title: Heterotypic cell adhesion assay for the study of cell adhesion inhibition.
abstract: CD2 is a cell adhesion molecule that mediates T-cell activation by binding to its ligand CD58 on antigen-presenting cells. Interaction between CD2 and CD58 or leukocyte function-associated antigen-3 (LFA-3) helps to optimize immune recognition facilitating contact between T lymphocytes and antigen-presenting cells. Modulation or inhibition of this interaction has been shown to be therapeutically useful in the treatment of autoimmune diseases. Antibodies and small molecules including peptides have been designed to modulate or disrupt the cell adhesion interactions due to CD2 and CD58. E-rosetting assay is a widely used method applied in the study of the modulation of CD2-CD58 interaction, which is either labor-intensive or radio-hazardous. In this chapter, we describe two methods that are used to study cell adhesion inhibition: (a) E-rosetting Assay and (b) Lymphocyte-epithelial assay. The second method, lymphocyte-epithelial assay, is a rapid and sensitive heterotypic cell adhesion assay for studying cell adhesion inhibition. The method relies on the CD2 expression on the surface of Jurkat cells and the CD58 expression on the surface of Caco-2 cells, which were confirmed by flow cytometry and ELISA studies respectively. This heterotypic cell adhesion assay described typically takes less than 4 h to perform, allows the evaluation of inhibitory activity of peptides/small molecules to modulate CD2-CD58 interaction in real cell system.
authors.affiliation: Department of Basic Pharmaceutical Sciences, College of Pharmacy, University of Louisiana at Monroe, Monroe, LA, USA. jois@ulm.edu
authors.forename: Seetharama D
authors.initials: SD
authors.lastname: Satyanarayanajois
authors.forename: Sharon
authors.initials: S
authors.lastname: Ronald
authors.forename: Jining
authors.initials: J
authors.lastname: Liu
mesh_terms: D000818:Animals; D018801:CD2 Antigens / Q000276:immunology*; D018968:CD58 Antigens / Q000276:immunology*; D018938:Caco-2 Cells; D002448:Cell Adhesion / Q000187:drug effects*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D019169:Jurkat Cells; D008213:Lymphocyte Activation / Q000187:drug effects; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D018613:Microscopy, Confocal / Q000379:methods; D010455:Peptides / Q000494:pharmacology; D054852:Small Molecule Libraries / Q000494:pharmacology
chemical_list: D018801:CD2 Antigens; D018968:CD58 Antigens; D010455:Peptides; D054852:Small Molecule Libraries