SOPs for the cytometric assessment of Th-cell immunity based on antigen-induced CD154 expression
We have worked out SOPs for the cytometric assessment of Th-cell immunity based on antigen-induced CD154 expression, a technology developed by us in the frame of previous DC-THERA activities. Now the entire antigen-specific Th-cells specific for defined antigens can be monitored simultaneously with respect to quantity and quality. Tests can either be performed on standard 4 colour cytometers or on 12 colour high-end cytometric analysers. Tests that can be either performed to analyse fixed or isolate live antigen-specific Th-cells have been adapted also to be used with murine cells (Kirchhoff et al., 2007).
protocol
Standard operating procedure
Identification and isolation of murine antigen-reactive T cells according to CD154 expression.
Proteomic assessment of CD4+ Th-cells specific for defined antigens
Alexander Scheffold
Andreas Thiel
CD4-positive T helper cell
CD40 ligand
CD154 receptor
Evaluation of different stimulation conditions for the cytometric assessment of antigen-specific Th-cells