anti-mouse cd69 (h1.2f3) pe- cy7  (Thermo Fisher)

Journal: Frontiers in Immunology

Article Title: T cell kinetics reveal expansion of distinct lung T cell subsets in acute versus in resolved influenza virus infection

doi: 10.3389/fimmu.2022.949299

Figure Lengend Snippet: Formation of lung-resident CD4 + T cells in lungs after influenza virus infection. CD4 + Trm express CD69 and CD11a and to verify their localization to the lung tissue anti-CD45.2 antibodies were injected intravenously three minutes prior to euthanasia (A) . In (B) representative flow cytometry plots gated from CD4 + Tem with up to 8000 events per plot from indicated time points after infection are shown. Frequency and numbers of lung-resident CD4 + CD69 + CD11a + T cells (C) . Frequency and numbers of antigen-specific, lung-resident CD4 + CD69 + CD11a + T cells (D) . Data are composites of two separate experiments with five mice per group. Data are shown as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001.

Article Snippet: The cells were stained for 30 minutes at +4°C with the following monoclonal antibodies diluted 1:50 in FACS buffer: PerCP hamster anti-mouse CD3e (145-2C11) (BD Biosciences Cat# 553067, RRID : AB_394599), V500 rat anti-mouse CD4 (RM4-5) (BD Biosciences Cat# 560782, RRID : AB_1937315)/FITC rat anti-mouse CD4 (GK1.5) (BD Biosciences Cat# 553729, RRID : AB_395013), APC rat anti-mouse CD8α (53-6.7) (BD Biosciences Cat# 553035, RRID : AB_398527), BV510 rat anti-mouse CD8α (53-6.7) (BD Biosciences Cat# 563068, RRID : AB_2687548), FITC rat anti-mouse CD8α (53-6.7) (BD Biosciences Cat# 553031, RRID : AB_394569), PE rat anti-mouse CD44 (IM7) (BD Biosciences Cat# 553134, RRID : AB_394649), FITC rat-anti mouse CD44 (IM7) (BD Biosciences Cat# 553133, RRID : AB_2076224), APC rat anti-mouse CD62L (MEL-14) (BD Biosciences Cat# 553152, RRID : AB_398533), APC-Cy7 rat anti-mouse CD62L (MEL-14) (BD Biosciences Cat# 560514, RRID : AB_10611861), V450 rat anti-mouse CD19 (1D3) (BD Biosciences Cat# 560375, RRID : AB_1645269), BV510 rat anti-mouse CD103 (M290) (BD Biosciences Cat# 563087, RRID : AB_2721775), PerCP-Cy5.5 hamster anti-rat/mouse CD49a (Ha31/8) (BD Biosciences Cat# 564862, RRID : AB_2734135), PerCP-Cy5.5 rat anti-mouse CD11a (2D7) (BD Biosciences Cat# 562809, RRID : AB_2737809), and PE-Cy7 hamster anti-mouse CD69 (H1.2F3) (BD Biosciences Cat# 552879, RRID : AB_394508), all from BD Biosciences (BD Biosciences, RRID : SCR_013311).

Techniques: Infection, Injection, Flow Cytometry

Journal: BioMed Research International

Article Title: Reevaluation of NOD/SCID Mice as NK Cell-Deficient Models

doi: 10.1155/2021/8851986

Figure Lengend Snippet: The expression of cytotoxicity-related molecules and inhibitory receptors in/on splenic NK cells from naïve mice. (a) Representative FCM data showing the CD69, FasL, TRAIL, perforin, granzyme B, NKp46, NKG2D, NKG2A/C/E, and Ly49A expression gated on splenic DX5 + NK cells. (b) Histogram shows the statistical analysis of FCM data for CD69, FasL, TRAIL, perforin, granzyme B, NKp46, NKG2D, NKG2A/C/E, and Ly49A expression. Results are presented as mean ± SD. n = 6–8 mice per group. Values of ∗∗ P < 0.01 were considered statistically significant. Three independent experiments were performed, and representative data are shown.

Article Snippet: PE/Cy7 anti-mouse CD69 (clone H1.2F3), PE/Cy7 anti-mouse NKp46 (clone 29A1.4), and FITC anti-mouse NKG2A/C/E (clone 20d5) were from eBioscience (San Diego, CA, USA).

Techniques: Expressing

Journal: Cell Reports Medicine

Article Title: Programming Multifaceted Pulmonary T Cell Immunity by Combination Adjuvants

doi: 10.1016/j.xcrm.2020.100095

Figure Lengend Snippet: Effector CD4 T Cell Response to Adjuvanted Vaccines Groups of C57BL/6 mice were vaccinated IN, as in Figure 1 . At day 8 PV, cells from lungs and BAL were stained with I-A b /NP311 tetramers along with antibodies to cell surface molecules and transcription factors. (A) FACS plots show the percentages of I-A b /NP311 tetramer-binding cells among CD4 T cells. (B) Percentages of the indicated cell population among NP311-specific, tetramer-binding CD4 T cells. (C) FACS plots are gated on I-A b /NP311 tetramer-binding cells, and the numbers in each quadrant are the percentages of cells among the gated population; MFIs for transcription factors in NP311-specific CD4 T cells are plotted in the adjoining graphs. (D) FACS plots in (C) were used to quantify the percentages of T-bet LO EOMES HI cells (quadrant 4) among NP311-specific CD4 T cells. (E) Percentages of CD103 HI and CD69 HI cells among NP311-specific CD4 T cells. Data are representative of two independent experiments. Comparisons were made using one-way ANOVA test with Tukey-corrected multiple comparisons; ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: Hamster Anti-Mouse CD69-PE-Cy7-conjugated (H1.2F3) , BD PharMingen , Cat# 552879.

