Quantification of specific antibody response to Cryptosporidium antigens by laser densitometry

Abstract

Cryptosporidium spp. is a protozoan parasite with worldwide distribution associated with diarrhea in immunocompromised patients (particularly those with acquired immunodeficiency syndrome [AIDS]) and in immunocompetent humans. Immunoglobulin M (IgM) and IgG antibody responses are readily detected by an enzyme-linked immunosorbent assay. To determine which Cryptosporidium antigens invoke antibody responses in humans, we performed polyacrylamide gel electrophoresis using purified oocysts, followed by Western blots with human sera from various populations. Of 40 sera from persons with cryptosporidiosis (24 AIDS and 16 non-AIDS patients), in 37 (93%) a 23,000-dalton antigen measured quantitatively by laser densitometry was recognized. Of 63 sera from IgM- or IgG-positive individuals, as determined by enzyme-linked immunosorbent assay, in 58 (92%) this same antigen was recognized. Up to three additional bands between 125,000 and 175,000 daltons were identified by some of these sera. These results suggest that most persons infected with Cryptosporidium spp. produce antibodies which recognize at least one common low-molecular-weight antigen. Isolation of this antigen will be useful in development of diagnostic tests and may be important in the study of immunity.