Mechanisms involved in the binding of thymocytes to rat thymic dendritic cells.
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 degrees C than at 4 degrees C. 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 TCR alpha beta hi subset. The binding of thymocytes to TDC at 37 degrees C 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 degrees C 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 alpha beta TCR (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.
Colić
M
M
Institute of Medical Research, Military Medical Academy, Belgrade, Yugoslavia.
Ilić
V
V
Pavlović
M D
MD
Tamatani
T
T
Miyasaka
M
M
D000818:Animals; D002413:Cations, Divalent / Q000494:pharmacology; D002448:Cell Adhesion; D005434:Flow Cytometry; D010641:Phenotype; D051381:Rats; D012397:Rosette Formation; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000378:metabolism*; D013950:Thymus Gland / Q000166:cytology*
D002413:Cations, Divalent