Characterization of mutant strains of Candida albicans deficient in expression of a surface determinant.
Monoclonal antibody (MAb) 17E4 reacts with a surface carbohydrate determinant and agglutinates cells of Candida albicans. Using this MAb, we have isolated N-methyl-N'-nitro-N-nitrosoguanidine-induced nonagglutinating mutants. Eleven of these were characterized for the presence and expression of the surface antigen recognized by MAb 17E4 by immunoblot analysis of whole cells and by fluorescence flow cytometry. Soluble cell wall extracts from five mutant strains were negative by immunoblot analysis. The reactivities of the strains with several other MAbs and commercial antisera (Candida Check; Iatron Laboratories, Tokyo, Japan) which also recognize carbohydrate determinants were examined by immunoblot analysis of whole cells. Mutant strains showed no or reduced expression of the MAb 17E4 antigen and could be placed into at least two distinct phenotypic classes. Recognition by the other MAbs tested showed a similar pattern, while recognition by the commercial antisera was unchanged in the mutant strains. 1H and 13C nuclear magnetic resonance spectral analysis of mannan prepared from the wild type and nonexpressing mutant-strain 4A showed that the spectra from the mutant strain were simpler than those of the wild type. Most of the beta-1,2 linkages and all of the C-1 phosphate linkages were absent in the 4A mannan spectra, which suggested that the mutant mannan lacked the phosphate-bound beta-1,2-linked mannooligosaccharides. The effect of the surface defect on the ability of mutant strain 4A to adhere and to invade host tissue was examined in two murine models. In ex vivo binding assays, strain 4A showed reduced binding to the marginal zone and increased binding to the white pulp of splenic tissue, decreased binding to kidney tissue, and no change in binding to liver tissue compared with the wild type. In vivo, no difference was observed in translocation of the wild type or strain 4A to liver following immuno-compromising treatment of infant mice which had been challenged with either strain by the oral-intragastric route.
Chaffin
W L
WL
Department of Microbiology, School of Medicine, Texas Tech University Health Sciences Center, Lubbock 79430.
Collins
B
B
Marx
J N
JN
Cole
G T
GT
Morrow
K J
KJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000946:Antigens, Fungal / Q000032:analysis*; D000954:Antigens, Surface / Q000032:analysis; D001692:Biological Transport; D002176:Candida albicans / Q000276:immunology* / Q000502:physiology; D005434:Flow Cytometry; D007668:Kidney / Q000382:microbiology; D008099:Liver / Q000382:microbiology; D008351:Mannans / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation
D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal; D000954:Antigens, Surface; D008351:Mannans