Figure 1.

B cell responses occur in the central nervous system of TH17-EAE. (A) Representative flow cytometry plots of the percentage of viable B220⁺CD19⁺ B cells in the spinal cords of TH17-experimental autoimmune encephalomyelitis (EAE) at 5, 9, and 15 days post-transfer of TH17 cells. Gated on total viable cells. (B) Percentage of total viable cells and absolute number of B cells infiltrating the spinal cord and brain of TH17-EAE. Statistical significance from Day 5 to Day 15 was determined using Kruskal–Wallis test (**p < 0.01, *p < 0.05). Data are from one experiment representative of two experiments (N = 3 mice per group/experiment). (C) Representative image of a spinal cord section from TH17-EAE stained with anti-CD3 Ab (red), anti-B220 Ab (green), and DAPI (blue). (D) Expression levels of CXCR5 (measured by mean fluorescent intensity) on viable B220⁺CD19⁺ B cells in spleen and spinal cord tissue was measured by flow cytometry. Tissues were collected at the peak of disease. Statistical significance was determined using Student’s t-test (****p < 0.0001) and N = 5. (E) Representative flow cytometry plots of CD19⁺IgM⁻IgD⁻ class-switched B cells in the spleen and spinal cord of mouse with TH17-EAE. Gated on viable B220⁺CD19⁺ B cell population. Percentage of class-switched B cells of the total CD19⁺ B cell population in the spleens and spinal cords of mice with TH17-EAE. Statistical significance from was determined using Student’s t-test (**p < 0.01) and N = 4. (F) Representative flow cytometry plots of CD19⁺GL7⁺PNA⁺ germinal center B cells in the spleen and spinal cord of mouse with TH17-EAE. Gated on viable CD19⁺ B cell population in the spleens and spinal cords of mice with TH17-EAE. Statistical significance was determined using Student’s t-test (**p < 0.01) and N = 4. (G) Correlation between CD19⁺IgM⁻IgD⁻ class-switched B cells and decrease in body weight at the peak of disease. Statistically significant correlations were determined using linear regression and N = 8.