Mapping of B cell epitopes in an immunodominant antigen of Trypanosoma cruzi using fusions to the Escherichia coli LamB protein.
The JL8 protein antigen from Trypanosoma cruzi, a dominant immunogen in man, has been characterized as containing tandem amino acid repeats. Here, we describe the use of the LamB protein of Escherichia coli as a carrier of JL8 derived sequences in order to map the immunodominant B cell epitopes in this antigen. Five different sequences of JL8 were inserted in the LamB protein and the JL8-LamB fusion proteins were tested by ELISA with human chronic chagasic sera. The fusion carrying the sequence AEKQKAAEATKVAE was recognized by most sera. This protein was also capable of inhibiting the binding of human chagasic antibodies to GST-JL8 in competitive ELISA suggesting that it contains an immunodominant B cell epitope of JL8.
Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, UNIFESP, São Paulo, Brazil.
C M
CM
Pereira
L M
LM
Yamauchi
M J
MJ
Levin
J F
JF
da Silveira
B A
BA
Castilho
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000913:Antibodies, Protozoan / Q000097:blood; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000032:analysis*; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D006801:Humans; D015151:Immunoblotting; D016056:Immunodominant Epitopes / Q000276:immunology / Q000302:isolation & purification; D021382:Protein Sorting Signals; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D014349:Trypanosoma cruzi / Q000276:immunology*
D000907:Antibodies, Bacterial; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D001426:Bacterial Proteins; D000939:Epitopes; D029968:Escherichia coli Proteins; D016056:Immunodominant Epitopes; C085044:LamB signal peptide, E coli; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins