A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates

A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates

Abstract We present a non-human primate mycobacterial growth inhibition assay (MGIA) using in vitro blood or cell co-culture with the aim of refining and expediting early tuberculosis vaccine testing. We have taken steps to optimise the assay using cryopreserved peripheral blood mononuclear cells, t...

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Autores principales: Rachel Tanner, Andrew D. White, Charelle Boot, Claudia C. Sombroek, Matthew K. O’Shea, Daniel Wright, Emily Hoogkamer, Julia Bitencourt, Stephanie A. Harris, Charlotte Sarfas, Rachel Wittenberg, Iman Satti, Helen A. Fletcher, Frank A. W. Verreck, Sally A. Sharpe, Helen McShane
Formato: article
Lenguaje:EN
Publicado: Nature Portfolio 2021
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Immunologic diseases. Allergy
RC581-607
Neoplasms. Tumors. Oncology. Including cancer and carcinogens
RC254-282
Acceso en línea:https://doaj.org/article/68af0e0e83b24984a10595f180e34dba
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spelling oai:doaj.org-article:68af0e0e83b24984a10595f180e34dba2021-12-02T15:08:21ZA non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates10.1038/s41541-020-00263-72059-0105https://doaj.org/article/68af0e0e83b24984a10595f180e34dba2021-01-01T00:00:00Zhttps://doi.org/10.1038/s41541-020-00263-7https://doaj.org/toc/2059-0105Abstract We present a non-human primate mycobacterial growth inhibition assay (MGIA) using in vitro blood or cell co-culture with the aim of refining and expediting early tuberculosis vaccine testing. We have taken steps to optimise the assay using cryopreserved peripheral blood mononuclear cells, transfer it to end-user institutes, and assess technical and biological validity. Increasing cell concentration or mycobacterial input and co-culturing in static 48-well plates compared with rotating tubes improved intra-assay repeatability and sensitivity. Standardisation and harmonisation efforts resulted in high consistency agreements, with repeatability and intermediate precision <10% coefficient of variation (CV) and inter-site reproducibility <20% CV; although some systematic differences were observed. As proof-of-concept, we demonstrated ability to detect a BCG vaccine-induced improvement in growth inhibition in macaque samples, and a correlation between MGIA outcome and measures of protection from in vivo disease development following challenge with either intradermal BCG or aerosol/endobronchial Mycobacterium tuberculosis (M.tb) at a group and individual animal level.Rachel TannerAndrew D. WhiteCharelle BootClaudia C. SombroekMatthew K. O’SheaDaniel WrightEmily HoogkamerJulia BitencourtStephanie A. HarrisCharlotte SarfasRachel WittenbergIman SattiHelen A. FletcherFrank A. W. VerreckSally A. SharpeHelen McShaneNature PortfolioarticleImmunologic diseases. AllergyRC581-607Neoplasms. Tumors. Oncology. Including cancer and carcinogensRC254-282ENnpj Vaccines, Vol 6, Iss 1, Pp 1-11 (2021)
institution DOAJ
collection DOAJ
language EN
topic Immunologic diseases. Allergy
RC581-607
Neoplasms. Tumors. Oncology. Including cancer and carcinogens
RC254-282
spellingShingle Immunologic diseases. Allergy
RC581-607
Neoplasms. Tumors. Oncology. Including cancer and carcinogens
RC254-282
Rachel Tanner
Andrew D. White
Charelle Boot
Claudia C. Sombroek
Matthew K. O’Shea
Daniel Wright
Emily Hoogkamer
Julia Bitencourt
Stephanie A. Harris
Charlotte Sarfas
Rachel Wittenberg
Iman Satti
Helen A. Fletcher
Frank A. W. Verreck
Sally A. Sharpe
Helen McShane
A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates
description Abstract We present a non-human primate mycobacterial growth inhibition assay (MGIA) using in vitro blood or cell co-culture with the aim of refining and expediting early tuberculosis vaccine testing. We have taken steps to optimise the assay using cryopreserved peripheral blood mononuclear cells, transfer it to end-user institutes, and assess technical and biological validity. Increasing cell concentration or mycobacterial input and co-culturing in static 48-well plates compared with rotating tubes improved intra-assay repeatability and sensitivity. Standardisation and harmonisation efforts resulted in high consistency agreements, with repeatability and intermediate precision <10% coefficient of variation (CV) and inter-site reproducibility <20% CV; although some systematic differences were observed. As proof-of-concept, we demonstrated ability to detect a BCG vaccine-induced improvement in growth inhibition in macaque samples, and a correlation between MGIA outcome and measures of protection from in vivo disease development following challenge with either intradermal BCG or aerosol/endobronchial Mycobacterium tuberculosis (M.tb) at a group and individual animal level.
format article
author Rachel Tanner
Andrew D. White
Charelle Boot
Claudia C. Sombroek
Matthew K. O’Shea
Daniel Wright
Emily Hoogkamer
Julia Bitencourt
Stephanie A. Harris
Charlotte Sarfas
Rachel Wittenberg
Iman Satti
Helen A. Fletcher
Frank A. W. Verreck
Sally A. Sharpe
Helen McShane
author_facet Rachel Tanner
Andrew D. White
Charelle Boot
Claudia C. Sombroek
Matthew K. O’Shea
Daniel Wright
Emily Hoogkamer
Julia Bitencourt
Stephanie A. Harris
Charlotte Sarfas
Rachel Wittenberg
Iman Satti
Helen A. Fletcher
Frank A. W. Verreck
Sally A. Sharpe
Helen McShane
author_sort Rachel Tanner
title A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates
title_short A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates
title_full A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates
title_fullStr A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates
title_full_unstemmed A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates
title_sort non-human primate in vitro functional assay for the early evaluation of tb vaccine candidates
publisher Nature Portfolio
publishDate 2021
url https://doaj.org/article/68af0e0e83b24984a10595f180e34dba
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