TRYPANOSOMA-CRUZI - CELLULAR AND ANTIBODY-RESPONSE AGAINST THE PARASITE IN MICE IMMUNIZED WITH A 19-AMINO ACID SYNTHETIC PEPTIDE

被引:9
作者
BUA, J
BONTEMPI, EJ
LEVIN, M
ORN, A
VELASCO, D
MORENO, M
LEVIYEYATI, P
ENGSTROM, A
SEGURA, EL
RUIZ, AM
机构
[1] INST NACL DIAGNOST & INVEST ENFERMEDAD CHAGAS DR M, PASEO COLON 568, RA-1063 BUENOS AIRES, ARGENTINA
[2] KAROLINSKA INST, DEPT IMMUNOL, S-10401 STOCKHOLM 60, SWEDEN
[3] BIOMED CTR, DEPT IMMUNOL, S-75123 UPPSALA, SWEDEN
[4] INST INVEST INGN GENET & BIOL MOLEC, BUENOS AIRES, ARGENTINA
关键词
TRYPANOSOMA-CRUZI; EPIMASTIGOTES FLAGELLAR FRACTION; MONOCLONAL ANTIBODY; SYNTHETIC PEPTIDE; IMMUNE RESPONSE; BETA-GALACTOSIDASE (BETA-GAL); BORDETELLA-PERTUSSIS (BP); BOVINE SERUM ALBUMIN (BSA); DELAYED-TYPE HYPERSENSITIVITY (DTH); IMMUNOFLUORESCENCE ASSAY (IFA); ISOPROPYL-D-THIOGALACTOPYRANOSIDE (IPTG); INTRAPERITONEAL (IP); KEYHOLE LIMPET HEMOCYANINE (KLH); MONOCLONAL ANTIBODY (MOAB); MOLECULAR WEIGHT (MW); PHOSPHATE-BUFFERED SALINE (PBS); NORMAL MOUSE SERUM (NMS); PHENYLMETHYLSULFONYL FLUORIDE (PMSF); ROOM TEMPERATURE (RT); SODIUM DODECYL SULFATE POLYACRYLAMIDE GEL ELECTROPHORESIS (SDS-PAGE); TRIS-BUFFERED SALINE (TBS); 5-BROMO-4-CHLORO-3-INDOLYL-BETA-D-GALACTOSIDE (X-GAL); WHOLE HOMOGENATE OF TRYPANOSOMA-CRUZI EPIMASTIGOTES (WH);
D O I
10.1016/0014-4894(91)90120-L
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
Several monoclonal antibodies were prepared against the flagellar fraction of Trypanosoma cruzi epimastigotes (Tulahuén strain, stock Tul 2). One of them, FCH-F8-4, has previously shown biologic activity against the parasite (complement-mediated lysis and neutralization of the trypomastigote infectivity). Immunopurified antigens using this monoclonal antibody elicited a protective immune response in mice. Two recombinant cDNA clones were detected with this anti-flagellar fraction monoclonal antibody on a λgt11 expression library prepared from T. cruzi epimastigote mRNA. The insert of one of these cDNA clones, λ(FCH-F8-4)1 (150 bp) coded for a 19-amino acid peptide (PAFLGCSSRFSGSFSGVEP). This insert hybridized with a 5.0-kb mRNA from epimastigotes. The β-galactosidase fusion protein was produced in lysogenic bacteria. The monoclonal antibody recognized the epitope present in the fusion protein after western blotting of the crude lysate. A synthetic peptide (SP4) containing the complete sequence of λ(FCH-F8-4)1 was constructed on solid phase. This peptide was able to inhibit the ELISA reactivity (in a range from 13 to 52%) of flagellar fraction immunized mouse sera and when administered (coupled to KLH or alone) to BALB/c mice with Bordetella pertussis as adjuvant, it induced a humoral and cellular immune response which was detected by ELISA, immunofluorescence, blotting, and DTK reactions against T. cruzi antigens. The immune response obtained indicates that this synthetic peptide resembles the parasite antigen conformation and could be useful for diagnosis purposes or be able to elicit immunoprotection against T. cruzi infection. © 1991.
引用
收藏
页码:54 / 62
页数:9
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