US 7,514,232 B2
Method for detecting T cell response to specific antigens in whole blood
Vernon C. Maino, Los Altos, Calif. (US); Maria Suni, Los Gatos, Calif. (US); and Louis J. Picker, Plano, Tex. (US)
Assigned to Becton, Dickinson and Company, Franklin Lakes, N.J. (US)
Filed on Feb. 21, 1997, as Appl. No. 8/803,702.
Application 08/803702 is a continuation in part of application No. 08/760447, filed on Dec. 06, 1996, abandoned.
Prior Publication US 2001/0006789 A1, Jul. 05, 2001
Int. Cl. G01N 33/00 (2006.01)
U.S. Cl. 435—7.24 41 Claims
 
1. A method of detecting T lymphocytes that are specific for a nominal antigen, comprising:
culturing a sample containing peripheral blood mononuclear cells with a nominal antigen;
adding to said sample an inhibitor of cytokine secretion;
permeabilizing said cells;
adding to said sample at least one cytokine-specific antibody and at least one T lymphocyte subset-defining antibody; and then
flow cytometrically detecting the intracellular binding of said cytokine-specific antibody by cells in the defined T lymphocyte subset.