Overview
Our laboratory investigates the genetic control mechanisms allowing the immune system to recognize and destroy foreign pathogens, but not normal constituents of the body. Defects in these mechanisms underlie many autoimmune diseases, including type 1 (juvenile onset, insulin dependent) diabetes. Using NOD (non-obese diabetic) inbred mice, we focus on the process through which genes that normally elicit immune responses to foreign intruders can sometimes trigger autoimmune responses against the body's own cells, such as the pancreatic cells that make insulin. We have shown that some genes play a role in type 1 diabetes even when they do not contain deleterious mutations. Instead they are normal genes that only contribute to type 1 diabetes pathology when collected together in a specific fashion. We are also investigating defects in the differentiation of a particular type of leukocyte in NOD mice that subsequently allows for the development of autoimmune responses. This work may help identify the mechanisms and compounds that normally prevent autoimmunity, and hence reveal strategies for pharmacological interventions in humans at risk for type 1 diabetes.
Research details
Genetic Basis of Immunological Tolerance Defects Underlying Type 1 Diabetes
Our primary research interest is understanding the genetic basis for immune tolerance to endogenous proteins. Defects in these mechanisms lead to many debilitating autoimmune diseases, of which type 1 diabetes (T1D) is one of the most serious. In both humans and NOD mice, T1D results when insulin-producing pancreatic ß-cells are destroyed by an autoreactive T-cell response. Thus, insights into the genetic mechanisms responsible for the normal maintenance of immunological tolerance can be gained by identifying the pathogenic basis of T1D in NOD mice.
T1D is controlled by a large number of susceptibility (Idd) genes. However, genes within particular major histocompatibility complexes (MHC) provide the primary component for T1D susceptibility in both humans and NOD mice. The MHC encodes two primary types of gene products, termed class I and class II. Class I molecules present peptides derived from intracellular proteins to CD8 T-cells, which usually exert cytotoxic functions. Virtually all cells express MHC class I molecules. In contrast, MHC class II expression is largely limited to a specialized subset of hematopoietically derived antigen presenting cells (APC) that include B-lymphocytes, macrophages, and dendritic cells (DC). MHC class II molecules expressed by APC display peptides derived from internalized extracellular proteins to CD4 T-cells, which produce cytokine molecules that amplify other components of the immune response, including cytotoxic CD8 T-cells.
T-cells specifically recognize a particular peptide/MHC antigenic complex through expression of clonally distributed T-cell receptor (TCR) molecules generated by somatically rearranged gene sequences. In addition to those derived from infectious pathogens, APC also display to T-cells MHC class I- and class II-bound peptides generated from endogenous "self" proteins. A normal consequence of expressing a TCR that engages "self antigenic" complexes on APC at a critically high threshold level is the induction of signals triggering the deletion or permanent inactivation of potentially autoreactive T-cells. Defects in this process of immunological tolerance induction manifested by both T-cells and APC underlie susceptibility to T1D.
In both humans and NOD mice, unusual MHC class II genes clearly contribute to T1D by inducing pathogenic CD4 T-cell responses. However, while representing common variants shared by many strains lacking autoimmune proclivity, the MHC class I molecules expressed by NOD mice also exert pathogenic functions essential to T1D development. These common class I variants aberrantly exert diabetogenic functions in NOD mice through interactions with some of this strain's multiple Idd genes located outside the MHC. The strongest such interactive effect is provided by a gene(s) within the Idd7 locus. The NOD Idd7 variant appears to prevent the TCR of autoreactive T-cells from being expressed at the level necessary to elicit a signaling response that is sufficiently strong to trigger the deletion of such pathogenic effectors upon their engagement of self antigen.
In humans, epidemiological studies also suggested that when co-expressed with particular combinations of other genes, some common MHC class I variants such as HLA-A2.1 could contribute to T1D development. To test this possibility, we generated NOD mice that transgenically express human HLA-A2.1, but no murine class I molecules. The human HLA-A2.1 molecules in these mice were found to mediate diabetogenic T-cell responses. Together with collaborators at The Albert Einstein College of Medicine, we identified three ß -cell autoantigenic peptides that are presented to diabetogenic T-cells by human HLA-A2.1 class I molecules in our transgenic mice. Subsequent work by another collaborator at Leiden University in The Netherlands revealed that at least one of these peptides is recognized in an HLA-A2.1-dependent fashion by CD8 T-cells isolated from a human T1D patient.
