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Ryan Santiago Salvador, Reiko Horai, Carlos Zárate-Bladés, Yingyos Jittayasothorn, Rachel R Caspi; Alteration of gut microbiota by antibiotics impacts spontaneous ocular autoimmunity. Invest. Ophthalmol. Vis. Sci. 2017;58(8):546.
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© ARVO (1962-2015); The Authors (2016-present)
The R161H spontaneous uveitis model permits to study natural triggers of uveitis. We have previously shown that elimination of commensals from R161H mice by oral antibiotic treatment or germ-free conditions attenuated uveitis and reduced Th17 cells in the gut. Because oral antibiotics do not completely deplete commensals, we set out to associate modulation of gut microbiota by alternative antibiotic treatments with spontaneous disease and immune responses.
R161H mice and WT littermates were treated with antibiotics (ampicillin, metronidazole, neomycin and vancomycin, either individually or as a mix (AMNV), or with gentamicin added (AMNV/G)) in drinking water from before birth. Antibiotic-treated mice were compared to age-/genotype-matched specific-pathogen-free (SPF) mice that received normal water. Uveitis was evaluated by fundoscopy and histology. Proteinaceous extracts of cecal contents were used to stimulate R161H lymphocytes in vitro; activation was assessed by induction of CD69 positivity (flow cytometry) and IL-2 secretion into the supernatant (ELISA). Fecal pellets were collected for 16S rRNA sequencing. Intestinal lamina propria lymphocytes were analyzed for the Th17 phenotype and other activation markers.
Either AMNV or AMNV/G treatment attenuated spontaneous uveitis in R161H mice, whereas single antibiotic treatments did not. Fewer proliferating and Th17 cells were detected in the intestines of AMNV- or AMNV/G-treated mice. In contrast to cecal extracts from SPF mice, extracts from antibiotic-treated mice failed to upregulate CD69 expression on R161H T cells and did not induce secretion of IL-2. Interestingly, cecal contents from R161H SPF mice appeared to activate the autoreactive T cells more strongly than extracts from WT SPF mice.
Depletion of gut microbiota through antibiotic treatment impacts spontaneous uveitis by abrogating the ability of intestinal contents to activate retina-specific T cells in the gut. Interestingly, R161H SPF mice may harbor altered microbiota that can stimulate retina-specific T cells better than microbiota of WT mice. Further work, including 16S rRNA sequencing to compare the microbial communities between WT and R161H mice, as well as between antibiotic-treated vs. non-treated mice, is needed to identify specific organisms and molecular stimuli that help to trigger spontaneous uveitis.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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