Genetic manipulation with the T -cell receptor (TCR) and major histocompatibility complex (MHC) bind...

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" Genetic manipulation with the T -cell receptor (TCR) and major histocompatibility complex (MHC) binding sites of arthritogenic epitopes of the G1 domain of human aggrecan "
Yanal Muhammad Murad T. T. Glant

Document Type	:	Latin Dissertation
Language of Document	:	English
Record Number	:	53213
Doc. No	:	TL23167
Call number	:	‭3159080‬
Main Entry	:	Yanal Muhammad Murad
Title & Author	:	Genetic manipulation with the T -cell receptor (TCR) and major histocompatibility complex (MHC) binding sites of arthritogenic epitopes of the G1 domain of human aggrecan\ Yanal Muhammad Murad
College	:	Rush University
Date	:	2005
Degree	:	Ph.D.
student score	:	2005
Page No	:	119
Abstract	:	Systemic immunization of BALB/c mice with human cartilage proteoglycan (PG) aggrecan induces progressive polyarthritis (PGIA). Most of the T cell epitopes of cartilage PG, including those that induce dominant T cell responses, are present in the poorly glycosylated G1 domain of the PG aggrecan molecule. The GI domain contains at least three immunodominant and six subdominant T cell epitopes. The aim of this study is to investigate the immune pathological function of these three dominant epitopes when disrupted (deleted) individually or in combination, and which dominant epitope is critical for arthritis induction. A cDNA clone encoding for the entire 360 amino acids of the G1 domain of the human cartilage PG aggrecan molecule was used for in vitro targeted disruption of three epitopes found to be dominant in preliminary experiments. The cDNA was cloned in a Baculovirus Expression System, and the recombinant human G1 proteins (rhG1) were purified by a combination of ion exchange and metal affinity chromatography. BALB/c mice were immunized with rhG1 protein wild-type or mutated, immune response monitored during the immunization periods and at the end of experiments. Histology of joints of immunized BALB/c mice was assayed at the end of experiment. Close to 100% incidence of the disease was achieved after 5 injections of 50 μg wild-type (non-mutated) rhG1 domain, which was lower in the absence of 5/4E8 epitope and very low incidence (<10%) in the absence of all three dominant T-cell epitopes. The lack of T cell response to the deleted epitope(s) was specific, but the overall immune response against the wild-type rhG1 domain of human PG was not significantly affected. This observation indicates that the other (at least 6 additional subdominant T cell epitopes) can indeed induce T- and B-cell responses, but the arthritis induction is fundamentally linked to these three dominant epitopes. Histology of the inflamed paws (joints) was highly comparable with those induced with intact cartilage PG aggrecan. This study will help us to understand the dynamics and immune-regulatory mechanisms of the PGIA model, and confirms the immune function of a single, or the combination of two-three dominant/arthritogenic epitopes in arthritis induction.
Subject	:	Health and environmental sciences; Pure sciences; Biological sciences; Aggrecan; Arthritogenic; MHC; T-cell receptor; Molecular biology; Biochemistry; Immunology; 0982:Immunology; 0487:Biochemistry; 0307:Molecular biology
Added Entry	:	T. T. Glant
Added Entry	:	Rush University