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. Author manuscript; available in PMC: 2013 Aug 1.
Published in final edited form as: J Immunol. 2013 Jan 2;190(3):977–986. doi: 10.4049/jimmunol.1201331

Figure 2. Cellular infiltration at the site of VZV injection is Ag-specific.

Figure 2

Skin suction blisters were induced on day 7 following VZV injection. (A) PBMCs and blister cells were stimulated with VZV lysate for 15hrs in the presence of Brefeldin A and stained for intracellular expression of IFN-γ and IL-2. Representative FACS plots are shown on the left. The percentage IFN-γ producing antigen specific CD4+ T cells was significantly increased compared to peripheral blood (n=15 for IFN-γ; Wilcoxon paired test ***P=0.0002, *P=0.015). (B) IL-2 producing antigen specific CD4+ T cells were also significantly increased compared to peripheral blood (n=8, Mann Whitney test **P=0.001, *P=0.016). (C) A Large proportion of VZV specific cells produce both cytokines. Representative dot plot shows day 7 blister cells (gated on CD4+) secreting IL-2 and IFN-γ. (D) The number of antigen specific CD4+ cells was also determined by class II tetramer staining. Blister cells and PBMC were stained with HLA-DRB1*1501 restricted IE63 tetramer (18). Left panel shows representative FACS plots for tetramer staining. Graph shows cumulative data, each data point represents one individual, horizontal bars represent the mean (n=11, Wilcoxon paired test **P=0.0046).