<named-content content-type="genus-species">Pneumocystis jirovecii</named-content> Can Be Productively Cultured in Differentiated CuFi-8 Airway Cells

ABSTRACT Although Pneumocystis jirovecii is a well-known and serious pathogen, all previous attempts to isolate, cultivate, and propagate this fungus have failed. This serious challenge in microbiology was addressed in the present study. We examined whether P. jirovecii could be cultured in a perman...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Verena Schildgen, Stephanie Mai, Soumaya Khalfaoui, Jessica Lüsebrink, Monika Pieper, Ramona L. Tillmann, Michael Brockmann, Oliver Schildgen
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/c5bd7c36d1bc4bd08ae61b4ee498b446
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:c5bd7c36d1bc4bd08ae61b4ee498b446
record_format	dspace
spelling	oai:doaj.org-article:c5bd7c36d1bc4bd08ae61b4ee498b4462021-11-15T15:47:38Z<named-content content-type="genus-species">Pneumocystis jirovecii</named-content> Can Be Productively Cultured in Differentiated CuFi-8 Airway Cells10.1128/mBio.01186-142150-7511https://doaj.org/article/c5bd7c36d1bc4bd08ae61b4ee498b4462014-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01186-14https://doaj.org/toc/2150-7511ABSTRACT Although Pneumocystis jirovecii is a well-known and serious pathogen, all previous attempts to isolate, cultivate, and propagate this fungus have failed. This serious challenge in microbiology was addressed in the present study. We examined whether P. jirovecii could be cultured in a permanent three-dimensional air-liquid interface culture system formed by CuFi-8 cells, a differentiated pseudostratified airway epithelial cell line. Cultured pseudostratified cells were inoculated with bronchoalveolar fluid that had been confirmed to be positive for P. jirovecii using PCR. Five days later, the cells and basal medium were harvested and tested for P. jirovecii using quantitative PCR (qPCR), commercially available immunofluorescence detection assays, and Grocott staining of formalin-fixed, paraffin-embedded thin sections of infected-cell cultures. We successfully productively cultivated and propagated P. jirovecii from these P. jirovecii-positive bronchoalveolar lavage fluid (BALF) samples. Furthermore, we provide evidence that P. jirovecii induced cytopathic effects on lung epithelial cells and was even invasive in cell culture. To the best of our knowledge, the cell culture system developed herein represents the first methodology to enable molecular analyses of this pathogen’s life cycle and further in vitro studies of P. jirovecii, such as assessments of drug sensitivity and resistance as well as investigations of the pathogen’s stability against environmental factors and disinfectants. IMPORTANCE This is the first report of the successful productive cultivation and propagation of Pneumocystis jirovecii, a human-pathogenic fungus of major clinical significance. These findings are groundbreaking because they will influence the field of diagnostic microbiology, facilitate the testing of antibiotics against P. jirovecii, and enable stability studies of this pathogen when exposed to the environmental factors and chemicals that hospitals are required to use for disinfection. Because productively culturing P. jirovecii has been attempted unsuccessfully for several decades, this study represents a breakthrough in this field.Verena SchildgenStephanie MaiSoumaya KhalfaouiJessica LüsebrinkMonika PieperRamona L. TillmannMichael BrockmannOliver SchildgenAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 3 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Verena Schildgen Stephanie Mai Soumaya Khalfaoui Jessica Lüsebrink Monika Pieper Ramona L. Tillmann Michael Brockmann Oliver Schildgen <named-content content-type="genus-species">Pneumocystis jirovecii</named-content> Can Be Productively Cultured in Differentiated CuFi-8 Airway Cells
