Localisation of mycobacterial DNA and mRNA in human tuberculous granulomas

Gael Fenhalls, Liesel Stevens-Muller, Rob Warren, Nora Carroll, Juanita Bezuidenhout, Paul Van Helden, Philip Bardin

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Abstract

In situ hybridisation was used to detect the presence of Mycobacterium tuberculosis in paraffin-embedded lung tissue of nine patients diagnosed with tuberculosis (TB). Mycobacterial DNA was found in all nine patients and in 175 out of 191 granulomas examined. A combination of in situ hybridisation and immunohistochemistry techniques demonstrated that mycobacterial DNA was associated with CD68-positive cells with the morphology of macrophages and giant cells. Mycobacterial DNA was also found within the necrotic regions of some granulomas. mRNA for the mycobacterial RNA polymerase β subunit (rpoB) was detected by RNA:RNA in situ hybridisation. The rpoB mRNA was also localised to CD68-positive cells with the morphology of macrophages and to giant cells of certain necrotic granulomas. No rpoB mRNA was found in the necrotic regions of granulomas. Mycobacterial DNA was detected in 92% of patient granulomas of which 8% were positive for rpoB mRNA. The ability to identify mycobacterial RNA transcripts within human tuberculous granulomas affords us the opportunity to analyse the interplay between pathogen gene expression and the human immune response and should provide valuable insight into the mechanisms used by M. tuberculosis to persist within the human host.

Original languageEnglish
Pages (from-to)197-208
Number of pages12
JournalJournal of Microbiological Methods
Volume51
Issue number2
DOIs
Publication statusPublished - 3 Aug 2002

Keywords

  • Granuloma
  • Human lung
  • mRNA
  • Mycobacterial DNA
  • Tuberculosis

Cite this

Fenhalls, G., Stevens-Muller, L., Warren, R., Carroll, N., Bezuidenhout, J., Van Helden, P., & Bardin, P. (2002). Localisation of mycobacterial DNA and mRNA in human tuberculous granulomas. Journal of Microbiological Methods, 51(2), 197-208. https://doi.org/10.1016/S0167-7012(02)00076-3