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Developmental Immunology
Volume 5, Issue 1, Pages 37-51
http://dx.doi.org/10.1155/1996/18312

Mechanisms Involved in the Binding of Thymocytes to Rat Thymic Dendritic Cells

1Institute of Medical Research, Military Medical Academy, Crnotravska 17, Belgrade, Serbia and Montenegro
2Department of Immunology, Tokyo Metropolitan Institute of Medical Science, 3-18-22, Hon-Komagome, Bun Kyo, Tokyo 113, Japan

Received 28 June 1995; Accepted 4 October 1995

Copyright © 1996 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The effects of monoclonal antibodies (mAbs) to cell-surface molecules, divalent cations, and various cell-signaling and metabolic inhibitors on the binding of thymocytes to rat thymic dendritic cells (TDC) were studied using a rosette assay. It was found that TDC/thymocyte adhesion was stronger and faster at 37℃ than at 4℃. Flow cytometric analysis demonstrated that bound thymocytes were predominantly CD4+CD8+ and CD4+CD8-, but in comparison to the phenotype of whole thymocytes, they were enriched in the mature TCRαβhi subset. The binding of thymocytes to TDC at 37℃ was almost completely dependent on Ca2+ and Mg2+ and partly on an intact cytoskeleton and calmodulin-dependent protein kinase. The adhesion was independent of new protein synthesis and the activities of protein kinases A and C, tyrosine kinases, as well as phosphotyrosine protein phosphatases. The TDC/thymocyte adhesion at 37℃ was partly blocked by anti-LFA-1 (WT.1), anti-CD18 (WT.3), and anti-ICAM-1 (1A29) mAb. MAbs to class II MHC (OX-3 and OX-6), CD4 (W3/25), CD8 (OX-8), and αβTCR (R73) stimulated the adhesion via an LFA-1-dependent pathway, whereas an anti-CD45 mAb (G3C5) stimulated the rosette formation independently of LFA-1. MAbs to CD2 (OX-34), CD11b (ED7), CD11b/c (OX-42), and class I MHC (OX-18) were without significant effects on the adhesion process.