NLRC3 negatively regulates CD4+ T cells and impacts protective immunity during Mycobacterium tuberculosis infection
Fig 3
T cell activation in vitro requires NLRC3.
(A) Relative expression of Nlrc3 in purified macrophages (CD11b+ Gr-1-), dendritic cells (CD11c+ MHCIIhi), polymorphonuclear leukocytes (PMNs) (CD11b+ Gr-1+), CD4+ T cells (CD3+ CD4+) and CD8+ T cells (CD3+ CD8+). (B) Purified naïve T cells isolated from WT and Nlrc3-/- mice were stimulated with plate bound anti-CD3 (increasing concentrations) and anti-CD28 (1 μg/ml) for 48 hr and the incorporation of thymidine was measured during the final 8 hr. (C) Purified WT and Nlrc3-/- naïve CD4+ T cells were labeled with CFSE and stimulated with anti-CD3 (1 μg/ml) and anti-CD28 (1 μg/ml) for 3 d. (D) Concentrations of IL-2 in supernatants of purified WT and Nlrc3-/- naïve CD4+ T cells stimulated for 0–80 h with anti-CD3 (1 μg/ml) and anti-CD28 (1 μg/ml) were detected by ELISA. (E) Thymidine incorporation in purified WT and Nlrc3-/- naive CD4+ T cells first primed with anti-CD3 and CD28 and then cultured with various concentrations of IL-2. (F) Purified WT and Nlrc3-/- naïve CD4+ T cells were polarized in Th1 or Th17 culture conditions for 4 days. Data shown in (B, C, E, F) are the mean ±SD. *P < 0.05 and **P < 0.01. Data are representative of three independent experiments with similar results.