Identification of the C-terminal region of 70 kDa heat shock protein from Leishmania (Viannia) braziliensis as a target for the humoral immune response

Identification of the C-terminal region of 70 kDa heat shock protein from Leishmania (Viannia) braziliensis as a target for the humoral immune response

Autor Amorim, A. G. Google Scholar
Carrington, M. Google Scholar
Miles, M. A. Google Scholar
Barker, D. C. Google Scholar
deAlmeida, MLC Google Scholar
Instituição UNIV CAMBRIDGE
UNIV LONDON LONDON SCH HYG & TROP MED
Universidade Federal de São Paulo (UNIFESP)
Resumo A Leishmania (Viannia) braziliensis (Lb) promastigote cDNA library in lambda g11 was screened with patients' sera with the aim of identifying antigens specifically related to mucocutaneous leishmaniasis (MCL). One of the clones isolated, 133P, consistently reacted with MCL sera; it was sequenced and found to encode the C-terminal three-quarters of a protein belonging to the highly conserved Hsp70 family. Since Hsp70 proteins from different species tend to be less conserved through the C-terminal end, it was predicted that this region would be more antigenic and was likely to bear the discriminatory epitopes. in order to test this hypothesis, the N- and C-terminal halves of the polypeptide encoded by 133P, 133P-N and 133P-C, respectively, were expressed in Esherichia coli Immunoblotting analysis demonstrated that 133P-C reacted more strongly with a pool of MCL sera than 133P-N, and both recombinant proteins reacted faintly with pools of cutaneous (CL) and visceral (VL) leishmaniasis sera. These results confirmed the predicted epitope location in the C-terminal region. the 133P-C fragment was also expressed as a fusion protein with glutathione-S-transferase (GST-133P-C), affinity-purified with glutathione-agarose and tested by ELISA with individual sera. From 46 Lb-infected patients, 41 sera(89%) were positive, no cross-reactivity was observed with healthy, Trypanosoma cruzior L. amazonensis-infected individuals. Despite a relatively high cross-reactivity with VL sera, the enhanced humoral response of MCL as compared with CL patients might be interesting for studies on disease aggravation.
Idioma Inglês
Data de publicação 1996-09-01
Publicado em Cell Stress & Chaperones. Edinburgh: Churchill Livingstone, v. 1, n. 3, p. 177-187, 1996.
ISSN 1355-8145 (Sherpa/Romeo, fator de impacto)
Publicador Churchill Livingstone
Extensão 177-187
Fonte http://www.ncbi.nlm.nih.gov/pmc/articles/PMC248477/
Direito de acesso Acesso aberto Open Access
Tipo Artigo
Web of Science WOS:A1996VY14300004
Endereço permanente http://repositorio.unifesp.br/handle/11600/25620

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