Spectophotometric quantification of cell-cell adherence by an enzyme-linked immuno-cell adhesion assay

J Immunol Methods. 1990 Aug 28;132(1):51-6. doi: 10.1016/0022-1759(90)90397-e.

Abstract

In the present study a spectophotometric method was developed to determine the number of T cells binding to human epidermal keratinocytes (KC). In this cell adhesion assay the adherent T cells were detected with a cell-specific monoclonal antibody conjugated to horseradish peroxidase and the coloured substrate quantified in an ELISA reader at 492 nm. A correlation was demonstrated between the number of T cells and the extinction values measured. The enzyme-linked immuno-cell adhesion assay (ELICAA) was used to quantify KC/T lymphocyte adherence in a series of experiments designed to evaluate its reliability and reproducibility. Compared with the 51Cr-labelled adherence assay, the ELICAA was a safe, rapid and accurate method avoiding the use of radioactive material.

Publication types

  • Comparative Study

MeSH terms

  • Antibodies, Monoclonal
  • Antigens, Differentiation, T-Lymphocyte / physiology
  • CD4 Antigens / physiology
  • CD8 Antigens
  • Cell Adhesion*
  • Cell Communication
  • Chromium
  • Dose-Response Relationship, Drug
  • Female
  • HLA-DR Antigens / physiology
  • Horseradish Peroxidase
  • Humans
  • Immunoenzyme Techniques*
  • Interferon-gamma / pharmacology
  • Keratinocytes / physiology
  • Male
  • Spectrophotometry
  • T-Lymphocytes / physiology*
  • Tetradecanoylphorbol Acetate / pharmacology
  • Time Factors

Substances

  • Antibodies, Monoclonal
  • Antigens, Differentiation, T-Lymphocyte
  • CD4 Antigens
  • CD8 Antigens
  • HLA-DR Antigens
  • Chromium
  • Interferon-gamma
  • Horseradish Peroxidase
  • Tetradecanoylphorbol Acetate