Home » Volumes » Volume 24 January/March 1991 » Immunocytochemical identification of leishmania and Trypanosoma cruzi amastigotes in situ with homologous and heterologous polyclonal antibodies

Immunocytochemical identification of leishmania and Trypanosoma cruzi amastigotes in situ with homologous and heterologous polyclonal antibodies

A.J.A. Barbosa; C.A. da Costa; M.S.M. Michalick; W. Mayrink; R.T. Gazzinelli; W.L. Tafuri.

DOI: 10.1590/S0037-86821991000100002


The unlabelled antibody peroxidase-antiperoxidase method was used to study the immunocytochemical properties of Leishmania and Trypanosoma cruzi amastigotes in situ after tissues had been submitted to different fixation procedures. Antisera were obtained from rabbits chronically infected with different strains of T. cruzi or immunized with L. mexicana amazonensis and L. braziliensis guyanensis, and were applied on 5 µm thick sections. T. cruziantigens were well stained by the three anti-T. cruzi sera and the two anti-heis.hmama.sera at optimum dilution between 1:1,000 and 1:2,000, regardless the parasite strain. Differently, the leishmanial antigens were revealed by Leishmania sera only at low dilutions (between 1:60 -1:160), whereas the anti-T. cruzi sera, at these low dilutions, gave rather weak stainings. Although there is no clear explanation for this immunocytochemical “reverse-monodirectional” cross-reactivity between Leishmania and T. cruzi, the present results show that polyclonal antibodies agains Leishmania species, when used for immunocytochemical detection of these parasites in situ, react more strongly with T. cruzi amastigotes than with the homologous amastigotes.

Keywords: LeishmaniaTrypanosoma cruzi. Antigenicity of amastigotes. Peroxidase- antiperoxidase method.


O método daperoxidase-antiperoxidase foi utilizado para estudar as propriedades imunocitoquimicas de Leishmanias e de amastigotas do Trypanosoma cruziin situ, após os tecidos terem sido submetidos a diferentes tipos de fixação. Anti-soros foram obtidos de coelhos cronicamente infectados com três cepas de T. cruzi ou imunizados com L. mexicana ámazonensis e L. braziliensis guyanensis e aplicados nos cortes histológicos de 5 µm de espessura. Os antígenos de T. cruzi foram corados muito bem pelos três soros anti-T. cruzi e pelos dois soros anti-Leishmania com diluições entre 1:1.000 e 1:2.000. Diferentemente, os antígenos dç Leishmania foram revelados pelos soros anti- Leishmania somente em baixas diluições, ou seja, entre 1:60 e 1:160 enquanto que os soros anti-T. cruzi, mesmo nestas diluições baixas, proporcionaram colorações fracas e irregulares auando usados para revelar Leishmania. Embora não haja explicação clara para esta reação imunocitoquimica cruzada “reversa-monodirecional” entre Leishmania e amastigotas de T. cruzi os resultados do presente trabalho mostram que anticorpos policlonais contra diferentes espécies de Leishmania, quando usados para detecção imunocitoquímica de Leishmania e T. cruzi in situ, reagem mais fortemente com amastigotas de T. cruzi do que com espécies homólogas.

Palavras-chave: Leishmania. Trypanosoma cruzi. Antigenicidade de amastigotas. Método peroxidase-antiperoxidase.



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 Adress to correspondence:
Dr. Alfredo J. A. Barbosa
Facuidade de Medicina da UFMG
Av. Alfredo Balena 190
Belo Horizonte, MG, Brasil.



Recebido para publicação em 28/08/90.



Trabalho realizado nos Departamentos de Anatomia Pato-lógica e Medicina Legal e de Parasitologia da Universidade Federal de Minas Gerais e no Centro de Pesquisas René Rachou, Belo Horizonte, MG, Brasil.
Auxílio financeiro da FINEP, CNPq e FAPEMIG.