Techniques: Staining, Binding Assay

Journal: Cell Reports Medicine

Article Title: Programming Multifaceted Pulmonary T Cell Immunity by Combination Adjuvants

doi: 10.1016/j.xcrm.2020.100095

Figure Lengend Snippet: Mucosal CD8 and CD4 T Cell Memory in Vaccinated Mice At 100 days after booster vaccination, NP366-specific memory CD8 T cells (A–D) and NP311-specific CD4 T cells (E–H) were characterized in lungs, airways (BAL), and spleen. To stain for vascular cells, mice were injected intravenously with fluorescent-labeled anti-CD45.2 antibodies, 3 min prior to euthanasia. Cells from lungs and BAL were stained with D b /NP366 tetramers, I-A b /NP311 tetramers, and anti-CD4, anti-CD8, anti-CD44, anti-CD103, and anti-CD69 antibodies. (A) Percentages and total numbers of NP366-specific CD8 T cells in lungs, BAL, and spleen. (B) FACS plots are gated on NP366-specific, tetramer-binding CD8 T cells; numbers are the percentages of vascular and non-vascular cells in the gated population. (C) Percentages of CD69 +ve CD103 +ve T RM cells among NP366-specific CD8 T cells. (D) Total numbers of vascular and non-vascular CD103 +ve NP366-specific CD8 T cells in lungs. (E) Percentages and total numbers of NP311-specific CD4 T cells in lungs. (F) Percentages of vascular and non-vascular cells among NP311-specific CD4 T cells in lungs. (G) Percentages of IFN-γ- or IL-17-producing cells among CD4 T cells. (H) Calculated percentages of IFN-γ- and/or IL-17-producing CD4 T cells among total NP311-specific, cytokine-producing (IFN-γ + IL-17) peptide-stimulated CD4 T cells. Data are pooled from two independent experiments. Comparisons were made using one-way ANOVA test with Tukey-corrected multiple comparisons; ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: Hamster Anti-Mouse CD69-PE-Cy7-conjugated (H1.2F3) , BD PharMingen , Cat# 552879.

Techniques: Staining, Injection, Labeling, Binding Assay

Journal: Cell Reports Medicine

Article Title: Programming Multifaceted Pulmonary T Cell Immunity by Combination Adjuvants

doi: 10.1016/j.xcrm.2020.100095

Figure Lengend Snippet:

Article Snippet: Hamster Anti-Mouse CD69-PE-Cy7-conjugated (H1.2F3) , BD PharMingen , Cat# 552879.

Techniques: Blocking Assay, Derivative Assay, Recombinant, Protein Extraction, Staining, Electron Microscopy, Cell Culture, Enzyme-linked Immunosorbent Assay, Software

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

Article Title: Activation-induced iron flux controls CD4 T cell proliferation by promoting proper IL-2R signaling and mitochondrial function

doi: 10.4049/jimmunol.1901399

Figure Lengend Snippet: Iron chelation does not affect IL-2 production by activated T cells. (A-C) Enriched CD4 T cells were stimulated for 3 days in the presence (5, 10, 20 µM) or absence (0 µM) of DFO. (A) The graph represents the fold change in mean fluorescence intensity (MFI) of CD25 and CD69 at each of the indicated DFO concentrations relative to untreated cells. Statistics in (A) were performed by comparing each DFO concentration to untreated cells (0 µM). (n=3) (B) Graphs show IL-2 levels (µg/mL) in media collected from CD4 T cells as measured by ELISA. (n=3) (C) Representative dot plots show the % IL-2+ CD4 T cells after 3 days of stimulation followed by re-stimulation with PMA and Ionomycin for 4 h. The bar graph shows the cumulative percentages of IL-2+ cells from 3 independent experiments. (D) Enriched CD4 T cells were stained with CellTrace Violet (CTV) and stimulated in the absence (0 µM) or presence (5, 10, 20 µM) of DFO either with or without 50 units of recombinant IL-2. Representative histograms show cell proliferation at day 3 post-activation as measured by CTV dilutions. (n=3) E) Enriched CD4 T cells were stimulated for 48 h in the presence (10 µM, 20 µM) or absence (0 µM) of DFO and stained for pSTAT5. The pooled graph shows pSTAT5 levels at each treatment condition. Statistics in (E) were performed by comparing DFO-treated conditions to the untreated (0 µM) condition. (n=3). Error bars represent Mean ± SEM. *p<0.05, **p<0.005, ****p<0.0001.

Article Snippet: The fluorescently-conjugated antibodies used for surface and intracellular staining in the presence of anti-FcγR mAb (2.4G2) were: anti-mouse TCR-β (H57–597) Pacific Blue/APC, anti-mouse CD4 (GK1.5) APC-Cy7, anti-mouse CD8 (53–6.7) PE-Cy7, anti-mouse CD71 ( {"type":"entrez-nucleotide","attrs":{"text":"R17217","term_id":"770827","term_text":"R17217"}} R17217 ) FITC/PerCP-Cy5.5, anti-mouse CD25 (PC61.5) PerCP-Cy5.5/PE-Cy7, anti-mouse CD69 (H1.2F3) PE-Cy7, and IL-2 (JES6–5H4) PE (all from eBioscience).

Techniques: Fluorescence, Concentration Assay, Enzyme-linked Immunosorbent Assay, Staining, Recombinant, Activation Assay

Journal: Immunity

Article Title: KLRG1 + Effector CD8 + T Cells Lose KLRG1, Differentiate into All Memory T Cell Lineages, and Convey Enhanced Protective Immunity

doi: 10.1016/j.immuni.2018.03.015

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-mouse CD69 (H1.2F3) PE-Cy7 , BD Biosciences , Cat#552879, RRID:AB_394508.

Techniques: Expressing, Recombinant, Purification, Cell Isolation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Software