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首页> 外文期刊>Journal of Clinical Microbiology >Immunologic characterization of a 35-kilodalton recombinant antigen of Mycobacterium tuberculosis.
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Immunologic characterization of a 35-kilodalton recombinant antigen of Mycobacterium tuberculosis.

机译：结核分枝杆菌的35千克多尔顿重组抗原的免疫学表征。

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摘要

A 35-kilodalton (kDa) recombinant antigen (35-kDa antigen) produced by Escherichia coli JM107 carrying DNA from Mycobacterium tuberculosis was purified and immunologically examined by in vivo and in vitro methods. A monoclonal antibody (2B2) was produced against the 35-kDa antigen. The protein was purified from the insoluble fraction of the recombinant E. coli strain by either affinity chromatography with the 2B2 monoclonal antibody or preparative isoelectric focusing. In enzyme-linked immunosorbent assay and Western blot (immunoblot) analyses, antibody to 2B2 reacted with whole-cell sonic extracts of M. tuberculosis and other slowly growing mycobacteria but not with two rapid growers, M. chelonae and M. fortuitum. An injection series totaling less than 1 mg of purified protein without adjuvant elicited a humoral response in guinea pigs. In one guinea pig, 10 micrograms of purified protein injected intradermally elicited both a humoral and a cell-mediated response. Results of these studies suggest that the 35-kDa antigen is a membrane-associated protein that stimulates both humoral and cell-mediated immune responses and should be evaluated as a vaccine candidate.
机译：纯化由携带来自结核分枝杆菌DNA的大肠杆菌JM107产生的35-千达尔顿（kDa）重组抗原（35-kDa抗原），并通过体内和体外方法进行免疫学检查。产生了针对35 kDa抗原的单克隆抗体（2B2）。通过使用2B2单克隆抗体的亲和色谱或制备性等电聚焦，从重组大肠杆菌菌株的不溶级分中纯化蛋白质。在酶联免疫吸附测定和蛋白质印迹（免疫印迹）分析中，针对2B2的抗体与结核分枝杆菌和其他缓慢生长的分枝杆菌的全细胞声波提取物发生了反应，但与两个快速生长的植物，M。chelonae和Fortuitum没有反应。总共少于1 mg的纯化蛋白而没有佐剂的注射系列在豚鼠中引起体液反应。在一只豚鼠中，皮内注射10毫克的纯化蛋白可引起体液反应和细胞介导的反应。这些研究结果表明，35 kDa抗原是一种膜相关蛋白，可刺激体液和细胞介导的免疫反应，应作为候选疫苗进行评估。

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