Pathogenesis of Staphylococcus haemolyticus on primary human skin fibroblast cells

Staphylococcus haemolyticus (S. haemolyticus) is one of the Coagulase-negative staphylococci (CoNS) that inhabits the skin as a commensal. It is increasingly implicated in opportunistic infections, including diabetic foot ulcer (DFU) infections. In contrast to the abundance of information available...

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Autores principales:	Hala O. Eltwisy, Medhat Abdel-Fattah, Amani M. Elsisi, Mahmoud M. Omar, Ahmed Aly Abdelmoteleb, Mohamed A. El-Mokhtar
Formato:	article
Lenguaje:	EN
Publicado:	Taylor & Francis Group 2020
Materias:	staphylococcus haemolyticus diabetic foot ulcer primary human skin fibroblast cells bacterial invasion pathogenesis Infectious and parasitic diseases RC109-216
Acceso en línea:	https://doaj.org/article/a27df34e79b94c4dadd3c639da3d878c
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spelling	oai:doaj.org-article:a27df34e79b94c4dadd3c639da3d878c2021-11-17T14:21:59ZPathogenesis of Staphylococcus haemolyticus on primary human skin fibroblast cells2150-55942150-560810.1080/21505594.2020.1809962https://doaj.org/article/a27df34e79b94c4dadd3c639da3d878c2020-12-01T00:00:00Zhttp://dx.doi.org/10.1080/21505594.2020.1809962https://doaj.org/toc/2150-5594https://doaj.org/toc/2150-5608Staphylococcus haemolyticus (S. haemolyticus) is one of the Coagulase-negative staphylococci (CoNS) that inhabits the skin as a commensal. It is increasingly implicated in opportunistic infections, including diabetic foot ulcer (DFU) infections. In contrast to the abundance of information available for S. aureus and S. epidermidis, little is known about the pathogenicity of S. haemolyticus, despite the increased prevalence of this pathogen in hospitalized patients. We described, for the first time, the pathogenesis of different clinical isolates of S. haemolyticus isolated from DFU on primary human skin fibroblast (PHSF) cells. Virulence-related genes were investigated, adhesion and invasion assays were carried out using Giemsa stain, transmission electron microscopy (TEM), MTT and flowcytometry assays. Our results showed that most S. haemolyticus carried different sets of virulence-related genes. S. haemolyticus adhered to the PHSF cells to variable degrees. TEM showed that the bacteria were engulfed in a zipper-like mechanism into a vacuole inside the cell. Bacterial internalization was confirmed using flowcytometry and achieved high intracellular levels. PHSF cells infected with S.haemolyticus suffered from amarked decrease in viability and increased apoptosis when treated with whole bacterial suspensions or cell-free supernatants but not with heat-treated cells. After co-culture with PBMCs, S. haemolyticus induced high levels of pro-inflammatory cytokines. This study highlights the significant development of S. haemolyticus, which was previously considered a contaminant when detected in cultures of clinical samples. Their high ability to adhere, invade and kill the PHSF cells illustrate the severe damage associated with DFU infections. Abbreviations CoNS, coagulase-negative staphylococci; DFU, diabetic foot ulcer; DM, diabetes mellitus; DMEM, Dulbecco’s Modified Eagle Medium; MTT, 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide; PBMCs,peripheral blood mononuclear cells; PHSF, primary human skin fibroblast; CFU, colony-forming unit.Hala O. EltwisyMedhat Abdel-FattahAmani M. ElsisiMahmoud M. OmarAhmed Aly AbdelmotelebMohamed A. El-MokhtarTaylor & Francis Grouparticlestaphylococcus haemolyticusdiabetic foot ulcerprimary human skin fibroblast cellsbacterial invasionpathogenesisInfectious and parasitic diseasesRC109-216ENVirulence, Vol 11, Iss 1, Pp 1142-1157 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	staphylococcus haemolyticus diabetic foot ulcer primary human skin fibroblast cells bacterial invasion pathogenesis Infectious and parasitic diseases RC109-216
spellingShingle	staphylococcus haemolyticus diabetic foot ulcer primary human skin fibroblast cells bacterial invasion pathogenesis Infectious and parasitic diseases RC109-216 Hala O. Eltwisy Medhat Abdel-Fattah Amani M. Elsisi Mahmoud M. Omar Ahmed Aly Abdelmoteleb Mohamed A. El-Mokhtar Pathogenesis of Staphylococcus haemolyticus on primary human skin fibroblast cells
description	Staphylococcus haemolyticus (S. haemolyticus) is one of the Coagulase-negative staphylococci (CoNS) that inhabits the skin as a commensal. It is increasingly implicated in opportunistic infections, including diabetic foot ulcer (DFU) infections. In contrast to the abundance of information available for S. aureus and S. epidermidis, little is known about the pathogenicity of S. haemolyticus, despite the increased prevalence of this pathogen in hospitalized patients. We described, for the first time, the pathogenesis of different clinical isolates of S. haemolyticus isolated from DFU on primary human skin fibroblast (PHSF) cells. Virulence-related genes were investigated, adhesion and invasion assays were carried out using Giemsa stain, transmission electron microscopy (TEM), MTT and flowcytometry assays. Our results showed that most S. haemolyticus carried different sets of virulence-related genes. S. haemolyticus adhered to the PHSF cells to variable degrees. TEM showed that the bacteria were engulfed in a zipper-like mechanism into a vacuole inside the cell. Bacterial internalization was confirmed using flowcytometry and achieved high intracellular levels. PHSF cells infected with S.haemolyticus suffered from amarked decrease in viability and increased apoptosis when treated with whole bacterial suspensions or cell-free supernatants but not with heat-treated cells. After co-culture with PBMCs, S. haemolyticus induced high levels of pro-inflammatory cytokines. This study highlights the significant development of S. haemolyticus, which was previously considered a contaminant when detected in cultures of clinical samples. Their high ability to adhere, invade and kill the PHSF cells illustrate the severe damage associated with DFU infections. Abbreviations CoNS, coagulase-negative staphylococci; DFU, diabetic foot ulcer; DM, diabetes mellitus; DMEM, Dulbecco’s Modified Eagle Medium; MTT, 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide; PBMCs,peripheral blood mononuclear cells; PHSF, primary human skin fibroblast; CFU, colony-forming unit.
format	article
author	Hala O. Eltwisy Medhat Abdel-Fattah Amani M. Elsisi Mahmoud M. Omar Ahmed Aly Abdelmoteleb Mohamed A. El-Mokhtar
author_facet	Hala O. Eltwisy Medhat Abdel-Fattah Amani M. Elsisi Mahmoud M. Omar Ahmed Aly Abdelmoteleb Mohamed A. El-Mokhtar
author_sort	Hala O. Eltwisy
title	Pathogenesis of Staphylococcus haemolyticus on primary human skin fibroblast cells
title_short	Pathogenesis of Staphylococcus haemolyticus on primary human skin fibroblast cells
title_full	Pathogenesis of Staphylococcus haemolyticus on primary human skin fibroblast cells
title_fullStr	Pathogenesis of Staphylococcus haemolyticus on primary human skin fibroblast cells
title_full_unstemmed	Pathogenesis of Staphylococcus haemolyticus on primary human skin fibroblast cells
title_sort	pathogenesis of staphylococcus haemolyticus on primary human skin fibroblast cells
publisher	Taylor & Francis Group
publishDate	2020
url	https://doaj.org/article/a27df34e79b94c4dadd3c639da3d878c
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Pathogenesis of Staphylococcus haemolyticus on primary human skin fibroblast cells

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