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Research Article Free access | 10.1172/JCI110181

Responses of Canine Lymphocytes to Allogeneic and Autologous Islets of Langerhans in Mixed Cell Cultures

Alexander Rabinovitch, Laphalle Fuller, Daniel Mintz, Walter Severyn, Jack Noel, Cathy Flaa, George Kyriakides, and Joshua Miller

Department of Medicine, University of Miami School of Medicine, Miami, Florida 33101

Department of Microbiology, University of Miami School of Medicine, Miami, Florida 33101

Department of Surgery, University of Miami School of Medicine, Miami, Florida 33101

Miami Veterans Administration Hospital, Miami, Florida 33101

Find articles by Rabinovitch, A. in: JCI | PubMed | Google Scholar

Department of Medicine, University of Miami School of Medicine, Miami, Florida 33101

Department of Microbiology, University of Miami School of Medicine, Miami, Florida 33101

Department of Surgery, University of Miami School of Medicine, Miami, Florida 33101

Miami Veterans Administration Hospital, Miami, Florida 33101

Find articles by Fuller, L. in: JCI | PubMed | Google Scholar

Department of Medicine, University of Miami School of Medicine, Miami, Florida 33101

Department of Microbiology, University of Miami School of Medicine, Miami, Florida 33101

Department of Surgery, University of Miami School of Medicine, Miami, Florida 33101

Miami Veterans Administration Hospital, Miami, Florida 33101

Find articles by Mintz, D. in: JCI | PubMed | Google Scholar

Department of Medicine, University of Miami School of Medicine, Miami, Florida 33101

Department of Microbiology, University of Miami School of Medicine, Miami, Florida 33101

Department of Surgery, University of Miami School of Medicine, Miami, Florida 33101

Miami Veterans Administration Hospital, Miami, Florida 33101

Find articles by Severyn, W. in: JCI | PubMed | Google Scholar

Department of Medicine, University of Miami School of Medicine, Miami, Florida 33101

Department of Microbiology, University of Miami School of Medicine, Miami, Florida 33101

Department of Surgery, University of Miami School of Medicine, Miami, Florida 33101

Miami Veterans Administration Hospital, Miami, Florida 33101

Find articles by Noel, J. in: JCI | PubMed | Google Scholar

Department of Medicine, University of Miami School of Medicine, Miami, Florida 33101

Department of Microbiology, University of Miami School of Medicine, Miami, Florida 33101

Department of Surgery, University of Miami School of Medicine, Miami, Florida 33101

Miami Veterans Administration Hospital, Miami, Florida 33101

Find articles by Flaa, C. in: JCI | PubMed | Google Scholar

Department of Medicine, University of Miami School of Medicine, Miami, Florida 33101

Department of Microbiology, University of Miami School of Medicine, Miami, Florida 33101

Department of Surgery, University of Miami School of Medicine, Miami, Florida 33101

Miami Veterans Administration Hospital, Miami, Florida 33101

Find articles by Kyriakides, G. in: JCI | PubMed | Google Scholar

Department of Medicine, University of Miami School of Medicine, Miami, Florida 33101

Department of Microbiology, University of Miami School of Medicine, Miami, Florida 33101

Department of Surgery, University of Miami School of Medicine, Miami, Florida 33101

Miami Veterans Administration Hospital, Miami, Florida 33101

Find articles by Miller, J. in: JCI | PubMed | Google Scholar

Published May 1, 1981 - More info

Published in Volume 67, Issue 5 on May 1, 1981
J Clin Invest. 1981;67(5):1507–1516. https://doi.org/10.1172/JCI110181.
© 1981 The American Society for Clinical Investigation
Published May 1, 1981 - Version history
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Abstract

Because successful allotransplantation of islets of Langerhans isolated by collagenase digestion has been difficult in many animal species, we asked whether isolated islet preparations might have tissue specific determinants conferring amplified immunogenicity in vitro. Lymphocyte proliferative responses ([3H]thymidine uptake) were studied in beagle dogs in mixed culture combinations of lymphocyte vs. lymphocyte (MLC) and lymphocyte vs. islet (MLIC). In five MLC responder and five nonresponder pairs, peripheral blood lymphocytes of dogs A and B were used as responding cells, and dog B x-irradiated lymphocytes (Bx), x-irradiated (or nonirradiated) islets (BI), or hepatic cells (BH) were used as stimulating cells in primary and secondary reactions. For the secondary reactions, A + Bx, A + BI, or B + BI were incubated for 9 d (A′B, A′BI, B′BI, respectively) before addition of new stimulating cells.

The results showed that islets were autostimulatory, eliciting a tissue-specific lymphoproliferative response in a primary MLIC. Thus, B + BI reactivity was evident at 3,5, and 7 d in primary culture, whereas collagenase-digested liver cells, or lymphocytes obtained from collagenase-digested lymph nodes did not stimulate autologous lymphocytes. A separate reactivity was observed in the allogeneic A + BI combination in MLC responder pairs, and the peak response of A + BI at 9 d was markedly greater than that of B + BI, suggesting the presence of major histocompatibility complex lymphocyte-defined locus determinants in the islet preparations, in addition to islet-specific determinants. A secondary reaction was observed if lymphocytes were primed with islets and challenged with islets (A′BI + BI or B′BI + BI), but not if they were challenged with lymphocytes (A′BI + Bx, B′BI + Bx) or hepatic cells (A′BI + BH, B′BI + AH). Furthermore, priming of lymphocytes with autologous islets (B′BI) led to exclusion of any reactivity against allogeneic lymphocytes, i.e., B′BI suppressed A + Bx, and B′BI also markedly suppressed phytohemagglutinin-stimulated lymphoproliferative responses. Experiments were performed that excluded the possibility that the insulin levels present in the MLIC, the presence of passenger lymphocytes in the islets, or the maintenance of islets in tissue culture for 1-7 d affected the observations. These results provide evidence for the existence of alloantigens as well as tissue-specific antigens on collagenase-isolated islets of Langerhans.

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