Gating procedures of flow cytometry analyses to determine protein candidate specific T cell proportions.

<p>Representative analyses from a patient with Tb (A) and an LTBI (B) are shown. Open red circles and dot plot connected by red arrows indicate the sequence of analysis steps. First, lymphocytes were gated using size (forward scatter; FSC) and granularity (side scatter, SSC). These cells were then analyzed for CD4 expression. CD4⁺ T cells were analyzed for IFNγ CD45RO expression for each stimulation (without stimulus, w/o; proteine 3; protein 11; SEB). Proportions of CD45RO_high IFNγ expressing CD4⁺ T cells (upper right quadrants) were determined. The background of non-stimulated T cells (w/o) was subtracted for analyses.