Figure 3. Recruitment of 8.8 CD8⁺ T cells enhances chemokine and cytokine gene expression as well as the numbers of donor CD4⁺ T cells, MdCs, and monocytes in the brain.

EAE was induced by transfer of Thy1.1⁺ MOG-specific CD4⁺ T cells into Thy1.2⁺ WT mice that had received Thy1.2⁺ WT or 8.8 CD8⁺ T cells. (A and B) Brain and spinal cord (SC) tissues were harvested 6 days after CD4⁺ T cell transfer (WT, n = 9; 8.8, n = 10). Chemokine (A) and cytokine (B) gene expression were analyzed directly ex vivo by quantitative PCR. All data were normalized to GAPDH. The fold change was calculated relative to gene expression values in irradiated healthy control mice (n = 2). (C) The numbers of CD45^hiCD11b⁺Ly6C^hiMHCII^– monocytes and CD45^hiCD11b⁺Ly6C^+/–MHCII⁺ MdCs were determined on days 5 (WT, n = 9; 8.8, n = 10) and 7 (WT, n = 12; 8.8, n = 13) after CD4⁺ T cell transfer for the brain and spinal cord by flow cytometry. (D) The number of Thy1.1⁺ donor CD4⁺ T cells was determined on days 5 (n = 8 per group) and 7 (WT, n = 19; 8.8, n = 23) after CD4⁺ T cell transfer for the brain and spinal cord by flow cytometry. (E) Brain and spinal cord mononuclear cells isolated from WT (n = 10) and 8.8 (n = 9) recipients on day 7 after CD4⁺ T cell transfer were stimulated with MOG_97–114 before intracellular cytokine staining. Percentages of Thy1.1⁺ donor CD4⁺ T cells producing the indicated cytokines are shown. Gating strategies for C–E are shown in Supplemental Figure 4. Graphs show mean + SEM (1 mouse per symbol) and are compiled from at least 2 independent experiments. Statistical significance was determined using a Mann-Whitney U test. *P < 0.05, **P < 0.01.