Cytokines are soluble, antigen non‐specific, non‐immunoglobulin mediators produced and secreted by blood mononuclear cells interacting in the cellular immune‐response. To test the possibility that cytokines participate in the autoimmune destruction of the pancreatic beta‐cells leading to insulin‐dependent diabetes mellitus, isolated human or rat islets of Langerhans were incubated for 7 days with cytokine‐rich, cell‐free supernatants of blood mononuclear cells from healthy human donors stimulated with or without purified protein derivative of tuberculin or phytohaemagglutinin. Glucose stimulated insulin‐release, and contents of insulin and glucagon in islets incubated with cytokine‐rich supernatants were markedly reduced. This impairment of islet function was due to a cytotoxic effect of cytokine‐rich supernatants as judged by disintegration of normal light‐microscopic morphology.

The morphologic and metabolic effect of a single intracutaneous injection of homologous endocrine pancreas in Freund's complete adjuvant (CFA) was studied in 100 mice and compared with control groups which had been (1) immunized with murine insulin in CFA, (2) injected with CFA alone, or had (3) received no treatment. There were no differences between the control groups as regards the morphology of the pancreatic islets, and the glucose tolerance was normal. Mice immunized with islet homogenate exhibited morphological changes in the form of degranulation and cytoplasmic disintegration. These changes involved B-cells as well as A2-cells and were present from 7 to 18 days after the immunization. A significant reduction in glucose tolerance was observed 14 days after the immunization. Another characteristic finding in the islets from the immunized mice was the extra-vascular presence of mononuclear, agranular cells which on the basis of their morphological criteria appeared to represent lymphocytes.

Mice developed degenerative changes of B-cells with loss of insulin granules, extravascularly situated mononuclear cell infiltration of the islets of Langerhans, and glucose intolerance eight and fifteen days after immunization with homologous islets in complete Freund's adjuvant. Murine insulin and adjuvants in comparable preparations did not cause similar changes. The condition was transient, but has the following features in common with juvenile diabetes mellitus in man: glucose intolerance, mononuclear cell infiltration of the islets and organ specific, anti-pancreatic cellular hypersensitivity. It may therefore serve as a useful model in diabetes research.

AllergyVolume 28, Issue 4 p. 223-230 ANTI-PANCREATIC, CELLULAR HYPERSENSITIVITY IN DIABETES MELLITUS. ANTIGENIC ACTIVITY OF FETAL CALF PANCREAS AND CORRELATION WITH CLINICAL TYPE OF DIABETES JØSRN NERUP, JØSRN NERUP From Medical Department TA, Laboratory of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorOLE ORTVED ANDERSEN, OLE ORTVED ANDERSEN From Medical Department TA, Laboratory of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorGUNNAR BENDIXEN, GUNNAR BENDIXEN From Medical Department TA, Laboratory of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorJØRN EGEBERG, JØRN EGEBERG From Medical Department TA, Laboratory of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorJACOB E. POULSEN, JACOB E. POULSEN From Medical Department TA, Laboratory of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this author JØSRN NERUP, JØSRN NERUP From Medical Department TA, Laboratory of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorOLE ORTVED ANDERSEN, OLE ORTVED ANDERSEN From Medical Department TA, Laboratory of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorGUNNAR BENDIXEN, GUNNAR BENDIXEN From Medical Department TA, Laboratory of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorJØRN EGEBERG, JØRN EGEBERG From Medical Department TA, Laboratory of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorJACOB E. POULSEN, JACOB E. POULSEN From Medical Department TA, Laboratory of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this author First published: October 1973 https://doi.org/10.1111/j.1398-9995.1973.tb01443.xCitations: 33AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume28, Issue4October 1973Pages 223-230 RelatedInformation