B-lymphocytes represent a critical subset of APC for activating the MHC class II-dependent T-cell responses contributing to T1D in NOD mice. The preferential ability of NOD B-lymphocytes to serve as diabetogenic APC results from the fact that they, unlike other types of APC, express plasma membrane-bound immunoglobulin (Ig) molecules which specifically capture and internalize ß -cell proteins. Mechanisms are normally in place to prevent the development of B-lymphocytes expressing autoreactive Ig molecules. We found that several of these mechanisms are defective in NOD mice. Further studies indicated that genes within the Idd5 and Idd9/11 loci contribute to the defective ability of NOD mice to inactivate B-lymphocytes expressing autoreactive Ig molecules.
The DC subset of APC in NOD mice is characterized by developmental defects preventing them from presenting self antigens in a sufficiently vigorous fashion to induce T-cell tolerogenic processes. Human T1D patients share similar DC developmental defects. Impaired DC differentiation in NOD mice is in turn linked to numerical deficiencies in the CD4-expressing subset of immunoregulatory natural killer (NK) T-cells that also characterize this strain. The loss of this NKT-cell subset appears to result from the fact that they are highly sensitive to apoptotic deletion when the cell surface enzyme ADP-ribosyltransferase 2 (ART2) is activated by its substrate NAD. Thus, blocking ART2 activity may have T1D protective effects in NOD mice by enhancing the survival of NKT-cells that in turn drive development of tolerogenic DC. However, due to its small size, and the fact that it is deeply imbedded in the plasma membrane, the catalytic site of ART2 cannot be targeted by conventional two-chain antibodies. However, llamas have the unusual feature of being able to generate single-chain antibodies that can target small, not readily accessible molecules. Early results from studies done in collaboration with investigators at the University of Hamburg in Germany indicate that treatment with a llama single-chain antibody can block ART2 activity in NOD mice resulting in enhanced numbers of NKT-cells and potentially tolerogenic DC. Further studies are ongoing to determine if blocking ART2 activity with this llama single-chain antibody has T1D protective effects in NOD mice.
Lab staff
Principal Investigator: David V. Serreze, Ph.D.
Research Scientist: Yi-Guang Chen, Ph.D.
Postdoctoral Fellows: John P. Driver, Ph.D., Marijke Niens, Ph.D., Felix Scheuplein, Ph.D.
Professional Assistant: Harold D. Chapman, B.S.
Research Assistants: Alexandra E. Grier, B.A., Deanna J. Lamont, B.S.
Predoctoral Student: Elisa I. Rivas, M.S.
Laboratory Technician: Brenice E. Briggs
Shared Service Technician: Eleanor Dewey
Research Administrative Assistant: Christina Gagliardi
Publication listings
(2004-present)
Serreze DV, Choisy-Rossi C, Grier A, Holl TM, Chapman HD, Gahagan JR, Osborne MA, Zhang W, King BL, Brown A, Roopenian D, Marron MP . 200_. Through regulation of TCR expression levels, an Idd7 region gene(s) interactively contributes to the impaired thymic deletion of autoreactive diabetogenic CD8 T-cells in NOD mice. J Immunol, (in press).Chen Y-G, Silveira PA, Osborne MA, Chapman HD, Serreze DV. 2007. Cellular expression requirements for inhibition of type 1 diabetes by a dominantly protective major histocompatibility complex haplotype. Diabetes 56(2):424-430.
Chen YG, Driver JP, Silveira PA, Serreze DV. 2007. Subcongenic analysis of genetic basis for impaired development of invariant NKT cells in NOD mice. Immunogenetics 59(9):705-712.
Driver JP, Foreman O, Mathieu C, van Etten E, Serreze DV. 2007. Comparative therapeutic effects of orally administered 1,25-dihydroxyvitamin D(3) and 1alpha-hydroxyvitamin D(3) on type-1 diabetes in non-obese diabetic mice fed a normal-calcaemic diet. Clin Exp Immunol 151(1):76-85.
Jarchum I, Baker JC, Yamada T, Takaki T, Marron MP, Serreze DV, DiLorenzo TP. 2007. In vivo cytotoxicity of insulin-specific CD8+ T-cells in HLA-A*0201 transgenic NOD mice. Diabetes 56(10):2551-2560.
