anti cd127 pe  (BioLegend)

 
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    Name:
    PE anti human CD127 IL 7Rα
    Description:
    PE anti human CD127 IL 7Rα A019D5 Isotype Mouse IgG1 κ Reactivity Human Apps FC Size 25 tests
    Catalog Number:
    351303
    Price:
    115
    Category:
    Human Immunology
    Source:
    Mouse
    Applications:
    FC
    Conjugate:
    PE
    Immunogen:
    Recombinant human CD127
    Size:
    25 tests
    Quantity:
    1
    Preparation:
    The antibody was purified by affinity chromatography and conjugated with PE under optimal conditions The solution is free of unconjugated PE and unconjugated antibody
    Buy from Supplier


    Structured Review

    BioLegend anti cd127 pe
    PE anti human CD127 IL 7Rα
    PE anti human CD127 IL 7Rα A019D5 Isotype Mouse IgG1 κ Reactivity Human Apps FC Size 25 tests
    https://www.bioz.com/result/anti cd127 pe/product/BioLegend
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd127 pe - by Bioz Stars, 2021-07
    92/100 stars

    Images

    1) Product Images from "Peptide Processing Is Critical for T-Cell Memory Inflation and May Be Optimized to Improve Immune Protection by CMV-Based Vaccine Vectors"

    Article Title: Peptide Processing Is Critical for T-Cell Memory Inflation and May Be Optimized to Improve Immune Protection by CMV-Based Vaccine Vectors

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006072

    Gene expression context does not define the quality of CD8 responses to MHC-I restricted epitopes of MCMV. 129/Sv mice were infected intraperitoneally (i.p.) with 2x10 5 PFU of MCMV M45SL . Blood leukocytes were stimulated with the SSIEFARL or the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. Cells were surface-stained for CD3, CD4, CD8, CD11a, CD44, KLRG1, CD127 and intracellularly for IFNγ expression and analyzed by flow cytometry. ( A ) Cells responding to the SSIEFARL (gB K b response) or the HGIRNASFI (M45D b response) peptide. ( B ) Upper graph—epitope specific cells with the EM phenotype (CD127 - KLRG1 + ). Lower graph—epitope specific cells with the CM phenotype (CD127 + KLRG1 - ). The experiment was performed three times independently, at 5 mice per group in each experiment, and grouped averages +/- SEM from all three experiments are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. **** p
    Figure Legend Snippet: Gene expression context does not define the quality of CD8 responses to MHC-I restricted epitopes of MCMV. 129/Sv mice were infected intraperitoneally (i.p.) with 2x10 5 PFU of MCMV M45SL . Blood leukocytes were stimulated with the SSIEFARL or the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. Cells were surface-stained for CD3, CD4, CD8, CD11a, CD44, KLRG1, CD127 and intracellularly for IFNγ expression and analyzed by flow cytometry. ( A ) Cells responding to the SSIEFARL (gB K b response) or the HGIRNASFI (M45D b response) peptide. ( B ) Upper graph—epitope specific cells with the EM phenotype (CD127 - KLRG1 + ). Lower graph—epitope specific cells with the CM phenotype (CD127 + KLRG1 - ). The experiment was performed three times independently, at 5 mice per group in each experiment, and grouped averages +/- SEM from all three experiments are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. **** p

    Techniques Used: Expressing, Mouse Assay, Infection, Staining, Flow Cytometry, Cytometry, MANN-WHITNEY

    Epitope localization defines the quality of CD8 T-cell responses. ( A ) Representative dot plots of intracellular IFNγ expression at 7 and 180 dpi upon HGIRNASFI peptide stimulation. ( B ) Grouped means +/- SEM of cells responding to the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. ( C ) Representative dot plots of the surface expression of CD127 and KLRG1 on HGIRNASFI specific CD8 T cells at 7 and 180 dpi with MCMV WT or MCMV M45Cterm . The staining was used to define the CM (CD127 + KLRG1 - ) and the EM (CD127 - KLRG1 + ) subsets. ( D, E ) Grouped means +/- SEM of the percentage of EM ( D ) or CM ( E ) cells in the HGIRNASFI-responding subset at indicated time points p.i‥ The experiment was performed twice, at 5 mice per group in each experiment, and pooled results are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. *p
    Figure Legend Snippet: Epitope localization defines the quality of CD8 T-cell responses. ( A ) Representative dot plots of intracellular IFNγ expression at 7 and 180 dpi upon HGIRNASFI peptide stimulation. ( B ) Grouped means +/- SEM of cells responding to the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. ( C ) Representative dot plots of the surface expression of CD127 and KLRG1 on HGIRNASFI specific CD8 T cells at 7 and 180 dpi with MCMV WT or MCMV M45Cterm . The staining was used to define the CM (CD127 + KLRG1 - ) and the EM (CD127 - KLRG1 + ) subsets. ( D, E ) Grouped means +/- SEM of the percentage of EM ( D ) or CM ( E ) cells in the HGIRNASFI-responding subset at indicated time points p.i‥ The experiment was performed twice, at 5 mice per group in each experiment, and pooled results are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. *p

    Techniques Used: Expressing, Staining, Mouse Assay, MANN-WHITNEY

    2) Product Images from "Plasma-Activated Medium Potentiates the Immunogenicity of Tumor Cell Lysates for Dendritic Cell-Based Cancer Vaccines"

    Article Title: Plasma-Activated Medium Potentiates the Immunogenicity of Tumor Cell Lysates for Dendritic Cell-Based Cancer Vaccines

