The focus of this project is to investigate resident corneal cell interaction with the immunoregulatory cytokine IL-10. Studies will be carried out with cultured human and murine corneal epithelial cells and fibroblasts. RT- PCR analysis will be used to determine precisely which resident corneal cells express IL-10 receptor mRNA. Flow cytometry and immunohistology will be used to search for the protein product on the different corneal cell types. The consequences of IL-10 binding to its receptor on corneal cells will be analyzed from the perspective of determining whether ligand binding leads to suppressed cytokine/chemokine mRNA and/or protein synthesis. Prior RT-PCR and ELISA studies in our laboratory have shown that IL-10 suppresses IL-6 mRNA and protein synthesis. The applicant hypothesizes that IL-10 inhibits cytokine synthesis at the level of gene transcription or interferes with mRNA stability. Preliminary data indicate that human and mouse corneas possess mRNA for IL-10. The applicant will test whether resident corneal cells make IL-10 protein. Positive results would suggest that IL-10 may function as an autoregulatory factor for resident corneal cells. The basic information generated by these studies may prove useful for devising new approaches to control ocular inflammation.
|Molesworth-Kenyon, Sara J; Yin, Rong; Oakes, John E et al. (2008) IL-17 receptor signaling influences virus-induced corneal inflammation. J Leukoc Biol 83:401-8|
|Fenton, Robin R; Molesworth-Kenyon, Sara; Oakes, John E et al. (2002) Linkage of IL-6 with neutrophil chemoattractant expression in virus-induced ocular inflammation. Invest Ophthalmol Vis Sci 43:737-43|
|Yan, X T; Zhuang, M; Oakes, J E et al. (2001) Autocrine action of IL-10 suppresses proinflammatory mediators and inflammation in the HSV-1-infected cornea. J Leukoc Biol 69:149-57|
|Tumpey, T M; Cheng, H; Yan, X T et al. (1998) Chemokine synthesis in the HSV-1-infected cornea and its suppression by interleukin-10. J Leukoc Biol 63:486-92|
|Yan, X T; Tumpey, T M; Kunkel, S L et al. (1998) Role of MIP-2 in neutrophil migration and tissue injury in the herpes simplex virus-1-infected cornea. Invest Ophthalmol Vis Sci 39:1854-62|
|Tumpey, T M; Cheng, H; Cook, D N et al. (1998) Absence of macrophage inflammatory protein-1alpha prevents the development of blinding herpes stromal keratitis. J Virol 72:3705-10|