We recently reported the isolation of a new retrovirus, termed human immunodeficiency virus type 2 (HIV-2), from two West African patients with the acquired immunodeficiency syndrome (AIDS). This virus is related to but distinct from the well-characterized AIDS retrovirus, human immunodeficiency virus type 1 (HIV-1). We report here evidence of infection with HIV-2 in 30 patients, almost all from West Africa. Seventeen of them had a clinical syndrome indistinguishable from AIDS (7 of these 17 died). Others had either the AIDS-related complex or no HIV-related symptoms. All patients had serum antibodies reacting with HIV-2 in an indirect immunofluorescence assay. All serum tested contained antibodies reacting with the envelope glycoprotein of the virus in an immunoprecipitation assay. Cross-reactivity of serum antibodies with HIV-1 was detected in a minority of patients and varied according to the assay used. Retroviral isolates were obtained from the blood lymphocytes of 11 patients and were all identified as HIV-2 by nucleic acid hybridization; none hybridized with an HIV-1 probe. These findings indicate that some cases of AIDS in West Africa may be caused by HIV-2, but the extent of the spread of this virus and its clinical correlates will require careful epidemiologic investigation.
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 investigation was made of the antigenic composition of oocyst isolates of Cryptosporidium parvum by immunoblotting using rabbit polyclonal or murine monoclonal antibodies (MoAbs) developed against this parasite. Using the polyclonal antibodies in blots, a common antigenic profile was obtained from a number of human oocyst isolates from AIDS patients and immunocompetent children in the UK and Portugal. Antigenic differences were observed, however, between a human isolate from Turkey and these other human isolates. The antigenic profiles of oocyst isolates from deer and cattle were similar, but the profiles of the animal and human isolates differed to some extent. Two MoAbs which, in immunofluorescence microscopy, reacted with the surface of the C. parvum sporozoite were also used in blots. A major antigen(s) from 9 of 11 human oocyst isolates recognized by these MoAbs had a molecular weight of 47 kD, but the sizes of the corresponding antigens of the remaining 2 human isolates, one from Turkey (same as above) and one from the UK, were 45.5 and 51 kD, respectively. The equivalent antigen(s) from 4 bovine and 4 ovine isolates was 48 kD. One of the MoAbs failed to react in blots with 2 of the isolates, 1 human and 1 bovine.
A comparison was made of the antigenic composition of oocyst walls and sporozoites from Cryptosporidium baileyi from turkeys, C. muris from rodents, and C. parvum from ruminants, employing immunoblotting and immunofluorescence. In immunoblotting, oocyst antigens were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting (immunoblotting) and detected with rabbit polyclonal anti-C. muris or-C. parvum antibodies or murine monoclonal antibodies developed against C. parvum. Immunofluorescence was used to investigate the reactivity of these monoclonal antibodies with air-dried excystation mixtures of sporozoites and oocysts of the different species. The results from both types of experiment indicated that the three Cryptosporidium species could be differentiated immunologically. In comparison, few antigenic differences were found between a number of isolates of C. parnum in immunoblotting. There was also evidence to suggest that C. parvum and C. bailkyi were more closely related antigenically to one another than to C. muris.
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