    Journal: Cancers

    doi: 10.3390/cancers13071626

    Treg induction capacity of DCs treated with A375 lysates. DCs either treated or not with the A375 lysate (+A375lys) or the PAM-treated A375 lysate (+PAM-A375lys) were co-cultivated with MACS-purified allogeneic T cells (1 × 10 5 /well) at 1:40 (DC: T cell ratio) for 5 days, in the presence of 2 ng/mL of IL-2, followed by the stimulation of the co-cultures with PMA/ Ca Ionophore/ Monensin for the last 3 h prior to their staining for flow cytometry. ( a ) Representative dot-plots are shown on total gated CD4 + T cells after staining to CD25, CD127 and intracellular FoxP3. The summarized results on the % of CD4 Tregs are shown as % ± SD of 3 independent experiments. ( b ) Representative dot-plots are shown on total gated CD8 + T cells after staining to CD25, and intracellular IFN-γ and IL-10. The summarized results on the % of CD8 Tregs are shown as % ± SD of 3 independent experiments. * p
    Figure Legend Snippet: Treg induction capacity of DCs treated with A375 lysates. DCs either treated or not with the A375 lysate (+A375lys) or the PAM-treated A375 lysate (+PAM-A375lys) were co-cultivated with MACS-purified allogeneic T cells (1 × 10 5 /well) at 1:40 (DC: T cell ratio) for 5 days, in the presence of 2 ng/mL of IL-2, followed by the stimulation of the co-cultures with PMA/ Ca Ionophore/ Monensin for the last 3 h prior to their staining for flow cytometry. ( a ) Representative dot-plots are shown on total gated CD4 + T cells after staining to CD25, CD127 and intracellular FoxP3. The summarized results on the % of CD4 Tregs are shown as % ± SD of 3 independent experiments. ( b ) Representative dot-plots are shown on total gated CD8 + T cells after staining to CD25, and intracellular IFN-γ and IL-10. The summarized results on the % of CD8 Tregs are shown as % ± SD of 3 independent experiments. * p

    Techniques Used: Magnetic Cell Separation, Purification, Staining, Flow Cytometry

    3) Product Images from "Transient Expression of GATA3 in Hematopoietic Stem Cells Facilitates Helper Innate Lymphoid Cell Differentiation"

    Article Title: Transient Expression of GATA3 in Hematopoietic Stem Cells Facilitates Helper Innate Lymphoid Cell Differentiation

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2019.00510

    GATA3 induces the development of ILC progenitors. Five-day expanded UCB CD34 + α4β7 − HSCs were transfected with either GATA3 mRNA (GATA3) or scrambled mRNA (control) and differentiated under conditions that favor the development of ILCs (A) Gating strategy, 5-days expanded CD34 + α4β7 − HSCs were sorted by FACS. (B) CD34 + α4β7 − HSCs were stained for the surface α4β7 at day 7 post electroporation and dot plot is shown ( n = 4). Values represent the percentage of α4β7 + cells. (C,D) CD34 + α4β7 − HSCs were stained for the surface α4β7 at day 7 post electroporation and the percentage (C) and MFI (D) of α4β7 + cells is shown ( n = 4/group). (E) CD34 + α4β7 − HSCs were stained for the surface CD127 at day 14 post electroporation and dot plot is shown ( n = 4). Values represent the percentage of CD127 + cells. (F) CD34 + α4β7 − HSCs were stained for the surface CD127 at day 14 post electroporation and the percentage of CD127 + cells is shown in bar graph ( n = 4/group). (G) CD34 + α4β7 − HSCs were stained for the surface CD11a and CD117 at day 14 post electroporation and dot plot is shown ( n = 4). Values represent the percentage of CD117 + CD11a − ILCPs. (H) CD34 + α4β7 − HSCs were stained for the surface CD11a and CD117 at day 14 post electroporation and the percentage of CD117 + CD11a − ILCPs is shown in bar graph ( n = 4/group). (I) Total counts of transfected and differentiating HSCs were performed at day 14 of culture and live cells in 1 mL of culture media is depicted ( n = 4/group). (J) Absolute counts of CD117 + CD11a − ILCPs in 1 mL of culture media is depicted ( n = 4/group). (C,D,F,H–J) , Data are shown as means ± SD, and p -values are depicted and paired two-tailed t -tests.
    Figure Legend Snippet: GATA3 induces the development of ILC progenitors. Five-day expanded UCB CD34 + α4β7 − HSCs were transfected with either GATA3 mRNA (GATA3) or scrambled mRNA (control) and differentiated under conditions that favor the development of ILCs (A) Gating strategy, 5-days expanded CD34 + α4β7 − HSCs were sorted by FACS. (B) CD34 + α4β7 − HSCs were stained for the surface α4β7 at day 7 post electroporation and dot plot is shown ( n = 4). Values represent the percentage of α4β7 + cells. (C,D) CD34 + α4β7 − HSCs were stained for the surface α4β7 at day 7 post electroporation and the percentage (C) and MFI (D) of α4β7 + cells is shown ( n = 4/group). (E) CD34 + α4β7 − HSCs were stained for the surface CD127 at day 14 post electroporation and dot plot is shown ( n = 4). Values represent the percentage of CD127 + cells. (F) CD34 + α4β7 − HSCs were stained for the surface CD127 at day 14 post electroporation and the percentage of CD127 + cells is shown in bar graph ( n = 4/group). (G) CD34 + α4β7 − HSCs were stained for the surface CD11a and CD117 at day 14 post electroporation and dot plot is shown ( n = 4). Values represent the percentage of CD117 + CD11a − ILCPs. (H) CD34 + α4β7 − HSCs were stained for the surface CD11a and CD117 at day 14 post electroporation and the percentage of CD117 + CD11a − ILCPs is shown in bar graph ( n = 4/group). (I) Total counts of transfected and differentiating HSCs were performed at day 14 of culture and live cells in 1 mL of culture media is depicted ( n = 4/group). (J) Absolute counts of CD117 + CD11a − ILCPs in 1 mL of culture media is depicted ( n = 4/group). (C,D,F,H–J) , Data are shown as means ± SD, and p -values are depicted and paired two-tailed t -tests.

