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

Article Title: SUBSET-SPECIFIC REDUCTIONS IN LUNG LYMPHOCYTE ACCUMULATION FOLLOWING INTRATRACHEAL ANTIGEN CHALLENGE IN ENDOTHELIAL SELECTIN-DEFICIENT MICE

doi:

Figure Lengend Snippet: Peripheral blood analysis indicates a lymphocytosis in E-P- mice. Peripheral blood lymphocytes were analyzed by flow cytometry. Black bars, CD4 cells; light gray bars, CD8 cells; dark gray bars, CD19 cells; white bars, γ† cells. Data represent mean ± SEM of six mice per group (untreated) or 11-12 mice per group (day 7) assayed individually in at least two separate experiments. **, p<0.01, unpaired Student t test.

Article Snippet: mAbs The following epitopes were assessed by three-color flow cytometry analyses using directly-conjugated mAbs (PharMingen; San Diego, CA) (clone, isotype and fluorochrome are indicated in parentheses): CD4 (RM4-4; rat IgG2a; cychrome), CD8 (53-6.72; rat IgG2a; FITC), CD19 (1D3; rat IgG2a; phycoerythrin), CD45 (30-F-11; rat IgG2b; cychrome), γ/†-TCR (GL3; hamster IgG2; cychrome), Gr-1 (RB6-8C5; rat IgG2b; phycoerythrin), pan-NK cell (DX-5; rat IgM; FITC), CD14 (rmC5-3; rat IgG1; FITC), and Mac-3 (M3/84; rat IgG1; FITC).

Techniques: Flow Cytometry

Journal: European Thyroid Journal

Article Title: Alemtuzumab-induced thyroid eye disease successfully treated with a single low dose of rituximab

doi: 10.1530/ETJ-23-0236

Figure Lengend Snippet: Immunophenotyping by flow cytometry. Lymphocyte analysis was conducted in our patient at the time of thyroidectomy (PT), three healthy donors (HD; n = 3), and four patients with spontaneous Graves’ disease, of whom two also had thyroid eye disease (GD/TED; n = 4). Upper bar plots show the absolute lymphocyte counts of CD45+ (A), CD19+ B (B), CD3+CD4+ (C), and CD3+CD8+ T (D) cells obtained from peripheral blood-derived mononuclear cells (PBMCs). Bottom bar plots represent the frequencies of regulatory (Treg; CD45 + CD3 + CD4 + IL7R low/- CD25 + ) T cells on total CD4+ T cells (E), CD45 + CD19 + B cells on total CD45+ cells (F), CD45 + CD3 + CD4 + Helper) T cells on total CD45+ T cells (G), and CD45 + CD3 + CD4 - Cytotoxic T cells on total CD45+ T cells (H) obtained from paired PBMC and thyroid-infiltrating mononuclear cells (TMCs).

Article Snippet: The absolute lymphocyte counts (number of cells/μL) were calculated at several time points on fresh blood using fluorescent microspheres (Beckman Coulter Life Science, Brea, CA, USA) and labeling cells with the following surface markers: CD45 (Clone 2D1; BioLegend, San Diego, CA, USA), CD3 (Clone OKT3; BioLegend), CD4 (Clone RPA-T4; BioLegend), CD8 (Clone RPA-T8; BD Pharmingen, Franklin Lakes, NJ, USA), and CD19 (1D3; ImmunoTools, Frieosythe, Germany).

Techniques: Flow Cytometry, Derivative Assay

Journal: Nature microbiology

Article Title: The targets of acute norovirus infection are immune cells in the gut-associated lymphoid tissue

doi: 10.1038/s41564-017-0057-7

Figure Lengend Snippet: a–b) Duplicate wells of the indicated cell lines were infected with mock inoculum or MNV-1 at MOI 5. Virus titers were determined by TCID 50 assay on culture supernatants (a) and cell viability was assessed by propidium iodide staining (b) at the indicated time points. Error bars denote SEM. c) The frequency and abundance of cells expressing the MNV-1 receptor CD300lf were determined on naïve cells by flow cytometric staining using a matched isotype antibody as a negative control. d) Peyer’s patches cells from B6 mice (n=6) were stained with two panels of antibodies (see Methods). The frequency and abundance of DC/Mφ (CD45 + CD11c + ), B cells (CD45 + CD19 + B220 + ), T cells (CD45 + CD3 + ), and intestinal epithelial cells (CD45 − EpCAM + ) expressing CD300lf were determined. For all panels, experiments were performed three times.

Article Snippet: The following antibodies were used in one multicolor panel alongside CD300lf (panel 1): anti-mouse CD19 PE (clone 1D3, eBioscience, 12-0193), anti-mouse B220 PE-Cy7 (clone RA3-6B2, BioLegend, 103221), anti-mouse CD11c FITC (clone N418, BioLegend, 117305), and anti-mouse CD45 Pacific Blue (clone 30-F11, BioLegend, 103125).

Techniques: Infection, Virus, Staining, Expressing, Negative Control