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Exp Parasitol ; 124(2): 153-8, 2010 Feb.
Article in English | MEDLINE | ID: mdl-19735658

ABSTRACT

A 500 bp fragment encoding an isoform of cysteine proteinase from Leishmania (Leishmania) amazonensis was subcloned and expressed in the pHis vector, resulting in a recombinant protein of 24 kDa, rLacys24. In Western blots of L. (L.) amazonensis extracts, antibodies directed to rLacys24 recognized a cysteine proteinase isoform of 30 kDa. Analysis by fluorescence-activated cell sorter showed a significantly higher expression of CD8(+) lymphocytes in animals immunized with rLacys24 plus CFA, whereas a low expression of CD4(+) lymphocytes was observed in these animals. The cytotoxicity of lymphocytes isolated from mice immunized with rLacys24 plus CFA on L. (L.) amazonensis-infected macrophages was significantly higher than that observed in the presence of lymphocytes from control animals. Immunization of BALB/c mice with rLacys24 plus CFA resulted in a low but significant decrease of foot lesions after challenge with L. (L.) amazonensis compared to those exhibited by control mice.


Subject(s)
Cysteine Proteases/immunology , Leishmania mexicana/enzymology , Leishmania mexicana/immunology , Leishmaniasis, Cutaneous/immunology , Leishmaniasis, Cutaneous/prevention & control , Animals , Blotting, Western , Cricetinae , Cysteine Proteases/genetics , Cysteine Proteases/isolation & purification , DNA, Protozoan/chemistry , Disease Models, Animal , Electrophoresis, Polyacrylamide Gel , Female , Gene Expression Regulation, Enzymologic , Leishmania mexicana/genetics , Lymph Nodes/cytology , Lymph Nodes/immunology , Mesocricetus , Mice , Mice, Inbred BALB C , Polymerase Chain Reaction , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/isolation & purification , T-Lymphocytes/immunology
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