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首页> 外文期刊>Infection and immunity >Whole-Genome DNA Array Analysis of the Response of Borrelia burgdorferi to a Bactericidal Monoclonal Antibody
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Whole-Genome DNA Array Analysis of the Response of Borrelia burgdorferi to a Bactericidal Monoclonal Antibody

机译：伯氏疏螺旋体对杀菌单克隆抗体应答的全基因组DNA阵列分析

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Identification and characterization of genes that contribute to infection with Borrelia burgdorferi and, of those, genes that are targets of host responses is important for understanding the pathogenesis of Lyme disease. The complement-independent bactericidal monoclonal antibody (MAb) CB2 recognizes a carboxy-terminal, hydrophilic epitope of the outer surface protein B (OspB). CB2 kills B. burgdorferi by an unknown bactericidal mechanism. Upon binding of CB2 to OspB, differentially expressed gene products may be responsible for, or associated with, the death of the organism. A time course of the response of B. burgdorferi to CB2 was completed to analyze the differential gene expression in the bacteria over a period of visual morphological changes. Bacteria were treated with a sublethal concentration in which spirochetes were visibly distressed by the antibody but not lysed. Preliminary whole-genome DNA arrays at various time points within 1 h of incubation of B. burgdorferi with the antibody showed that most significant changes occurred at 25 min. Circular plasmid 32 (cp32)-encoded genes were active in this period of time, including the blyA homologs, phage holin system genes. DNA array data show that three blyA homologs were upregulated significantly, ≥2 standard deviations from the mean of the log ratios, and a P value of ≤0.01. Quantitative real-time PCR analysis verified blyA and blyB upregulation over an 18- to 35-min time course. The hypothesis to test is whether the killing mechanism of CB2 is through uncontrolled expression of the blyA and blyB phage holin system.
机译：鉴定和鉴定导致伯氏疏螺旋体感染的基因，其中，作为宿主反应靶标的基因对于理解莱姆病的发病机理很重要。不依赖补体的杀菌单克隆抗体（MAb）CB2识别外表面蛋白B（OspB）的羧基末端亲水表位。 CB2杀死 B。 burgdorferi 具有未知的杀菌机制。 CB2与OspB结合后，差异表达的基因产物可能导致生物死亡或与生物死亡有关。 B响应的时间过程。 burgdorferi 到CB2的构建完成了分析在视觉形态变化期间细菌中的差异基因表达。用亚致死浓度处理细菌，其中螺旋体明显地被抗体困扰，但没有被溶解。 B孵育1小时内各个时间点的初步全基因组DNA阵列。带有该抗体的burgdorferi 表明，最显着的变化发生在25分钟时。环状质粒32（cp32）编码的基因在这段时间内很活跃，包括 blyA 同源物，噬菌体holin系统基因。 DNA阵列数据显示，三个 blyA 同源物显着上调，与对数比的平均值相差≥2个标准差，并且 P 值≤0.01。实时定量PCR分析证实了在18至35分钟的时间过程中 blyA 和 blyB 的上调。要检验的假设是CB2的杀伤机制是否是通过 blyA 和 blyB 噬菌体霍林系统的不受控制的表达来实现的。

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