Muramidase-mediated damage to Candida yeast cells. Histochemical and immunochemical characterization of accumulating wall-like material.
Hen egg-white lysozyme is known to be fungicidal to blastoconidia of Candida albicans under defined in vitro conditions. This lethal action leads to changes in the layering of cell wall and to plasmolysis, caused by unremitting accumulation of wall-like material between the yeast cell wall and cytoplasmic membrane. Here, several methods were applied on ultrathin sections to define the nature of wall-like material: histochemical staining with periodic acid-thiocarbohydrazide-silver proteinate, periodic acid-alkaline bismuth, and phosphotungstic acid at low pH; the localization of the carbohydrate residues with lectin-gold complex; immunocytochemical staining with monospecific antibodies, factor 1 and 6, which recognized major cell wall antigens. The wall-like material was almost uniformly highlighted with periodic acid-thiocarbohydrazide-silver proteinate, factor 1 antibody, concanavalin A-gold and wheat germ agglutinin-ovomucoid-gold, indicating the presence of mannoproteins and chitin. The serotype A-specific epitope recognized by factor 6 antibody was not detected in the wall-like material, although it was demonstrated in the outer cell wall layers after 2 h of exposure to lysozyme.
Marquis
G
G
Department of Microbiology and Immunology, University of Montreal, Quebec, Canada.
Garzon
S
S
Strykowski
H
H
Auger
P
P
Benhamou
N
N
D000911:Antibodies, Monoclonal / Q000276:immunology; D000941:Antigens / Q000032:analysis / Q000276:immunology; D002176:Candida albicans / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure; D002241:Carbohydrates / Q000032:analysis; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D002686:Chitin / Q000032:analysis / Q000276:immunology; D017979:Gold Colloid; D006651:Histocytochemistry; D006834:Hydrazines; D007150:Immunohistochemistry; D008562:Membrane Glycoproteins / Q000032:analysis; D008854:Microscopy, Electron; D009113:Muramidase / Q000494:pharmacology / Q000502:physiology*; D010504:Periodic Acid; D010772:Phosphotungstic Acid; D012836:Silver Proteins; D014909:Wheat Germ Agglutinins
D000911:Antibodies, Monoclonal; D000941:Antigens; D002241:Carbohydrates; D017979:Gold Colloid; D006834:Hydrazines; D008562:Membrane Glycoproteins; D012836:Silver Proteins; D014909:Wheat Germ Agglutinins; C025363:mannoproteins; C055835:periodic acid-thiocarbohydrazide-silver proteinate; D010504:Periodic Acid; D010772:Phosphotungstic Acid; D002686:Chitin; D009113:Muramidase