Figure 10

CD8 T cell response to column-activated malignant T cell loaded DC. Fig. 10A: CD8 T cells were magnetic-bead enriched (≥96% CD8⁺ T cells) from the leukapheresis of a CTCL patient and added to column-activated DC with and without apoptotic malignant T cell loading. The percentage of CD8 T cells was identified by immunophenotyping and flow cytometry and the results presented as 1-color histograms. a: Percentage of CD8 T cells found after overnight culture with column-activated DC pulsed with CD3-magnetic bead rendered apoptotic CTCL cells. b: Percentage of CD8 T cells identified after overnight culture with DC co-cultivated with viable CTCL cells. c: Absolute number of CD8 T cells after overnight cultivation with DC loaded with apopototic malignant T cells or DC co-incubated with viable CD4-bead isolated CTCL cells. Horizontal line indicates the initial number of CD8 T cells added to the co-cultures on Day 0. Fig. 10B: The percentage of apoptotic cells was determined in the co-cultures by staining for APO2.7 and flow cytometry. The quadstats are gated on the lymphocyte population by side scatter (SS) and represent 100% of the lymphocyte population. Percent apoptotic cells found in co-cultures of a: CD8 T cells and DC loaded with apoptotic malignant T cells; b: CD8 T cells, DC and viable CTCL cells; c: DC loaded with apoptotic CTCL in the absence of CD8 T cells; d: DC cultured with viable CTCL without CD8 T cells. e: Culture supernatants were obtained from CD8 T cells cultured overnight alone, or in the presence of DC loaded with apoptotic CTCL cells or DC cultured with viable CTCL cells and the secretion of TNF-α determined in an ELISA assay. The results are presented as pg/ml and significance determined with a student's t test.