December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Expression Of Interleukin-1 Receptors In Human Trabecular Meshwork
Author Affiliations & Notes
  • AS Czuk
    Casey Eye Institute Oregon Health & Science University Portland OR
  • JR Samples
    Casey Eye Institute Oregon Health & Science University Portland OR
  • Footnotes
    Commercial Relationships   A.S. Czuk, None; J.R. Samples, None. Grant Identification: none
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 1041. doi:
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      AS Czuk, JR Samples; Expression Of Interleukin-1 Receptors In Human Trabecular Meshwork . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1041.

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Abstract

Abstract: : Purpose: To evaluate receptors to interleukin-1 (IL-1) in human trabecular meshwork (HTM) cell cultures obtained from young eyes and organ cultures obtained from adult eyes, and to determine whether or not IL-1 modulates production of receptor in HTM. Methods: HTM cells from young donors and anterior segments from adult donors were obtained from the Oregon Lion's Eye Bank, and the protocols were approved by the institutional human subjects committee and conformed to the Declaration of Helsinki. Briefly, cells were prepared by establishing cultures from HTM explants. Third or fourth passage cells were incubated in serum free Dulbecco's medium for 48 hours, treated with IL-1alpha (4 ng/ml; R&D Systems) or phorbol ester (10 ng/ml; Sigma) for 24 hours, and then fixed in paraformaldehyde. Anterior segment organ cultures were incubated serum free for 7 days prior to 24 hour treatment with or without IL-1alpha. Paraformaldehyde-fixed tissue was cut into wedges, embedded in agarose blocks, and cut into 100-150 micron sections on a vibratome. Tissues and cells were incubated with monoclonal (R&D Systems and Serotec) and polyclonal (R&D Systems and Santa Cruz) antibodies during indirect immunohistochemistry protocols; negative controls were processed without primary antibody. Data were collected with a confocal microscope. Results: Localization of IL-1R1 with several different antibodies was seen in cultured HTM cells. The Santa Cruz polyclonal yielded more intense staining for this receptor than the other antibodies. Treatment with IL-1 alpha increased IL-1R1 staining more than the phorbol ester upon comparison with untreated controls. Not as much receptor staining was seen in the trabecular meshwork of organ culture sections. Use of several different antibodies did not demonstrate the presence of any IL-1R2 whatsoever. Conclusion: Considering prior experiments demonstrating expression of IL-1 in the trabecular meshwork and the effects of IL-1 on the expression of matrix metalloproteinases, all of the elements are present to demonstrate an IL-1 autocrine loop in the trabecular meshwork that modulates matrix metalloproteinases and their inhibitor. Subsequently, activation of the IL-1 receptors in meshwork is likely to have important effects upon the extracellular matrix of trabecular cells.

Keywords: 601 trabecular meshwork • 380 cytokines/chemokines • 434 immunohistochemistry 
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