RESUMO
Macrophages are the primary phagocytes in the lungs and a part of the host defense system against Mycobacterium tuberculosis (Mtb), involved in the primary immune response. While several studies have assessed the effects of resistance to rifampin on Mtb physiology, the consequences of mutations in genes encoding the beta subunit of RNA polymerase (rpoB) for host-pathogen interactions remain poorly understood. In this study, rifampin-monoresistant (RMR) Mtb and H37Rv strains were used to infect the THP-1-derived macrophages. Real-time quantitative reverse transcription PCR assay was carried out to determine mRNA expression in 84 cytokine and chemokine genes. Production of specific cytokines and chemokines was measured by ELISA assay. In conclusion, the current study shed more light on the fitness cost of RMR strain and the potential effects of rpoB gene mutations on Mtb-host interactions. These results initially demonstrate that the Mtb carrying the rpoB-S450L can modulate macrophage responses to mediate bacterial survival.
Assuntos
Quimiocinas/genética , Citocinas/genética , Farmacorresistência Bacteriana/genética , Macrófagos/metabolismo , Mycobacterium tuberculosis/genética , Rifampina/farmacologia , Proteínas de Bactérias/genética , Linhagem Celular , RNA Polimerases Dirigidas por DNA/genética , Interações Hospedeiro-Patógeno/genética , Humanos , Mutação/genética , Mycobacterium tuberculosis/efeitos dos fármacos , Células Th1RESUMO
BACKGROUND: The increasing prevalence of TB drug resistant strains in absence of recent transmission evidence, highlights the need for an improved control program, coupled with a need to improve detection rate and early diagnosis. IS6110-RFLP is a means of genotyping TB clinical samples. In this study IS6110- RFLP was used for specification and quick tracking of TB infection source, transmission and reactivation of infection, in Iran. MATERIALS AND METHODS: This study was carried out on 258 TB patients from Tehran, Mashhad, Isfahan, Shiraz and Ahwaz. DNA from positive cultures was extracted and digested by PVUII restriction enzyme. The digested sequences were separated based on the size on agar gel and then southern Blot was transferred on the membrane. IS6110 probe was marked by HRD and hybridized to the target parts along genome. RESULTS: Sixty-one strains (24%) showed similar patterns (Recent transmission) and 197 strains (76%) showed different IS6110 patterns (Reactivation). Average number of IS6110 copies was between 10-11 bands. Frequency of IS6110 similar pattern was 11.46 in Afghan immigrants and 10.68 in Iranians. CONCLUSION: High diversity of IS6110, indicates that 76% of the patients have been infected through reactivation by different sources, while 24% have been infected due to recent transmission. Observing different antibiogram patterns in patients infected with the same strain indicated vast transmission of a single strain in the society. A susceptible strain can be changed into mono drug resistant and MDR strain in the transition period.
