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首页> 外文期刊>Infection and immunity >Molecular Characterization and Human T-Cell Responses to a Member of a Novel Mycobacterium tuberculosis mtb39Gene Family
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Molecular Characterization and Human T-Cell Responses to a Member of a Novel Mycobacterium tuberculosis mtb39Gene Family

机译：新型结核分枝杆菌mtb39基因家族成员的分子表征和人类T细胞反应。

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We have used expression screening of a genomic Mycobacterium tuberculosis library with tuberculosis (TB) patient sera to identify novel genes that may be used diagnostically or in the development of a TB vaccine. Using this strategy, we have cloned a novel gene, termed mtb39a, that encodes a 39-kDa protein. Molecular characterization revealed that mtb39a is a member of a family of three highly related genes that are conserved among strains of M. tuberculosis and Mycobacterium bovis BCG but not in other mycobacterial species tested. Immunoblot analysis demonstrated the presence of Mtb39A in M. tuberculosis lysate but not in culture filtrate proteins (CFP), indicating that it is not a secreted antigen. This conclusion is strengthened by the observation that a human T-cell clone specific for purified recombinant Mtb39A protein recognized autologous dendritic cells infected with TB or pulsed with purified protein derivative (PPD) but did not respond to M. tuberculosis CFP. Purified recombinant Mtb39A elicited strong T-cell proliferative and gamma interferon responses in peripheral blood mononuclear cells from 9 of 12 PPD-positive individuals tested, and overlapping peptides were used to identify a minimum of 10 distinct T-cell epitopes. Additionally, mice immunized with mtb39a DNA have shown increased protection from M. tuberculosis challenge, as indicated by a reduction of bacterial load. The human T-cell responses and initial animal studies provide support for further evaluation of this antigen as a possible component of a subunit vaccine for M. tuberculosis.
机译：我们已经使用结核病（TB）患者血清对基因组结核分枝杆菌（emembacterium tuberculosis）库进行表达筛选，以鉴定可用于诊断或开发TB疫苗的新基因。使用这种策略，我们克隆了一个新基因，称为 mtb39a ，该基因编码39 kDa的蛋白质。分子鉴定表明， mtb39a 是三个高度相关基因家族的成员，这些基因在 M菌株中保守。结核分枝杆菌和牛分枝杆菌BCG，但其他分枝杆菌中没有。免疫印迹分析表明Memb39A在 M中存在。结核病的溶解产物，而不是培养滤液蛋白（CFP），表明它不是一种分泌的抗原。通过观察发现，对纯化的重组Mtb39A蛋白具有特异性的人T细胞克隆可识别感染了TB或被纯化的蛋白衍生物（PPD）脉冲的自体树突状细胞，但对 M无反应，从而进一步证实了这一结论。结核病 CFP。纯化的重组Mtb39A在测试的12位PPD阳性个体中的9位的外周血单个核细胞中引起强烈的T细胞增殖和γ干扰素反应，并且使用重叠的肽来鉴定至少10个不同的T细胞表位。此外，用 mtb39a DNA免疫的小鼠显示出对 M的增强保护。细菌载量减少表明结核菌感染。人类T细胞反应和初步的动物研究为进一步评估这种抗原作为em M亚单位疫苗的可能成分提供了支持。结核病。

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