Serreze DV, Marron MP, Dilorenzo TP. 2007. "Humanized" HLA transgenic NOD mice to identify pancreatic beta cell autoantigens of potential clinical relevance to type 1 diabetes. Ann N Y Acad Sci 1103:103-111.
Unger WW, Pinkse GG, Mulder-van der Kracht S, van der Slik AR, Kester MG, Ossendorp F, Drijfhout JW, Serreze DV, Roep BO. 2007. Human clonal CD8 autoreactivity to an IGRP islet epitope shared between mice and men. Ann N Y Acad Sci 1103:192-195.
Yamanouchi J, Rainbow D, Serra P, Howlett S, Hunter K, Garner VE, Gonzalez-Munoz A, Clark J, Veijola R, Cubbon R, Chen SL, Rosa R, Cumiskey AM, Serreze DV, Gregory S, Rogers J, Lyons PA, Healy B, Smink LJ, Todd JA, Peterson LB, Wicker LS, Santamaria P. 2007. Interleukin-2 gene variation impairs regulatory T cell function and causes autoimmunity. Nat Genet 39(3):329-337.
Chen J, Chen Y-G, Reifsnyder PC, Schott WH, Lee C-H, Scheuplein F, Haag F, Koch-Nolte F, Serreze DV, Leiter EH. 2006. Targeted disruption of CD38 accelerates autoimmune diabetes in NOD/Lt mice by enhancing autoimmunity in an ADP-ribosyltransferase 2-dependent fashion. J Immunol 176:4590-4599.
Chen Y-G, Chen J, Osborne MA, Chapman HD, Besra GS, Porcelli SA, Wilson SB, Leiter EH, Serreze DV. 2006. CD38 is required for the peripheral survival of immunotolerogenic CD4+ invariant NK T cells in nonobese diabetic mice. J. Immunol 177:2939-2947.
Lee C-H, Chen Y-G, Chen J, Reifsnyder PC, Serreze DV, Clare-Salzler M, Rodriguez M, Wasserfall C, Atkinson MA, Leiter EH. 2006. Novel leptin receptor mutation in NOD/LtJ mice suppresses type 1 diabetes progression. II. Immunologic analysis. Diabetes 55:171-178.
Razavi R, Chan Y, Afifiyan FN, Liu XJ, Wan X, Yantha J, Tsui H, Tang L, Tsai S, Santamaria P, Driver JP, Serreze DV, Salter MW, Dosch HM. 2006. TRPV1+ sensory neurons control beta cell stress and islet inflammation in autoimmune diabetes. Cell 127:1123-1135.
Serreze DV, Osborne MA, Chen Y-G, Chapman HD, Pearson T, Greiner DL. 2006. Partial versus full allogeneic hematopoietic chimerization is a preferential means to inhibit type 1 diabetes as the latter induces generalized immunosuppression. J Immunol 177:6675-6684.
Silveira PA, Chapman HD, Stolp J, Johnson E, Cox SL, Hunter K, Wicker L, Serreze DV. 2006. Genes within the Idd5 and Idd9/11 diabetes susceptibility loci affect the pathogenic activity of B-cells in nonobese diabetic mice. J Immunol 177:7033-7041.
Takaki T, Marron MP, Mathews CE, Guttman ST, Bottino R, Trucco M, DiLorenzo TP, Serreze DV. 2006. HLA-A*201-restricted T cells from humanized NOD mice recognize autoantigens of potential clinical relevance to type 1 diabetes. J Immunol 176:3257-3265.
Chen Y-G, Choisy-Rossi C-M, Holl TM, Chapman HD, Besra GS, Porcelli SA, Shaffer DJ, Roopenian D, Wilson SB, Serreze DV. 2005. Activated NKT cells inhibit autoimmune diabetes through tolerogenic recruitment of dendritic cells to pancreatic lymph nodes. J Immunol 174:1196-1204.
DiLorenzo TP, Serreze DV. 2005. The good turned ugly: immunopathogenic basis for diabetogenic CD8+ T cells in NOD mice. Immunol Rev 204:250-263.
Gordon EJ, Wicker LS, Peterson LB, Serreze DV, Markees TG, Shultz LD, Rossini AA, Greiner DL, Mordes JP. 2005. Autoimmune diabetes and resistance to xenograft transplantation tolerance in NOD mice. Diabetes 54:107-115.
