Journal: Molecular Oncology

Article Title: Breast cancer metastasis: immune profiling of lymph nodes reveals exhaustion of effector T cells and immunosuppression

doi: 10.1002/1878-0261.13047

Figure Lengend Snippet: The topological distribution of CD4‐ and CD8‐positive cells in lymph nodes with no, small, intermediate, and large metastatic tumor burden. (A–D) One SNneg (non‐metastatic sentinel node) (patient 11) and three SNmets (metastatic sentinel nodes) (patients 37,39, and 27) were IHC stained with AE1/AE3, CD4, and CD8. Magnified area of extratumoral (orange) and intratumoral (green) regions stained with CD8 and CD4 of lymph nodes with small (B), intermediate (C), and large (D) tumor burden. Scales and magnifications are shown in images. (E, F) A scoring system, ImmunoPath (with field fraction function measuring positive and negative pixels was used to calculate the percentage positive areas) was used to objectively score IHC‐positive areas. (E) Histograms comparing tumor cell percentage and CD4/CD8 ratio measured by IHC (left axis) and mass cytometry (right axis) performed on cells from the same lymph nodes. (F) Comparison of CD4/CD8 ratio in intratumoral and extratumoral areas of the metastatic lymph node analyzed separately by ImmunoPath. (G) Immunofluorescence staining with CD8 (green) and Treg (FoxP3, red) and counterstained with DAPI on non‐metastatic sentinel node and metastatic sentinel nodes (same samples as A‐D) Scales and magnifications are shown in images.

Article Snippet: Immunofluorescence was performed with high PH (Envision TM FLEX target retrieval solution, DM828, DAKO) antigen retrieval and stained with CD8 ((C8/144B, cat.nr: 372902 BioLegend, San Diego, CA, USA)+ biotin (Gt anti‐mouse, M30115, Invitrogen, Thermo Fisher Scientific, MA, USA)+ streptavidin‐conjugated AF488 (S11223, Invitrogen)) and FoxP3 (236A/E7,ab20034 Abcam + goat‐anti‐Mouse AF594 (A21125, Invitrogen)) and counterstained with DAPI (InvitrogenTM ProLongTM Gold Antifade Mountant with DAPI, P36941, Fisher Scientific).

Techniques: Staining, Mass Cytometry, Comparison, Immunofluorescence

Journal: Cell

Article Title: Suppression of Exosomal PD-L1 Induces Systemic Anti-tumor Immunity and Memory

doi: 10.1016/j.cell.2019.02.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Prior imaging, sample were mounted with VECTASHIELD Antifade Mounting Medium with DAPI (Fisher Scientific, cat. NC9524612).

Techniques: Western Blot, Flow Cytometry, Control, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Recombinant, SYBR Green Assay, cDNA Synthesis, Red Blood Cell Lysis, Software