The role of resuscitation promoting factors in pathogenesis and reactivation of Mycobacterium tuberculosis during intra-peritoneal infection in mice.
BACKGROUND
Mycobacterium tuberculosis can enter into a dormant state which has resulted in one third of the world's population being infected with latent tuberculosis making the study of latency and reactivation of utmost importance. M. tuberculosis encodes five resuscitation promoting factors (Rpfs) that bear strong similarity to a lysozyme-like enzyme previously implicated in reactivation of dormant bacteria in vitro. We have developed an intraperitoneal infection model in mice, with immune modulation, that models chronic infection with similar properties in mouse lungs as those observed in the murine aerosol infection model. We have assessed the behavior of mutants that lack two or three rpf genes in different combinations in our intraperitoneal model.


METHODS
C57Bl/6 mice were intraperitonealy infected with H37Rv wild type M. tuberculosis or mutant strains that lacked two or three rpf genes in different combinations. After 90 days of infection aminoguanidine (AG) or anti-TNFalpha antibodies were administrated. Organ bacillary loads were determined at various intervals post infection by plating serial dilutions of organ homogenates and enumerating bacteria.


RESULTS
We found that the rpf triple and double mutants tested were attenuated in their ability to disseminate to mouse lungs after intraperitoneal administration and were defective in their ability to re-grow after immunosuppression induced by administration of aminoguanidine and anti-TNFalpha antibodies.


CONCLUSIONS
Rpf proteins may have a significant physiological role for development of chronic TB infection and its reactivation in vivo.
Biketov
Sergey
S
Bakh Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia. biketov@obolensk.org
Potapov
Vasilii
V
Ganina
Elena
E
Downing
Katrina
K
Kana
Bavesh D
BD
Kaprelyants
Arseny
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002908:Chronic Disease; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006146:Guanidines / Q000494:pharmacology; D008168:Lung / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000472:pathogenicity* / Q000502:physiology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D010537:Peritoneum / Q000382:microbiology*; D017384:Sequence Deletion; D014397:Tuberculosis, Pulmonary / Q000276:immunology / Q000382:microbiology; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D016207:Cytokines; D004791:Enzyme Inhibitors; D006146:Guanidines; D014409:Tumor Necrosis Factor-alpha; C113704:resuscitation-promoting factor, bacteria; D019001:Nitric Oxide Synthase; C004479:pimagedine