Optimisation of a functional mycobacterial growth-inhibition assay to improve its suitability for infant TB vaccine studies

The development of vaccines against tuberculosis continues to be hindered by the lack of correlates of protection. Immunity to Mycobacterium tuberculosis (M.tb) infection relies predominantly on cell mediated response, which is routinely measured using a read-out of host cytokine profiles. However,...

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Bibliographic Details
Published in:	Journal of immunological methods Vol. 394; no. 1-2; pp. 121 - 124
Main Authors:	Burl, S., Holder, B.S., Lo, B.K.M., Kampmann, B.
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 30-08-2013 Elsevier
Subjects:	Adult BCG Vaccine - immunology BCG-lux Humans Infant Microbiological Techniques - methods Mycobacterium tuberculosis Mycobacterium tuberculosis - growth & development Paediatric tuberculosis Technical Note Vaccine immunogenicity Vaccine immunogenicity Mycobacterium tuberculosis BCG-lux Paediatric tuberculosis
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Summary:	The development of vaccines against tuberculosis continues to be hindered by the lack of correlates of protection. Immunity to Mycobacterium tuberculosis (M.tb) infection relies predominantly on cell mediated response, which is routinely measured using a read-out of host cytokine profiles. However, to date none of the cytokine profiles have been found to predict protection. A number of functional in vitro approaches that measure growth of mycobacteria pre- and post-vaccination as a potential functional surrogate marker for vaccine take have been developed. The use of a reporter-gene tagged BCG-lux assay measuring the viability of mycobacteria in whole blood samples has previously been described by our group to assess vaccine immunogenicity. Since only very small blood samples are usually available in paediatric studies, we now report a modification of the BCG-lux assay to reduce the volume required and make it more field-friendly. Our results show that a 2-fold reduction in blood volume made no significant difference to bacterial growth ratios, used as the main read-out. These results confirm the suitability of the BCG-lux assay for functional studies of vaccine immunogenicity and immunopathogenesis in young children and could play a role in late-phase TB vaccine trials of novel candidates. •A 2-fold reduction in blood volume can now be used in a functional BCG-lux assay.•This growth-restriction assay is a useful tool for vaccine assessment.•Reduction in blood volumes will aid its use in trials of TB vaccines in infants.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0022-1759 1872-7905
DOI:	10.1016/j.jim.2013.05.006