    Techniques Used: Transfection, FACS, Staining, Electroporation, Two Tailed Test

    4) Product Images from "Peptide Processing Is Critical for T-Cell Memory Inflation and May Be Optimized to Improve Immune Protection by CMV-Based Vaccine Vectors"

    Article Title: Peptide Processing Is Critical for T-Cell Memory Inflation and May Be Optimized to Improve Immune Protection by CMV-Based Vaccine Vectors

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006072

    Gene expression context does not define the quality of CD8 responses to MHC-I restricted epitopes of MCMV. 129/Sv mice were infected intraperitoneally (i.p.) with 2x10 5 PFU of MCMV M45SL . Blood leukocytes were stimulated with the SSIEFARL or the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. Cells were surface-stained for CD3, CD4, CD8, CD11a, CD44, KLRG1, CD127 and intracellularly for IFNγ expression and analyzed by flow cytometry. ( A ) Cells responding to the SSIEFARL (gB K b response) or the HGIRNASFI (M45D b response) peptide. ( B ) Upper graph—epitope specific cells with the EM phenotype (CD127 - KLRG1 + ). Lower graph—epitope specific cells with the CM phenotype (CD127 + KLRG1 - ). The experiment was performed three times independently, at 5 mice per group in each experiment, and grouped averages +/- SEM from all three experiments are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. **** p
    Figure Legend Snippet: Gene expression context does not define the quality of CD8 responses to MHC-I restricted epitopes of MCMV. 129/Sv mice were infected intraperitoneally (i.p.) with 2x10 5 PFU of MCMV M45SL . Blood leukocytes were stimulated with the SSIEFARL or the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. Cells were surface-stained for CD3, CD4, CD8, CD11a, CD44, KLRG1, CD127 and intracellularly for IFNγ expression and analyzed by flow cytometry. ( A ) Cells responding to the SSIEFARL (gB K b response) or the HGIRNASFI (M45D b response) peptide. ( B ) Upper graph—epitope specific cells with the EM phenotype (CD127 - KLRG1 + ). Lower graph—epitope specific cells with the CM phenotype (CD127 + KLRG1 - ). The experiment was performed three times independently, at 5 mice per group in each experiment, and grouped averages +/- SEM from all three experiments are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. **** p

    Techniques Used: Expressing, Mouse Assay, Infection, Staining, Flow Cytometry, Cytometry, MANN-WHITNEY

    Epitope localization defines the quality of CD8 T-cell responses. ( A ) Representative dot plots of intracellular IFNγ expression at 7 and 180 dpi upon HGIRNASFI peptide stimulation. ( B ) Grouped means +/- SEM of cells responding to the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. ( C ) Representative dot plots of the surface expression of CD127 and KLRG1 on HGIRNASFI specific CD8 T cells at 7 and 180 dpi with MCMV WT or MCMV M45Cterm . The staining was used to define the CM (CD127 + KLRG1 - ) and the EM (CD127 - KLRG1 + ) subsets. ( D, E ) Grouped means +/- SEM of the percentage of EM ( D ) or CM ( E ) cells in the HGIRNASFI-responding subset at indicated time points p.i‥ The experiment was performed twice, at 5 mice per group in each experiment, and pooled results are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. *p
    Figure Legend Snippet: Epitope localization defines the quality of CD8 T-cell responses. ( A ) Representative dot plots of intracellular IFNγ expression at 7 and 180 dpi upon HGIRNASFI peptide stimulation. ( B ) Grouped means +/- SEM of cells responding to the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. ( C ) Representative dot plots of the surface expression of CD127 and KLRG1 on HGIRNASFI specific CD8 T cells at 7 and 180 dpi with MCMV WT or MCMV M45Cterm . The staining was used to define the CM (CD127 + KLRG1 - ) and the EM (CD127 - KLRG1 + ) subsets. ( D, E ) Grouped means +/- SEM of the percentage of EM ( D ) or CM ( E ) cells in the HGIRNASFI-responding subset at indicated time points p.i‥ The experiment was performed twice, at 5 mice per group in each experiment, and pooled results are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. *p

    Techniques Used: Expressing, Staining, Mouse Assay, MANN-WHITNEY

    5) Product Images from "Off-target effects of Cre recombinase reveal limits of adoptive T-cell transfers and persistent proliferation of effector CD8 T-cells"

    Article Title: Off-target effects of Cre recombinase reveal limits of adoptive T-cell transfers and persistent proliferation of effector CD8 T-cells

    Journal: bioRxiv

    doi: 10.1101/502666

    The activated Cre-Recombinase induces cell death in Ki67 hi effector T-cells in blood and spleen. R26CreER T2 mice were infected with 10 6 PFU MCMV. At 94 dpi the mice received 80mg/kg BW Tamoxifen by oral gavage for 5 consecutive days, followed by a three day break, and one more day of treatment, before mice were sacrificed. Cell proliferation in CD8 T-cell subsets was characterized by CD8, CD44, CD127, and Ki67 expression as well as by the DNA marker FXcycle. (A) Representative dot plots for CD8 T-cell proliferation in blood and splenocytes in treated and untreated animals. (B) Percentages of cell cycle subsets Ki67 - , Ki67 int , Ki67 hi FX - , and Ki67 hi FX + in total CD8 T-cells. (C) Percentages of cell cycle subsets Ki67 - , Ki67 int , Ki67 hi FX - , and Ki67 hi FX + in naïve (CD127 + CD44 - ), memory (CD127 + CD44 + ), and effector (CD127 - CD44 + ) CD8 T-cell subsets. (D) The percentage of dead cells (7-AAD + ) was determined in the naïve (CD62L + CD44 - ), central-memory (CD62L + CD44 + ), and effector-memory (CD62L - CD44 + ) CD8 T-cell subsets in blood and spleen. Pooled data from two independent experiments (12 mice in total) are shown. Bars indicate means, error bars indicate SEM. Significant differences were determined by Mann-Whitney test, *=p
    Figure Legend Snippet: The activated Cre-Recombinase induces cell death in Ki67 hi effector T-cells in blood and spleen. R26CreER T2 mice were infected with 10 6 PFU MCMV. At 94 dpi the mice received 80mg/kg BW Tamoxifen by oral gavage for 5 consecutive days, followed by a three day break, and one more day of treatment, before mice were sacrificed. Cell proliferation in CD8 T-cell subsets was characterized by CD8, CD44, CD127, and Ki67 expression as well as by the DNA marker FXcycle. (A) Representative dot plots for CD8 T-cell proliferation in blood and splenocytes in treated and untreated animals. (B) Percentages of cell cycle subsets Ki67 - , Ki67 int , Ki67 hi FX - , and Ki67 hi FX + in total CD8 T-cells. (C) Percentages of cell cycle subsets Ki67 - , Ki67 int , Ki67 hi FX - , and Ki67 hi FX + in naïve (CD127 + CD44 - ), memory (CD127 + CD44 + ), and effector (CD127 - CD44 + ) CD8 T-cell subsets. (D) The percentage of dead cells (7-AAD + ) was determined in the naïve (CD62L + CD44 - ), central-memory (CD62L + CD44 + ), and effector-memory (CD62L - CD44 + ) CD8 T-cell subsets in blood and spleen. Pooled data from two independent experiments (12 mice in total) are shown. Bars indicate means, error bars indicate SEM. Significant differences were determined by Mann-Whitney test, *=p

