Expression in fibroblasts and in live animals of Entamoeba histolytica polypeptides EhCP112 and EhADH112.
EhCPADH is an immunogenic, heterodimeric protein that is formed by EhCP112 (cysteine protease) and EhADH112 (adhesin), polypeptides involved in Entamoeba histolytica's cytopathic effect, target-cell adherence and phagocytosis. The EhCPADH complex is located in the plasma membrane and cytoplasmic vacuoles. Here, the independent expression of EhCP112 and EhADH112 in fibroblasts and hamsters was analysed. Also investigated was the immunological response in animals independently inoculated with plasmid pcDNA-Ehcp112, which carries the complete cysteine protease-encoding gene, or with plasmid pcDNA-Ehadh112, which carries the C terminus of the adhesin-encoding gene, or with a mixture of both. Both proteins were expressed in the plasma membranes of the transfected fibroblasts. EhCP112 was toxic for the mammalian cells. Proteins were also independently expressed in hamsters after inoculation with the plasmids. Their expression was indirectly evaluated by the presence of antibodies in the inoculated animals. Remarkably, co-immunization of the animals with the two DNA plasmids resulted in an earlier and higher anti-E. histolytica IgG induction than immunization with separate plasmids. In contrast, the cellular immune response was not noticeably improved by the plasmid mixture. Interestingly, protection against liver abscesses was detected only in animals that received the plasmid mixture and no protection was observed in hamsters independently inoculated with plasmid pcDNA-Ehcp112 or pcDNA-Ehadh112.
Madriz
Xochil
X
División de Biología Molecular, Instituto Finlay, AP 16017 Ciudad de la Habana, C.P. 11600, Cuba.|Escuela Nacional de Medicina y Homeopatía, IPN, Guillermo Massieu Helguera #239, DF 07320, Mexico.
Martínez
Máximo B
MB
División de Biología Molecular, Instituto Finlay, AP 16017 Ciudad de la Habana, C.P. 11600, Cuba.
Rodríguez
Mario A
MA
Departamento de Patología Experimental, CINVESTAV, IPN, AP 14-740, DF 07000, Mexico.
Sierra
Gustavo
G
División de Biología Molecular, Instituto Finlay, AP 16017 Ciudad de la Habana, C.P. 11600, Cuba.
Martínez-López
Carolina
C
Escuela Nacional de Medicina y Homeopatía, IPN, Guillermo Massieu Helguera #239, DF 07320, Mexico.
Riverón
Ana M
AM
Departamento de Biología Molecular, CENIC, AP 6690 Ciudad de La Habana, Cuba.
Flores
Leopoldo
L
Departamento de Patología Experimental, CINVESTAV, IPN, AP 14-740, DF 07000, Mexico.
Orozco
Esther
E
Departamento de Patología Experimental, CINVESTAV, IPN, AP 14-740, DF 07000, Mexico.
D018829:Adhesins, Bacterial / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D004748:Entamoeba histolytica / Q000254:growth & development / Q000276:immunology* / Q000378:metabolism / Q000472:pathogenicity*; D004749:Entamoebiasis / Q000469:parasitology / Q000503:physiopathology / Q000517:prevention & control*; D004912:Erythrocytes / Q000276:immunology / Q000469:parasitology; D005347:Fibroblasts / Q000378:metabolism*; D007106:Immune Sera / Q000276:immunology; D007114:Immunization; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D010957:Plasmids; D016052:Protozoan Vaccines / Q000008:administration & dosage / Q000276:immunology; D014162:Transfection; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000276:immunology; D014774:Virulence
D018829:Adhesins, Bacterial; D000913:Antibodies, Protozoan; D001426:Bacterial Proteins; C122005:EhADH112 protein, Entamoeba histolytica; D007106:Immune Sera; D016052:Protozoan Vaccines; D019444:Vaccines, DNA; D003546:Cysteine Endopeptidases; C122004:EhCP112 protein, Entamoeba histolytica