Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Heterosubtypic influenza infection antagonizes elicitation of immunological reactivity to HA

doi: 10.4049/jimmunol.1203520

Figure Lengend Snippet: B10 CD8 T cells were depleted in vivo throughout the duration of secondary infection (days −2, 0, 2, 4, and 6). Eight days post infection, CD4 T cell reactivity to various epitopes was examined by IFNγ Elispot assay. Panel A depicts data obtained from MACS purified CD4 T cells isolated from the spleen. Panel B shows IFNγ Elispots obtained from unpurified MLN cells. Data are presented as spots per 106 CD4 T cells, with MLN data normalized for the percent of CD4 T cells present. Flow cytometric analysis of both spleen and MLN cells indicated complete depletion of CD8 T cells.

Article Snippet: CD8 antibody (Clone 2.43, BioXCell, West Lebanon, NH) was used for in vivo depletion of CD8 T cells.

Techniques: In Vivo, Infection, Enzyme-linked Immunospot, Purification, Isolation

Journal: Molecular oncology

Article Title: Targeting histone deacetylase 6 mediates a dual anti-melanoma effect: Enhanced antitumor immunity and impaired cell proliferation

doi: 10.1016/j.molonc.2015.04.002

Figure Lengend Snippet: Anti‐tumor activity of HDAC6 in vivo is dependent on intact immunity. (A) In vivo growth of B16–F10 WT melanoma in SCID mice treated or not with the HDAC6 inhibitor Nexturastat compared with control treatment. (B) In vivo growth of B16–F10 HDAC6KD melanoma cells and control non‐target cells in C57BL/6 mice. Mice were treated with antibodies to deplete CD4, CD8, or CD4 and CD8 cells.

Article Snippet: For the T‐cell depletion experiments we used Monoclonal rat anti‐mouse CD4 (GK1.5 clone, BioXcell #BE003‐1) and rat anti‐mouse CD8 (2.43 clone, BioXcell #BE0061).

Techniques: Activity Assay, In Vivo, Control

Journal: PLoS ONE

Article Title: Resident Cardiac Immune Cells and Expression of the Ectonucleotidase Enzymes CD39 and CD73 after Ischemic Injury

doi: 10.1371/journal.pone.0034730

Figure Lengend Snippet: A: Gating strategy used to identify different leukocyte subpopulations by multiparametric flow cytometry. CD45 + cells were gated and DAPI-staining was used to exclude dead and apoptotic immune cells. Living CD45 + cells were then divided in subleukocyte populations using a panel of cell-specific fluorochrome-labeld antibodies. Lymphocytes were gated into CD45R(B220) + cells (B-cells) and CD3 + cells (T-cells). T-cells were subdivided in CD4 + cells (T-helper cells) and CD8 + cells (cytotoxic T-cells). Myeloid cells were characterized as CD11b + cells and further subdivided in CD11c + cells (APCs) and Ly6g + cells (granulocytes). B: Leukocyte subpopulations in the unstressed heart. Values are means ± SD of n = 5 experiments.

Article Snippet: Cells were resuspended in MACS buffer, preincubated with FcR Blocking Reagent (Miltenyi Biotech) and stained with the following antibodies: anti- CD45 -APC, 30-F11 (BD Bioscience), - CD11b -PE, M1/70 (BD Bioscience), - CD8 a-APC-H7, 53-6.7 (BD Bioscience), - CD25 -PE-Cy7, PC61 (BD Bioscience), - Ly-6C -FITC, AL-21 (BD Bioscience), TER-119 -PE, TER119 (BD Bioscience), - CD45 -PE, 30F11 (Miltenyi Biotech), - CD11b -APC, M1/70.15 (Miltenyi Biotech), - CD3 -APC, 145-2C11 (Miltenyi Biotech), CD49b -APC, DX5 (Miltenyi Biotech), NKp46 -APC, 29A1.4 (Miltenyi Biotech), - CD39 -PE-Cy7, 24DMS1 (eBioscience), - CD4 -PerCP-Cy5.5, RM4-5 (eBioscience), - Ly-6G (Gr-1) -PerCP-Cy5.5, RB6-8C5 (eBioscience), - CD45R(B220) -APC-eFluor780, RA3-6B2 (eBioscience), - F4/80 -APC-eFluor780, BM8 (eBioscience), - CD11c -PE-Cy7, N418 (eBioscience), - CD73 -FITC, 496406 (R&D Systems), - CD39 -FITC, 495826 (R&D Systems), - CD31 -APC, 390 (Biolegend), FoxP3 -PE, MF23 (BD Bioscience), Ly6c -APC-Cy7, AL-21 (BD Bioscience), MHCII -FITC, M5/114.15.2 (Miltenyi Biotech).

Techniques: Flow Cytometry, Staining