Long-term culture of islets abrogates cytokine-induced or lymphocyte-induced increase of antigen expression on beta cells.

Abstract:

BACKGROUND:Immunogenicity of a graft depends on its expression of major histocompatability complex (MHC) antigens and adhesion molecules and on the amount of intragraft leukocytes, the so-called passenger leukocytes. Although long-term culture reduces passenger leukocytes, permanent acceptance is not necessarily observed after allogeneic transplantation. Because little is known about antigen expression on the surface of islet cells after long-term culture of islets, we investigated whether antigen expression of pancreatic beta cells is influenced by long-term culture and whether long-term culture can counteract the increase of antigen expression induced by cytokines or by allogeneic lymphocytes. We also investigated whether long-term cultured islets were able to stimulate allogeneic lymphocytes to produce cytokines. METHODS:Isolated LEW.1A (RT1a) rat islets of Langerhans were cultured for 14 and 28 days. Precultured and freshly-isolated islets were then incubated for 2 days with rat recombinant interferon (rIFN)-gamma (1,000 IU/mL), or were co-cultured for 4 days with LEW.1W (RT1u) splenic lymphocytes. RESULTS:Long-term culture significantly reduced CD45 leukocytes within the islets and decreased the amount of beta cells expressing intercellular adhesion molecule (ICAM)-1, whereas MHC antigen expression remained unchanged. After incubation of freshly isolated islets with IFN-gamma induction of MHC class II antigens on beta cells, an increase of MHC class I antigen density and an enhancement of ICAM-1+ beta cells were observed. Similar results were found after co-culturing with allogeneic lymphocytes. Using precultured islets, the induction of MHC class II on beta cells by IFN-gamma was still present but significantly lower and was absent after co-culture with allogeneic lymphocytes. Enhancement of ICAM-1+ beta cells by IFN-gamma or by allogeneic lymphocytes was markedly lowered because of preculturing. The proportion of MHC class I beta cells remained unchanged; however, antigen density of long-term cultured islets (28 days) could not be enhanced by allogeneic lymphocytes. Precultured islets were not able to stimulate allogeneic lymphocytes to produce and release normal amounts of cytokines (IFN-gamma or interleukin [IL]-2). CONCLUSIONS:In conclusion, in addition to reduction-depletion of passenger leukocytes, long-term culturing of islets also is able to counteract the IFN-gamma-induced or allogeneic lymphocyte-induced increase of antigen expression. Therefore, initiation of rejection and generation of cytotoxic cells might be altered or timely delayed when long-term cultured islets are transplanted. The variable and conflicting in vivo results after transplantation of long-term cultured islets might be explained by the possible indirect antigen presentation, which is not influenced by islet preculture.

journal_name

Transplantation

journal_title

Transplantation

authors

Kuttler B,Hartmann A,Wanka H

doi

10.1097/00007890-200208270-00003

subject

Has Abstract

pub_date

2002-08-27 00:00:00

pages

440-5

issue

4

eissn

0041-1337

issn

1534-6080

journal_volume

74

pub_type

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