Activation Induced Marker (AIM) assay and experimental design
Evaluations of T cell responses were based on previously described Activation Induced Marker (AIM) ex vivo assays30,35,36, utilizing the CD154 (CD40L) and CD137 (4-1BB) markers, combined with intracellular cytokine staining (ICS), using the antibodies described in Supplementary Table 2 and performed as detailed in the online supporting information. The cytokines IL-4, IFNγ, and IL-10 were included as representative of Th2, Th1, and Tr1/T Regulatory (Tregs) CD4 helper T cell subsets, respectively 24,37. The phenotypic CD127 and CD25 Tregs markers were also included in the cytometry analysis panel30,38 and the several T cell parameters determined as described in the supporting information.