Title

Candida albicans induces internalization of the Na + -K + -2Cl – cotransporter 1 expressed in Madin-Darby Canine Cells

Faculty Sponsor

Dr. Patrice Bouyer

Discipline(s)

Cell Biology Research

ORCID Identifier(s)

0000-0001-7691-5432

Presentation Type

Poster Presentation

Symposium Date

Spring 4-28-2022

Abstract

Candida albicans (C. albicans) is one of the most common human fungal pathogens, causing life threatening systemic infections in critically ill and immunocompromised patients. In the gut, fluid secretion participates in the body defense by flushing microorganisms and by maintaining the surface mucus hydrated. It is still unknown whether C. albicans decreases the host defense mechanism prior to invading intestinal cells. Previous work in the lab has demonstrated that the basolateral Na+ -K+ -2Cl - cotransporter 1 (NKCC1) plays a key role in the regulation of fluid secretion, and internalization of NKCC1 represents a potent pathway to blunt fluid secretion. We hypothesize that C. albicans, before invading intestinal cells, decreases fluid secretion by causing NKCC1 internalization. To test our hypothesis, we utilized Madin-Darby Canine Kidney (MDCK) cells stably expressing eGFP-NKCC1. MDCK were cultured on coverslips in 6 well plate. On the day of the experiment, MDCK were exposed to 50,000 C. albicans, for 15, 30 min,1 and 2 h hours. MDCK were also exposed to 100 nM phorbol 12-myristate 13-acetate (PMA) for 15 min as a positive control for NKCC1 internalization. Coverslips were fixed in 1% paraformaldehyde for 30 min and mounted for fluorescence microscopy. Fluorescent images were acquired using an inverted IX83 Olympus microscope equipped with a CCD camera. Exposing MDCK cells to C. albicans for 15 and 30 min did not result in NKCC1 internalization as compared to 100 nM PMA. Conversely, after 1- and 2-hours exposure to C. albicans, NKCC1 internalization was observed in some cells. Our preliminary experiments suggest that C. albicans induces NKCC1 internalization and may represent one mechanism by which C. albicans lower the intestinal defense mechanism. Gaining a better understanding of how C. albicans invades the mucosal barrier will greatly help developing new strategies for fighting candidiasis.

Biographical Information about Author(s)

Idalia Zachara is a student in Valparaiso University's Physician Assistant Graduate Program. She has been working with Dr. Bouyer and C. albicans since her undergraduate days at Valpo. The skills learned in the research lab have been very impactful on the pathophysiology taught in PA school. She hopes purse research in her future career outside of being a clinician.

George Gundelach is a student at Ivy Tech Community College. He is interested in nursing and biological research. He joined Dr. Bouyer's lab in 2021 and has greatly contributed to its advancements.

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