Whole blood T cell assay for NHPs, containment protocol v1

This is a protocol used to perform T cell assays using whole blood (collected in EDTA tubes) or cells from bronchoalveolar lavage (BAL). We have successfully used this protocol for rhesus macaques, cynomolgus macaques, and African green monkeys (although antibody mix provided was titrated on macaques). It allows the characterization of T cells into Th1 and Th2 (or Tc1 and Tc2) phenotypes. It technically offers 64 different activation profiles (4 cytokines + CD107a + CD69). The nature and concentration of the stimulant is left unspecified as we have used different ones depending on reagent availability. This protocol was designed to deal with samples coming from containment labs (> CL2) at our facility. If you are using this protocol at a different facility please ensure that proper testing and approvals are in place. The protocol can be used for experiments completed entirely in CL2, simply go from step 17 directly to 31.