    Techniques Used: Mouse Assay, Infection, Expressing, Marker, MANN-WHITNEY

    MCMV-specific CD8 T-cells are depleted by Tamoxifen treatment in R26 CreER T2 mice. (A, B) R26CreER T2 mice were infected with 10 6 PFU MCMV or 200μl PBS (mock). 60 days post infection mice received food pellets with 400 mg/kg Tamoxifen for 4 weeks (indicated by the grey rectangle). Blood was collected and analysed at 0, 7, 14, 28, 60, 90, 120, 150, 180, and 270 dpi. Blood leukocytes were stimulated with the M38 peptide (SSPPMFRV) or the gB peptide (SSIEFARL) and stained for CD8, CD44, CD127, and IFNy expression. (A) Kinetic of M38-specific CD8 T-cells before, during, and after Tamoxifen treatment. (B) Kinetic of gB-specific CD8 T-cells before, during, and after Tamoxifen treatment. Data are pooled from three independent experiments with up to 15 mice per group in total. Error bars indicate SEM. Significance was assessed by Mann-Whitney test. ** p
    Figure Legend Snippet: MCMV-specific CD8 T-cells are depleted by Tamoxifen treatment in R26 CreER T2 mice. (A, B) R26CreER T2 mice were infected with 10 6 PFU MCMV or 200μl PBS (mock). 60 days post infection mice received food pellets with 400 mg/kg Tamoxifen for 4 weeks (indicated by the grey rectangle). Blood was collected and analysed at 0, 7, 14, 28, 60, 90, 120, 150, 180, and 270 dpi. Blood leukocytes were stimulated with the M38 peptide (SSPPMFRV) or the gB peptide (SSIEFARL) and stained for CD8, CD44, CD127, and IFNy expression. (A) Kinetic of M38-specific CD8 T-cells before, during, and after Tamoxifen treatment. (B) Kinetic of gB-specific CD8 T-cells before, during, and after Tamoxifen treatment. Data are pooled from three independent experiments with up to 15 mice per group in total. Error bars indicate SEM. Significance was assessed by Mann-Whitney test. ** p

    Techniques Used: Mouse Assay, Infection, Staining, Expressing, MANN-WHITNEY

    Proliferating CD8 T-cells in latently MCMV infected mice are largely effector cells. (A) and (B): 129/Sv mice were infected with 2 × 10 5 PFU MCMV or VACV. At 7 and 120 dpi, blood was taken and proliferating T-cell subsets were analyzed with antibodies directed against CD8, Ki67, CD44, and CD127. (A) Representative dot plots demonstrating the determination of the immune phenotype using CD44 and CD127. (B) Percentage of TM (CD44 + CD127 + ) and TE (CD44 + CD127 - ) in cycling (Ki67 int and Ki67 hi ) subsets of CD8 T-cells. Bars indicate means, error bars indicate SEM. (C) and (D): 129/Sv mice were infected with 2 × 10 5 PFU MCMV. From 120 dpi onward they were given D 2 O for 28 days. Mice were sacrificed at indicated time points and CD8 + tetramer-negative splenocytes were sorted on CD44 and CD62L expression to isolate DNA from TEM (CD62L - CD44 + ), TCM (CD62L + CD44 + ), and naïve (CD62L + CD44 - ) CD8 T-cells. The deuterium content in the DNA of these CD8 T-cell subsets was determined by mass spectrometry. Three independent experiments with 5-6 mice per time-point were performed, and data from a representative experiment are shown. (C) Best fits of the mathematical model (solid curves) to the deuterium enrichment levels in the indicated cell subsets(circles for individual mice). The gray-shaded area indicates the window of D 2 O administration. Parameters corresponding to the best fits are summarized in Table 1 . (D) Comparison of the percentage of deuterium labelled DNA in the three T-cell subsets of each mouse at the indicated time points during label administration. Each symbol represents a T-cell subset from one mouse and lines connect data points from the same mouse. (E) Blood leukocytes from C57BL/6 mice gated by CD62L and CD44 expression into naïve, TCM, and TEM subsets at 100 dpi with MCMV were labelled intracellularly with Ki67 and FXcycle (see Fig 3A for representative gating) and mean frequencies of Ki67 hi populations in indicated subsets are shown. Experiments were performed twice (total n=12). Error bars are SEM.
    Figure Legend Snippet: Proliferating CD8 T-cells in latently MCMV infected mice are largely effector cells. (A) and (B): 129/Sv mice were infected with 2 × 10 5 PFU MCMV or VACV. At 7 and 120 dpi, blood was taken and proliferating T-cell subsets were analyzed with antibodies directed against CD8, Ki67, CD44, and CD127. (A) Representative dot plots demonstrating the determination of the immune phenotype using CD44 and CD127. (B) Percentage of TM (CD44 + CD127 + ) and TE (CD44 + CD127 - ) in cycling (Ki67 int and Ki67 hi ) subsets of CD8 T-cells. Bars indicate means, error bars indicate SEM. (C) and (D): 129/Sv mice were infected with 2 × 10 5 PFU MCMV. From 120 dpi onward they were given D 2 O for 28 days. Mice were sacrificed at indicated time points and CD8 + tetramer-negative splenocytes were sorted on CD44 and CD62L expression to isolate DNA from TEM (CD62L - CD44 + ), TCM (CD62L + CD44 + ), and naïve (CD62L + CD44 - ) CD8 T-cells. The deuterium content in the DNA of these CD8 T-cell subsets was determined by mass spectrometry. Three independent experiments with 5-6 mice per time-point were performed, and data from a representative experiment are shown. (C) Best fits of the mathematical model (solid curves) to the deuterium enrichment levels in the indicated cell subsets(circles for individual mice). The gray-shaded area indicates the window of D 2 O administration. Parameters corresponding to the best fits are summarized in Table 1 . (D) Comparison of the percentage of deuterium labelled DNA in the three T-cell subsets of each mouse at the indicated time points during label administration. Each symbol represents a T-cell subset from one mouse and lines connect data points from the same mouse. (E) Blood leukocytes from C57BL/6 mice gated by CD62L and CD44 expression into naïve, TCM, and TEM subsets at 100 dpi with MCMV were labelled intracellularly with Ki67 and FXcycle (see Fig 3A for representative gating) and mean frequencies of Ki67 hi populations in indicated subsets are shown. Experiments were performed twice (total n=12). Error bars are SEM.

