Rambukkana A, Burggraaf J D, Faber W R, Harboe M, Teeling P, Krieg S, Das P K
Department of Dermatology, Academic Medical Center, University of Amsterdam, The Netherlands.
Infect Immun. 1993 May;61(5):1835-45. doi: 10.1128/iai.61.5.1835-1845.1993.
The granulomatous skin lesions in leprosy are thought to be initiated by the immune response to certain antigens of the causative agent, Mycobacterium leprae. The antigen 85 complex is one of the major targets in the immune response to M. leprae infection. In the present study, a panel of previously characterized monoclonal antibodies (MAbs) (3A8, Rb2, A4g4, A2h11, Pe12, and A3c12) reacting with different epitopes of the 85 complex proteins of Mycobacterium tuberculosis and M. leprae was employed in a comparative immunohistological analysis to demonstrate the in situ expression of 85 complex antigenic epitopes in leprosy lesions across the clinical spectrum and in M. leprae-infected armadillo liver tissues. These MAbs showed a heterogeneous staining pattern in a given leprosy lesion. In highly bacilliferous borderline and lepromatous leprosy lesions, MAbs Rb2, A4g4, A2h11, and Pe12 stained clear rod-shaped M. leprae bacilli within macrophages, and the degree of staining correlated with the bacillary index of the lesion. On the other hand, MAbs 3A8 and A3c12 staining was mostly seen as a diffuse staining pattern within interstitial spaces and on the membranes of the infiltrated cells but not the bacilli. In paucibacillary borderline and tuberculoid leprosy lesions, only 3A8, Rb2, and A3c12 showed distinct staining in association with infiltrates in the granuloma. None of these MAbs showed any detectable reaction with control nonleprosy skin lesions, while MAb A3c12 positively stained the granulomas of both leprosy and control specimens. In situ reactivity of these MAbs with M. leprae-infected armadillo liver tissues also showed a heterogeneous staining pattern. Interestingly, a clear difference in expression of these epitopes was observed between armadillo tissues and human leprosy lesions. By immunogold ultracytochemistry, we further showed the differential localization of these MAb-reactive epitopes on the cell surface, in the cytosol, and at the vicinity of M. leprae within Kupffer cells of armadillo liver tissues. Our results indicate that these antigenic epitopes of the antigen 85 complex are differentially expressed in leprosy lesions and infected armadillo tissues and that they could be target determinants in the immunopathological responses during M. leprae infection.
麻风病中的肉芽肿性皮肤病变被认为是由对病原体麻风分枝杆菌某些抗原的免疫反应引发的。抗原85复合体是对麻风分枝杆菌感染免疫反应的主要靶点之一。在本研究中,一组先前已鉴定的单克隆抗体(MAbs)(3A8、Rb2、A4g4、A2h11、Pe12和A3c12),它们与结核分枝杆菌和麻风分枝杆菌的85复合体蛋白的不同表位发生反应,被用于比较免疫组织学分析,以证明85复合体抗原表位在整个临床谱的麻风病病变以及麻风分枝杆菌感染的犰狳肝脏组织中的原位表达。这些单克隆抗体在给定的麻风病病变中呈现出异质性染色模式。在高菌量的界线类和瘤型麻风病病变中,单克隆抗体Rb2、A4g4、A2h11和Pe12对巨噬细胞内清晰的杆状麻风分枝杆菌进行染色,染色程度与病变的菌指数相关。另一方面,单克隆抗体3A8和A3c12的染色大多表现为间质空间内以及浸润细胞的细胞膜上的弥漫性染色模式,而非杆菌染色。在少菌量的界线类和结核样型麻风病病变中,只有3A8、Rb2和A3c12与肉芽肿中的浸润相关呈现出明显染色。这些单克隆抗体与对照非麻风病皮肤病变均未显示出任何可检测到的反应,而单克隆抗体A3c12对麻风病和对照标本的肉芽肿均呈阳性染色。这些单克隆抗体与麻风分枝杆菌感染的犰狳肝脏组织的原位反应性也呈现出异质性染色模式。有趣的是,在犰狳组织和人类麻风病病变之间观察到这些表位表达的明显差异。通过免疫金超微细胞化学,我们进一步显示了这些单克隆抗体反应性表位在犰狳肝脏组织库普弗细胞的细胞表面、胞质溶胶以及麻风分枝杆菌附近的差异定位。我们的结果表明,抗原85复合体的这些抗原表位在麻风病病变和感染的犰狳组织中差异表达,并且它们可能是麻风分枝杆菌感染期间免疫病理反应中的靶标决定因素。