Extended Data Fig. 2: Neomycin treatment outside the lung does not affect CNS autoimmunity.
a, b, Oral treatment with neomycin does not ameliorate EAE. a, Left, PCA of the microbiota composition of faecal samples from rats that were orally treated with neomycin (1 or 10 mg) or PBS for 7 days. Middle, corresponding Shannon and phylogenetic diversity indices. Right, quantification of bacterial abundance based on tuf gene expression via 16S rRNA-based quantitative PCR. Mean ± s.e.m. Cumulative data of 2 independent experiments. n = 6 (all groups). b, Clinical parameters observed in lung EAE of rats pre-treated orally with PBS or neomycin (1 or 10 mg) for 7 days. Mean ± s.e.m. Cumulative data of 2 independent experiments. n = 8 (PBS, 1 mg Neo); n = 7 (10 mg Neo). c, d, I.tr. neomycin treatment does not affect TMBP cell proliferation and effector function. c, Quantification of TMBP cells cultured in presence of neomycin at the indicated concentrations. Flow cytometry on D2, D3 and D4 after antigen challenge. Representative data of 2 independent experiments. n = 3 (all groups). d, Clinical outcome of EAE induced by transfer of TMBP cells previously stimulated in vitro in presence of neomycin at the indicated concentrations. Clinical parameters. Mean ± s.e.m. Representative data of 3 independent experiments. n = 3 (both groups). e, f, Subcutaneous (s.c.) treatment with neomycin does not affect EAE. e, Lung EAE was induced in rats s.c. pre-treated for 7 days with PBS or neomycin. Clinical parameters. Mean ± s.e.m. Representative data of 3 independent experiments. n = 5 (both groups). f, Rats s.c. pre-treated for 7 days with PBS or neomycin were i.v. transferred with resting TMBP cells. 6 h later, they were s.c. immunized with MBP. Clinical parameters. Mean ± s.e.m. Representative data of 3 independent experiments. n = 3 (both groups). a–d, Statistical significance determined by one-way ANOVA with Tukey’s multiple comparisons test (Gaussian distribution) and Kruskal–Wallis test with Dunn’s multiple comparisons test (non-Gaussian distribution). e, f, Statistical significance determined by unpaired two-tailed t-test (Gaussian distribution) and Mann–Whitney test (non-Gaussian distribution). *P < 0.05, **P < 0.01, ***P < 0.001.
