Polarized Th1 and Th17 Cells Derived From a Mouse Model of Dry Eye Induce Ocular Surface Disease

Abstract

Purpose: : To determine the role of polarized Th1 and Th17 mouse CD4+ T cells in inducing ocular inflammation in the mouse model of dry eye.

Methods: : C57BL/6 wild type (WT) female mice were exposed to a desiccating environmental stress (DS; subcutaneous scopolamine injections, humidity <40%, and air flow across wire meshed screened cages) for 10 days. Control or 10 day DS C57BL/6 mouse spleen and superficial cervical lymph node naïve CD4+ T cells were incubated in the presence of the appropriate polarizing cytokines, antibodies, and anti-CD3/CD28 antibodies for Th1 or Th17. After four days in culture, the supernatants of these cells were analyzed by Luminex for cytokine levels. Following polarization, cells were IP injected into nude T cell deficient mouse recipients.

Results: : Control and DS Th1 polarized cell supernatants had significantly high levels of IFN-γ (10 ng/ml) and IL-12(p70) (700 pg/ml) as compared to Th17 polarized cells (*P<0.001). Polarized control and DS Th17 cell supernatants had high levels of IL-17 (11 ng/ml (*P<0.001)). IL-17 was not detectable in Th1 supernatants. Adoptive transfer of polarized 10 Day DS CD4⁺ Th1 cells resulted in significant IFN-γ tear levels (83+/-21 pg/ml; *P<0.0486) and CD4+ cellular infiltration (31+/-7) as compared to polarized control cells (11+/-1.5) into the recipient conjunctiva. Adoptive transfer of in vitro polarized DS Th17 cells resulted in a more robust cellular infiltration into the conjunctiva of nude recipient mice (147+/-70) as compared to control Th17 polarized cells (11.3+/-5). Goblet cells in mice receiving DS Th17 polarized cells were not significantly inhibited which corroborates previous data by De Paiva et al. demonstrating that IFN-γ is responsible for goblet cell loss in the mouse model of dry eye.

Conclusions: : These data strongly indicate that both Th1 and Th17 play important roles in the immunopathogenesis of experimental dry eye.

Keywords: cytokines/chemokines • inflammation • conjunctiva