Extended Data Fig. 9: Experimental autoimmune encephalomyelitis in mice harbouring cross-genotype transferred complex microbiotas.
From: Gut microbial factors predict disease severity in a mouse model of multiple sclerosis
Microbiota from SPF-housed Muc2−/− (KO) and Muc2+/+ (WT) mice was transferred to germ-free (GF) C57BL/6J and Muc2−/− mice (Fig. 6a). EAE was induced after 21 d exposure to the donor microbiota. a, Faecal microbiota composition in EAE-induced mice on the day of EAE induction (d 0; ‘pre-EAE’), ordered by donor–recipient combination. Microbiota composition was determined on a species level by full-length 16S rRNA gene sequencing. The top 5 most abundant species per phylum are shown. In the case of Verrucomicrobiota, Akkermansia muciniphila was the only detected feature. b, Shannon index as a measure of α-diversity of the pre-EAE microbiota compositions. One-way ANOVA with donor–recipient combinations as tested variable. c, β-Diversity of pre-EAE microbiota compositions from a Bray–Curtis distance matrix of arcsine square root-transformed relative abundance data, ordinated using a principal coordinate analysis. Upper panel, mice coloured by genotype of the microbiota donor. Lower panel, mice coloured by genotype of the microbiota recipient. PERMANOVA using the genotype of either donor (upper panel) or recipient (lower panel) as tested variable. d, EAE disease course of mice (same mice as in panel a), separated by donor–recipient combinations, visualized as EAE disease score as a function of time (day after EAE induction). Dots represents daily mean EAE score; dashed lines represent SD. e, Evaluation of EAE disease course using AUC of the individual disease course (panel d) or the maximum achieved EAE disease score per mouse (Max). Left, mice coloured by donor genotype. Right, mice coloured by recipient genotype. Wilcoxon rank-sum test with donor (left) or recipient (right) genotype as tested variable. Boxplots show median, quartiles, and 1.5 × interquartile range. f, Distribution of individual AUC and Max values to determine ‘Severe’ and ‘Mild’ EAE; accordingly, AUC = 30 and Max = 2 were selected as thresholds to establish a binary categorization of individual EAE phenotypes. g, Individual pre-EAE microbiota composition. Same underlying Bray–Curtis distance matrix and PCoA-ordination as panel c, coloured by binary phenotype classification (panel f) based on individual AUC (left) or Max (right). PERMANOVA with the indicated phenotype classification as tested variable.
