Antigenic, genetic, and regulatory basis of autoimmune diabetes mellitus in vitro and in vivo

Persistent Link:
http://hdl.handle.net/10150/282436
Title:
Antigenic, genetic, and regulatory basis of autoimmune diabetes mellitus in vitro and in vivo
Author:
Wilson, Stephen Stewart, 1970-
Issue Date:
1997
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
This work introduces and explores a novel model which incorporates Fetal Thymus Organ Culture (FTOC) from non-obese Diabetic (NOD) mice to replicate thymic development and function of diabetogenic T cells in vitro. NOD FTOC is shown to posses a predictable diabetogenic activity measured in vitro, and a protective, regulatory activity when adoptively transferred to genetically IDDM-predisposed NOD mice. This in vitro IDDM (ivIDDM) activity is unique to NOD FTOC, and can be abbrogated by co-culture of developing NOD FTOC with FT from immunologically incompetent C.B-17 scid/scid mice. Additionally early exposure of NOD T cell precursors to islet antigens by co-culture with NOD Fetal Pancreas can negatively select for diabetogenic T cells or activate immuno-regulatory cells that can suppress diabetogenic T cell activity. The addition of blocking F(ab' )2 fragments of anti-CD3epsilon monoclonal antibody to NOD FTOC/FP co-cultures prevented insulin reduction, implicating a role for TcR-mediated recognition in this "in vitro IDDM" model. Transfer of unprimed syngeneic FTOC cells to pre-diabetic NOD mice prevents the onset of IDDM while transfer of islet-cell primed FTOC/FP cells slightly increased disease incidence. Spontaneous proliferation to peptides of Glutamate Decarboxylase (GAD) was not detected in NOD FTOC in contrast to reports of such responses in pre-diabetic NOD mice. A marked response to GAD peptides is induced by priming NOD FTOC, and increases ivIDDM. Proliferation is significantly diminished by tolergenic early treatment of FTOC, as is ivIDDM activity. Offspring of GAD peptide-treated NOD mice have a lower incidence of IDDM indicating potential beneficial tolerance to islet antigens by in utero exposure. Injection of identical GAD peptides to pre-diabetic NOD mice enhances the incidence of IDDM demonstrating the deleterious effects of inappropriate in vivo administration of autoantigenic peptides. NOD FTOC is shown to readily integrate and express retrovirus-delivered class II MHC I-E(α)d as measured by PCR and flow cytometry, respectively. The contribution to understanding the antigenic, genetic and regulatory basis of IDDM in NOD mice and humans is discussed.
Type:
text; Dissertation-Reproduction (electronic)
Keywords:
Health Sciences, Immunology.
Degree Name:
Ph.D.
Degree Level:
doctoral
Degree Program:
Graduate College; Microbiology and Immunology
Degree Grantor:
University of Arizona
Advisor:
DeLuca, Dominick

Full metadata record

DC FieldValue Language
dc.language.isoen_USen_US
dc.titleAntigenic, genetic, and regulatory basis of autoimmune diabetes mellitus in vitro and in vivoen_US
dc.creatorWilson, Stephen Stewart, 1970-en_US
dc.contributor.authorWilson, Stephen Stewart, 1970-en_US
dc.date.issued1997en_US
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractThis work introduces and explores a novel model which incorporates Fetal Thymus Organ Culture (FTOC) from non-obese Diabetic (NOD) mice to replicate thymic development and function of diabetogenic T cells in vitro. NOD FTOC is shown to posses a predictable diabetogenic activity measured in vitro, and a protective, regulatory activity when adoptively transferred to genetically IDDM-predisposed NOD mice. This in vitro IDDM (ivIDDM) activity is unique to NOD FTOC, and can be abbrogated by co-culture of developing NOD FTOC with FT from immunologically incompetent C.B-17 scid/scid mice. Additionally early exposure of NOD T cell precursors to islet antigens by co-culture with NOD Fetal Pancreas can negatively select for diabetogenic T cells or activate immuno-regulatory cells that can suppress diabetogenic T cell activity. The addition of blocking F(ab' )2 fragments of anti-CD3epsilon monoclonal antibody to NOD FTOC/FP co-cultures prevented insulin reduction, implicating a role for TcR-mediated recognition in this "in vitro IDDM" model. Transfer of unprimed syngeneic FTOC cells to pre-diabetic NOD mice prevents the onset of IDDM while transfer of islet-cell primed FTOC/FP cells slightly increased disease incidence. Spontaneous proliferation to peptides of Glutamate Decarboxylase (GAD) was not detected in NOD FTOC in contrast to reports of such responses in pre-diabetic NOD mice. A marked response to GAD peptides is induced by priming NOD FTOC, and increases ivIDDM. Proliferation is significantly diminished by tolergenic early treatment of FTOC, as is ivIDDM activity. Offspring of GAD peptide-treated NOD mice have a lower incidence of IDDM indicating potential beneficial tolerance to islet antigens by in utero exposure. Injection of identical GAD peptides to pre-diabetic NOD mice enhances the incidence of IDDM demonstrating the deleterious effects of inappropriate in vivo administration of autoantigenic peptides. NOD FTOC is shown to readily integrate and express retrovirus-delivered class II MHC I-E(α)d as measured by PCR and flow cytometry, respectively. The contribution to understanding the antigenic, genetic and regulatory basis of IDDM in NOD mice and humans is discussed.en_US
dc.typetexten_US
dc.typeDissertation-Reproduction (electronic)en_US
dc.subjectHealth Sciences, Immunology.en_US
thesis.degree.namePh.D.en_US
thesis.degree.leveldoctoralen_US
thesis.degree.disciplineGraduate Collegeen_US
thesis.degree.disciplineMicrobiology and Immunologyen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.contributor.advisorDeLuca, Dominicken_US
dc.identifier.proquest9806825en_US
dc.identifier.bibrecord.b37555819en_US
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