Desiccating Stress Stimulates Expression of Th-1 Immune Response Associated Chemokines and Chemkine Receptors on the Ocular Surface of C57BL/6 Mice

Abstract

Purpose:: To evaluate the effects of desiccating ocular surface stress on expression of chemokines and their receptors in the corneal and conjunctival epithelia of C57BL/6 (Th-1 preferred) and BALB/c mice (Th-2 preferred).

Methods:: Experimental dry eye was created in C57BL/6 and BALB/c mice by subcutaneous scopolamine injection and exposure to an air draft for 5 and 10 days. The protein concentrations of macrophage inflammatory protein 1α (MIP-1α), MIP-1ß, monokine induced by interferon-γ (MIG) and interferon-γ-inducible protein 10 (IP-10) in the corneal and conjunctival epithelia were measured by a multiplex immunobead assay. Expression of CCR5, CXCR3, CCR3, MIP-1α, MIP-1ß, regulated upon activation, normal T-cell expressed and secreted (RANTES), MIG, IP-10, monocyte chemoattractant protein 3 (MCP3) and eotaxin-1 transcripts were evaluated by real time PCR, immunofluorescent staining and immunohistochemistry in the corneal and conjunctival epithelia.

Results:: Desiccating stress significantly increased protein levels of MIP-1α, MIP-1ß, IP-10 and MIG in the corneal and conjunctival epithelia in C57BL/6 mice compared to BALB/c mice. Dry eye increased the Th-1 chemokine receptor CCR5 and its ligands MIP-1α and MIP-1ß transcripts in the cornea and conjunctiva and RANTES, MIG and IP-10 transcripts in the conjunctiva of C57BL/6 mice. On the other hand, levels of transcripts encoding the Th-2 chemokine receptor CCR3 and its ligands MCP-3 and eotaxin-1 increased in both cornea and conjunctiva of BALB/c mice. The pattern of expression of chemokines and their receptors detected by immunofluorescent staining and immunohistochemistry was similar to their gene expression for most chemokines and chemokine receptors.

Conclusions:: Specific Th-1 and Th-2 immune responses were induced after desiccating stress in a strain-related fashion. Desiccating stress stimulated expression of Th-1 cell attracting chemokines and chemokine receptors on the ocular surface of C57BL/6 mice.

Keywords: cytokines/chemokines • cornea: epithelium