Journal: Human Gene Therapy Methods
Article Title: Potential Limitations of the NSG Humanized Mouse as a Model System to Optimize Engineered Human T cell Therapy for Cancer
Figure Lengend Snippet: Effects of IL-7 and IL-15 on the proliferation, function, and phenotype of DMF5 T cells. (A) Proliferation of T cells cultured in IL-2, IL-7, and IL-15 after being activated with anti-CD3 and anti-CD28 antibodies, and transduced with the retroviral vector encoding the DMF5 TCR. (B) Viability of DMF5 T cells cultured in IL-2, IL-7, and IL-15 measured by flow cytometry according to the percentage of cells within the live lymphocyte gate. (C–F) Expression of costimulatory molecules CD27 and CD28 (C and D) , and markers of homing to central lymphoid organs CCR7 and CD62L (E and F) on CD8 + DMF5 T cells after being cultured in IL-2, IL-7, and IL-15. (G) Representative flow cytometry plots showing production of IFN-γ and IL-2 by CD8 + DMF5 T cells cultured in IL-2, IL-7, or IL-15 and stimulated with T2 cells loaded with MART-1 peptide. (H) Cytotoxicity of DMF5 T cells cultured in IL-2, IL-7, or IL-15 by CD107a expression after coculture with HLA-A*02 + cell lines Mel624, Mel501, and WM2664, and HLA-A*02 − cell line Mel888. p -Values
Article Snippet: For retroviral transduction, non-tissue-culture six-well plates (BD Biosciences) were coated with retronectin (10 μg/ml; Takara, Invitrogen) and incubated at 4°C overnight.
Techniques: Cell Culture, Transduction, Plasmid Preparation, Flow Cytometry, Cytometry, Expressing