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Antigenic characterization of human isolates of cryptosporidia
O Ditrich1, P Kopácek, Z Kucerová
1Institute of Parasitology, Academy of Sciences of the Czech Republic, Ceské Budĕjovice.
Insights
Cryptosporidium antigen analysis revealed distinct membrane proteins in isolates from immunodeficient and immunocompetent individuals. Missing antigens correlated with reduced infectivity in mice, suggesting species-specific differences in Cryptosporidium strains.
Area of Science:
- Parasitology
- Immunology
- Molecular Biology
Background:
- Cryptosporidiosis is a significant cause of diarrhea, particularly in immunocompromised individuals and young children.
- Understanding antigenic variations in Cryptosporidium isolates is crucial for diagnosis and control.
Purpose of the Study:
- To compare the antigenic profiles of Cryptosporidium isolates from diverse patient groups.
- To investigate the correlation between antigenic differences and infectivity in animal models.
Main Methods:
- Antigen preparation from 21 Cryptosporidium isolates (19 children, 2 AIDS patients).
- Enzyme-linked immunosorbent assay (ELISA) for serological examination.
- Polyacrylamide gel electrophoresis and electroimmunotransfer blots for antigen analysis.
- Infectivity studies in neonatal mice, chickens, and guinea pigs.
Main Results:
- No differences observed in soluble antigens between isolates.
- Two membrane-enriched antigens (15.5 and 33 kD) were absent in some isolates.
- Isolates lacking these bands showed reduced or no infectivity in neonatal mice.
- One isolate with missing bands infected chickens (potentially C. baileyi), while another infected guinea pigs (uncertain species).
Conclusions:
- Membrane-enriched antigens appear to be key discriminators between Cryptosporidium isolates.
- Antigenic profiles, particularly membrane proteins, correlate with host infectivity and may aid in species identification.
- Further research is needed to clarify the species classification of isolates with uncertain antigenic patterns.
Abstract:
Isolates of cryptosporidia from immunodeficient patients and from immunocompetent children suffering from diarrhoea were sources for preparation of antigens. Altogether, antigens from 21 isolates were obtained, 19 from children and 2 from AIDS patients. With one exception, all children were under 4 years old and most of them were between 1 and 2 years old. The probable source of infection was found in 11 cases. In 8 cases, as well as in both AIDS patients, the source of infection was not determined. The same groups of persons and the control group of healthy blood donors were examined serologically using ELISA with the antigen of bovine origin and positive sera were used for following experiments. Soluble and membrane-enriched antigens from oocysts were compared using polyacrylamide gel electrophoresis and electroimmunotransfer blots developed by human immune sera. While no differences were recorded in soluble antigens, two of the membrane-enriched antigens were missing (15.5 and 33 kD bands). While all isolates with a pattern typical for Cryptosporidium parvum were infective for neonatal mice, both isolates with missing bands were not infective for newborn mice in repeated experiments. The first of them was infective for chickens and was originally determined to be Cryptosporidium baileyi (Ditrich et al. 1991). However, the antigenic pattern differs from pattern of this species. The second isolate was infective for guinea pigs and its species classification remains uncertain.
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