Year

2021

Season

Spring

Paper Type

Master's Thesis

College

College of Arts and Sciences

Degree Name

Master of Science in Biology (MS)

Department

Biology

NACO controlled Corporate Body

University of North Florida. Department of Biology

First Advisor

Dr. Judith Ochrietor

Second Advisor

Dr. Terri Ellis

Rights Statement

http://rightsstatements.org/vocab/InC/1.0/

Third Advisor

Dr. Elizabeth Stotz-Potter

Department Chair

Dr. Cliff Ross

College Dean

Dr. George Rainbolt

Abstract

The neural retina is considered an immune privileged area, in that the eye attempts to suppress the inflammatory response to preserve vision. If there is damage to the blood-retina barrier, such as in diabetes or macular degeneration, it is possible for monocytes to travel out of the blood vessels and into the retina. While studying retinoblastoma, a research group determined that the protein, Basigin, through its extracellular domain, elicits an immune response by enhancing transcription of the pro-inflammatory cytokine interleukin-6 (IL-6). Basigin is cell adhesion molecule that belongs to the immunoglobulin superfamily. The Basigin gene has two main transcripts, that produce the proteins Basigin-variant-1 and Basigin-variant-2, that differ in the extracellular region. The expression of Basigin variants differ, in that variant-1 is specifically expressed by photoreceptor cells in the neural retina, and variant-2 is expressed throughout the body, including monocytes of immune cells and Müller glial cells of the retina. Because a previous study by this laboratory indicates that the two Basigin gene products interact via their immunoglobulin (Ig) extracellular domains in the retina, the aim of the present study was to determine if the binding region in the Ig0 domain of Basigin-variant-1 is responsible for eliciting the immune response observed in monocytes. In addition, specific amino acids within the binding region in the Ig0 domain were tested for their ability to bind to Basigin-variant-2, and their ability to induce an immune response in monocytes. The data indicate that recombinant versions of the Basigin-variant-1 Ig0 domain do not stimulate production of IL-6 in mouse RAW 264.7 monocytes. The results from the binding assay show the mutated proteins had reduced binding to Basigin-variant-2 when compared to the Ig0 protein, however, no significant difference in binding was observed. These data suggest that the Ig0 domain of Basigin-variant-1 does not stimulate a proinflammatory response in monocytes. The results of the current study contradict the data of a previous study, which suggests that the true receptor for Basigin-variant-1 Ig0 domain is not expressed in the mouse cell line used in this current study. It remains to be determined if the ability of Basigin-variant-1 to induce an immune response is species-specific.

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