Usefulness of acr expression for monitoring latent Mycobacterium tuberculosis bacilli in 'in vitro' and 'in vivo' experimental models.
Real-time RT-PCR was used to quantify the expression of genes possibly involved in Mycobacterium tuberculosis latency in in vitro and murine models. Exponential and stationary phase (EP and SP) bacilli were exposed to decreasing pH levels (from 6.5 to 4.5) in an unstirred culture, and mRNA levels for 16S rRNA, sigma factors sigA,B,E,F,G,H and M, Rv0834c, icl, nirA, narG, fpbB, acr, rpoA, recA and cysH were quantified. The expression of acr was the one that best correlated with the CFU decrease observed in SP bacilli. In the murine model, the expressions of icl, acr and sigF tended to decrease when bacillary counts increased and vice versa. Values from immunodepressed mice (e.g. alpha/beta T cells, TNF, IFN-gamma and iNOs knock out strains), with accelerated bacillary growth rate, confirmed this fact. Finally, the expression of acr was maintained in mice following long-term treatment with antibiotics. The quantification of acr expression could be useful for monitoring the presence of latent bacilli in some murine models of tuberculosis.
Gordillo
S
S
Unitat de Tuberculosi Experimental, Department of Microbiology, Fundació Institut per a la Investigació en Ciències de la Salut Germans Trias i Pujol, Badalona, Spain.
Guirado
E
E
Gil
O
O
Díaz
J
J
Amat
I
I
Molinos
S
S
Vilaplana
C
C
Ausina
V
V
Cardona
P-J
PJ
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D004195:Disease Models, Animal; D005260:Female; D015964:Gene Expression Regulation, Bacterial; D005819:Genetic Markers; D007522:Isocitrate Lyase / Q000235:genetics; D051379:Mice; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000254:growth & development*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012808:Sigma Factor / Q000235:genetics; D013047:Specific Pathogen-Free Organisms; D014376:Tuberculosis / Q000382:microbiology*; D038201:alpha-Crystallins / Q000235:genetics*
D001426:Bacterial Proteins; C064971:FliA protein, Bacteria; D005819:Genetic Markers; D012808:Sigma Factor; D038201:alpha-Crystallins; C402427:icl protein, Mycobacterium; D007522:Isocitrate Lyase