Figure 6.

Cold exposure reduces monocyte and T cell pathogenicity during EAE

(A) Volcano plot identifying the up- and downregulated transcripts after RNA sequencing of monocytes that were MACS (anti-PE) and FACS (CD115-PE⁺) sorted from blood of mice at onset of EAE (as in Figure 4A) and exposed to cold (10°C) or room temperature for 2 weeks before and continued during EAE.

(B and C) Top deregulated and enriched MetaCore Metabolic Networks (B, top), GO Cellular Components (B, middle), Reactome pathways (B, bottom), and GO processes (C). Genes were considered when p < 0.05 and for Reactome pathways when p < 0.05 and FC > 2. Data of sequencing as in (A).

(D and E) Flow cytometry analysis of blood cells of mice as in (A) at EAE onset. Percentage of Ly6C high (left), intermediate (middle), and low (right) monocytes of total, single CD45⁺ alive cells (D). Percentage of MHCII⁺ cells of Ly6C^hi monocytes (left), MHCII⁺ Ly6C⁺ monocytes of total MHCII⁺ cells (middle), and total MHCII⁺ cells of CD45⁺ cells (E). Pool of 3 experiments.

(F–H) Flow cytometry analysis of CNS cells from mice as in (A) at EAE onset. Percentage of Ly6C^hi monocytes/monocyte-derived cells of total, single CD45⁺ alive cells (F, left). Percentage of MHCII, arginase 1 (Arg1), and iNOS expression of Ly6C^hi monocytes/monocyte-derived cells (F, right). Percentage of microglia of total, single CD45⁺ alive cells and percentage of MHCII of microglia (G). Percentage of corresponding cytokine expression as indicated in CD4⁺ T cells (H).

(I) Flow cytometry analysis of dLN cells from mice as in (A) at EAE onset. Percentage of cytokine expression in CD4⁺ T cells. Observed in 2 out of 3 experiments.

(J) Flow cytometry analysis of dLN cells from mice as in (A) on day 2 after EAE induction. Percentage of Ly6C^hi monocytes of total, single CD45⁺ alive cells (upper panel). Percentage of MHCII on Ly6C^hi monocytes (lower panel).

(K) Flow cytometry analysis of lymph node (LN) dendritic cells (DCs) and their MFI of MHCII on different days after immunization.

(D–K) Shown is mean ± SD; significance was calculated using Student’s t test, ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001. Shown is 1 representative example of 3 (F), a pool of 2 experiments (H), or one representative experiment of two out of three that were similar (I).