Intracellular IFNγ production in CD4+ and CD8+ cells in response to HER2-ICD. PBMC from patient Her2-I-01 were stimulated with HER2-ICD (50 μg/ml), KLH (50 μg/ml), Tetanus toxoid (5 Lf/ml) or media alone and analyzed for intracellular IFNγ at weeks 0, 1, 3, 6, and 10. Stimulated cells were fixed, permeabilized and stained with αIFNγ-FITC, αCD69-PE, αCD8-PerCP and αCD4-APC. The percent of CD4+ and CD8+ T cells that are IFNγ and CD69 double positive are represented for each antigen. Cells were gated by forward and side scatter for lymphocytes and positively for CD8 or CD4. Arrows represent when the patient was given each vaccination: week 0, 3, 6 and 9. The x-axis represents week of analysis and y-axis represents percent CD4+ or CD8+ cells that are CD69+IFNγ+ in response to each antigen.