Characteristics and specificity of hybridoma antibodies against oocyst antigens of Cryptosporidium parvum from man

Abstract

Hybridoma antibodies (HAbs) against oocyst antigens of a human isolate of Cryptosporidium parvum were developed by fusion of SP2/0 mouse myeloma cells and spleen cells from BALB/c mice immunized with oocyst homogenates. In an indirect immunofluorescence antibody test (IFAT), using as antigen a mixture of air-dried sporozoites and oocysts, HAbs labelled either the oocyst wall or areas of the sporozoite, including the whole organism, the entire surface, a polar region or the interior. Most of the HAbs were specific for the sporozoite surface, and few of them recognized the oocyst wall. In Western blot analysis of oocyst antigens, sporozoite surface-reactive monoclonal antibodies (MoAbs) recognized one or more of seven polypeptide bands with molecular weights in the range 47- greater than 200 kD, and all reacted with the 47 kD band. Each of four heterologous parasite isolates had a unique recognition pattern with a panel of MoAbs in IFAT, suggesting antigenic differences may exist between strains of C. parvum. The ability to differentiate between parasite isolates by immunological methods might be of value in epidemiological studies of cryptosporidiosis.

An Immunoglobulin G1 Monoclonal Antibody Highly Specific to the Wall of Cryptosporidium Oocysts

“…The presence of sporozoites in this preparation may have also reduced the immungenicity. McDonald et al (12) showed that the sporo-…”

“…In previous studies, immunization with whole oocysts produced MAbs predominantly to the sporozoite and a few to the oocyst wall (12). Removal of sporozoites from antigen preparations may reduce this predominant IgM response.…”

A common oocyst surface antigen of Cryptosporidium recognized by monoclonal antibodies

“…Anusz et al (1990) described mAb 18.280.2, which recognized a 40-kDa molecule located in the exterior surface of C. parvum oocysts. Additionally, mAb 181B5 reacted with 41-and 44-kDa molecules of C. parvum (McDonald et al 1991;Nina et al 1992). MAb OW-IGO also recognized a 40-kDa antigen of the C. parvum oocyst wall (Bonnin et al 1991).…”

“…Many kinds of mAbs against C. parvum have been reported (Anusz et al 1990;Bonnin et al 1991Bonnin et al , 1995McDonald et al 1991). However, no mAbs generated against C. muris have been described to date.…”

More productive in vitro culture of Cryptosporidium parvum for better study of the intra- and extracellular phases

“…It is known that different Cryptosporidium isolates from various regions have different antigens, virulence, infectivity, and sensitivity to drugs as well as disinfectants (McDonald et al 1991, Griffin et al 1992, Fayer et al 2000. Therefore, it is necessary to characterize the parasites molecularly in each region and know the parasites to be used in the in vitro culture.…”