NLRC3 negatively regulates CD4+ T cells and impacts protective immunity during Mycobacterium tuberculosis infection
Fig 5
NLRC3 deficiency promotes the antibacterial ability in vivo via regulating CD4+ T cells.
Purified WT or Nlrc3-/- naïve CD4+ T cells were adoptively transferred into Rag2-/- mice. Then recipient mice were infected with M. tuberculosis and parts of mice were harvested at 3w.p.i.. (A) Bacterial burdens were determined in lungs and spleen at 3w.p.i.. (B) Survival of mice every other day from 0–150 day post-infection (d.p.i.). (C) Lung cells were restimulated with M. tuberculosis lysate directly ex vivo and the intracellular production of IFN-γ, IL-2, and TNF-α by CD4+ T cells was determined. (D) Expression of activation markers by lung CD4+ T cells. (E) Concentration of IFN-γ, IL-2 and TNF-α in lungs (homogenized in 2 ml PBS and 0.05% Tween 80) as detected by ELISA. (F) ROS production by monocyte-macrophages (CD11b+ Gr-1-) were detected assessed as mean fluorescence intensity (MFI) of intracellular CFDA. (G) Concentrations of nitrate were measured by nitrate reductase assay and concentrations of IL-6 and IL-1β in lungs (homogenized in 2 ml PBS and 0.05% Tween 80) were detected by ELISA. Data shown in (A, E, G) are the mean ±SD. *P < 0.05, **P < 0.01 and ***P < 0.001. Data are representative of three independent experiments with similar results.