We found that upon NK cell depletion there was a significant increase in the spleen NKT (CD3+/CD1d+) cell population compared with NKTDEPL, C and S (P = 0.014, P = 0.021 and P = 0.033, respectively). Interestingly, upon NKT cell depletion, spleen NK (CD3-/NK1.1+) cells increased significantly compared with NKDEPL, C and S (P < 0.0001, P < 0.0001 and P = 0.001, respectively). NKT depletion led to decreased lymphocyte apoptosis compared with C (P = 0.035), higher bacterial load in the lung compared with C and NKDEPL (P = 0.014 and P = 0.022 respectively) and in the liver compared with C (P = 0.012). In addition, serum levels of IFNγ were significantly increased and splenocytes from NKT depleted animals, incubated ex vivo in the presence or absence of IL-2, produced more IFNγ in comparison with all other groups. Furthermore, splenocyte miRNA analysis showed that miR-200c and miR-29a were downregulated, while miR-125a-5p was upregulated, in the NKT depleted animals compared with all other groups.