Staphylococcus aureus adherence to Candida albicans hyphae is mediated by the hyphal adhesin Als3p.
The bacterium Staphylococcus (St.) aureus and the opportunistic fungus Candida albicans are currently among the leading nosocomial pathogens, often co-infecting critically ill patients, with high morbidity and mortality. Previous investigations have demonstrated preferential adherence of St. aureus to C. albicans hyphae during mixed biofilm growth. In this study, we aimed to characterize the mechanism behind this observed interaction. C. albicans adhesin-deficient mutant strains were screened by microscopy to identify the specific receptor on C. albicans hyphae recognized by St. aureus. Furthermore, an immunoassay was developed to validate and quantify staphylococcal binding to fungal biofilms. The findings from these experiments implicated the C. albicans adhesin agglutinin-like sequence 3 (Als3p) in playing a major role in the adherence process. This association was quantitatively established using atomic force microscopy, in which the adhesion force between single cells of the two species was significantly reduced for a C. albicans mutant strain lacking als3. Confocal microscopy further confirmed these observations, as St. aureus overlaid with a purified recombinant Als3 N-terminal domain fragment (rAls3p) exhibited robust binding. Importantly, a strain of Saccharomyces cerevisiae heterologously expressing Als3p was utilized to further confirm this adhesin as a receptor for St. aureus. Although the parental strain does not bind bacteria, expression of Als3p on the cell surface conferred upon the yeast the ability to strongly bind St. aureus. To elucidate the implications of these in vitro findings in a clinically relevant setting, an ex vivo murine model of co-infection was designed using murine tongue explants. Fluorescent microscopic images revealed extensive hyphal penetration of the epithelium typical of C. albicans mucosal infection. Interestingly, St. aureus bacterial cells were only seen within the epithelial tissue when associated with the invasive hyphae. This differed from tongues infected with St. aureus alone or in conjunction with the als3 mutant strain of C. albicans, where bacterial presence was limited to the outer layers of the oral tissue. Collectively, the findings generated from this study identified a key role for C. albicans Als3p in mediating this clinically relevant fungal-bacterial interaction.
Department of Microbial Pathogenesis, University of Maryland - Baltimore, Dental School, 650 W. Baltimore Street, Baltimore, MD 21201, USA.|Graduate Program in Life Sciences, Molecular Microbiology and Immunology Program, University of Maryland - Baltimore, 660 W. Redwood Street, Baltimore, MD 21201, USA.
Brian M
BM
Peters
Department of Biomedical Engineering, W. J. Kolff Institute, University Medical Center Groningen and University of Groningen, Antonius Deusinglaan 1, 9713 AV, Groningen, The Netherlands.
Ekaterina S
ES
Ovchinnikova
Department of Preventive Dentistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and Free University Amsterdam, Gustav Mahlerlaan 3004, 1081 LA Amsterdam, The Netherlands.|Department of Biomedical Engineering, W. J. Kolff Institute, University Medical Center Groningen and University of Groningen, Antonius Deusinglaan 1, 9713 AV, Groningen, The Netherlands.
Bastiaan P
BP
Krom
Department of Restorative Dentistry and Periodontology, Ludwig Maximilian University of Munich, Goethestrasse 70, 80336 Munich, Germany.
Lisa Marie
LM
Schlecht
West China College of Stomatology, Sichuan University, PR China.|Department of Microbial Pathogenesis, University of Maryland - Baltimore, Dental School, 650 W. Baltimore Street, Baltimore, MD 21201, USA.
Han
H
Zhou
Department of Pathobiology, University of Illinois, 2001 S. Lincoln Ave., Urbana, IL 61802, USA.
Lois L
LL
Hoyer
Department of Biomedical Engineering, W. J. Kolff Institute, University Medical Center Groningen and University of Groningen, Antonius Deusinglaan 1, 9713 AV, Groningen, The Netherlands.
Henk J
HJ
Busscher
Department of Biomedical Engineering, W. J. Kolff Institute, University Medical Center Groningen and University of Groningen, Antonius Deusinglaan 1, 9713 AV, Groningen, The Netherlands.
Henny C
HC
van der Mei
Department of Pathology, School of Medicine, University of Maryland, Baltimore, USA.|Department of Oncology and Diagnostic Sciences, Dental School, University of Maryland - Baltimore, 650 W Baltimore Street, Baltimore, MD 21201, USA.|Department of Microbiology and Immunology, School of Medicine, University of Maryland - Baltimore, 660 W. Redwood Street, Baltimore, MD 21201, USA.
Mary Ann
MA
Jabra-Rizk
Department of Microbiology and Immunology, School of Medicine, University of Maryland - Baltimore, 660 W. Redwood Street, Baltimore, MD 21201, USA.|Department of Microbial Pathogenesis, University of Maryland - Baltimore, Dental School, 650 W. Baltimore Street, Baltimore, MD 21201, USA.
Mark E
ME
Shirtliff
D000818:Animals; D001422:Bacterial Adhesion; D002176:Candida albicans / Q000235:genetics / Q000502:physiology*; D002177:Candidiasis / Q000382:microbiology / Q000473:pathology; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D004848:Epithelium / Q000382:microbiology / Q000473:pathology; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D055786:Gene Knockout Techniques; D025301:Hyphae / Q000502:physiology*; D051379:Mice; D018625:Microscopy, Atomic Force; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology; D013211:Staphylococcus aureus / Q000502:physiology*; D014059:Tongue / Q000382:microbiology / Q000473:pathology
C109455:ALA1 protein, Candida albicans; C113620:ALS3 protein, Candida albicans; D015815:Cell Adhesion Molecules; D005656:Fungal Proteins