Journal: PLoS ONE

Article Title: Neutrophil Gelatinase-Associated Lipocalin Increases HLA-G + /FoxP3 + T-Regulatory Cell Population in an In Vitro Model of PBMC

doi: 10.1371/journal.pone.0089497

Figure Lengend Snippet: A) Flow cytometry analysis of surface HLA-G on CD4 + and CD8 + cells. PBMCs were treated with increasing concentrations of NGAL:Enterobactin:Iron (40 ng/ml, 80 ng/ml, 160 ng/ml, and 320 ng/ml). A dose-dependent rise in the percentage of CD4 + HLA-G + cells was evident. The percentage of CD8 + HLA-G + cells did not show any marked changes. Data are representative of 8 independent experiments. The number in each quadrant represents the percentage of the total population. B) Data are expressed as percentages of CD4 + HLA-G + cells. Means ± SD; n = 8. * p<0.05.

Article Snippet: Detection of CD4+ CD25+ FoxP3+ regulatory T cells on PBMC was performed with the Treg Detection Kit, CD4/CD25/FoxP3 (Miltenyi, Bergish Gladbach, Germany) as described in the manufacturer's protocol.

Techniques: Flow Cytometry

Journal: PLoS ONE

Article Title: Neutrophil Gelatinase-Associated Lipocalin Increases HLA-G + /FoxP3 + T-Regulatory Cell Population in an In Vitro Model of PBMC

doi: 10.1371/journal.pone.0089497

Figure Lengend Snippet: Flow cytometry analysis of HLA-G expression on CD4 + cells. Iron treatment (without NGAL) did not increase the percentage of HLA-G + cells. Treatment with the NGAL:enterobactin complex (without iron) reduced HLA-G expression in contrast to stimulation with NGAL:Enterobactin:Iron. Incubation with anti-NGAL antibody reduced the percentage of HLAG + cells. Data are representative of 8 independent experiments. The number in each quadrant represents the percentage of the total population. B) Data are expressed as percentages of CD4 + HLA-G + cells. Means ± SD; n = 8. * p<0.05.

Article Snippet: Detection of CD4+ CD25+ FoxP3+ regulatory T cells on PBMC was performed with the Treg Detection Kit, CD4/CD25/FoxP3 (Miltenyi, Bergish Gladbach, Germany) as described in the manufacturer's protocol.

Techniques: Flow Cytometry, Expressing, Incubation

Journal: PLoS ONE

Article Title: Neutrophil Gelatinase-Associated Lipocalin Increases HLA-G + /FoxP3 + T-Regulatory Cell Population in an In Vitro Model of PBMC

doi: 10.1371/journal.pone.0089497

Figure Lengend Snippet: A). The percentage of CD4 + /CD25 + /FoxP3 + cells in PHA-activated PBMCs after stimulation with increasing concentrations of NGAL: Enterobactin:Iron (40 ng/ml, 80 ng/ml, 160 ng/ml, and 320 ng/ml), raised in a dose dependent manner. Data are representative of 8 independent experiments. The number in each quadrant represents the percentage of CD25 + /FoxP3 + cells on gated CD4 + cells of the total population. B) Data are expressed as percentages of CD4 + /CD25 + /FoxP3 + cells. Means ± SD; n = 8. *p<0.05.

Article Snippet: Detection of CD4+ CD25+ FoxP3+ regulatory T cells on PBMC was performed with the Treg Detection Kit, CD4/CD25/FoxP3 (Miltenyi, Bergish Gladbach, Germany) as described in the manufacturer's protocol.

Techniques:

Journal: PLoS ONE

Article Title: Neutrophil Gelatinase-Associated Lipocalin Increases HLA-G + /FoxP3 + T-Regulatory Cell Population in an In Vitro Model of PBMC

doi: 10.1371/journal.pone.0089497

Figure Lengend Snippet: Flow cytometry analysis of CD25 + /FoxP3 + expression on CD4 + cells in PHA-activated PBMCs. Iron treatment (without NGAL) did not increase the percentage of CD25 + /FoxP3 + cells. Treatment with NGAL:enterobactin complex (without iron) reduced the percentage of CD25 + /FoxP3 + cells in contrast to stimulation with NGAL:Enterobactin:Iron. Incubation with anti-NGAL antibody reduced the percentage of CD25 + /FoxP3 + cells. Data are representative of 8 independent experiments. The number in each quadrant represents the percentage of CD25+foxP3 cells on gated CD4 + cells of the total population. B) Data are expressed as percentages of CD4+/CD25+/FoxP3+ cells. Means ± SD; n = 8. * p<0.05.

