Journal: Frontiers in Immunology

Article Title: Targeting LAG-3 and PD-1 to Enhance T Cell Activation by Antigen-Presenting Cells

doi: 10.3389/fimmu.2018.00385

Figure Lengend Snippet: Effect of programmed cell death protein 1 (PD-1) and lymphocyte activation gene 3 (LAG-3) blockade on IFN-γ secretion of different T cell subpopulations after stimulation with TLR-3-DCs. MACS-enriched CD3 + T cells of 8 healthy donor (HDs) were sorted according to CCR7 and CD45 RA expression (A) . The various T cell subpopulations were cocultured with autologous CMV, EBV, influenza, tetanus (CEFT)-pulsed TLR-3-DCs in the presence or absence of α-PD-1 (B) and α-LAG-3 (C) antibody. IFN-γ secretion was determined by cytometric bead array (CBA) assay, and the ratio between concentration with and without blocking antibody was calculated. All data are presented as box-and-whisker plots, and statistical significance was calculated against a fold change of 1.0. * p < 0.05.

Article Snippet: Immunofluorescent staining of T-cell surface antigens was performed using the following fluorescence-conjugated monoclonal antibodies: CD244 (PE, C1.7; 329507 or APC, C1.7; 329511), PD-1 (Brilliant Violet 421, EH12.7H7; 329919), CD3 (FITC, UCHT1; 300406), CD45RA (Brilliant Violet 421, HI100; 304129; all BioLegend), CD160 (APC, 688327; FAB6700A), TIM-3 (PE, 344823; FAB2365P; both R&D Systems), CD8 (PerCP-eFluor 710, SK1; 8046-0087; eBioscience), CD4 (APC-H7, RPA-T4; 560158; BD Biosciences), LAG-3 (ATTO 647N, 17B4; AG-20B-0012TS AdipoGen), CCR7 (CD197, APC, FR 11-11E8; 130-098-125; Miltenyi Biotec).

Techniques: Activation Assay, Expressing, Concentration Assay, Blocking Assay, Whisker Assay

Journal: Immunology

Article Title: Lentiviral calnexin-modified dendritic cells promote expansion of high-avidity effector T cells with central memory phenotype.

doi: 10.1111/j.1365-2567.2009.03067.x

Figure Lengend Snippet: Figure 6. Increased CCR7 expression in calnexin–dendritic cell (CNX-DC)-activated T cells. (a) Flow cytometry analysis of memory markers in T cells after coculture with lentiviral vector (LV)-modi- fied DCs. T cells were cocultured with different LV-modified DCs as indicated, restimulated for 7 days and stained with GLC-pentamer together with anti-CD8 antibody and antibodies against CCR7, CD62L, CD28 or CD69. GLC pentamer-positive T cells were CD8- gated and analysed for the expression of differentiation markers. The percentages of different cell population are indicated in the flow graphs. (b) Quantitative analysis of CCR7+ cells in pentamer-positive T cells. Data are representative of four experiments.

Article Snippet: Other fluorochromeconjugated antibodies used in this study included FITC-CCR7 (R&D Systems Inc., Minneapolis, MN), FITC-conjugated human leucocyte antigen DR (HLA-DR) (Caltag Laboratories, Invitrogen, Carlsbad, CA), APC-DCSIGN and APC-CD28 (eBioscience, San Diego, CA).

Techniques: Expressing, Flow Cytometry, Plasmid Preparation, Staining

Journal: Discover Oncology

Article Title: Integrated WGCNA retrieval of T-cell exhaustion genes related to radiosensitivity in locally advanced cervical cancer and prediction of immunotherapy efficacy

doi: 10.1007/s12672-025-03673-y

Figure Lengend Snippet: IHC assay results of the protein expression levels of APIBEC3H, CCR7, PILRA, and SLC15A3 in LACC patients

Article Snippet: Subsequently, the sections were incubated at 4 °C overnight with the corresponding primary antibody against APOBEC3H (1:200, PA5-54426, Invitrogen, USA), CCR7 (1:100, 25898-1-AP, Proteintech, China), PLRA (1:100, PA5-147474, Invitrogen, USA), SLC15A3 (1:100, PA5-61614, Invitrogen, USA).

Techniques: Expressing

Journal: Virology

Article Title: Selective infection of CD4+ effector memory T lymphocytes leads to preferential depletion of memory T lymphocytes in R5 HIV-1-infected humanized NOD/SCID/IL-2Rgammanull mice.

doi: 10.1016/j.virol.2009.08.011

Figure Lengend Snippet: Fig. 5. Phenotype of productively infected p24+ cells in the spleen of R5 HIV-1-infected mice. (A) Representative profiles of flow cytometric p24 staining in splenic nucleated cells of mock-infected (n=4) and HIV-1JR-CSF-infected (n=6) mice. The numbers indicate the percentage of cells in splenic nucleated cells. (B and C) Staining of splenic nucleated cells of HIV-1JR-CSF-infected mice for p24, CD3, and CD4. Representative profiles are shown in B, and the numbers in each quadrant indicate the percentage of cells in p24+ cells. The percentages of each population in p24+ cells are shown in C. (D–F) Staining of splenic nucleated cells of HIV-1JR-CSF-infected mice for p24, CD45RA, CD45RO, and CCR7. Representative profiles are shown in D, and the numbers in each quadrant indicate the percentage of cells in p24+ cells (left) or in p24+CD45RA−CD45RO+ cells (right). The percentages of memory (CD45RA−CD45RO+) and naïve (CD45RA+CD45RO−) phenotyped cells in p24+ cells are shown in E. The percentages of TEM (CCR7−CD45RA−CD45RO+) and TCM (CCR7+CD45RA−CD45RO+) in p24+ cells are shown in F. The error bars in C, E, and F show standard deviations. Asterisks indicate statistical significance (Pb0.05).

Article Snippet: Fluorescein isothiocyanate-conjugated (FITC-conjugated) anti-human CD19 (HD37; Dako, Tokyo, Japan), CD8 (DK25; Dako), CD14 (TUK4; Miltenyi Biotec Inc), CD4 (L3T4; eBioscience, San Diego, CA), CD3 (UCHT1; BD Pharmingen, San Diego, CA), CCR5 (3A9; BD Pharmingen), and CD303/BDCA2 (AC144; Miltenyi Biotec Inc) mouse IgG monoclonal antibodies (mAb); phycoerythrinconjugated anti-human CD3 (UCHT1; Dako), CD4 (MT310; Dako), CD34 (AC136; Miltenyi Biotec Inc), CD11c (B-ly6; BD Pharmingen), CXCR4 (12G5; BD Pharmingen), CCR7 (FAB197; R&D systems, Abingdon, UK), and CCR5 (3A9; BD Pharmingen) mouse IgG mAb; biotinylated anti-human CD45 (H130; eBioscience), CD45RA (HI-100; BD Pharmingen), CD8 (RPA-T8; BD Pharmingen), CD4 (RPA-T4; BD Pharmingen), and mouse IgG mAb; peridinin-chlorophyll-conjugated (PerCP-conjugated) anti-human CD69 (L78; BD Immunocytometry Systems, San Jose, CA) mouse IgG mAb; PE-Cy5-conjugated antihuman HLA-DR (G46-6; BD Pharmingen) mouse IgG mAb; allophycocyamin-conjugated anti-human CD45RO (UCHL1; BD Pharmingen) and CD8 (DK25; Dako) mouse IgG mAb were used.

Techniques: Infection, Staining