An ex vivo culture model for screening drug activity against in vivo phenotypes of Mycobacterium tuberculosis

Abstract

Since the activity of drugs against Mycobacterium tuberculosis grown in microbiological culture can differ from their activity against bacteria present in infected tissues, compounds with optimal activity against in vivo phenotypes may be overlooked in drug-discovery programmes that rely on in vitro screens. The authors have investigated the use of an ex vivo cell-culture model to assess the action of drugs on M. tuberculosis in an environment resembling that encountered during infection. Mycobacterial viability in the ex vivo model was shown to be regulated by the cell-mediated immune system, with growth inhibited by CD4(+) T cells at an early stage of infection in BCG-vaccinated mice, and at a later stage after infection in naive mice. Screening of drugs in the ex vivo model demonstrated a window of pyrazinamide susceptibility that coincides with the onset of the T-cell-mediated immune response in naive or vaccinated mice. It is proposed that pyrazinamide acts on a population of bacteria that are exposed to an acidic environment as a result of immune activation. Clinically, administration of pyrazinamide during the initial phase of treatment reduces the risk of relapse after 6 months, suggesting that the early pyrazinamide-susceptible population may contribute to the later pool of mycobacteria that persist during prolonged chemotherapy.