The truncated Rv2820c of Mycobacterium tuberculosis Beijing family augments intracellular survival of M. smegmatis by altering cytokine profile and inhibiting NO generation.

Genetic variations among genes of Mycobacterium tuberculosis may be associated with antigenic variation and immune evasion, which complicates the pathogenesis of M. tuberculosis. The hyper-virulent M. tuberculosis Beijing strains harbored several large sequence deletions, among which RD207 attributed to the deletion of CRISPR loci and several Cas genes. RD207 also gave rise to a truncated gene Rv2820c-Bj with 60% deletion in length at the 3'-end and a new 3'-end of five amino acid mutations. It has been reported that Rv2820c-Bj correlated with enhanced intracellular survival of M. smegmatis in macrophages when compared to its full-length counterpart Rv2820c in M. tuberculosis, however, the respective contribution of the truncation and the new 3'-end of Rv2820c-Bj to this enhancement was unclear. Here, by infecting THP-1 macrophages with Ms_Rv2820c-Bj, Ms_Rv2820c and MS_Rv2820c-Tr (expressing the truncated Rv2820c without five amino acid mutations at 3'-end), we found only Ms_Rv2820c-Bj was responsible for the enhancement of survival of M. smegmatis in macrophages. Furthermore, we detected that Ms_Rv2820c-Tr and Ms_Rv2820c-Bj induced similar cytokine profile and NO production after infection of macrophages, which was distinctly different from Ms_Rv2820c. However, Ms_Rv2820c-Bj evoked higher levels of interleukin-10 (IL-10) and lower levels of interleukin- 6 (IL-6), interleukin-1ß (IL-1ß) and interleukin-12 (IL-12) in infected THP-1 macrophages than Ms_Rv2820c-Tr. Accordingly, we concluded that the new 3'-end of Rv2820c-Bj was important to dampen host defense and enhance the intracellular survival of M. smegmatis.

Origin, phylogeny, and spread of Mycobacterium tuberculosis Beijing lineage in the five provinces of northern China / 中华流行病学杂志

Objective Using methodology of molecular genetics to explore the origin,phylogen,and gene flow of Mycobacterium tuberculosis (MTB) Beijing lineage in the five provinces from northern China,including Heilongjiang,Jilin,Liaoning,Neimenggu and Ningxia.Methods 234 MTB Beijing lineage strains were genotyped by 24 Variable Number Tandem Repeat (VNTR),and the h (the allelic diversity) value of each VNTR locus was calculated.On individual level of phylogeny,it was constructed Neighbor-Joining (N-J) tree and minimum spanning tree (MST).Phylogenetic tree was built at the population level,and the most recent common ancestor (TMRCA)was estimated through Bayesian model.Molecular variance (AMOVA) was used to understand the gene flow among strains discovered from the five provinces.Results Allelic diversities of the 24VNTR loci were low (h:0.000-0.744).234 strains of MTB Beijing lineage were dispersed in individual branch of the N-J tree,with 62.0％ (145) of them grouped to the same "colonial complexes" in MST.At the population level,the evolution relationship of 234 strains appeared the closest to Beijing lineage,which was from MIRU-VNTRplus database,and the bootstrap was 100.The TMRCA was 5308 (95％ CI:4263-6470) years.Differences of pairwise Fst values acquired by AMOVA between Jilin and Heilongjiang,Liaoning,Neimenggu and Ningxia,were not statistically significant (P＞0.05).Conclusion The genetic similarity of Beijing lineage MTB from the five provinces of northern China was high.The phylogeny branches had no characteristic dispersal in each province.It was speculated that these strains showed an evolution from a clone of MTB Beijing lineage (about 5000 years ago).The gene flow was taking place between neighboring zones.