title: Characterization of Innate Immunity in an Extended Whole Blood Model of Human Islet Allotransplantation.
abstract: The instant blood-mediated inflammatory reaction (IBMIR) has been studied in whole blood models of human allo-islet transplantation for short periods (<6 h). Beyond this time frame the innate response to intraportally transplanted islets is less well described. A novel whole blood model was applied to study blood-islet-graft interactions up to 48 h. Heparinized polyvinyl chloride tubing was sealed into small bags containing venous blood together with allogeneic human islets and exocrine tissue, respectively. The bags were attached to a rotating wheel (37°C). Concentrated glucose and sodium hydrogen carbonate were added every 12 h to maintain physiological limits for sustained immune cell functions. Plasma was collected at repeated time points for analyses of coagulation/complement activation and chemokine/cytokine production. Immune cell infiltration was analyzed using immunohistochemistry. Coagulation and platelet activation markers, thrombin-antithrombin complex (TAT) and soluble CD40 ligand (sCD40L) showed early high concentrations (at 6-12 h). sC5b-9 steadily increased over 48 h. At 6 h neutrophils and monocytes surrounded the clotted cellular grafts with a following massive infiltration of neutrophils. High and increasing concentrations of CXCR1/2 ligands [IL-8 and growth-regulated oncogene α/β/γ (Gro-α/β/γ)] and IL-6 were produced in response to human islets and exocrine tissue. The CCR2 ligand monocyte chemoattractant protein 1 (MCP-1) exhibited increasing concentrations in response to exocrine tissue. The CXCR3 ligand interferon-inducible T cell α chemoattractant (I-TAC) was produced in response to both human islets and exocrine tissue from 6 h. Monokine induced by γ interferon (Mig) and interferon γ-induced protein 10 (IP-10) showed a later response, preferentially to exocrine tissue and with larger variations among preparations. An extended blood model of clinical islet transplantation allowed characterization of early immune activation in response to human islets and exocrine tissue. Increased production of chemokines targeting CXCR1/2, CCR2, and CXCR3 was observed, accompanied by massive intraislet neutrophil infiltration over 48 h. The model proved to be useful in exploring early blood-mediated reactions to cellular transplants and has relevance for evaluation of pharmacological interventions to prevent graft loss.
authors.affiliation: Department of Immunology, Genetics and Pathology, Clinical Immunology, The Rudbeck Laboratory, Uppsala University, Uppsala, Sweden.
authors.forename: Maria
authors.initials: M
authors.lastname: Hårdstedt
authors.forename: Susanne
authors.initials: S
authors.lastname: Lindblom
authors.forename: Alex
authors.initials: A
authors.lastname: Karlsson-Parra
authors.forename: Bo
authors.initials: B
authors.lastname: Nilsson
authors.forename: Olle
authors.initials: O
authors.lastname: Korsgren
mesh_terms: D000990:Antithrombin III / Q000276:immunology; D001777:Blood Coagulation; D019013:CD40 Antigens / Q000097:blood / Q000276:immunology; D002478:Cells, Cultured; D018925:Chemokines / Q000097:blood / Q000276:immunology; D003167:Complement Activation; D006801:Humans; D007111:Immunity, Cellular; D007113:Immunity, Innate; D007249:Inflammation / Q000097:blood* / Q000276:immunology; D007378:Interleukins / Q000097:blood / Q000276:immunology; D016381:Islets of Langerhans Transplantation / Q000276:immunology*; D010447:Peptide Hydrolases / Q000097:blood / Q000276:immunology; D015539:Platelet Activation; D014184:Transplantation, Homologous
chemical_list: D019013:CD40 Antigens; D018925:Chemokines; D007378:Interleukins; C046193:antithrombin III-protease complex; D000990:Antithrombin III; D010447:Peptide Hydrolases