RESUMO
Nestes estudos foram investigados se o Mycobacterium leprae (M. leprae) e o homem compartilham determinantes antigenicos que podem estar localizados nas Proteinas de Choque Termico (HSPs) e que podem ser responsaveis pela destruicao tecidual. Usando-se tecnica de coloracao unica pela imunoperoxidase em cortes feitos com criostato, observaram-se tres anticorpos que eram dirigidos contra HSP-60 (anticorpos policlonais SPA-804, SP-805 e o anticorpo monoclonal SPA-807, que provavelmente reagiram especificamente com macrofagos e celular epitelioides em biopsia cutaneas de pacientes com hanseniase. No Western Blot foi observado que todos os anticorpos contra HSP-60 humana a anticorpos monoclonais (MoAbs) contra HSP-65 do M. leprae (F47-10, F67-18, F88-1) reagiram intensamente com as proteinas do M. leprae sonicado com peso molecular de 65 kDa, indicando semelhanca de alguns determinantes antigenicos entre HSP-60 humana e HSP-65 do M. leprae. Subsequentemente, um estudo imunohistoquimico comparativo dos padroes de coloracao de anticorpos contra HSP-60 humana e anticorpos contra HSP-65 do M. leprae, usando cortes cutaneos feito em criostato de hanseniase paucibacilar (PB), multibacilar (MB) e outras doencas granulomatosas, revelaram que os MoAbs F47-10 e F67-18 reagiram somente fracamente com os granulomas em hanseniase PB e em outras doencas granulomatosas cutaneas, mas coravam o granuloma da hanseniase MB intensamente. O MoAb F88-1 e os anticorpos policlonais SPA-804, SPA-805 e o MoAb SPA 807 coraram os granulomas dos pacientes PB e de outros doencas cutaneas granulomatosas com a mesma intensidade daquela nos pacientes MB. Utilizando-se uma tecnica de dupla coloracao, observou-se que os determinantes antigenicos reconhecidos pelo MoAb contra HSP-60 humana (SPA-807) e os MoBbs contra a HSP-65 do M. leprae (F67-18, F47-10, F88-1) estavam, na maioria das vezes, localizados nos macrofagos. Esses achados nao contradizem nossa hipotese de que semelhancas entre determinantes antigenicos nas HSPs no M. leprae e no hospedeiro humano podem ser, no minimo em parte, responsaveis pela inducao de uma reacao autoimune na hanseniase causando formacao de granuloma com subsequente dano tecidual. Os resultados deste estudo tambem indicaram que alguns destes determinantes estao provavelmente localizados na HSP-60. Uma explicacao similar possivelmente se aplique aos achados em...
Assuntos
Imuno-Histoquímica , Mycobacterium leprae , Hanseníase/fisiopatologia , Hanseníase/imunologia , Hanseníase/microbiologia , Imunoeletroforese , Resposta ao Choque Térmico/imunologiaRESUMO
In these studies, it was investigated whether M. leprae and man share antigenic determinants which may be located on Heat Shock Proteins (HSPs), and which may be responsible for tissue destruction. Using immunoperoxidase single-staining technique on cryostat sections it was observed that three antibodies which are directed against HSP 60 (polyclonal antibodies SPA 804 and SPA 805 and monoclonal antibody SPA 807) probably reacted specifically with macrophages and epitheloid cells in leprosy skin sections. On Western Blotting, it was observed that the antibodies againts human HSP 60 nomoclonal antibodies (MoAbs) against M. leprae HSP 65 (F 47-10, F 67-18, F 88-1) all reacted strongly with sonicated M. leprae proteins with a molecular mass of 65 kDa undicating similarity of some antigenic determinants between human HSP 60 and M. leprae HSP 65. Subsequently, a comparative immunohistochemical study of the staining patterns of antibodies against human HSP 60 and antibodies against M. leprae HSP 65 using cryostat skin section of paucibacillary (PB) leprosy multibacillary (MB) leprosy and other granulomatous skin disorders revealed that the MoAbs F 47-10 and F 67-18 reacted only weakly with the granulomas in PB leprosy and in other granulomatous skin diseases, but stained MB leprosy granuloma strongly. The MoAb F 88-1 and the polyclonal antibodies SPA 804, SPA 805 and the MoAb SPA 807 stained granulomas of PB patients and of other granulomatous skin disorders with the same intensity as that MB patients. Using a double-staining technique, it was observed that the antigenic determinants recognized by the MoAb against human HSP 60 (SPA 807) and the MoAbs against M. leprae HSP 65 (F 67-18, F 47-10, F 88-1) were mostly located in the macrophages. These findings do not contradict our suggestion, Heath Shock Proteins of M. leprae and the human host may be at least in part responsible for the induction of an autoimmune reaction causing granuloma formation with subsequent tissue damage in leprosy. The results of this study also indicated that some of these determinats are probably located on HSP 60. A similar explanation possibly applies to the findings in the other granulomatous disease e.g. sarcoidosis probably micobacterial induced and necrobiosis lipoidica related to diabetis, in which antigenic similarities between bacterial HSP 65 and human HSP 60 are considered to play a part.
Assuntos
Imuno-Histoquímica , Mycobacterium leprae , Hanseníase/fisiopatologia , Hanseníase/imunologia , Hanseníase/microbiologia , Imunoeletroforese , Resposta ao Choque Térmico/imunologiaAssuntos
Antígenos de Bactérias/biossíntese , Antígenos de Bactérias/imunologia , Análise de Sequência de DNA , Biblioteca Gênica , Dados de Sequência Molecular , Escherichia coli/genética , Genes Bacterianos/genética , Hanseníase/sangue , Imunoeletroforese Bidimensional , Mycobacterium leprae/genética , Mycobacterium leprae/imunologia , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/imunologia , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/imunologia , Sequência Conservada , Sequência de Aminoácidos , Sequência de Bases , Tuberculose Pulmonar/sangueAssuntos
Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/genética , Antígenos de Bactérias/química , Clonagem Molecular , DNA Bacteriano/genética , Dados de Sequência Molecular , Fertilização in vitro , Genes Bacterianos , Hanseníase Dimorfa/imunologia , Hanseníase Virchowiana/imunologia , Linfócitos T/imunologia , Mycobacterium leprae/genética , Mycobacterium leprae/imunologia , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/imunologia , Peso Molecular , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/química , Sequência de Aminoácidos , Sequência de BasesRESUMO
A panel of 17 mouse monoclonal antibodies (MoAb) raised against Mycobacterium leprae (M. leprae) antigens was used to detect antigenic determinants in normal human skin. An indirect immunoperoxidase technique was used. Eight of the MoAb detected epidermal antigens similar to patterns well known for human sera. Five of these MoAb detected determinants in the dermis, too. These observations may indicate a certain degree of similarity between the antigenic determinants occurring in M. leprae and in the human host. We propose that such a similarity on the one hand may facilitate the survical of M. leprae in the human host when the antigens are not recognized as "non-self", a situation which seems to ocuur in lepromatous leprosy, when the patients' tissues are loaded with bacteria virtually without any immune response. On the other hand, M. leprae antigens which mimic host antigens may induce an auto-immune reaction against the host's own antigens, which could explain the immune reaction in tuberculod leprosy and during a "reversal reaction" when M. leprae is not observed in the host tissues, but extensive granuloma formation occurs