title: Interleukins increase surface ganglioside expression of pancreatic islet cells in vitro.
abstract: This experiment was conducted in order to investigate whether expression of gangliosides on islet cell surface in vitro is influenced by cytokines, especially interleukin 1. Islets from adult Lewis rats were incubated with different concentrations of recombinant-derived human cytokines. Following dispase treatment, the single cells were labeled with monoclonal antiganglioside antibodies A2B5 or R2D6, and conjugate. Both are directed against beta cells; A2B5 is recognized to bind specifically to pancreatic islet cells, while R2D6 is shown to bind no other pancreatic cells than beta cells. Surface labeling was evaluated in blind trials using a fluorescence microscope and a fluorescence-activated cell sorter (FACS). A2B5 staining demonstrated a significantly higher number of labeled cells after incubation with interleukin 1 alpha (14.9% +/- 2.8; p less than 0.005), interleukin 1 beta (23.2% +/- 4.2; p less than 0.0005) or TNF alpha (16.1% +/- 4.0; p = 0.005) compared to endotoxin controls (4.1% +/- 1.1). Interleukin 1 beta (9.5% +/- 1.5; p less than 0.005) showed a significantly increased number of R2D6-stained cells (control: 2.3% +/- 1.3). A similar but not significant effect was seen with interleukin 1 alpha and TNF alpha. Interleukin 6 had no effect on the antigen expression. The intensity of labeling was elevated among interleukin 1 beta-incubated cells compared to control samples. Thus, treatment of islets with different cytokines, especially interleukin 1 beta, increases surface antigen expression. We suggest that this mechanism of action in vitro may be of importance for the putative diabetogenic effect of interleukin 1.
authors.affiliation: Bartholin Instituttet, Kommunehospitalet, Copenhagen, Denmark.
authors.forename: T W
authors.initials: TW
authors.lastname: Kjaer
authors.forename: J
authors.initials: J
authors.lastname: Rygaard
authors.forename: K
authors.initials: K
authors.lastname: Bendtzen
authors.forename: K
authors.initials: K
authors.lastname: Josefsen
authors.forename: T
authors.initials: T
authors.lastname: Bock
authors.forename: K
authors.initials: K
authors.lastname: Buschard
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000737:chemistry; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005732:Gangliosides / Q000032:analysis*; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D007378:Interleukins / Q000494:pharmacology*; D007515:Islets of Langerhans / Q000737:chemistry* / Q000166:cytology* / Q000276:immunology; D008297:Male; D008856:Microscopy, Fluorescence; D051381:Rats; D011917:Rats, Inbred Lew; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D000954:Antigens, Surface; D005732:Gangliosides; D007375:Interleukin-1; D015850:Interleukin-6; D007378:Interleukins; D014409:Tumor Necrosis Factor-alpha