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Transcriptional profiling of stress response in cultured porcine islets
University of Minnesota.
Diabetes Institute of Immunology and Transplantation, Department of Surgery, University of Minnesota, Minneapolis, USA.
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2007 (English)In: Biochemical and Biophysical Research Communications - BBRC, ISSN 0006-291X, E-ISSN 1090-2104, Vol. 357, no 1, 118-125 p.Article in journal (Refereed) Published
Abstract [en]

Cell-based diabetes therapy may be achieved through xenotransplantation of adult porcine islets, but tissue quality and immunoreactivity barriers need to be overcome. Early identification and exclusion of irreversibly stressed and dying islets may improve transplant outcomes. We used oligonucleotide microarray and quantitative RT-PCR to identify molecular markers of physiological and immunological stress in porcine islets cultured under stress conditions of elevated glucose (16.7 mM), inflammatory cytokine addition (IL-1beta, TNF-alpha, and IFN-gamma), or both, for 48 h. Hyperglycemic conditions were associated with increased thioredoxin interacting protein and metabolic process mRNAs, as observed in rodent and primate species. Cytokine treatment increased expression of JAK-STAT pathway components, oxidative stress (transglutaminase 2), and beta cell dysfunction genes. Transglutaminase 2 induction is unique to porcine islets. Biomarkers involved in hyperglycemia and islet inflammation may serve as novel targets for improving and monitoring isolated porcine islet function and viability.

Place, publisher, year, edition, pages
2007. Vol. 357, no 1, 118-125 p.
National Category
Immunology in the medical area
URN: urn:nbn:se:uu:diva-232861DOI: 10.1016/j.bbrc.2007.03.101PubMedID: 17407763OAI: oai:DiVA.org:uu-232861DiVA: diva2:750009
Available from: 2014-09-25 Created: 2014-09-25 Last updated: 2014-10-20Bibliographically approved
In thesis
1. Studies of Innate and Adaptive Immunity in Islet Transplantation
Open this publication in new window or tab >>Studies of Innate and Adaptive Immunity in Islet Transplantation
2014 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Clinical islet transplantation is today an established alternative treatment for a selected group of type 1 diabetes patients. The predominant technique for transplantation is infusion of islets in the liver via the portal vein. Obstacles to advancing islet transplantation include limited engraftment resulting from an immediate blood-mediated inflammatory reaction (IBMIR), a life-long need for immunosuppression and the shortage of organs available.

In this thesis, innate and adaptive immunity were explored in allogeneic and xenogeneic settings, with the long-term goal of preventing islet graft destruction. Methods for studying immune responses to islets in blood and engrafted islets in liver tissue (intragraft gene expression) were developed and refined. The innate response to human islets and exocrine tissue in ABO-compatible blood was characterized up to 48 h using a novel whole-blood model. Physiological changes in the blood during incubations were explored and adjusted to allow prolonged experiments. Increased production of chemokines targeting CXCR1/2, CCR2 and CXCR3 was observed, accompanied by massive intra-islet neutrophil infiltration. Notably, endocrine and exocrine tissue triggered a similarly strong innate immune response.

Two studies of adult porcine islet transplantation to non-human primates (NHPs) were performed. Expression of immune response genes induced in liver tissue of non-immunosuppressed NHPs (≤72 h) was evaluated after porcine islet transplantation. Up-regulation of CXCR3 mRNA, together with IP-10, Mig, MIP-1α, RANTES, MCP-1 and cytotoxic effector molecule transcripts, was associated with T-cell and macrophage infiltration at 48-72 h. Long-term survival (>100 days) of adult porcine islets in a NHP model was later demonstrated using T-cell-based immunosuppression, including co-stimulatory blockade (anti-CD154 mAb). Graft failure was associated with increased levels of circulating, indirectly activated T cells, non-Gal pig-specific IgG and gene transcripts of inflammatory cytokines. Microarray analysis of the response to inflammatory cytokines in cultured porcine islets identified genes involved in cell death, immune responses and oxidative stress; this gene pattern coincided with physiological changes (decrease in insulin and ATP content).

In summary, allogeneic whole-blood experiments and xenogeneic in vivo studies underscored the importance of preventing early inflammation and cell-recruitment to avoid islet graft loss in islet transplantation. Long-term survival of porcine islets in NHPs was shown to be feasible using T-cell-directed immunosuppression, including anti-CD154 mAb.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2014. 117 p.
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 1030
diabetes, islet transplantation, islets of Langerhans, xenotransplantation, nonhuman primate, blood, whole blood model, innate immunity, adaptive immunity, IBMIR, chemokines
National Category
Immunology in the medical area
urn:nbn:se:uu:diva-232863 (URN)978-91-554-9046-1 (ISBN)
Public defence
2014-11-07, Fåhreussalen, Rudbecklaboratoriet, Dag Hammarskjölds väg 20, Uppsala, 10:15 (English)
Available from: 2014-10-17 Created: 2014-09-25 Last updated: 2015-01-23

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