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

Painter H., Prabowo SA., Cia F., Stockdale L., Tanner R., Willcocks S., Reljic R., Fletcher HA., Zelmer A.

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.

DOI

10.1038/s41598-020-60223-y

Type

Journal article

Journal

Scientific reports

Publication Date

02/2020

Volume

10

Addresses

Department of Infection Biology, London School of Hygiene and Tropical Medicine, Keppel Street, London, WC1E 7HT, UK. hannah.painter1@lshtm.ac.uk.

Keywords

Lung, Cells, Cultured, Animals, Mice, Inbred C57BL, Mycobacterium tuberculosis, BCG Vaccine, Immunization, Biological Assay, Cell Count

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