A simple plaque assay method for antibody-dependent cell-mediated cytotoxicity using Cunningham's chamber.
An improved and simpler plaque assay method for antibody-dependent cell-mediated cytotoxicity using Cunningham's chamber has been developed. Effector lymphocytes, target sheep red blood cells (SRBC) and anti-SRBC serum were incorporated together into a Cunningham's chamber and, immediately after making monolayers, incubated at 37 degrees C for 20 h. A hemolytic plaque of SRBC was formed around a lymphocyte in the presence of anti-SRBC serum, which was inhibited specifically by addition of aggregated IgG or Fc-fragment of human IgG in the culture. A hemolytic plaque formed around a monocyte-like cell was clearly differentiated from those around a lymphocyte (K-cell). The percentage of plaques in the purified lymphocytes from healthy individuals correlated significantly with ADCC activity measured by 51Cr-release. The method is a significant improvement on the earlier plaque assay methods with regard to: (1) making the monolayer, (2) observation and counting of plaques, and (3) shortening the time required for assay. Thus, the method should be valuable for clinical studies of cytotoxic activity in ADCC systems in healthy and diseased persons.
D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity; D002452:Cell Count; D006462:Hemolytic Plaque Technique; D006801:Humans; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G / Q000096:biosynthesis; D007694:Killer Cells, Natural / Q000276:immunology; D011817:Rabbits; D012756:Sheep; D013997:Time Factors
D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G