description	ABSTRACT Although Pneumocystis jirovecii is a well-known and serious pathogen, all previous attempts to isolate, cultivate, and propagate this fungus have failed. This serious challenge in microbiology was addressed in the present study. We examined whether P. jirovecii could be cultured in a permanent three-dimensional air-liquid interface culture system formed by CuFi-8 cells, a differentiated pseudostratified airway epithelial cell line. Cultured pseudostratified cells were inoculated with bronchoalveolar fluid that had been confirmed to be positive for P. jirovecii using PCR. Five days later, the cells and basal medium were harvested and tested for P. jirovecii using quantitative PCR (qPCR), commercially available immunofluorescence detection assays, and Grocott staining of formalin-fixed, paraffin-embedded thin sections of infected-cell cultures. We successfully productively cultivated and propagated P. jirovecii from these P. jirovecii-positive bronchoalveolar lavage fluid (BALF) samples. Furthermore, we provide evidence that P. jirovecii induced cytopathic effects on lung epithelial cells and was even invasive in cell culture. To the best of our knowledge, the cell culture system developed herein represents the first methodology to enable molecular analyses of this pathogen’s life cycle and further in vitro studies of P. jirovecii, such as assessments of drug sensitivity and resistance as well as investigations of the pathogen’s stability against environmental factors and disinfectants. IMPORTANCE This is the first report of the successful productive cultivation and propagation of Pneumocystis jirovecii, a human-pathogenic fungus of major clinical significance. These findings are groundbreaking because they will influence the field of diagnostic microbiology, facilitate the testing of antibiotics against P. jirovecii, and enable stability studies of this pathogen when exposed to the environmental factors and chemicals that hospitals are required to use for disinfection. Because productively culturing P. jirovecii has been attempted unsuccessfully for several decades, this study represents a breakthrough in this field.
format	article
author	Verena Schildgen Stephanie Mai Soumaya Khalfaoui Jessica Lüsebrink Monika Pieper Ramona L. Tillmann Michael Brockmann Oliver Schildgen
author_facet	Verena Schildgen Stephanie Mai Soumaya Khalfaoui Jessica Lüsebrink Monika Pieper Ramona L. Tillmann Michael Brockmann Oliver Schildgen
author_sort	Verena Schildgen
title	<named-content content-type="genus-species">Pneumocystis jirovecii</named-content> Can Be Productively Cultured in Differentiated CuFi-8 Airway Cells
title_short	<named-content content-type="genus-species">Pneumocystis jirovecii</named-content> Can Be Productively Cultured in Differentiated CuFi-8 Airway Cells
title_full	<named-content content-type="genus-species">Pneumocystis jirovecii</named-content> Can Be Productively Cultured in Differentiated CuFi-8 Airway Cells
title_fullStr	<named-content content-type="genus-species">Pneumocystis jirovecii</named-content> Can Be Productively Cultured in Differentiated CuFi-8 Airway Cells
title_full_unstemmed	<named-content content-type="genus-species">Pneumocystis jirovecii</named-content> Can Be Productively Cultured in Differentiated CuFi-8 Airway Cells
title_sort	<named-content content-type="genus-species">pneumocystis jirovecii</named-content> can be productively cultured in differentiated cufi-8 airway cells
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/c5bd7c36d1bc4bd08ae61b4ee498b446
work_keys_str_mv	AT verenaschildgen namedcontentcontenttypegenusspeciespneumocystisjiroveciinamedcontentcanbeproductivelyculturedindifferentiatedcufi8airwaycells AT stephaniemai namedcontentcontenttypegenusspeciespneumocystisjiroveciinamedcontentcanbeproductivelyculturedindifferentiatedcufi8airwaycells AT soumayakhalfaoui namedcontentcontenttypegenusspeciespneumocystisjiroveciinamedcontentcanbeproductivelyculturedindifferentiatedcufi8airwaycells AT jessicalusebrink namedcontentcontenttypegenusspeciespneumocystisjiroveciinamedcontentcanbeproductivelyculturedindifferentiatedcufi8airwaycells AT monikapieper namedcontentcontenttypegenusspeciespneumocystisjiroveciinamedcontentcanbeproductivelyculturedindifferentiatedcufi8airwaycells AT ramonaltillmann namedcontentcontenttypegenusspeciespneumocystisjiroveciinamedcontentcanbeproductivelyculturedindifferentiatedcufi8airwaycells AT michaelbrockmann namedcontentcontenttypegenusspeciespneumocystisjiroveciinamedcontentcanbeproductivelyculturedindifferentiatedcufi8airwaycells AT oliverschildgen namedcontentcontenttypegenusspeciespneumocystisjiroveciinamedcontentcanbeproductivelyculturedindifferentiatedcufi8airwaycells
_version_	1718427548545712128

<named-content content-type="genus-species">Pneumocystis jirovecii</named-content> Can Be Productively Cultured in Differentiated CuFi-8 Airway Cells

Ejemplares similares