Environmental triggers, such as viral infection and environmental toxins, have been proposed to
initiate the autoimmune disease of Type 1 Diabetes (T1D), however, the mechanism is unknown.
The identification of novel autoantigens may provide insight to the mechanism of environmental
triggers and pathogenesis of T1D. I identified the antigen recognized by the diabetogenic BDC-
2.5 T cell clone using a novel in vivo reconstitution system, Restricted Immune System via
Adoptive Transfer (RISAT). In RISAT, immunodeficient mice are adoptive transferred with a
single T cell clone and an open repertoire of B cells. Reconstituted mice are immunized with an
antigenic protein preparation. This system will drive an antibody response to the cognate antigen
for the T and B cell through the co-stimulatory pathways involved in linked recognition. For the
BDC-2.5 RISAT, non-obese diabetic (NOD).Rag-/- mice were adoptive transferred with the
diabetogenic BDC-2.5 T cells and NOD B cells and then immunized with an antigenic beta cell
membrane preparation (βmem) to drive an antibody response. The resulting antibodies
recognized the endoplasmic reticulum (ER) stress associated protein glucose regulated protein 78
(GRP78) from βmem. To determine if ER stress plays a role in the antigenic response of the
BDC-2.5 T cell clone, the non-antigenic NOD insulinoma cell line, NIT-1, were treated with
thapsigargin, which induces ER stress. The treatment of NIT-1 with thapsigargin led to
increased GRP78 synthesis, correlating with antigenic recognition by the BDC-2.5 T cell clone. The antibodies from the BDC-2.5 TCR-Tg recognizes a subset of GRP78 which is modified with
phosphoserine. The data presented in this thesis demonstrates a mechanistic link between ER
stress and environmental triggers leading to the initiation of TID through the novel autoantigen,
GRP78. Also the technique, RISAT, can be used to identify additional potential autoantigens of
isolated T cell clones in both T1D and other autoimmune diseases.
| Identifer | oai:union.ndltd.org:PITT/oai:PITTETD:etd-12102007-204111 |
| Date | 17 December 2007 |
| Creators | Schreiner, Sheila Marie |
| Contributors | Jon D. Piganelli, Ph.D., Tim D. Oury, M.D./Ph.D., Massimo Trucco, M.D., Nick Giannoukakis, Ph.D., Wendy M. Mars, Ph.D. |
| Publisher | University of Pittsburgh |
| Source Sets | University of Pittsburgh |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.pitt.edu/ETD/available/etd-12102007-204111/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University of Pittsburgh or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
Page generated in 0.002 seconds