AllergyVolume 28, Issue 4 p. 231-249 ANTI-PANCREATIC, CELLULAR HYPERSENSITIVITY IN DIABETES MELLITUS. EXPERIMENTAL INDUCTION OF ANTI-PANCREATIC, CELLULAR HYPERSENSITIVITY AND ASSOCIATED MORPHOLOGICAL B-CELL CHANGES IN THE RAT JØSRN NERUP, JØSRN NERUP From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorOLE ORTVED ANDERSEN, OLE ORTVED ANDERSEN From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorGUNNAR BENDIXEN, GUNNAR BENDIXEN From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorJØRN EGEBERG, JØRN EGEBERG From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorJACOB E. POULSEN, JACOB E. POULSEN From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorMOGENS VILIEN, MOGENS VILIEN From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorMIKKEL WESTRUP, MIKKEL WESTRUP From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this author JØSRN NERUP, JØSRN NERUP From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorOLE ORTVED ANDERSEN, OLE ORTVED ANDERSEN From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorGUNNAR BENDIXEN, GUNNAR BENDIXEN From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorJØRN EGEBERG, JØRN EGEBERG From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorJACOB E. POULSEN, JACOB E. POULSEN From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorMOGENS VILIEN, MOGENS VILIEN From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this authorMIKKEL WESTRUP, MIKKEL WESTRUP From Medical Department TA, Division of Clinical Immunology, Rigshospitalet University Hospital, Copenhagen; Steno Memorial Hospital, Gentofte; and Anatomy Department B, University of Copenhagen, Denmark.Search for more papers by this author First published: October 1973 https://doi.org/10.1111/j.1398-9995.1973.tb01444.xCitations: 25AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL REFERENCES 1 Bendixen, G. & Søborg, M. (1969): A leucocyte migration technique for in vitro detection of cellular (delayed type) hypersensitivity in man. Danish Med. Bull. 16, 1– 6. 2 B. R. Bloom & P. R. Glade, (Eds.) (1971): In vitro methods in cell-mediated immunity. Academic Press. New York, London . 3 Deckert, T. (1967): Autoimmunological aspects of diabetes mellitus. Acta Med. Scand., Suppl. 476, 29– 41. 4 Federlin, K. (1971): Immunopathology of insulin. Springer, Berlin, Heidelberg, New York p. 129. 5 Freytag, G, Mitschke, H. & Klöppel, G. (1970): Immunopathologische untersuchungen èur experimentellen insulitis. Verh. Dtsch. Ges. Path. 54, 290– 297. 6 Gepts, W. (1965): Pathologic anatomy of the pancreas in juvenile diabetes mellitus. Diabetes 14, 619– 633. 7 Grodsky, G. M., Feldman, R., Toreson, W. E. & Lee, J. C. (1966): Diabetes mellitus in rabbits immunized with insulin. Diabetes 15, 579– 585. 8 Kåresen, R. (1970): Experimental allergic thyroiditis in the guinea-pig. A light, fluorescence and electron microscopic study, with particular reference to the migration of lymphocytes through the vessel walls. Acta path, microbiol. scand. Section A. 78, 625– 648. 9 Luft, J. H. (1961): Improvement in epoxy resin embedding methods. J. Biophys. Biochem. Cytol. 9, 404– 414. 10 Nerup, J., Ortved Andersen, O., Bendixen, G., Egeberg, J. & Poulsen, J. E. (1971): Anti-pancreatic cellular hypersensitivity in diabetes mellitus. Diabetes 20, 424– 427. 11 Nerup, J., Ortved Andersen, O., Bendixen, G., Egeberg, J. & Poulsen, J. E. (1973): Anti-pancreatic, cellular hypersensitivity in diabetes mellitus. Antigenic activity of fetal calf pancreas and correlation with clinical type of diabetes. Acta allergol. 28, —. 12 Palade, G. E. (1952): A study of fixation for electron microscopy. J. Exp. Med. 95, 285– 297. 13 Renold, A. E., Soeldener, J. S. & Steinke, J. (1964): Immunological studies with homologous and heterologous pancreatic insulin in the cow. Ciba foundation Colloq. Endocr. p. 122, Churchill, London . 14 Reynolds, E. S. (1963): The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. Cell. Biol. 77, 208– 212. 15 Sabatini, D. D., Bensch, K. G. & Barrnett, R. J. (1963): Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J. Cell Biol. 17, 19– 58. Citing Literature Volume28, Issue4October 1973Pages 231-249 ReferencesRelatedInformation

Leucocyte migration cultures for detection of cellular hypersensitivity in man were prepared for light and electron microscopy 2 and 4 hours after the onset of migration. Antigen‐containing as well as control cultures were studied. The cell types found were polymorphonuclear granulocytes (PMN), eosinophilic granulocytes (EO) and lymphocytes (LYM). In the central part of the cultures differential countings showed values comparable to those of peripheral blood. The peripheral monolayer of the migration area contained a large proportion of EO and comparatively few LYM. The cell number of the monolayer increased between the second and fourth hours of migration. Ultrastructurally the PMN presented a more or less disintegrated cytoplasm, but their ability to phagocytize and hydrolyse the corpuscular antigen (brucella bacteria) seemed undisturbed. The EO and LYM appeared less active, although their cytoplasm also contained signs of phagocytic activity. The LYM contained sparse elements of granular endoplasmic reticulum. Apart from the bacterial phagocytosis in the PMN no morphological differences were noted between the cells in the control and antigen‐containing cultures.

The occurrence of organ-specific, cellular hypersensitivity against pancreatic components was examined in twenty-two diabetics by means of the leucocyte migration test. An extract was prepared from pooled porcine pancreatic glands, in which atrophy of the exocrine tissue had been induced by ligation of the pancreatic duct. A specifically altered in vitro reactivity to the pancreatic preparation, consistent with a state of organ-specific, cellular hypersensitivity, was demonstrated in the diabetic group as compared to a control group. Intracutaneous injection of the same preparation in six diabetics with a positive in vitro reaction induced a typical delayed type reaction in four.