Spectophotometric quantification of cell-cell adherence by an enzyme-linked immuno-cell adhesion assay.
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.
Bruynzeel
I
I
Department of Dermatology, Free University Hospital, Amsterdam, The Netherlands.
Van der Raaij
L M
LM
Boorsma
D M
DM
De Haan
P
P
Scheper
R J
RJ
Kraal
G
G
Willemze
R
R
D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte / Q000502:physiology; D015704:CD4 Antigens / Q000502:physiology; D016827:CD8 Antigens; D002448:Cell Adhesion; D002450:Cell Communication; D002857:Chromium; D004305:Dose-Response Relationship, Drug; D005260:Female; D006684:HLA-DR Antigens / Q000502:physiology; D006735:Horseradish Peroxidase; D006801:Humans; D007124:Immunoenzyme Techniques; D007371:Interferon-gamma / Q000494:pharmacology; D015603:Keratinocytes / Q000502:physiology; D008297:Male; D013053:Spectrophotometry; D013601:T-Lymphocytes / Q000502:physiology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors
D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte; D015704:CD4 Antigens; D016827:CD8 Antigens; D006684:HLA-DR Antigens; D002857:Chromium; D007371:Interferon-gamma; D006735:Horseradish Peroxidase; D013755:Tetradecanoylphorbol Acetate