Identification of a 35 kDa glycoprotein from Entamoeba histolytica by sera from patients with amoebic liver abscess and with mouse monoclonal antibody.
In this work, we explored the relevance of a 35 kDa glycoprotein (Gm) of the outer membrane from E. histolytica in the diagnosis of the amoebic liver abscess (ALA) through ELISA and immunoblotting. We were interested in defining the relevance of this antigen in the immune response in patients with amoebic liver abscess and in exploring whether the mouse monoclonal antibody against this 35 kDa glycoprotein recognises the same epitope. We found that 87% of ALA patients had raised antibody levels to Gm antigen, whereas none of the healthy control subjects presented this same increase. We also found 90% sensitivity, 100% specificity, 100% positive predictive value, 90% negative predictive value, and 90% prevalence value for this Gm antigen. Nonetheless, we did not find any statistically significant differences in the levels of immunoglobulins against Gm, although IgG showed a tendency to increase, probably because we are dealing with a secondary immune response. Using electroimmunotransfer blot assay, we found that sera from ALA patients recognise the 35 kDa Gm protein in the same way as it is recognised by the mouse monoclonal antibody, suggesting that is a relevant molecule for the diagnosis of amebiasis, and eventually could lead to its use as protection against the disease.
Departamento de Bioquimica, Facultad de Medicina, Universidad Nacional Autonoma de Mexico, Mexico D.F.
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C
Agundis-Mata
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Blanco
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Toledo
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Reta
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Kumate
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Isibasi
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000913:Antibodies, Protozoan / Q000032:analysis; D000953:Antigens, Protozoan / Q000032:analysis / Q000276:immunology; D004748:Entamoeba histolytica / Q000737:chemistry / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007106:Immune Sera / Q000737:chemistry*; D007132:Immunoglobulin Isotypes / Q000096:biosynthesis / Q000097:blood; D007075:Immunoglobulin M / Q000737:chemistry; D008101:Liver Abscess, Amebic / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D015800:Protozoan Proteins / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D007106:Immune Sera; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M; D008562:Membrane Glycoproteins; D015800:Protozoan Proteins