Gillis T P, Job C K
Int J Lepr Other Mycobact Dis. 1987 Mar;55(1):54-62.
The cell wall-associated protein of Mycobacterium gordonae (Mr = 65,000) was purified by affinity chromatography using a murine monoclonal antibody produced in response to the crossreactive 65 kD protein of M. leprae. The affinity-purified material was analyzed for purity by protein and carbohydrate analyses, SDS-PAGE, and immunoblotting. The final preparation contained a major protein band on SDS-PAGE analysis (Mr = 65,000) with no detectable carbohydrates. The affinity fraction was prepared at 250 micrograms/ml (protein) in sterile saline and 0.1 ml injected intradermally into guinea pigs immunized 30 days earlier. Gross changes at 48 hr were consistent with the characteristics of a delayed hypersensitivity skin reaction measuring 2.5 mm, 3.4 mm, and 2.7 mm in animals which had been immunized with M. leprae, M. gordonae, or M. bovis (BCG), respectively. Histologically, all 65 kD protein skin-test sites showed marked edema and infiltration by numerous lymphocytes, macrophages, and scattered neutrophils. Animals injected with Freund's incomplete adjuvant showed a minimal or no reaction (1.8 mm) to the purified protein. These results further define the immunogenicity of the 65 kD protein of M. gordonae and by inference M. leprae, and demonstrate the ability of crossreactive epitopes of the 65 kD protein to sensitize lymphocytes involved in delayed-type hypersensitivity reaction to M. leprae, M. gordonae, and M. bovis (BCG).
戈登分枝杆菌的细胞壁相关蛋白（分子量=65,000），通过亲和层析法进行纯化，所用的鼠单克隆抗体是针对麻风分枝杆菌的交叉反应性65 kD蛋白产生的。通过蛋白质和碳水化合物分析、SDS-PAGE及免疫印迹法对亲和纯化的物质进行纯度分析。SDS-PAGE分析显示，最终制品含有一条主要蛋白带（分子量=65,000），未检测到碳水化合物。亲和级分以250微克/毫升（蛋白质）的浓度用无菌生理盐水配制，将0.1毫升皮内注射到30天前免疫的豚鼠体内。48小时时的大体变化符合迟发型超敏皮肤反应的特征，在用麻风分枝杆菌、戈登分枝杆菌或牛分枝杆菌（卡介苗）免疫的动物中，反应分别为2.5毫米、3.4毫米和2.7毫米。组织学检查显示，所有65 kD蛋白皮肤试验部位均出现明显水肿，并有大量淋巴细胞、巨噬细胞和散在的中性粒细胞浸润。注射弗氏不完全佐剂的动物对纯化蛋白的反应极小或无反应（1.8毫米）。这些结果进一步明确了戈登分枝杆菌65 kD蛋白的免疫原性，由此推断也明确了麻风分枝杆菌65 kD蛋白的免疫原性，并证明了65 kD蛋白的交叉反应表位能够使参与对麻风分枝杆菌、戈登分枝杆菌和牛分枝杆菌（卡介苗）迟发型超敏反应的淋巴细胞致敏。