    Techniques Used: Infection, Mouse Assay, Expressing, Transmission Electron Microscopy, Mass Spectrometry

    MCMV-specific CD8 T-cells are depleted by Tamoxifen treatment in R26 CreER T2 mice. (A, B) R26 CreER T2 mice were infected with 10 6 PFU MCMV or 200 μl PBS (mock). 4 months post infection half of the mice received food pellets containing 400 mg/kg Tamoxifen for 4 weeks (indicated with a grey rectangle). Blood was collected and analysed at 0, 7, 14, 28, 60, 90, 120, 150, 180, and 270 dpi. Blood leukocytes were stimulated with the IE3 peptide (RALEYKNL) and stained for CD8, CD44, CD127, and IFNγ expression. (A) Kinetics of effector (CD44 + CD127 - , TE) CD8 T-cells before, during, and after Tamoxifen treatment. (B) Percentages of IFNγ + CD8 T-cells specific for the IE3 peptide before, during, and after Tamoxifen treatment. Data are pooled from three independent experiments with up to 15 mice per group in total. Error bars indicate SEM. Significance was assessed by Mann-Whitney test. ** p
    Figure Legend Snippet: MCMV-specific CD8 T-cells are depleted by Tamoxifen treatment in R26 CreER T2 mice. (A, B) R26 CreER T2 mice were infected with 10 6 PFU MCMV or 200 μl PBS (mock). 4 months post infection half of the mice received food pellets containing 400 mg/kg Tamoxifen for 4 weeks (indicated with a grey rectangle). Blood was collected and analysed at 0, 7, 14, 28, 60, 90, 120, 150, 180, and 270 dpi. Blood leukocytes were stimulated with the IE3 peptide (RALEYKNL) and stained for CD8, CD44, CD127, and IFNγ expression. (A) Kinetics of effector (CD44 + CD127 - , TE) CD8 T-cells before, during, and after Tamoxifen treatment. (B) Percentages of IFNγ + CD8 T-cells specific for the IE3 peptide before, during, and after Tamoxifen treatment. Data are pooled from three independent experiments with up to 15 mice per group in total. Error bars indicate SEM. Significance was assessed by Mann-Whitney test. ** p

    Techniques Used: Mouse Assay, Infection, Staining, Expressing, MANN-WHITNEY

    Supplemental Tamoxifen data Fraction of CD8 subsets in the CD8 pool upon Tam treatment. Mice were MCMV infected and Tam treated as shown in figure 3 . Blood leukocytes were stained for CD8 and the indicated surface markers and gated in the CD44-CD62L+CD127+ (naïve) CD44+CD62L+CD127+ (central memory), CD44+CD62L-CD127+ (effector memory) and CD44+CD62L-CD127-(Effector) subsets. Shown are group means + standard deviations from two pooled experiments. (B) Tamoxifen induces cell death in splenic TEM subsets after a life of latency. R26 CreER T2 mice were infected with 10 6 PFU MCMV. 21 months post infection the mice received food pellets containing 400 mg/kg Tamoxifen for 2 weeks. Splenocytes were stained with antibodies against CD8, CD44, CD62L, and 7AAD. Percentage of 7AAD+ cells in immune phenotype subsets are shown. 4 mice per group; significance was assessed by Mann-Whitney test. * = p
    Figure Legend Snippet: Supplemental Tamoxifen data Fraction of CD8 subsets in the CD8 pool upon Tam treatment. Mice were MCMV infected and Tam treated as shown in figure 3 . Blood leukocytes were stained for CD8 and the indicated surface markers and gated in the CD44-CD62L+CD127+ (naïve) CD44+CD62L+CD127+ (central memory), CD44+CD62L-CD127+ (effector memory) and CD44+CD62L-CD127-(Effector) subsets. Shown are group means + standard deviations from two pooled experiments. (B) Tamoxifen induces cell death in splenic TEM subsets after a life of latency. R26 CreER T2 mice were infected with 10 6 PFU MCMV. 21 months post infection the mice received food pellets containing 400 mg/kg Tamoxifen for 2 weeks. Splenocytes were stained with antibodies against CD8, CD44, CD62L, and 7AAD. Percentage of 7AAD+ cells in immune phenotype subsets are shown. 4 mice per group; significance was assessed by Mann-Whitney test. * = p

    Techniques Used: Mouse Assay, Infection, Staining, Transmission Electron Microscopy, MANN-WHITNEY

