M. tuberculosis H37Ra were taken as a control. The study indicated that, cultured in vitro, MBT strains belonging to different genotypic clusters differed in the level of 5,6-[3H]-uracil inclusion. When grown in MP, the MBT population of all genotypes showed a diminished viability as compared with that cultured without MP. The MBT clusters of W and AI clusters, unclustered strains, and M. tuberculosis H37Rv displayed a higher inclusion of 5,6-[3H]-uracil and the strains of KQ and HD clusters and M. tuberculosis H37Ra exhibited a significantly lower inclusion of 5,6-[3H]-uracil. W-cluster strains, the unclustered strain R807, and M. tuberculosis H37Rv showed the highest fitness (adaptability when grown in MP). The virulent strain M. tuberculosis H37Rv and avirulent strain M. tuberculosis H37Ra differed in MP viability by almost 5 times. Evaluation of the cytopathogenic effect indicated that the clinical MBT strains led to a specific MP lysis greater than 40%, the highest effect was produced by the MBT strains of the W cluster (more than 93%) and the HD cluster (96.45%). The control laboratory strains M. tuberculosis H37Rv and M. tuberculosis H37Ra contrasted sharply in their induced specific MP lysis (93.35 and 5.93%, respectively).

摘要

为了在MBT感染的C57B1/6小鼠腹膜巨噬细胞(MP)的体外模型上研究W簇结核分枝杆菌(MBT)菌株在巨噬细胞(MP)中的复制特性,作者通过5,6-[3H]-尿嘧啶掺入分枝杆菌细胞的情况来评估6株对6种抗结核药物敏感的W簇MBT菌株的活力,并从乳酸脱氢酶(LDH)水平评估其诱导的特异性MP。选取8株通过限制性片段长度多态性(PDRF)IS6110聚类的其他基因型敏感MBT菌株以及2株实验室菌株结核分枝杆菌H37Rv和结核分枝杆菌H37Ra作为对照。研究表明,在体外培养时,属于不同基因型簇的MBT菌株在5,6-[3H]-尿嘧啶掺入水平上存在差异。在MP中生长时,所有基因型的MBT群体与无MP培养时相比活力降低。W簇和AI簇的MBT簇、未聚类菌株以及结核分枝杆菌H37Rv显示出较高的5,6-[3H]-尿嘧啶掺入量,而KQ簇和HD簇的菌株以及结核分枝杆菌H37Ra表现出显著较低的5,6-[3H]-尿嘧啶掺入量。W簇菌株、未聚类菌株R807和结核分枝杆菌H37Rv显示出最高的适应性(在MP中生长时的适应性)。强毒株结核分枝杆菌H37Rv和无毒株结核分枝杆菌H37Ra在MP活力上相差近5倍。细胞致病效应评估表明,临床MBT菌株导致特异性MP裂解率大于40%,W簇MBT菌株产生的效应最高(超过93%),HD簇(96.45%)。对照实验室菌株结核分枝杆菌H37Rv和结核分枝杆菌H37Ra在诱导的特异性MP裂解方面形成鲜明对比(分别为93.35%和5.93%)。

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