Identification and characterization of a 100-kD ligand for CD6 on human thymic epithelial cells.
CD6 is a 130-kD glycoprotein expressed on the surface of thymocytes and peripheral blood T cells that is involved in TCR-mediated T cell activation. In thymus, CD6 mediates interactions between thymocytes and thymic epithelial (TE) cells. In indirect immunofluorescence assays, a recombinant CD6-immunoglobulin fusion protein (CD6-Rg) bound to cultured human TE cells and to thymic fibroblasts. CD6-Rg binding to TF and TE cells was trypsin sensitive, and 54 +/- 4% of binding was divalent cation dependent. By screening the blind panel of 479 monoclonal antibodies (mAbs) from the 5th International Workshop on Human Leukocyte Differentiation Antigens for expression on human TE cells and for the ability to block CD6-Rg binding to TE cells, we found one mAb (J4-81) that significantly inhibited the binding of CD6-Rg to TE cells (60 +/- 7% inhibition). A second mAb to the surface antigen identified by mAb J4-81, J3-119, enhanced the binding of CD6-Rg to TE cells by 48 +/- 5%. Using covalent cross-linking and trypsin digestion, we found that mAb J4-81 and CD6-Rg both bound to the same 100-kD glycoprotein (CD6L-100) on the surface of TE cells. These data demonstrate that a 100-kD glycoprotein on TE cells detected by mAb J4-81 is a ligand for CD6.
Patel
D D
DD
Department of Medicine, Duke University Medical Center, Durham, North Carolina 27710, USA.
Wee
S F
SF
Whichard
L P
LP
Bowen
M A
MA
Pesando
J M
JM
Aruffo
A
A
Haynes
B F
BF
D020410:Activated-Leukocyte Cell Adhesion Molecule; D000911:Antibodies, Monoclonal / Q000276:immunology; D015703:Antigens, CD / Q000378:metabolism*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism*; D000954:Antigens, Surface / Q000276:immunology; D002648:Child; D004848:Epithelium / Q000737:chemistry; D000939:Epitopes / Q000276:immunology; D005347:Fibroblasts / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D008970:Molecular Weight; D009928:Organ Specificity; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013950:Thymus Gland / Q000737:chemistry* / Q000166:cytology
D020410:Activated-Leukocyte Cell Adhesion Molecule; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D000954:Antigens, Surface; C062279:CD6 antigen; D000939:Epitopes; D006023:Glycoproteins; D011993:Recombinant Fusion Proteins