Article Snippet: Detection of CD4+ CD25+ FoxP3+ regulatory T cells on PBMC was performed with the Treg Detection Kit, CD4/CD25/FoxP3 (Miltenyi, Bergish Gladbach, Germany) as described in the manufacturer's protocol.

Techniques: Flow Cytometry, Expressing, Incubation

Journal: PLoS ONE

Article Title: Neutrophil Gelatinase-Associated Lipocalin Increases HLA-G + /FoxP3 + T-Regulatory Cell Population in an In Vitro Model of PBMC

doi: 10.1371/journal.pone.0089497

Figure Lengend Snippet: Flow cytometry analysis of HLA-G + /FoxP3 + expression on CD4 + cell PHA-activated PBMCs. PBMCs were treated with increasing concentrations of NGAL:Enterobactin:Iron (40 ng/ml, 80 ng/ml, 160 ng/ml, and 320 ng/ml). A dose-dependent raise in the percentage of HLA-G + /FoxP3 + cells was evident. Data are representative of 8 independent experiments. The number in each quadrant represents the percentage of HLA-G + /FoxP3 + cells on gated CD4 + cells of the total population. B) Data are expressed as percentages of CD4 + /HLA-G + /FoxP3 + cells. Means ± SD; n = 8. * p<0.05.

Article Snippet: Detection of CD4+ CD25+ FoxP3+ regulatory T cells on PBMC was performed with the Treg Detection Kit, CD4/CD25/FoxP3 (Miltenyi, Bergish Gladbach, Germany) as described in the manufacturer's protocol.

Techniques: Flow Cytometry, Expressing

Journal: Frontiers in Immunology

Article Title: LAMP1 targeting of the large T antigen of Merkel cell polyomavirus results in potent CD4 T cell responses and tumor inhibition

doi: 10.3389/fimmu.2023.1253568

Figure Lengend Snippet: ITI-3000 induces spatial immune T cell infiltration into tumors. Experimental design shown in (A) . C57BL/6 mice were given 5E4 B16-LT tumor cells subcutaneously in the right flank. Starting on day 3 post tumor injection, mice were vaccinated twice, weekly, with 40µg of ITI-3000 or control vector via intradermal injection followed by electroporation. Tumors were collected and formalin fixed on day 15 post tumor injection, followed by a sucrose gradient, paraffin embedding, and sectioning. The cellular composition of the tumor microenvironment on FFPE slides was determined by multiplex-IHC including percent of CD3 + (green), CD8 + (magenta) (B) , and CD4 + (red) (C) cells. One representative section (displayed both as whole tumor section (right) and magnified area of interest (left)) of each group is shown in (D) The total cell population was determined by nuclear DAPI staining (blue). Sections were analyzed by AI gating in the Biodock platform. N=5. Statistical significance was calculated by unpaired Student’s t-test. The "*" symbol represents a p-value of <0.05.

Article Snippet: A murine 5-color panel including CD3, CD4, CD8, FoxP3, and DAPI was done at Ultivue Inc. (Cambridge, MA).

Techniques: Injection, Control, Plasmid Preparation, Electroporation, Multiplex Assay, Staining

Journal: Cell and Tissue Research

Article Title: Lung development and immune status under chronic LPS exposure in rat pups with and without CD26/DPP4 deficiency

doi: 10.1007/s00441-021-03522-8

Figure Lengend Snippet: Primary antibodies used for FACS-Analysis

Article Snippet: Furthermore, earlier results exhibited a significantly increased influx of CD4 + CD25 + FoxP3 + into the lungs of CD26 − rats and increased IL-10-secretion/production by draining lymph node cells in culture, however, after OVA challenge (Schmiedl et al. ).

Techniques:

Journal: Cell and Tissue Research

Article Title: Lung development and immune status under chronic LPS exposure in rat pups with and without CD26/DPP4 deficiency

doi: 10.1007/s00441-021-03522-8

Figure Lengend Snippet: Absolute immune cell numbers in the lung during alveolarization

Article Snippet: Furthermore, earlier results exhibited a significantly increased influx of CD4 + CD25 + FoxP3 + into the lungs of CD26 − rats and increased IL-10-secretion/production by draining lymph node cells in culture, however, after OVA challenge (Schmiedl et al. ).

Techniques: Control, Cell Counting

Journal: Cell and Tissue Research

Article Title: Lung development and immune status under chronic LPS exposure in rat pups with and without CD26/DPP4 deficiency

doi: 10.1007/s00441-021-03522-8

Figure Lengend Snippet: Percentage of immunological cells in the lung during alveolarization

Article Snippet: Furthermore, earlier results exhibited a significantly increased influx of CD4 + CD25 + FoxP3 + into the lungs of CD26 − rats and increased IL-10-secretion/production by draining lymph node cells in culture, however, after OVA challenge (Schmiedl et al. ).

Techniques: Control