    6) Product Images from "Prolactin Acts on Myeloid Progenitors to Modulate SMAD7 Expression and Enhance Hematopoietic Stem Cell Differentiation into the NK Cell Lineage"

    Article Title: Prolactin Acts on Myeloid Progenitors to Modulate SMAD7 Expression and Enhance Hematopoietic Stem Cell Differentiation into the NK Cell Lineage

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-63346-4

    CD56 + lymphocytes generate from CD34 + PRLR − progenitors not CD34 + PRLR + cells. UCB-derived CD34 + HSCs were expanded for four days using FLT3L, LDL, SCF and TPO followed by FACS sorting into CD34 + PRLR + and CD34 + PRLR − subsets and differentiated for up to 28 days. ( A,B ) Cells were stained for ILCs surface markers at day 28 of culture and the percentage ( A ) and absolute number ( B ) of CD56 + cells are shown in bar graphs (n = 6). ( C ) Flow cytometry showing surface expression of integrin α4β7 by day 7 differentiating CD34 + PRLR + cells. Representative dot plots (n = 5). ( D ) Surface expression of CD56, CD94, CD127 and CD336 by differentiating CD34 + PRLR + cells (upper row) compared to CD34 + PRLR − cells (lower row) at day 28. Representative histograms and values represent the percentage (n = 6). ( A,B ) Data are shown as means ± SD, paired t-tests and significance is shown (**** = p
    Figure Legend Snippet: CD56 + lymphocytes generate from CD34 + PRLR − progenitors not CD34 + PRLR + cells. UCB-derived CD34 + HSCs were expanded for four days using FLT3L, LDL, SCF and TPO followed by FACS sorting into CD34 + PRLR + and CD34 + PRLR − subsets and differentiated for up to 28 days. ( A,B ) Cells were stained for ILCs surface markers at day 28 of culture and the percentage ( A ) and absolute number ( B ) of CD56 + cells are shown in bar graphs (n = 6). ( C ) Flow cytometry showing surface expression of integrin α4β7 by day 7 differentiating CD34 + PRLR + cells. Representative dot plots (n = 5). ( D ) Surface expression of CD56, CD94, CD127 and CD336 by differentiating CD34 + PRLR + cells (upper row) compared to CD34 + PRLR − cells (lower row) at day 28. Representative histograms and values represent the percentage (n = 6). ( A,B ) Data are shown as means ± SD, paired t-tests and significance is shown (**** = p

    Techniques Used: Derivative Assay, FACS, Staining, Flow Cytometry, Expressing

    CD34 + PRLR + progenitors enhance the generation of CD56 + lymphocytes from CD34 + PRLR − progenitors. UCB-derived CD34 + HSCs were expanded for four days using FLT3L, LDL, SCF and TPO. Cells were then differentiated with or without PRL and SR1 for 21 days. The CD34 + PRLR + and CD34 + PRLR − cells were sorted at day 4 by FACS followed by differentiation for 21 days. CD34 + PRLR − and CD34 + PRLR + co-culture was done in a 2:1 ratios. ( A–C ) CD56 staining by CD34 + PRLR − , CD34 + PRLR − + CD34 + PRLR + co-culture and CD34 + PRLR − + CD34 + PRLR + co-culture in transwell system at day 21. Representative histograms ( A ) percentage in bar graphs ( B ) and absolute number in bar graphs ( C ) of CD56 + cells are shown (n = 3). ( D,E ) Cells differentiating in the presence or absence of PRL and SR1 were stained for CD56 at day 21 of culture and the percentage ( D ) and absolute number ( E ) of CD56 + cells are shown in bar graphs (n = 7). ( F ) Surface expression of CD56, CD94, CD127 and CD336 by cells differentiating in the presence or absence of PRL and SR1 at day 21. Representative histograms and values represent the percentage (n = 7). ( B–E ) Data are shown as means ± SD, One-way ANOVA and significance is shown (* = p
    Figure Legend Snippet: CD34 + PRLR + progenitors enhance the generation of CD56 + lymphocytes from CD34 + PRLR − progenitors. UCB-derived CD34 + HSCs were expanded for four days using FLT3L, LDL, SCF and TPO. Cells were then differentiated with or without PRL and SR1 for 21 days. The CD34 + PRLR + and CD34 + PRLR − cells were sorted at day 4 by FACS followed by differentiation for 21 days. CD34 + PRLR − and CD34 + PRLR + co-culture was done in a 2:1 ratios. ( A–C ) CD56 staining by CD34 + PRLR − , CD34 + PRLR − + CD34 + PRLR + co-culture and CD34 + PRLR − + CD34 + PRLR + co-culture in transwell system at day 21. Representative histograms ( A ) percentage in bar graphs ( B ) and absolute number in bar graphs ( C ) of CD56 + cells are shown (n = 3). ( D,E ) Cells differentiating in the presence or absence of PRL and SR1 were stained for CD56 at day 21 of culture and the percentage ( D ) and absolute number ( E ) of CD56 + cells are shown in bar graphs (n = 7). ( F ) Surface expression of CD56, CD94, CD127 and CD336 by cells differentiating in the presence or absence of PRL and SR1 at day 21. Representative histograms and values represent the percentage (n = 7). ( B–E ) Data are shown as means ± SD, One-way ANOVA and significance is shown (* = p

    Techniques Used: Derivative Assay, FACS, Co-Culture Assay, Staining, Expressing

    7) Product Images from "TREM-1 links dyslipidemia to inflammation and lipid deposition in atherosclerosis"

    Article Title: TREM-1 links dyslipidemia to inflammation and lipid deposition in atherosclerosis

