Quantification and modeling of tripartite CD2-, CD58FC chimera (alefacept)-, and CD16-mediated cell adhesion.

Journal article

Alefacept is a chimeric protein combining CD58 immunoglobulin-like domain 1 with human IgG1 Fc. Alefacept mediates adhesion by bridging CD2 on T cells to activating Fc receptors on effector cells, but the equilibrium binding parameters have not been determined. Alefacept mediated T cell killing by NK cells and adhesion between CD2- and CD16-expressing cells at an optimum concentration of 100 nM. We introduce novel measurements with supported planer bilayers, from which key two-dimensional and three-dimensional parameters can be determined by data fitting. Alefacept competitively inhibited cell bilayer adhesion mediated by the CD2-CD58 interaction. Alefacept mediated maximal adhesion of CD2(+) T cells to CD16B, an Fc receptor, in planar bilayers at 500 nM. A mechanistic model for alefacept-mediated cell-bilayer adhesion allowed fitting of the data and determination of two-dimensional binding parameters. These included the density of bonds in the adhesion area, which grew to maintain a consistent average bond density of 200 molecules/microm(2) and two-dimensional association constants of 3.1 and 630 microm(2) for bivalently and monovalently bound forms of alefacept, respectively. The maximum number of CD16 bound and the fit value of 4,350 CD2 per cell are much lower than the 40,000 CD2 per cell measured with anti-CD2 Fab. These results suggest that additional information is needed to correctly predict Alefacept-mediated bridge formation.

Authors: Dustin, M; Starr, T; Coombs, D; Majeau, G; Meier, W

• Journal: Journal of biological chemistry
• Volume: 282
• Issue: 48
• Pages: 34748-34757
• Publication date: 2007-11-01
• DOI: 10.1074/jbc.m705616200
• EISSN: 1083-351X
• ISSN: 0021-9258
• Language: English
• Keywords: Protein Binding; Receptors, Fc; Lipid Bilayers; Cell Adhesion; Cricetulus; Kinetics; T-Lymphocytes; CHO Cells; Antigens, CD58; Animals; Cricetinae; Dermatologic Agents; Recombinant Fusion Proteins; Receptors, IgG; Antigens, CD2; Dose-Response Relationship, Drug; Humans; Jurkat Cells