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The Role of Tetraspanins in the Uptake of the Fungus Candida albicans by Host Cells

CSEF · 2018 Microbiology (General)

Chen E. Filler · Palos Verdes High School, Palos Verdes Estates, Los Angeles County

Overview

Objectives/Goals My hypothesis is that specific endothelial cell membrane proteins called tetraspanins are necessary for C. albicans to invade endothelial cells, and my goal was to discover which tetraspanins were important for invasion. Methods/Materials The HUVEC-TERT endothelial cell line was grown in tissue culture. The tetraspanins, CD9, CD63, and CD151 were detected by indirect immuno- fluorescence using specific mouse monoclonal antibodies. Each mouse antibody was detected with a fluorescent labeled goat-anti mouse antibody. Internalized organisms were identified by staining the endothelial cells with Alexafluor-labeled phalloidin which labels the endothelial cell actin microfilaments that are respon- sible for the endocytosis of the organism. The cells were imaged by confocal microscopy. The capacity of the anti-tetraspanin antibodies to inhibit the uptake of C. albicans by endothelial cells was determined using a differential fluorescence assay. Results By confocal microscopy, it was found that all three tetraspanins, CD9, CD63, and CD151, accumulated around C. albicans hyphae that were invading endothelial cells. Using blocking monoclonal antibodies, it was determined that only CD9 and CD63, but not CD151 were required for C. albicans to invade endothelial cells. Conclusions/Discussion The results indicate that the tetraspanins, CD9 and CD63 are required for C. albicans to invade endothelial cells, thus supporting my hypothesis. Blocking CD9 or CD63 is a potential approach to prevent C. albicans infections in patients.

Summary statement

I found that the tetraspanins CD9 and CD63 were required for endothelial cells to endocytose Candida albicans.

Help received

Dr. Hong Liu helped me design the experiments and taught me the experimental techniques. I performed all the experiments by myself.

Competition history

  • CSEF 2018 Microbiology (General) · Entry S1608

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