Journal: Cancer Immunology, Immunotherapy : CII
Article Title: Loss of type I IFN responsiveness impairs natural killer cell antitumor activity in breast cancer
doi: 10.1007/s00262-021-02857-z
Figure Lengend Snippet: Loss of NK cell IFNAR1 impairs NK cell function, may impact adaptive immune regulation of tumorigenesis and increases lung metastasis. 1 × 105 EO771.LMB cells were injected into the fourth mammary fat pad (IMFP) of controliCre and Ifnar−/−NKp46 C57BL/6 mice at day 0 and subsequent alterations to circulating and primary tumor cells assessed. Peripheral blood (PB) was taken at day 7 for FACS analysis and for intracellular staining (ICS) of monocytes was performed at endpoint. a Primary tumor weights (mg) of control (n = 7) and Ifnar−/−NKp46 mice (n = 7) mice at experimental endpoint. Flow cytometric analysis of b absolute number (cell #) of primary tumor NK cells (n = 6/group). c PB CD69+ NK (NK1.1+) cell frequency (%). Primary tumor d CD69+ (e. representative plots shown) and f IFNγ+ NK cell frequency (%) and g IFNAR1 expression, represented as MFI (n = 5/group). Flow cytometric analysis of h CD8+ T cell IFNAR1 (MFI; n = 5/group) and i. PB CD69+ CD8+ T cells, j primary tumor CD69+ CD8+ T cells, k CD8+ IFNγ+ T cells, l. PB CD69+ CD4+ T cells and m primary tumor CD4+ IFNγ+ T cells, expressed as frequency (%). n RT-qPCR of mCherry (EO771.LMB) DNA expression in the lungs post-intravenous (IV) injection of EO771.LMB cells into WT (n = 9), controliCre (n = 5) and Ifnar−/−NKp46 (n = 8) mice at day 24. Representative fluorescence imaging of mCherry (EO771.LMB) in lungs shown for each group. All PB analysis (n = 7/group) and primary tumor analysis (n = 6/group). p values * < 0.05, ** < 0.005, *** < 0.0005. **** < 0.0001 determined by Student’s t test. Errors bars, SEM
Article Snippet: When required, cells were transfected with poly I:C (10 μg/ml) overnight, prior to RNA extraction. cDNA was generated using the iScript Reverse Transcriptase Supermix cDNA for RT-qPCR kit (BioRad).
Techniques: Cell Function Assay, Injection, Staining, Expressing, Quantitative RT-PCR, IV Injection, Fluorescence, Imaging