Functional studies of stable L cell transfectants expressing intercellular adhesion molecule 1 [ICAM-1].
High molecular weight DNA isolated from human tonsil was transfected into mouse L cells to produce transfectants expressing the human intercellular adhesion molecule [ICAM-1]. The transfected ICAM-1 molecule was highly expressed on the cell membrane and our studies show that the transfected ICAM-1 molecule was a fully functional adhesion protein. All leucocyte subtypes showed specific binding to the ICAM-1 transfectants, their adhesion being inhibited by Fab1 fragments of W-CAM-1 antibody and LFA-1 MoAb (leucocyte function antigen). B cell lines (RAJI, Nalm1) showed the highest degree of ICAM-1 mediated adhesion. Normal lymphoblasts showed comparable levels of binding whilst normal neutrophils (both resting and activated by (N-formyl-methionyl-leucyl-phenylalanine) fMLP) showed the least ICAM-1-mediated adhesion. Despite a significant level of adhesion to ICAM-1 transfectants shown by T lymphoblasts generated in a two way MLR there was no evidence of cytolysis of ICAM-1 transfectants. These studies demonstrate the potential of ICAM-1 transfectants as tools for analysis of the role of ICAM-1 in lymphoid adhesion.
Wawryk
S O
SO
Lions Clinical Cancer Research Laboratory, Walter and Eliza Hall Institute of Experimental Medicine, PO Royal Melbourne Hospital, Vic., Australia.
Salvaris
E
E
Sandrin
M S
MS
Boyd
A W
AW
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000943:Antigens, Differentiation / Q000276:immunology; D002448:Cell Adhesion / Q000276:immunology*; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005470:Fluorometry; D006801:Humans; D015151:Immunoblotting; D018799:Intercellular Adhesion Molecule-1; D007739:L Cells; D007962:Leukocytes / Q000276:immunology; D016169:Lymphocyte Function-Associated Antigen-1; D008565:Membrane Proteins / Q000276:immunology; D051379:Mice; D009504:Neutrophils / Q000276:immunology; D014066:Palatine Tonsil / Q000648:ultrastructure; D011233:Precipitin Tests; D016028:Receptors, Leukocyte-Adhesion / Q000276:immunology; D012397:Rosette Formation; D014162:Transfection
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D015815:Cell Adhesion Molecules; D016169:Lymphocyte Function-Associated Antigen-1; D008565:Membrane Proteins; D016028:Receptors, Leukocyte-Adhesion; D018799:Intercellular Adhesion Molecule-1; D004247:DNA