Identification of two Mycobacterium tuberculosis H37Rv ORFs involved in resistance to killing by human macrophages.
BACKGROUND
The ability of Mycobacterium tuberculosis to survive and replicate in macrophages is crucial for the mycobacterium's ability to infect the host and cause tuberculosis. To identify Mycobacterium tuberculosis genes involved in survival in macrophages, a library of non-pathogenic Mycobacterium smegmatis bacteria, each carrying an individual integrated cosmid containing M. tuberculosis H37Rv genomic DNA, was passed through THP-1 human macrophages three times.


RESULTS
Two of the clones recovered from this enrichment process, sur2 and sur3, exhibited significantly increased survival relative to wild-type bacteria. In coinfection experiments, the ratio of sur2 colonies to wild-type colonies was 1:1 at 0 hours but increased to 20:1 at 24 hours post phagocytosis. The ratio of sur3 colonies to wild-type colonies was 1:1 at 0 hours and 5:1 at 24 hours. The M. tuberculosis ORFs responsible for increased survival were shown to be Rv0365c for the sur2 clone and Rv2235 for the sur3 clone. These ORFs encode proteins with as-of-yet unknown functions.


CONCLUSIONS
We identified two M. tuberculosis ORFs which may be involved in the ability of tubercle bacilli to survive in macrophages.
Miller
B H
BH
Department of Microbiology and Immunology, Emory University School of Medicine and Division of AIDS, STD, and TB Laboratory Research, Centers for Disease Control and Prevention, Atlanta, Georgia, USA. bhmiller@med.umich.edu
Shinnick
T M
TM
D006801:Humans; D008264:Macrophages / Q000382:microbiology / Q000502:physiology*; D006899:Mixed Function Oxygenases / Q000502:physiology*; D009169:Mycobacterium tuberculosis / Q000378:metabolism* / Q000502:physiology; D016366:Open Reading Frames; D010587:Phagocytosis / Q000502:physiology*
D006899:Mixed Function Oxygenases