Staphylococcus aureus adherence to Candida albicans hyphae is mediated by the hyphal adhesin Als3p

Brian Peters, Ekaterina S. Ovchinnikova, Bastiaan P. Krom, Lisa Marie Schlecht, Han Zhou, Lois L. Hoyer, Henk J. Busscher, Henny C. van der Mei, Mary Ann Jabra-Rizk, Mark E. Shirtliff

Research output: Contribution to journalArticle

87 Citations (Scopus)

Abstract

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 highmorbidity 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.

Original languageEnglish (US)
Pages (from-to)2975-2986
Number of pages12
JournalMicrobiology (United Kingdom)
Volume158
Issue number12
DOIs
StatePublished - Dec 1 2012

Fingerprint

Hyphae
Candida albicans
Staphylococcus aureus
Biofilms
Tongue
Epithelium
Bacteria
Agglutinins
Atomic Force Microscopy
Coinfection
Immunoassay
Critical Illness
Confocal Microscopy
Saccharomyces cerevisiae
Microscopy
Fungi
Yeasts
Mortality
Growth
Infection

All Science Journal Classification (ASJC) codes

  • Microbiology

Cite this

Peters, B., Ovchinnikova, E. S., Krom, B. P., Schlecht, L. M., Zhou, H., Hoyer, L. L., ... Shirtliff, M. E. (2012). Staphylococcus aureus adherence to Candida albicans hyphae is mediated by the hyphal adhesin Als3p. Microbiology (United Kingdom), 158(12), 2975-2986. https://doi.org/10.1099/mic.0.062109-0

Staphylococcus aureus adherence to Candida albicans hyphae is mediated by the hyphal adhesin Als3p. / Peters, Brian; Ovchinnikova, Ekaterina S.; Krom, Bastiaan P.; Schlecht, Lisa Marie; Zhou, Han; Hoyer, Lois L.; Busscher, Henk J.; van der Mei, Henny C.; Jabra-Rizk, Mary Ann; Shirtliff, Mark E.

In: Microbiology (United Kingdom), Vol. 158, No. 12, 01.12.2012, p. 2975-2986.

Research output: Contribution to journalArticle

Peters, B, Ovchinnikova, ES, Krom, BP, Schlecht, LM, Zhou, H, Hoyer, LL, Busscher, HJ, van der Mei, HC, Jabra-Rizk, MA & Shirtliff, ME 2012, 'Staphylococcus aureus adherence to Candida albicans hyphae is mediated by the hyphal adhesin Als3p', Microbiology (United Kingdom), vol. 158, no. 12, pp. 2975-2986. https://doi.org/10.1099/mic.0.062109-0
Peters, Brian ; Ovchinnikova, Ekaterina S. ; Krom, Bastiaan P. ; Schlecht, Lisa Marie ; Zhou, Han ; Hoyer, Lois L. ; Busscher, Henk J. ; van der Mei, Henny C. ; Jabra-Rizk, Mary Ann ; Shirtliff, Mark E. / Staphylococcus aureus adherence to Candida albicans hyphae is mediated by the hyphal adhesin Als3p. In: Microbiology (United Kingdom). 2012 ; Vol. 158, No. 12. pp. 2975-2986.
@article{d420789e13734be980c9db1fd5331eb2,
title = "Staphylococcus aureus adherence to Candida albicans hyphae is mediated by the hyphal adhesin Als3p",
abstract = "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 highmorbidity 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.",
author = "Brian Peters and Ovchinnikova, {Ekaterina S.} and Krom, {Bastiaan P.} and Schlecht, {Lisa Marie} and Han Zhou and Hoyer, {Lois L.} and Busscher, {Henk J.} and {van der Mei}, {Henny C.} and Jabra-Rizk, {Mary Ann} and Shirtliff, {Mark E.}",
year = "2012",
month = "12",
day = "1",
doi = "10.1099/mic.0.062109-0",
language = "English (US)",
volume = "158",
pages = "2975--2986",
journal = "Microbiology",
issn = "1350-0872",
publisher = "Society for General Microbiology",
number = "12",

}

TY - JOUR

T1 - Staphylococcus aureus adherence to Candida albicans hyphae is mediated by the hyphal adhesin Als3p

AU - Peters, Brian

AU - Ovchinnikova, Ekaterina S.

AU - Krom, Bastiaan P.

AU - Schlecht, Lisa Marie

AU - Zhou, Han

AU - Hoyer, Lois L.

AU - Busscher, Henk J.

AU - van der Mei, Henny C.

AU - Jabra-Rizk, Mary Ann

AU - Shirtliff, Mark E.

PY - 2012/12/1

Y1 - 2012/12/1

N2 - 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 highmorbidity 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.

AB - 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 highmorbidity 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.

UR - http://www.scopus.com/inward/record.url?scp=84870367328&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84870367328&partnerID=8YFLogxK

U2 - 10.1099/mic.0.062109-0

DO - 10.1099/mic.0.062109-0

M3 - Article

VL - 158

SP - 2975

EP - 2986

JO - Microbiology

JF - Microbiology

SN - 1350-0872

IS - 12

ER -