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Use of Image Cytometry for Quantification of Pathogenic Fungi in Association with Host Cells
Published on: June 19, 2013
Pathobiology of Candida auris infection analyzed by multiplexed imaging and single cell analysis
Chrystal Chadwick1, Magdia De Jesus2,3, Fiona Ginty1
1GE Research, Niskayuna, New York, United States of America.
Abstract:
Fungal organisms contribute to significant human morbidity and mortality and Candida auris (C. auris) infections are of utmost concern due to multi-drug resistant strains and persistence in critical care and hospital settings. Pathogenesis and pathology of C. auris is still poorly understood and in this study, we demonstrate how the use of multiplex immunofluorescent imaging (MxIF) and single-cell analysis can contribute to a deeper understanding of fungal infections within organs. We used two different neutrophil depletion murine models (treated with either 1A8-an anti-Ly6G antibody, or RB6-8C5-an anti-Ly6G/Ly6C antibody; both 1A8 and RB6-8C5 antibodies have been shown to deplete neutrophils) and compared to wildtype, non-neutropenic mice. Following pathologist assessment, fixed samples underwent MxIF imaging using a C. albicans antibody (shown to be cross-reactive to C. auris) and immune cell biomarkers-CD3 (T cells), CD68 (macrophages), B220 (B cells), CD45 (monocytes), and Ly6G (neutrophils) to quantify organ specific immune niches. MxIF analysis highlighted the heterogenous distribution of C. auris infection within heart, kidney, and brain 7 days post-infection. Size and number of fungal abscesses was greatest in the heart and lowest in brain. Infected mice had an increased count of CD3+, CD68+, B220+, and CD45+ immune cells, concentrated around C. auris abscesses. CD68+ cells were predominant in wildtype (non-neutropenic mice) and CD3+/CD45+ cells were predominant in neutropenic mice, with B cells being the least abundant. These findings suggest a Th2 driven immune response in neutropenic C. auris infection mice models. This study demonstrates the value of MxIF to broaden understanding of C. auris pathobiology, and mechanistic understanding of fungal infections.
Insights
Multiplex immunofluorescent imaging reveals how Candida auris infections affect organs and immune cells. This technique enhances understanding of fungal infections and host immune responses in neutropenic and non-neutropenic models.
Area of Science:
- Mycology and Immunology
- Infectious Diseases
- Medical Imaging and Pathology
Background:
- Candida auris (C. auris) poses a significant threat due to multidrug resistance and persistence in healthcare settings.
- The pathogenesis and pathology of C. auris infections remain poorly understood.
- Understanding organ-specific immune responses is crucial for combating C. auris.
Purpose of the Study:
- To investigate the organ-specific distribution and pathology of C. auris infections using multiplex immunofluorescent imaging (MxIF).
- To analyze the host immune cell response within infected organs in both neutropenic and non-neutropenic murine models.
- To demonstrate the utility of MxIF and single-cell analysis in studying fungal infections.
Main Methods:
- Utilized neutrophil-depleted (1A8 or RB6-8C5 antibody treatment) and wildtype murine models.
- Administered C. auris infection and collected organ samples (heart, kidney, brain) 7 days post-infection.
- Performed MxIF using a cross-reactive C. albicans antibody and immune cell biomarkers (CD3, CD68, B220, CD45, Ly6G) for single-cell analysis.
Main Results:
- MxIF revealed heterogeneous C. auris distribution across heart, kidney, and brain, with largest abscesses in the heart and smallest in the brain.
- Infected mice showed increased CD3+, CD68+, B220+, and CD45+ immune cells around C. auris abscesses.
- Wildtype mice had predominant CD68+ (macrophages) cells, while neutropenic mice showed more CD3+ (T cells) and CD45+ (monocytes) cells, suggesting a Th2-driven response in neutropenia.
Conclusions:
- MxIF and single-cell analysis provide valuable insights into the pathobiology of C. auris infections within organs.
- The study highlights distinct immune cell niches and responses in neutropenic versus non-neutropenic C. auris infections.
- Findings contribute to a mechanistic understanding of fungal infections and host immune dynamics.
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