# Simplified J774A.1 macrophage assay for fungal pathogenicity demonstrates non-clinical Nakaseomyces glabratus strains survive better than lab strains

## TL;DR

A simplified macrophage assay shows non-clinical strains of Nakaseomyces glabratus survive better in macrophages than lab strains.

## Contribution

A simplified macrophage assay using flow cytometry and antifungal treatment was developed to study fungal pathogenicity.

## Key findings

- Non-clinical N. glabratus strains survive better in macrophages than lab strains.
- Loss of NgTUP11 does not reduce the pathogenicity of N. glabratus.

## Abstract

Nakaseomyces glabratus
(formerly known as
Candida glabrata
) is the second most common cause of candidiasis, whereas the closely related yeast,
Saccharomyces cerevisiae,
causes few infections. Macrophages can control
N. glabratus
infections through phagocytosis, but in cell culture,
N. glabratus
is able to persist in macrophages better than non-pathogenic yeast. Using J774A.1 macrophages, we simplified a standard persistence/survival assay by counting yeast cells with flow cytometry and incorporating an antifungal treatment. These improvements minimized wash steps and variation so fewer replicates were needed. Here, we demonstrate that loss of
NgTUP11
does not lower pathogenicity, and that three non-clinical
N. glabratus
strains survive in macrophages better than a laboratory strain.