PMC

The FMD treatment promotes endogenous glucocorticoid production, increases T_reg cell numbers, blocks T-cell activation and promotes T-cell death. In the lesion area. FMD treatment reduces autoimmune T-cell and microglia infiltration, promotes oligodendrocyte precursor dependent regeneration and the differentiation of myelinating oligodendrocyte which engage with demyelinated axons to promote the formation of myelin sheaths.

(n=5–6 / group; mean ± S.E.M, * p < 0.05, ** p<0.01; *** p < 0.001; t test, 1-way ANOVA & Bonferroni Post Test).
a. Diagram for the adoptive transfer EAE model.
b–d. Quantification of the (b) TNFα, (c) IFNγ, and (d)IL-17 (pg/mL) of the supernatant from ex-vivo culture of splenocytes from naïve, EAE CTRL, and EAE FMD either with or without MOG_35-55 and IL-23 re-activation
e–f. Quantification of T_H1 or T_H17 (represented by % of CD4⁺) from lymphocytes culture of EAE CTRL and EAE FMD with or without MOG_35-55 and IL-23 re-activation
g. Incidence rate of adoptive transfer EAE groups.
h. EAE severity score of adoptive transfer EAE groups.

(mean ± S.E.M, * p < 0.05, ** p<0.01; *** p < 0.001; t test, 1-way or 2-way ANOVA & Bonferroni Post Test; Scale bar represents 200 μm).
a. Diagram displaying the time course of the immunization and the diet interventions.
b. The EAE severity scoresof the Control Diet (EAE CTRL; n=23), ketogenic diet (EAE KD); n=13, semi-therapeutic FMD cycles (EAE FMD(S); n=7) or therapeutic FMD cycles (FMD(T); n = 23).
c. Incidence rate of the EAE CTRL and EAE FMD (S) (n=7–23).
d. EAE severity score of the EAE FMD(T) mice that completely reversed the EAE severity and scored 0, no observable disease (n=5).
e. EAE severity score of the best-performing control mice (n=12) and the FMD(T) mice (n=12).
f. EAE severity score of the EAE CTRL mice that treated with FMD upon chronic EAE development(EAE CTRL-FMD) (n=6).
g–m. Spinal cord of the EAE CTRL and the EAE FMD (T) mice with quantification of (g) H&E staining, (h) solochrome cyanine staining, and (i) MBP (Myelin Basic Protein) and SMI32 staining of spinal cord sections isolated at day 14.

(At least 12 sections/ mouse were used for quantification; n=4; mean ± S.E.M, * p < 0.05, ** p<0.01; *** p < 0.001; 1-way ANOVA & Bonferroni Post Test).
a – c. Spinal cord sections isolated at Day 14 and quantification for (a) GST-π (mature oligodendrocyte) and BrdU, for (b) TUNEL and NG2 (oligodendrocyte precursor cells), and for (c) TUNEL and GST-π of the naive, EAE-CTRL or EAE-FMD.
f–h. Sections from the corpus callosum region and quantification of cuprizone treated brains, stained with Luxol Fast Blue of the naïve control, end of 5 weeks of cuprizone diet (week 0), cuprizone (5 weeks) + regular chow (2 weeks), cuprzione (5 weeks) + FMD cycle (2 weeks).
i–j. Section from the corpus callosum region and its quantification of the cuprizone treated brains stained with GST-π⁺ of cuprizone (5 weeks) + regular chow (2 weeks), cuprzione (5 weeks) + FMD (2 weeks). Quantification is normalized to % of the naïve GST-π⁺ level.
k–m. Change in the quality of life at 3 month of (k) overall quality of life, (l) change in health, (m) physical health composite, and (n) mental health composite. The dotted line represents a threshold that is thought to be clinically important (≥ 5 points) (mean ± SED; * p<0.05; Mann-Whitney-U test. Increase of ≥ 5 points are considered as clinically important).

(n=4–8 / group; mean ± S.E.M, * p < 0.05, ** p<0.01; *** p < 0.001; t test, 1-way ANOVA & Bonferroni Post Test; Scale bar represents 200 μm).
a. Total white blood cells (WBC), lymphocytes, monocytes and granulocyte counts of the naïve, EAE-CTRL, EAE-FMD, and after 3 days of the re-feeding (EAE-FMD: RF) mice after 3 cycles of the FMD and matched time point for the EAE-CTRL.
b–d. Spinal cord sections (D14) and quantification at D3 and D14 post the first EAE sign for (b)CD11b⁺, (c) CD4⁺, and (d) CD8⁺ (at least 6 sections / mouse).
e. CD11c⁺ isolated from the EAE CTRL or EAE FMD mice on D3 and the quantification of cells from the total isolated splenocyte.
f. CD4⁺ gated for CD44^L or CD44^H isolated from the EAE CTRL or EAE FMD and quantification of % splenocyte of CD4⁺ CD44^L (Inactive) or CD4⁺ CD44^H (Active) cells.
g. CD3⁺ lymphocytes gated for CD4 and MOG_35-55/IA^b from EAE CTRL or EAE FMD and quantification of the MOG specific CD4⁺ cells.
h. CD4⁺ CD25⁺ FoxP3⁺ isolated from EAE CTRL or EAE FMD and the quantification of CD25⁺ FoxP3⁺ of CD4⁺ cells.
i–j. Intracellular staining for either IFNγ (i) or IL17(j) after gated for CD4⁺ of the naïve, EAE CTRL, EAE FMD, EAE FMD:RF and quantification of cell counts.
k. Quantification of Annex in V⁺ apoptotic CD3⁺ MOG_35-55/IA^b cells.
l. Quantification of CD4⁺IFNγ⁺ of BrdU⁺ lymphocytes.
m–o. Serum (m)TNFα, (n)IFNγ, and (o)IL-17 level (pg/mL) of the naive, EAE CTRL and EAE FMD mice on D3 post first sign of EAE.
p. Serum corticosterone level (ng/mL) of before immunization, at the time of the symptom, 3 or 14 days after initial symptom of the control or FMD.