Enhanced immunogenicity of Mycobacterium bovis BCG through CRISPRi mediated depletion of AftC

Bala T S A Madduri, Lauren Allen, Stephen C Taylor, Gurdyal S Besra*, Luke J Alderwick

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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Abstract

Mycobacterium tuberculosis causes the disease tuberculosis and affects a third of the world's population. The recent COVID-19 pandemic exacerbated the situation with a projected 27% increase in tuberculosis related deaths. M. tuberculosis has an elaborate cell wall consisting of peptidoglycan, arabinogalactan and mycolic acids which shield the bacilli from the toxic bactericidal milieu within phagocytes. Amongst, the numerous glycosyltransferase enzymes involved in mycobacterial cell wall biosynthesis, arabinofuranosyltransferase C ( aftC) is responsible for the branching of the arabinan domain in both arabinogalactan and lipoarabinomannan. Using Clustered Regularly Interspaced Short Palindromic Repeats interference (CRISPRi) we have generated aftC knockdowns in Mycobacterium bovis BCG and demonstrated the generation of a truncated, immunogenic lipoarabinomannan within its cell envelope. The aftC depleted BCG mutants were unable to form characteristic mycobacterial pellicular biofilms and elicit a potent immunostimulatory phenotype compared to wild type M. bovis BCG in a THP1 cell line. This study paves the way to further explore novel BCG mutants as promising vaccine boosters in preventing pulmonary tuberculosis.

Original languageEnglish
Article number100088
Number of pages11
JournalThe Cell Surface
Volume8
Early online date11 Nov 2022
DOIs
Publication statusPublished - Dec 2022

Bibliographical note

© 2022 The Author(s).

Keywords

  • Mycobacterium bovis BCG
  • CRISPR interference
  • Transcriptional repression
  • Arabinofuranosytransferase C
  • Lipoarabinomannan

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