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

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 Sharpe; Helen McShane

doi:10.1038/s41541-020-00263-7

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

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 Sharpe, Helen McShane

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

9 Citations (Scopus)

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.

Original language	English
Article number	3
Number of pages	11
Journal	npj Vaccines
Volume	6
Issue number	1
DOIs	https://doi.org/10.1038/s41541-020-00263-7
Publication status	Published - 4 Jan 2021

Bibliographical note

Funding Information:
This work was funded by a grant awarded to RT by the National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) (grant number NC/R000905/1); and in part by a studentship awarded to RT by the Universities Federation for Animal Welfare (UFAW). Aspects of this work were conducted as part of the European Research Infrastructures for Poverty Related Diseases (EURIPRED), an EC seventh framework program (grant number 312661). HMcS is a Wellcome Trust Investigator (grant number WT 206331/Z/17/Z). We would like to thank Aeras for supporting the early MGIA development work and NHP Study 3, and for providing and distributing a standardised BCG Pasteur stock. We are grateful to Krista van Meijgaarden and Tom Ottenhoff (Leiden University Medical Centre) and to Marcus Morgan (Oxford University Hospitals NHS Trust) for use of their BD BACTEC MGIT machines for parts of this project. We would also like to thank Michael Andrews (University of California, Los Angeles) for advice on some of the statistical aspects.

Keywords

MYCOBACTERIAL GROWTH-INHIBITION
PERIPHERAL-BLOOD MONOCYTES
TUBERCULOSIS VACCINE
BCG-VACCINATION
WHOLE-BLOOD
PROTECTION
INFECTION
IMMUNITY
CELLS
BOVIS

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1038/s41541-020-00263-7

Cite this

Tanner, R., White, A. D., Boot, C., Sombroek, C. C., O’Shea, M. K., Wright, D., Hoogkamer, E., Bitencourt, J., Harris, S. A., Sarfas, C., Wittenberg, R., Satti, I., Fletcher, H. A., Verreck, F. A. W., Sharpe, S., & McShane, H. (2021). A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates. npj Vaccines, 6(1), Article 3. https://doi.org/10.1038/s41541-020-00263-7

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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.",

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author = "Rachel Tanner and White, {Andrew D.} and Charelle Boot and Sombroek, {Claudia C.} and O{\textquoteright}Shea, {Matthew K.} and Daniel Wright and Emily Hoogkamer and Julia Bitencourt and Harris, {Stephanie A.} and Charlotte Sarfas and Rachel Wittenberg and Iman Satti and Fletcher, {Helen A.} and Verreck, {Frank A.W.} and Sally Sharpe and Helen McShane",

note = "Funding Information: This work was funded by a grant awarded to RT by the National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) (grant number NC/R000905/1); and in part by a studentship awarded to RT by the Universities Federation for Animal Welfare (UFAW). Aspects of this work were conducted as part of the European Research Infrastructures for Poverty Related Diseases (EURIPRED), an EC seventh framework program (grant number 312661). HMcS is a Wellcome Trust Investigator (grant number WT 206331/Z/17/Z). We would like to thank Aeras for supporting the early MGIA development work and NHP Study 3, and for providing and distributing a standardised BCG Pasteur stock. We are grateful to Krista van Meijgaarden and Tom Ottenhoff (Leiden University Medical Centre) and to Marcus Morgan (Oxford University Hospitals NHS Trust) for use of their BD BACTEC MGIT machines for parts of this project. We would also like to thank Michael Andrews (University of California, Los Angeles) for advice on some of the statistical aspects.",

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Tanner, R, White, AD, Boot, C, Sombroek, CC, O’Shea, MK, Wright, D, Hoogkamer, E, Bitencourt, J, Harris, SA, Sarfas, C, Wittenberg, R, Satti, I, Fletcher, HA, Verreck, FAW, Sharpe, S & McShane, H 2021, 'A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates', npj Vaccines, vol. 6, no. 1, 3. https://doi.org/10.1038/s41541-020-00263-7

TY - JOUR

T1 - A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates

AU - Tanner, Rachel

AU - White, Andrew D.

AU - Boot, Charelle

AU - Sombroek, Claudia C.

AU - O’Shea, Matthew K.

AU - Wright, Daniel

AU - Hoogkamer, Emily

AU - Bitencourt, Julia

AU - Harris, Stephanie A.

AU - Sarfas, Charlotte

AU - Wittenberg, Rachel

AU - Satti, Iman

AU - Fletcher, Helen A.

AU - Verreck, Frank A.W.

AU - Sharpe, Sally

AU - McShane, Helen

N1 - Funding Information: This work was funded by a grant awarded to RT by the National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) (grant number NC/R000905/1); and in part by a studentship awarded to RT by the Universities Federation for Animal Welfare (UFAW). Aspects of this work were conducted as part of the European Research Infrastructures for Poverty Related Diseases (EURIPRED), an EC seventh framework program (grant number 312661). HMcS is a Wellcome Trust Investigator (grant number WT 206331/Z/17/Z). We would like to thank Aeras for supporting the early MGIA development work and NHP Study 3, and for providing and distributing a standardised BCG Pasteur stock. We are grateful to Krista van Meijgaarden and Tom Ottenhoff (Leiden University Medical Centre) and to Marcus Morgan (Oxford University Hospitals NHS Trust) for use of their BD BACTEC MGIT machines for parts of this project. We would also like to thank Michael Andrews (University of California, Los Angeles) for advice on some of the statistical aspects.

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Y1 - 2021/1/4

N2 - 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.

AB - 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.

KW - MYCOBACTERIAL GROWTH-INHIBITION

KW - PERIPHERAL-BLOOD MONOCYTES

KW - TUBERCULOSIS VACCINE

KW - BCG-VACCINATION

KW - WHOLE-BLOOD

KW - PROTECTION

KW - INFECTION

KW - IMMUNITY

KW - CELLS

KW - BOVIS

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U2 - 10.1038/s41541-020-00263-7

DO - 10.1038/s41541-020-00263-7

M3 - Article

AN - SCOPUS:85098629318

SN - 2059-0105

VL - 6

JO - npj Vaccines

JF - npj Vaccines

IS - 1

M1 - 3

ER -

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

Abstract

Bibliographical note

Keywords

UN SDGs

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