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Updated: Aug 8, 2026
Adhesion Frequency Assay for In Situ Kinetics Analysis of Cross-Junctional Molecular Interactions at the Cell-Cell Interface
Published on: November 2, 2011
A novel, rapid and sensitive heterotypic cell adhesion assay for CD2-CD58 interaction, and its application for
Jining Liu1, Vincent T K Chow, Seetharama D S Jois
1Department of Pharmacy, 18 Science Drive 4, National University of Singapore, Singapore 117543, Singapore.
Insights
Researchers developed a new assay to measure cell adhesion between CD2 and CD58, crucial for T-cell activation. This rapid, sensitive method aids in testing potential therapies targeting this interaction.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- CD2 is an immunoglobulin cell adhesion molecule mediating T-cell activation via binding to CD58 on antigen-presenting cells (APCs).
- Modulating the CD2-CD58 interaction shows therapeutic potential.
- Traditional E-rosetting assays are commonly used to study this interaction.
Purpose of the Study:
- To develop a novel, rapid, and sensitive heterotypic cell adhesion assay for the CD2-CD58 interaction.
- To validate the assay's utility in testing inhibitory peptides.
Main Methods:
- Confirmed CD2 expression on Jurkat cells and CD58 expression on Caco-2 cells using flow cytometry and ELISA.
- Fluorescently labeled Jurkat cells (BCECF-AM) were added to Caco-2 monolayers in 96-well plates.
- Quantified bound cells via fluorescence measurement after washing and cell lysis.
Main Results:
- A novel, sensitive, and rapid cell adhesion assay for CD2-CD58 interaction was successfully established.
- The assay confirmed CD2 and CD58 expression on the respective cell lines.
- The method effectively evaluated inhibitory peptides targeting the CD2-CD58 interaction.
Conclusions:
- The developed assay provides a robust platform for studying CD2-CD58 interactions.
- This method offers a sensitive and rapid alternative to traditional assays for evaluating modulators of T-cell activation.
- The assay is suitable for high-throughput screening of therapeutic agents targeting the CD2-CD58 pathway.
Abstract:
The immunoglobulin CD2 is a cell adhesion molecule that mediates T-cell activation by binding to its receptor CD58 on antigen-presenting cells (APCs). Modulation or inhibition of this interaction has been shown to be therapeutically useful. E-rosetting assay is usually applied in the study of the modulation of CD2-CD58 interaction. In this study, we demonstrated a novel, rapid and sensitive heterotypic cell adhesion assay for CD2-CD58 interaction. The CD2 expression on the surface of Jurkat cells and the CD58 expression on the Caco-2 cells were confirmed by flow cytometry and ELISA studies, respectively. Then Jurkat cells were fluorescent-labeled with 2 microM of BCECF-AM for 45 min at 37 degrees C before adding to confluent Caco-2 monolayers cultured in 96-well culture dishes. After 30 min, non-adherent Jurkat cells were removed by washing with PBS, while the monolayer-associated Jurkat cells were lysed with 0.5 ml of 2% Triton X-100 in 0.1 M NaOH. Fluorescence (FL) was quantitated using a microplate fluorescence analyzer with BCECF's excitation maximum of 485 nm and emission maximum of 535 nm. This method was successfully applied for testing inhibitory peptides to CD2-CD58 interaction.
