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Fig. 7 | Journal of Neuroinflammation

Fig. 7

From: An ocular Th1 immune response promotes corneal nerve damage independently of the development of corneal epitheliopathy

Fig. 7

Adoptive transfer of in vitro polarized Th1, Th2, or Th17 CD4+ T cells to T cell-deficient mice. A CD4+ T cells were isolated from the spleen and lymph nodes of OT-II [transgenic for an ovalbumin (OVA)-specific MHC II-restricted T cell receptor)]/recombination-activating gene 1 (RAG1)-deficient mice and cultured under Th1, Th2, and Th17-promoting conditions for 5 days. The resulting polarized cells were transferred i.p. to RAG1-deficient mice (T cell-deficient, 1 × 106 cells/mouse) that were given OVA eye drops daily for 4 weeks to induce an ocular immune response. A group of non-transferred (–) littermates was also included. B Representative dot plots of interferon-γ (IFN-γ), interleukin (IL)-4, and IL-17 production by CD4+ T cells after 5 days of in vitro polarization and before adoptive transfer. C Delayed-type hypersensitivity response to footpad OVA injection in recipient mice. Representative experiment (left) and images (right) of footpads. D Proportion of CD4+ T cells in the cervical lymph nodes of adoptively transferred mice (representative experiment). E Representative dot plots and F pooled data of IFN-γ, IL-4, and IL-17 production by CD4+ T cells obtained from cervical lymph nodes of mice 28 days after the adoptive transfer. G Representative photographs of transferred mouse eyes after 4 weeks of ocular challenge. * indicates a psoriasiform lesion. H Conjunctival CD4+ T cells and I neutrophils in transferred mice as assessed by flow cytometry (representative experiment). All experiments were performed twice or more with 5 mice/group/experiment. For all experiments, mean ± standard error of measurement is shown. To compare means, one-way ANOVA with post hoc testing was used. *p < 0.05, **p < 0.01, ***p < 0.001, and ns not significant

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