ADCC analysis with the Coulter counter: a highly sensitive, ultramicro assay suitable for clinical and experimental...

Antibody-dependent cellular cytotoxicity (ADCC) is an in vitro immune mechanism implicated in several in vivo phenomena such as transplant rejection, tumor immunity and parasite elimination. We developed a method for detecting ADCC using the Coulter Counter and the Coulter Channelyzer that circumvents many of the disadvantages associated with existing assays for ADCC. Effector mononuclear cells were incubated with chicken red blood cell (CRBC) targets and anti-target antibody for 1-1, 5h. Killing was quantified by the Coulter Counter on the basis of size differences between effector and target cell nuclei. Using a 4 microliter total volume we were able to detect cytotoxic levels of 55% when as few as 5000 effector cells were incubated with an equal number of target cells. This method for the detection of ADCC may be suitable for clinical and research application.