title: Perforin facilitates beta cell killing and regulates autoreactive CD8+ T-cell responses to antigen in mouse models of type 1 diabetes.
abstract: In type 1 diabetes, cytotoxic CD8(+) T lymphocytes (CTLs) directly interact with pancreatic beta cells through major histocompatibility complex class I. An immune synapse facilitates delivery of cytotoxic granules, comprised mainly of granzymes and perforin. Perforin deficiency protects the majority of non-obese diabetic (NOD) mice from autoimmune diabetes. Intriguingly perforin deficiency does not prevent diabetes in CD8(+) T-cell receptor transgenic NOD8.3 mice. We therefore investigated the importance of perforin-dependent killing via CTL-beta cell contact in autoimmune diabetes. Perforin-deficient CTL from NOD mice or from NOD8.3 mice were significantly less efficient at adoptive transfer of autoimmune diabetes into NODRag1(-/-) mice, confirming that perforin is essential to facilitate beta cell destruction. However, increasing the number of transferred in vitro-activated perforin-deficient 8.3 T cells reversed the phenotype and resulted in diabetes. Perforin-deficient NOD8.3 T cells were present in increased proportion in islets, and proliferated more in response to antigen in vivo indicating that perforin may regulate the activation of CTLs, possibly by controlling cytokine production. This was confirmed when we examined the requirement for direct interaction between beta cells and CD8(+) T cells in NOD8.3 mice, in which beta cells specifically lack major histocompatibility complex (MHC) class I through conditional deletion of β2-microglobulin. Although diabetes was significantly reduced, 40% of these mice developed diabetes, indicating that NOD8.3 T cells can kill beta cells in the absence of direct interaction. Our data indicate that although perforin delivery is the main mechanism that CTL use to destroy beta cells, they can employ alternative mechanisms to induce diabetes in a perforin-independent manner.
authors.affiliation: Immunology and Diabetes, St Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia.|Department of Medicine, St Vincent's Hospital, University of Melbourne, Fitzroy, Victoria, Australia.
authors.forename: Prerak
authors.initials: P
authors.lastname: Trivedi
authors.affiliation: Immunology and Diabetes, St Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia.|Department of Medicine, St Vincent's Hospital, University of Melbourne, Fitzroy, Victoria, Australia.
authors.forename: Kate L
authors.initials: KL
authors.lastname: Graham
authors.affiliation: Immunology and Diabetes, St Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia.|Department of Medicine, St Vincent's Hospital, University of Melbourne, Fitzroy, Victoria, Australia.
authors.forename: Balasubramaninan
authors.initials: B
authors.lastname: Krishnamurthy
authors.affiliation: Immunology and Diabetes, St Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia.
authors.forename: Stacey
authors.initials: S
authors.lastname: Fynch
authors.affiliation: Department of Immunology, Faculty of Medicine, Nursing and Health Sciences, Monash University, Alfred Hospital, Prahran, Victoria, Australia.
authors.forename: Robyn M
authors.initials: RM
authors.lastname: Slattery
authors.affiliation: Immunology and Diabetes, St Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia.|Department of Medicine, St Vincent's Hospital, University of Melbourne, Fitzroy, Victoria, Australia.
authors.forename: Thomas W H
authors.initials: TW
authors.lastname: Kay
authors.affiliation: Immunology and Diabetes, St Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia.|Department of Medicine, St Vincent's Hospital, University of Melbourne, Fitzroy, Victoria, Australia.
authors.forename: Helen E
authors.initials: HE
authors.lastname: Thomas
mesh_terms: D000818:Animals; D001324:Autoantigens / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology*; D004195:Disease Models, Animal; D006801:Humans; D050417:Insulin-Secreting Cells / Q000276:immunology*; D008213:Lymphocyte Activation / Q000235:genetics; D051379:Mice; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D019899:Paracrine Communication; D054353:Perforin / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
chemical_list: D001324:Autoantigens; D054353:Perforin