    Journal: Nature Communications

    doi: 10.1038/ncomms13151

    TREM-1 drives skewed monocyte differentiation and HFCD-dependent monocytosis. ( a ) Absolute numbers of peripheral blood myeloid cell subsets before HFCD feeding (week 0; n =4), after 4 weeks of HFCD-feeding ( n =4–5), after 16 weeks of HFCD-feeding ( n =13–15) and after 16 weeks of chow-feeding ( n =9–13) as determined by flow cytometry. Circles represent mean values+s.d. for each group. Data are pooled from three independent experiments for the 16-week time-point and from one experiment for the 0 and 4 week analyses. ( b ) Relative frequencies of LSK cells (Sca1 + cKit hi ), CMP (Sca1 − cKit hi FcgR lo CD34 + ) and GMP (Sca1 − cKit hi FcgR + CD34 + ) cells among lineage − (lin − ; Ter119 − , CD3e − , Gr1 − and B220 − ) and CD127 − BM cells isolated from 16-week-HFCD-fed or chow-fed mice. Bars show mean values+s.d. of n =6-9 HFCD-fed mice (two independent experiments) and n =4–5 chow-fed controls. ( c ) Myeloid colony-forming units per well of 5 × 10 3 plated lin − BM cells and ( d ) relative frequencies of colony subtypes (M: monocyte colonies; G: granulocyte colonies; GM: mixed monocyte/granulocyte colonies). Data show mean values+s.d. of n =6–9 HFCD-fed mice (from two independent experiments) and from n =4–5 chow-fed mice per group. ( e ) Representative examples of CFU-G, CFU-M and CFU-GM. ( f ) mRNA expression of Irf8 and Cebpe in GMP isolated from 16-week-HFCD-fed mice. ( g ) Representative histograms for TREM-1 surface expression (lines) versus isotype control staining (filled) on peripheral blood neutrophils, Ly6C hi and Ly6C lo monocytes from Trem1 +/+ Apoe −/− mice 16 weeks post chow or HFCD feeding. ( h ) Median fluorescence intensity (MFI) of TREM-1 surface expression (with subtracted MFI values of matched isotype control-stained cells) on peripheral blood myeloid cell subsets at the indicated time points post HFCD feeding in Trem1 +/+ Apoe −/− (black symbols) and Trem1 −/− Apoe −/− (open symbols) mice. Symbols indicate mean+s.d. of n =5 mice and data are representative of three independent experiments for Trem1 +/+ Apoe −/− mice. ( i ) serum soluble TREM-1 was determined by ELISA. * P
    Figure Legend Snippet: TREM-1 drives skewed monocyte differentiation and HFCD-dependent monocytosis. ( a ) Absolute numbers of peripheral blood myeloid cell subsets before HFCD feeding (week 0; n =4), after 4 weeks of HFCD-feeding ( n =4–5), after 16 weeks of HFCD-feeding ( n =13–15) and after 16 weeks of chow-feeding ( n =9–13) as determined by flow cytometry. Circles represent mean values+s.d. for each group. Data are pooled from three independent experiments for the 16-week time-point and from one experiment for the 0 and 4 week analyses. ( b ) Relative frequencies of LSK cells (Sca1 + cKit hi ), CMP (Sca1 − cKit hi FcgR lo CD34 + ) and GMP (Sca1 − cKit hi FcgR + CD34 + ) cells among lineage − (lin − ; Ter119 − , CD3e − , Gr1 − and B220 − ) and CD127 − BM cells isolated from 16-week-HFCD-fed or chow-fed mice. Bars show mean values+s.d. of n =6-9 HFCD-fed mice (two independent experiments) and n =4–5 chow-fed controls. ( c ) Myeloid colony-forming units per well of 5 × 10 3 plated lin − BM cells and ( d ) relative frequencies of colony subtypes (M: monocyte colonies; G: granulocyte colonies; GM: mixed monocyte/granulocyte colonies). Data show mean values+s.d. of n =6–9 HFCD-fed mice (from two independent experiments) and from n =4–5 chow-fed mice per group. ( e ) Representative examples of CFU-G, CFU-M and CFU-GM. ( f ) mRNA expression of Irf8 and Cebpe in GMP isolated from 16-week-HFCD-fed mice. ( g ) Representative histograms for TREM-1 surface expression (lines) versus isotype control staining (filled) on peripheral blood neutrophils, Ly6C hi and Ly6C lo monocytes from Trem1 +/+ Apoe −/− mice 16 weeks post chow or HFCD feeding. ( h ) Median fluorescence intensity (MFI) of TREM-1 surface expression (with subtracted MFI values of matched isotype control-stained cells) on peripheral blood myeloid cell subsets at the indicated time points post HFCD feeding in Trem1 +/+ Apoe −/− (black symbols) and Trem1 −/− Apoe −/− (open symbols) mice. Symbols indicate mean+s.d. of n =5 mice and data are representative of three independent experiments for Trem1 +/+ Apoe −/− mice. ( i ) serum soluble TREM-1 was determined by ELISA. * P

    Techniques Used: Flow Cytometry, Cytometry, Isolation, Mouse Assay, Expressing, Staining, Fluorescence, Enzyme-linked Immunosorbent Assay

    Related Articles

    FACS:

    Article Title: Sodium oligomannate therapeutically remodels gut microbiota and suppresses gut bacterial amino acids-shaped neuroinflammation to inhibit Alzheimer’s disease progression
    Article Snippet: The cells were resuspended in 100 μL PBS buffer, blocked with anti-CD16/32 (Biolegend, Cat No. 101320) for 10 min, and incubated with the antibody according to the manufacturers’ protocols at 4 °C for 30 min. .. The following antibodies were used in the FACS analysis: CD45(30-F11)-APC-Cy7(103116, Biolegend), CD11b(M1/70)-FITC (101205, Biolegend), CX3CR1(SA011F11)-PE-Dazzle 594 (149014, Biolegend), Siglec-H-BV421 (566581, BD), F4/80(BM8)-BV421 (123132, Biolegend), CD86(IT2.2)-PE (305438, Biolegend), CD206(15-2)-APC (321110, Biolegend), CD206(15-2)-BV785 (321142, Biolegend), CD11c(N418)-PE-Cy7 (117318, Biolegend), CD8(53-6.7)-Percp-Cy5.5 (100734, Biolegend), Ly-6C(HK1.4)-PE-Dazzle 594 (128044, Biolegend), Gr-1(RB6-8C5)-Percp-Cy5.5 (108428, Biolegend), B220(RA3-6B2)-BV421 (103240, Biolegend), CD19(6D5)-PE (115508, Biolegend), CD49b(DX5)-PE-Cy7 (108922, Biolegend), CD4(GK1.5)-PE-Cy7 (100422, Biolegend), CD4(GK1.5)-FITC (100406, Biolegend), CXCR3(CXCR3-173)-BV421 (126522, Biolegend), CCR4(L291H4)-PE-Cy7 (359410, Biolegend), CCR6(29-2L17)-APC (129814, Biolegend), CD127(A019D5)-PE (351304, Biolegend), CD25(3C7)-Percp-Cy5.5 (101912, Biolegend), Live/Dead (423104, Biolegend). ..

