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. 2022 Feb 7;12:805302. doi: 10.3389/fcimb.2022.805302

Figure 5.

Figure 5

Curdlan injection enhanced the production of interleukin (IL)-17 and interferon (IFN)-γ, but not antigen-specific lymphoproliferation or antibody responses in EAE. (A) PLP139-151-specific lymphoproliferative responses of lymph node cells from the control (EAE/PBS, black bar) and curdlan-treated (EAE/curdlan, white bar) EAE mice a few days before the disease onset, 8–9 days post sensitization. Inguinal lymph node cells were stimulated with the PLP139-151 peptide in the presence or absence of a blocking antibody against CD4 or CD8. Levels of lymphoproliferation were expressed as stimulation indexes (SI): (mean absorbance in PLP139-151 stimulation)/(mean absorbance without stimulation). (B) Enzyme-linked immunosorbent assays (ELISAs) of serum anti-PLP139-151 immunoglobulin (Ig) G1 and IgG2c antibodies in sera from the control EAE and curdlan-treated EAE mice 13–14 days post sensitization. The dotted line indicates the detection limit. (C, D) ELISAs of IL-17, IFN-γ (C), IL-4, and IL-10 (D) production from lymph node cells of the control EAE and curdlan-treated EAE mice harvested a few days before onset (Onset) or at the disease peak (Peak). Cells isolated from the inguinal lymph nodes were cultured with the PLP139-151 peptide. The four cytokines in the culture supernatants were quantified using ELISA kits. N.D., not detectable. (E, F) Real-time polymerase chain reaction (PCR) analyses of T cell-related (E) and immune cell migration/infiltration-related (F) genes in the spinal cord from the control and curdlan-treated EAE mice at the disease peak. Il17a (IL-17), Ifng (IFN-γ), Gzmb (granzyme B), Foxp3 (forkhead box P3, regulatory T cell marker), Il10 (IL-10), Igha (Ig heavy chain α, IgA marker), Vcam1 (vascular cell adhesion molecule 1), Icam1 (intracellular adhesion molecule 1), Itga4 (integrin α4), Itgb2 (integrin β2), Cxcl2 [chemokine (C-X-C motif) ligand 2, CXCL2], Cxcl9 (CXCL9), Cxcl0 (CXCL10, also known as IFN-γ-inducible protein 10), and Cxcr3 [chemokine (C-X-C motif) receptor 3, CXCR3]. Pgk1 expression was used as a housekeeping gene for normalization. (A–F) Values are the mean + SEM from six to eight mice per group. *P < 0.05, Student’s t-test.