Pierce MA, Svetlanov A, Horwitz MS, Serreze DV. 2005. Adenovirus early region 3 transgenes expressed in β cells prevent autoimmune diabetes in nonobese diabetic mice: effects of deleting the adenovirus death protein in 11.6K. J Virol 79:619-621.
Pomerleau DP, Bagley RJ, Holl TM, Serreze DV, Mathews CE, Leiter EH. 2005. Major histocompatibility complex-linked diabetes susceptibility in NOD/Lt mice: subcongenic analysis localizes a component of Idd16 at the H2-D end of the diabetogenic H2g7 complex. Diabetes 54:1603-1606.
Reifsnyder PC, Li R, Silveira P, Churchill G, Serreze DV, Leiter EH. 2005. Conditioning the genome identifies additional diabetes resistance loci in Type 1 diabetes resistant NOR/Lt mice. Genes Immun 6:528-538.
Serreze DV, Chen Y-G. 2005. Of mice and men: use of animal models to identify possible interventions for the prevention of autoimmune type 1 diabetes in humans. Trends Immunol 26:603-608.
Serreze DV, Wasserfall C, Ottendorfer EW, Stalvey M, Pierce MA, Gauntt C, O'Donnell B, Flanagan JB, Campbell-Thompson M, Ellis TM, Atkinson MA. 2005. Diabetes acceleration or prevention by a coxsackie B4 infection: critical requirements for both interleukin-4 and gamma interferon. J Virol 79:1045-1052.
Xu BY, Yang H, Serreze DV, MacIntosh R, Yu W, Wright JR. 2005. Rapid destruction of encapsulated islet xenografts by NOD mice is CD4-dependent and facilitated by B-cells: innate immunity and autoimmunity do not play significant roles. Transplantation 80:402-409.
Choisy-Rossi CM, Holl TM, Pierce MA, Chapman HD, Serreze DV. 2004. Enhanced pathogenicity of diabetogenic T cells escaping a non-MHC gene-controlled near death experience. J Immunol 173:3791-3800.
Fallarino F, Bianchi R, Orabona C, Vacca C, Belladonna ML, Fioretti MC, Serreze DV, Grohmann U, Puccetti P. 2004. CTLA-4ñIg activates forkhead transcription factors and protects dendritic cells from oxidative stress in nonobese diabetic mice. J Exp Med 200:1051-1062.
Lieberman SM, Takaki T, Han B, Santamaria P, Serreze DV, DiLorenzo TP. 2004. Individual nonobese diabetic mice exhibit unique patterns of CD8+ T cell reactivity to three islet antigens, including the newly identified widely expressed dystrophia myotonica kinase. J Immunol 173:6727-6734.
Mordes JP, Serreze DV, Greiner DL, Rossini AA. 2004. Animal models of autoimmune diabetes mellitus. In: Diabetes Mellitus: A Fundamental and Clinical Text, 3rd Edition,
LeRoith D (ed). Lippincott, Williams, & Wilkins Publishers, New York. p. 591.
Pearson T, Weiser P, Markees TG, Serreze DV, Wicker LS, Peterson LB, Cumisky AM, Shultz LD, Mordes JP, Rossini AA, Greiner DL. 2004. Islet allograft survival induced by costimulation blockade in NOD mice is controlled by allelic variants of Idd3. Diabetes 53:1972-1978.
Serreze DV, Holl TM, Marron MP, Graser RT, Johnson EA, Choisy-Rossi C, Slattery RM, Lieberman SM, DiLorenzo TP. 2004. MHC class II molecules play a role in the selection of autoreactive class-I restricted CD8 T cells that are essential contributors to type 1 diabetes development in nonobese diabetic mice. J Immunol 172:871-879.
Silveira PA, Dombrowsky J, Johnson E, Chapman HD, Nemazee D, Serreze DV. 2004. B cell selection defects underlie the development of diabetogenic APCs in nonobese diabetic mice. J Immunol 172:5086-5094.
Takaki T, Lieberman SM, Holl TM, Han B, Santamaria P, Serreze DV, DiLorenzo TP. 2004. Requirement for both H-2Db and H-2Kd for the induction of diabetes by the promiscuous CD8+ T cell clonotype AI4. J Immunol 173:2530-2541.
