Potential Use of a Monoclonal Antibody for the Detection of Candida Antigens in an Experimental Systemic Candidiasis Model

Abstract

The Candida species are the most common fungal pathogens of systemic candidiasis. The diagnosis of invasive candidiasis remains a laboratory and clinical challenge. Thus, development of diagnostic assays to detect systemic candidiasis and to identify Candida virulence factors and associated pathogenesis through immunohistochemistry using specific monoclonals and polyclonals will be useful. Inbred Balb/c mice were immunized with C. albicans antigens, and blood was checked for the presence of reactive antibodies using ELISA. Fusion was performed using the harvested spleen cells and NS1 myeloma cells, and the clones were screened for the presence of antibody producing hybrid cells by dot-blot. Western blot analysis showed that the L2D10 monoclonal antibody was reactive against the antigens with molecular weight of 20 kDa. Experimental systemic candidiasis in mice was induced through intravenous injection of C. albicans and all the vital organs were collected for immunohistochemistry study. The monoclonal antibody reacted to surface epitopes on the yeast cells, germ tubes, and hyphae, and to immune complexes. It was used with the polyclonal antibody in a sandwich ELISA for the detection of circulating antigens in experimental candiadiasis in mice. Antibody levels were also determined using the ELISA method, and the antibody levels of C. albicans infected mice were increased compared with uninfected animals. The monoclonal antibody was used in immunoperoxidase and immunofluorescence techniques for the detection of fungal infection in tissue sections and was found to be more sensitive than conventional periodic acid Schiff or silver staining techniques. This monoclonal antibody may serve as potential primary capture antibodies for the development of a rapid diagnostic test for human systemic fungal infection.

A comparative histopathological study of systemic candidiasis in association with experimentally induced breast cancer

“…histopathologically, favourable growth of the Candida colonies in the form of hyphae, yeast cells and pseudohyphae were discovered in the kidneys, pelvis and tubule region, but not in many of the other organs. this was attributed to the mild dose of 5x10 6 cells/ml Candida cells injected and the short duration of the experiment as also shown by Wong et al (16).…”

“…Information obtained from these studies on the necessary dosages, as well as previous observations were used to make comparisons with this study (15)(16)(17). Some of these studies have been dedicated to the observations of the correlation between systemic candidiasis and other forms of immunosuppression such as chemotherapy, steroid therapy, antibiotic therapy, as well as other types of malignancies such as leukemia and esophageal cancer.…”