TY - JOUR
T1 - Enumeration of cytokine‐secreting cells at the single‐cell level
AU - Skidmore, Barry J.
AU - Stamnes, Susan A.
AU - Townsend, Kay
AU - Glasebrook, Andrew L.
AU - Sheehan, Kathleen C.F.
AU - Schreiber, Robert D.
AU - Chiller, Jacques M.
PY - 1989/9
Y1 - 1989/9
N2 - A sensitive assay utilizing enzyme‐linked immunosorbent assay methodology has been developed for the quantitation of single cells secreting interferon (IFN)‐γ or tumor necrosis factor (TNF). Cloned T cells or cells from lymphoid organs were stimulated with antigen, concanavalin A, or phorbol myristate acetate and ionomycin in microwells coated with antibodies specific for IFN‐γ. Discrete “spots” overlying areas where cells secrete IFN‐γ were then developed by incubation with a second antibody to IFN‐γ, followed by an enzyme‐labeled antibody conjugate and substrate. Similarly, using TNF‐specific antibody reagents, TNF‐secreting cells were detected and quantitated in cell populations obtained from normal lymphoid tissues, bone marrow and peripheral blood, following activation with phorbol myristate acetate and ionomycin. Provided specific antibodies are available, this method has the potential to measure the frequency of cells secreting any cytokine.
AB - A sensitive assay utilizing enzyme‐linked immunosorbent assay methodology has been developed for the quantitation of single cells secreting interferon (IFN)‐γ or tumor necrosis factor (TNF). Cloned T cells or cells from lymphoid organs were stimulated with antigen, concanavalin A, or phorbol myristate acetate and ionomycin in microwells coated with antibodies specific for IFN‐γ. Discrete “spots” overlying areas where cells secrete IFN‐γ were then developed by incubation with a second antibody to IFN‐γ, followed by an enzyme‐labeled antibody conjugate and substrate. Similarly, using TNF‐specific antibody reagents, TNF‐secreting cells were detected and quantitated in cell populations obtained from normal lymphoid tissues, bone marrow and peripheral blood, following activation with phorbol myristate acetate and ionomycin. Provided specific antibodies are available, this method has the potential to measure the frequency of cells secreting any cytokine.
UR - http://www.scopus.com/inward/record.url?scp=0024438932&partnerID=8YFLogxK
U2 - 10.1002/eji.1830190911
DO - 10.1002/eji.1830190911
M3 - Article
C2 - 2507324
AN - SCOPUS:0024438932
SN - 0014-2980
VL - 19
SP - 1591
EP - 1597
JO - European Journal of Immunology
JF - European Journal of Immunology
IS - 9
ER -