Two Methods of Measuring Cryptococcus neoformans Fungal Burden in Macrophages

Emily Smith-Peavler; Linda M Sircy; David E Nelson; Erin E McClelland

doi:10.1007/978-1-0716-3722-7_14

Two Methods of Measuring Cryptococcus neoformans Fungal Burden in Macrophages

Methods Mol Biol. 2024:2775:211-221. doi: 10.1007/978-1-0716-3722-7_14.

Authors

Emily Smith-Peavler^#^{1

2}, Linda M Sircy^#^{1

3}, David E Nelson¹, Erin E McClelland⁴

Affiliations

¹ Department of Biology, Middle Tennessee State University, Murfreesboro, TN, USA.
² KIPP Collegiate High School, Nashville, TN, USA.
³ Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Center, Salt Lake City, UT, USA.
⁴ Department of Biomedical Sciences, Marian University College of Osteopathic Medicine, Indianapolis, IN, USA. [email protected].

^# Contributed equally.

PMID: 38758320
DOI: 10.1007/978-1-0716-3722-7_14

Abstract

The ability of C. neoformans to survive and replicate within host phagocytes enables it to evade the immune system and allows for persistence of the infection. As such, measuring fungal burden of C. neoformans strains-and indeed how drug treatments can influence fungal burden-provides important information about C. neoformans pathogenesis. In this chapter, we describe two methods that may be used to appraise fungal burden: a standard end-point colony-formation assay for calculating the average number of yeast per host cell and a fluorescence microscopy-based method that may be used to measure changes in fungal burden in individual living macrophages in real time.

Keywords: Cryptococcus neoformans; Fungal burden; Intracellular fungal pathogen; Live cell imaging; Macrophage cell lines; Phagocytosis assays.

MeSH terms

Animals
Colony Count, Microbial / methods
Cryptococcosis* / immunology
Cryptococcosis* / microbiology
Cryptococcus neoformans*
Humans
Macrophages* / immunology
Macrophages* / metabolism
Macrophages* / microbiology
Mice
Microscopy, Fluorescence* / methods