ABSTRACT
The protective role of humoral antibodies in the resolution of systemic candidiasis remains controversial. Investigation of the humoral immune responses in mouse strains of varying susceptibility to infection may demonstrate a link between mouse strain susceptibility, antibody production and specificity, and the ability to resolve an infection. The antibody response in five different strains of mice during primary immune response to systemic infection with Candida albicans was investigated. Immune sera were fractionated by protein A affinity chromatography to yield fractions containing IgG1, IgG2a and IgG2b immunoglobulins. BALB/c mice of low susceptibility to the infection and DBA/2J mice of high susceptibility produced increased levels of the IgG1 isotype and decreased levels of the IgG2a isotype. AKR, CBA/H and C57B1/6J mice of moderate susceptibility produced antibodies predominantly of the IgG2a isotype. The patterns of antigen recognition by antibodies in immune sera and in fractions obtained after protein A chromatography of immune sera were investigated by western blotting and immunostaining. Antibodies from AKR(H-2K) and CBA/H (H-2k) mice reacted strongly after immunoblotting with antigens of 87 and 96 kDa. In contrast, immune sera from both the highly susceptible DBA/2J (H-2d) mice and the resistant BALB/c (H-2d) mice reacted strongly with an antigen of 48 kDa. C57B1/6J (H-2b) mice produced variable antibody reactivity to antigens of 48, 65, 66 and 79 kDa depending on the IgG subclass tested. The IgG subclass responses and the patterns of antigen recognition in these mice suggest that humoral responses to C. albicans may be restricted by H-2 haplotype.(ABSTRACT TRUNCATED AT 250 WORDS)
Subject(s)
Antibodies, Fungal/blood , Candidiasis/immunology , Immunoglobulin Isotypes/blood , Mice, Inbred Strains/immunology , Animals , Antigens, Fungal/analysis , Antigens, Fungal/classification , Blotting, Western , Candidiasis/genetics , Disease Susceptibility/immunology , Electrophoresis, Polyacrylamide Gel , Female , Genetic Predisposition to Disease , Immunoglobulin G/blood , Immunoglobulin G/classification , Immunoglobulin Isotypes/classification , Immunoglobulin M/blood , Immunoglobulin M/classification , Mice , Mice, Inbred AKR , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred CBA , Mice, Inbred DBA , Mice, Inbred Strains/microbiology , Species SpecificityABSTRACT
A rapid assay was developed for determination of the viability of blastospores of Candida albicans utilizing flow cytometry to detect the accumulation of tetrabromofluorescein in non-viable yeast cells. Eosin Y was shown to stain non-viable C. albicans blastospores selectively without affecting the cellular viability of competent yeast cells or resulting in non-specific staining of viable cells. Results of these studies show that this flow cytometric method of determining cell viability of C. albicans is more accurate and more precise than the more common method of enumerating colony forming units.
Subject(s)
Candida albicans/isolation & purification , Colony Count, Microbial/methods , Eosine Yellowish-(YS) , Flow Cytometry/methods , Animals , Candida albicans/drug effects , Candida albicans/metabolism , Candidiasis/microbiology , Disease Models, Animal , Eosine Yellowish-(YS)/pharmacology , Evaluation Studies as Topic , Mice , Oxygen Consumption/drug effects , Spores, Fungal/drug effects , Spores, Fungal/isolation & purification , Spores, Fungal/metabolismABSTRACT
Reported targets of the specific immune responses to Candida albicans in human candidiasis include a 47-kDa breakdown product of a 90-kDa heat shock protein (HSP 90) (R. Matthews and J. Burnie, FEMS Microbiol. Lett. 60:25-30, 1989) and the 48-kDa enolase (K.M. Franklyn, J.R. Warmington, A.K. Ott, and R.B. Ashman, Immunol. Cell Biol. 68:173-178, 1990). These proteins are immunodominant antigens of C. albicans. Western blotting (immunoblotting) and immunoprecipitation were used to investigate the humoral response in a mouse model of systemic candidiasis. Resolution of systemic candidiasis in CBA/H mice is associated with a high level of antibody reactivity to C. albicans antigens. A significant antibody response against a non-HSP antigen of 96 kDa which was distinct from the C. albicans HSP 90 antigen was detected. Significant antibody reactivity against an HSP of 75 kDa was also detected. We concluded that resolution of C. albicans infections in CBA/H mice was associated with antibodies to an HSP and a non-HSP of 75 and 96 kDa, respectively.
Subject(s)
Antibodies, Fungal/biosynthesis , Antigens, Fungal/immunology , Candida albicans/immunology , Animals , CD4-Positive T-Lymphocytes/immunology , Female , Heat-Shock Proteins/immunology , Mice , Mice, Inbred CBA , Precipitin TestsABSTRACT
Mice injected with rat RBC produce RBC autoantibodies and spleen cells that delay autoantibody production. In this report it is shown that transfer of 5 X 10(6) blood leukocytes from mice producing autoantibodies to syngeneic recipients before the injections of rat RBC were started, significantly delayed autoantibody production. These circulating suppressor cells were effective if transferred up to 3 weeks before the first injection of rat RBC and were destroyed by 2,000 rad of irradiation before transfer.
