Fig. 1: Schematic representation of the analysis pipeline and approaches used to characterize the immune response after in vitro stimulation with Ag85B and TB10.4 or BCG in trial participants.

Lymphocytes expressing any cytokines, selected by conventional flow cytometry gating in stimulated and unstimulated samples, were subjected to unsupervised analyses (left) using tSNE and CITRUS, or to targeted analyses (right) where subsets were manually gated and cytokine-co-expression subsets were analyzed by comparing their frequencies directly, or by using COMPASS (functionality and polyfunctionality assessments) or MIMOSA2 (to assign vaccine responder and non-responder status). Double sided arrows show the analyses used in parallel to validate observations with multiple strategies. Single arrows indicate the flow of analysis.