Adaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells

Abstract In the absence of a correlate(s) of protection against human tuberculosis and a validated animal model of the disease, tools to facilitate vaccine development must be identified. We present an optimised ex vivo mycobacterial growth inhibition assay (MGIA) to assess the ability of host cells...

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Autores principales:	Hannah Painter, Satria A. Prabowo, Felipe Cia, Lisa Stockdale, Rachel Tanner, Samuel Willcocks, Rajko Reljic, Helen A. Fletcher, Andrea Zelmer
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8e428b49b409439e8ebb3565742e632f
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spelling	oai:doaj.org-article:8e428b49b409439e8ebb3565742e632f2021-12-02T10:59:52ZAdaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells10.1038/s41598-020-60223-y2045-2322https://doaj.org/article/8e428b49b409439e8ebb3565742e632f2020-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-60223-yhttps://doaj.org/toc/2045-2322Abstract In the absence of a correlate(s) of protection against human tuberculosis and a validated animal model of the disease, tools to facilitate vaccine development must be identified. We present an optimised ex vivo mycobacterial growth inhibition assay (MGIA) to assess the ability of host cells within the lung to inhibit mycobacterial growth, including Bacille Calmette–Guérin (BCG) and Mycobacterium tuberculosis (MTB) Erdman. Growth of BCG was reduced by 0.39, 0.96 and 0.73 log10 CFU following subcutaneous (s.c.) BCG, intranasal (i.n.) BCG, or BCG s.c. + mucosal boost, respectively, versus naïve mice. Comparatively, a 0.49 (s.c.), 0.60 (i.n.) and 0.81 (s.c. + mucosal boost) log10 reduction in MTB CFU was found. A BCG growth inhibitor, 2-thiophenecarboxylic acid hydrazide (TCH), was used to prevent quantification of residual BCG from i.n. immunisation and allow accurate MTB quantification. Using TCH, a further 0.58 log10 reduction in MTB CFU was revealed in the i.n. group. In combination with existing methods, the ex vivo lung MGIA may represent an important tool for analysis of vaccine efficacy and the immune mechanisms associated with vaccination in the organ primarily affected by MTB disease.Hannah PainterSatria A. PrabowoFelipe CiaLisa StockdaleRachel TannerSamuel WillcocksRajko ReljicHelen A. FletcherAndrea ZelmerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-9 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hannah Painter Satria A. Prabowo Felipe Cia Lisa Stockdale Rachel Tanner Samuel Willcocks Rajko Reljic Helen A. Fletcher Andrea Zelmer Adaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells
description	Abstract In the absence of a correlate(s) of protection against human tuberculosis and a validated animal model of the disease, tools to facilitate vaccine development must be identified. We present an optimised ex vivo mycobacterial growth inhibition assay (MGIA) to assess the ability of host cells within the lung to inhibit mycobacterial growth, including Bacille Calmette–Guérin (BCG) and Mycobacterium tuberculosis (MTB) Erdman. Growth of BCG was reduced by 0.39, 0.96 and 0.73 log10 CFU following subcutaneous (s.c.) BCG, intranasal (i.n.) BCG, or BCG s.c. + mucosal boost, respectively, versus naïve mice. Comparatively, a 0.49 (s.c.), 0.60 (i.n.) and 0.81 (s.c. + mucosal boost) log10 reduction in MTB CFU was found. A BCG growth inhibitor, 2-thiophenecarboxylic acid hydrazide (TCH), was used to prevent quantification of residual BCG from i.n. immunisation and allow accurate MTB quantification. Using TCH, a further 0.58 log10 reduction in MTB CFU was revealed in the i.n. group. In combination with existing methods, the ex vivo lung MGIA may represent an important tool for analysis of vaccine efficacy and the immune mechanisms associated with vaccination in the organ primarily affected by MTB disease.
format	article
author	Hannah Painter Satria A. Prabowo Felipe Cia Lisa Stockdale Rachel Tanner Samuel Willcocks Rajko Reljic Helen A. Fletcher Andrea Zelmer
author_facet	Hannah Painter Satria A. Prabowo Felipe Cia Lisa Stockdale Rachel Tanner Samuel Willcocks Rajko Reljic Helen A. Fletcher Andrea Zelmer
author_sort	Hannah Painter
title	Adaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells
title_short	Adaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells
title_full	Adaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells
title_fullStr	Adaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells
title_full_unstemmed	Adaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells
title_sort	adaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/8e428b49b409439e8ebb3565742e632f
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Adaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells

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