Programa de Inmunología Molecular Microbiana, Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México, México City, México.
Clin Exp Immunol. 2014 Aug;177(2):454-63. doi: 10.1111/cei.12309.
Tuberculosis is a disease caused by the Mycobacterium tuberculosis complex (MTb). In 2011, global mortality due to tuberculosis was 1·4 million individuals. The only available vaccine is the attenuated M. bovis [bacillus Calmette-Guérin (BCG)] strain, which confers variable protection against pulmonary tuberculosis. Some widely distributed non-tuberculous mycobacteria (NTM), such as M. avium and M. arupense, are also potential pathogens for humans. This work aimed to produce and characterize monoclonal antibodies against the M. bovis BCG Mexico strain of the MTb, M. avium subs. hominissuis and the M. arupense strain from NTM. Hybridomas were produced from splenocytes of BALB/c female mice immunized with radiation-inactivated mycobacteria, and the immunoglobulin (Ig)G2a antibody-producing clones with the highest antigenic recognition were selected. The selected clones, Mbv 2A10 for M. bovis BCG Mexico, Mav 3H1 for M. avium and Mar 2D10 for M. arupense, were used in further studies. Enzyme-linked immunosorbent assay (ELISA) and immune proteomics analyses characterized the clones as having the highest cross-reactivity with mycobacteria. Using mass spectrometry, a number of proteins recognized by the monoclonal antibody (mAb) clones were identified. These proteins had roles in metabolic processes, hypoxia, cell cycle and dormancy. In addition, a Clustal W and Immune Epitope Database (IEDB) in-silico analysis was performed in protein sequences that result in the conserved regions within probability epitopes that could be recognized for Mbv2A10 and Mav3H1 clones.
结核病是由结核分枝杆菌复合群（MTb）引起的疾病。2011 年，全球因结核病死亡的人数为 140 万人。唯一可用的疫苗是减毒牛分枝杆菌[卡介苗（BCG）]株，它对肺结核的保护作用各不相同。一些广泛分布的非结核分枝杆菌（NTM），如鸟分枝杆菌和马红球菌，也是人类的潜在病原体。本研究旨在生产和鉴定针对结核分枝杆菌 MTb 墨西哥牛分枝杆菌 BCG 株、鸟分枝杆菌亚种人型和 NTM 马红球菌株的单克隆抗体。用辐射灭活的分枝杆菌免疫 BALB/c 雌性小鼠脾细胞，产生杂交瘤，并选择具有最高抗原识别能力的 IgG2a 抗体产生克隆。选择的克隆 Mbv2A10 用于 M. bovis BCG Mexico，Mav3H1 用于 M. avium 和 Mar2D10 用于 M. arupense，用于进一步研究。酶联免疫吸附试验（ELISA）和免疫蛋白质组学分析表明，这些克隆与分枝杆菌具有最高的交叉反应性。使用质谱技术，鉴定出被单克隆抗体（mAb）克隆识别的多种蛋白质。这些蛋白质在代谢过程、缺氧、细胞周期和休眠中发挥作用。此外，还对蛋白质序列进行了 Clustal W 和免疫表位数据库（IEDB）的计算机分析，这些分析产生了 Mbv2A10 和 Mav3H1 克隆可能识别的保守区域内概率表位的序列。
