Figure 5.

MEx treatment reduces hyperoxia-induced T cell autoreactivity. (A) Schematics showing the experimental layout to test autoreactivity after hyperoxia exposure. Newborn mice were exposed to HYRX as previously described for 7 days with MEx treatments being performed as PN4. Mice were then moved to normoxia conditions for a further period of 14 days. HYRX-mice were compared to mice kept in room air (NMRX) for the 28-day period. At PN28, lungs, spleens and thymi were harvested and single cell suspensions were prepared. Splenocytes (SC) or thymocytes (ThyC) were co-cultured at a 1:1 ratio with autologous lung cells (LC). Co-cultures were maintained, and T cell proliferation was analyzed as described in the methods section. Splenocytes and thymocytes cultured in media alone or in the presence of anti-CD3/CD28 stimulator beads were used as negative and positive controls respectively. Representative graphs and quantitative CFSE dilution of frequencies of CD4⁺CD8^- (B, C) and CD4^-CD8⁺ (D, E) thymocytes from co-cultures with autologous LC (ThyC+LC; upper panels) and stimulator beads (ThyC+B; lower panels). Representative graphs and quantitative CFSE dilution frequencies of CD4⁺ (F, G) and CD8⁺ (H, I) splenocytes from co-cultures with autologous LCs (SC+LC; upper panels) and stimulator beads (SC+B; lower panels). Data derived from two independent experiments representing mean ± SEM of N = 6 and *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.