L-Selectin Serves as an E-Selectin Ligand on Cultured Human T Lymphoblasts

• Published in: The Journal of immunology (1950) Vol. 169; no. 4; pp. 1768 - 1773
• Main Authors: Jutila, Mark A, Kurk, Sandy, Jackiw, Larrisa, Knibbs, Randall N, Stoolman, Lloyd M
• Format: Journal Article
• Language: English
• Published: United States Am Assoc Immnol 15.08.2002
• Subjects: Antibodies, Monoclonal - pharmacology; Cell Movement; Cells, Cultured; E-Selectin - metabolism; Humans; Jurkat Cells; L-Selectin - metabolism; Ligands; Lymphocyte Activation - drug effects; P-Selectin - metabolism; Recombinant Fusion Proteins - metabolism; T-Lymphocytes - drug effects; T-Lymphocytes - immunology; T-Lymphocytes - physiology; Tetradecanoylphorbol Acetate - pharmacology
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.169.4.1768

Previous studies reported that L-selectin (CD62L) on human peripheral blood neutrophils serves as an E-selectin ligand. This study shows that CD62L acquired E-selectin-binding activity following phorbol ester (PMA) treatment of the Jurkat T cell line and anti-CD3/IL-2-driven proliferation of human T lymphocytes in vitro. The recombinant porcine E-selectin/human Ig chimera P11.4 showed neuraminidase-sensitive and calcium-dependent attachment to PMA-stimulated human Jurkat T cells in a flow cytometry assay. The anti-CD62L mAb (DREG 56) blocked this binding interaction by approximately 60% and P11.4 precipitated CD62L from detergent lysates of PMA-activated Jurkat cells. In contrast, P11.4 precipitated minimal amounts of CD62L from detergent lysates of nonactivated human PBL. As reported previously, P-selectin glycoprotein ligand 1 and a distinct 130-kDa glycoprotein were the major species in these precipitates. However, T cell activation on plate-immobilized anti-CD3 and growth in low-dose IL-2 increased the percentage of CD62L molecules with E-selectin-binding activity. After two cycles of activation and culture, approximately 60-70% of the CD62L was precipitated with the P11.4 chimera. These cultured T lymphoblasts rolled avidly on both E-selectin and P-selectin at physiologic levels of linear shear stress. The DREG 56 Ab partially blocked rolling on the E-selectin substrate, whereas no effect was seen on P-selectin. Thus, CD62L on human cultured T lymphoblasts is one of several glycoproteins that interacts directly with E-selectin and contributes to rolling under flow.