Fig. 5.

ChAT⁺ NK cells influence the distribution of CCR2⁺Ly6C^hi monocytes and the cytokine microenvironment in the CNS of EAE mice. (A and B) The percentages of CNS-infiltrated neutrophils (CD11b⁺Ly6G⁺), CD8⁺ T cells, B cells (CD3⁻CD19⁺), and microglia (CD11b⁺CD45^low) were not changed by ChAT⁺ NK cell implantation. However, the numbers of infiltrated monocytes and CD4⁺ T cells (P = 0.083) decreased in ChAT⁺ NK cell-implanted groups. n = 6 per group. (C and D) The percentage of CCR2⁺Ly6C^hi monocytes in the CNS and spleen of CX3CR1^−/−, CX3CR1^−/− with ChAT⁻ NK cells, and CX3CR1^−/− mice with ChAT⁺ NK cells. ChAT⁺ NK cell implantation significantly decreased the numbers of CCR2⁺Ly6C^hi monocytes in the CNS without altering their distribution in the spleen. n = 6 per group. (E–I) MBP staining with CCR2 revealed a decrease of infiltrated CCR2⁺ monocytes/macrophages and reduced demyelination from ChAT⁺ NK cell treatment, without much effect on resident microglia (CX3CR1-GFP⁺). n = 9 per group. (J) Relative mRNA expression of TNF-α, IL-1β, IL-6, IL-12, and IL-10 to HPRT-1 as the internal standard in CNS. n = 6 per group. Data are representative of three independent experiments. Mean ± SEM. *P < 0.05. (Scale bars, 40 μm.)