    Flow Cytometry:

    Article Title: Plasma-Activated Medium Potentiates the Immunogenicity of Tumor Cell Lysates for Dendritic Cell-Based Cancer Vaccines
    Article Snippet: For all mixed cell cultures, T cells cultivated without DCs and A375 cells cultivated without T cells were used as controls. .. Flow CytometryThe flow cytometry analysis of DCs and T cells was performed on the flow cytometers CyFlow Cube 6 (Sysmex, Kobe, Hyogo, Japan) and LSR II (BD) by staining the cells with the following directly conjugated antibodies: anti-CD83-FITC, anti-CD86-PE, CD86-PerCPCy5.5, anti-CD40-APC, anti-CCR7-FITC, anti-IL17A-Alexa Fluor 488, anti-CD25-PerCPcy5.5, anti-CD127-PE, anti-IL10-APC, anti-HLA-DR-APCCy7 (Biolegend Inc., San Diego, CA, USA), anti-HLA-DR-PerCP (Miltenyi Biotec, Bergisch Gladbach, Germany), anti-ILT4-PE, anti-IFNγ-FITC (R & D Systems Minneapolis, MN, USA), anti-IL-12p40/p70-PE, anti-IL-10-FITC (BioRad, Hercules, CA, USA), anti-CD4-APC, anti-CD8-PerCPCy5.5, anti-Granzyme A-PE (eBioscience, San Diego, CA, USA), anti-Foxp3-FITC, anti-IL-4-PerCP, anti-PDL1-PE (eBioscience, San Diego, CA, USA), anti-CD4-PE (Sysmex, Kobe, Hyogo, Japan), IgG1 negative control-PE, IgG1 negative control-FITC, IgG1 negative control-APC, IgG1 negative control PerCP, (Thermo Fisher, Waltham, MA, USA), IgG2 negative control APCCy7 (Millipore, Burlington, MA, USA). .. Surface staining with primary Abs was conducted in PBS/0.1% NaN3/0.5% FBS prior to the intracellular staining that was carried out using a flow cytometry fixation and permeabilization kit (Biolegend, San Diego, CA, USA).

    Article Title: Targeting IL-21 to tumor-reactive T cells enhances memory T cell responses and anti-PD-1 antibody therapy
    Article Snippet: .. Flow cytometryThe following antibodies were used for the flow cytometry analysis: phycoerythrin (PE)- and Cy7-conjugated anti-CD8 (clone 53-6.7; eBioscience), allophycocyanin (APC)-conjugated anti-CD3 145-2C11; BioLegend), fluorescein isothiocyanate (FITC)-conjugated-CD25 (clone PC 61; eBioscience), APC-anti-CD44 (clone IM7; eBioscience), PE-anti-CD62L (clone M1L-14; Tonbo), PE-anti-CD122 (clone TM-b1; eBioscience), PE-anti-CD127 (clone A7R34; BioLegend), PerCP- and Cy5.5-conjugated-ScaI (clone D7; eBioscience), APC-anti-KLRG1 (clone 2F1; BioLegend), PE-anti-NK1.1 (clone PK136, eBioscience), FITC-anti-PD-1 (clone J43; eBioscience), PE-anti-B7H1 (clone 10F.9G2), PerCP-Cy5.5-anti-CD45 (clone 30-F11; BioLegend), FITC-anti-CD90.1 (clone HIS51; eBioscience), APC-anti-Flag (clone L5; BioLegend), PE-Cy7-anti-Bcl-2 (clone 10C4; eBioscience), PE-anti-TCF1 (clone 14456S; Cell Signaling Technology), FITC-anti-CD95 (clone SA367H8, BioLegend), PE-anti-IFN-γ (clone XMG1.2; eBioscience), and APC-anti-IL-2 (clone JES6-5H4; eBioscience). .. For the CFSE dilution assay, T cells were labeled with 5 μM CFSE before being cultured.

    Staining:

    Article Title: Plasma-Activated Medium Potentiates the Immunogenicity of Tumor Cell Lysates for Dendritic Cell-Based Cancer Vaccines
    Article Snippet: For all mixed cell cultures, T cells cultivated without DCs and A375 cells cultivated without T cells were used as controls. .. Flow CytometryThe flow cytometry analysis of DCs and T cells was performed on the flow cytometers CyFlow Cube 6 (Sysmex, Kobe, Hyogo, Japan) and LSR II (BD) by staining the cells with the following directly conjugated antibodies: anti-CD83-FITC, anti-CD86-PE, CD86-PerCPCy5.5, anti-CD40-APC, anti-CCR7-FITC, anti-IL17A-Alexa Fluor 488, anti-CD25-PerCPcy5.5, anti-CD127-PE, anti-IL10-APC, anti-HLA-DR-APCCy7 (Biolegend Inc., San Diego, CA, USA), anti-HLA-DR-PerCP (Miltenyi Biotec, Bergisch Gladbach, Germany), anti-ILT4-PE, anti-IFNγ-FITC (R & D Systems Minneapolis, MN, USA), anti-IL-12p40/p70-PE, anti-IL-10-FITC (BioRad, Hercules, CA, USA), anti-CD4-APC, anti-CD8-PerCPCy5.5, anti-Granzyme A-PE (eBioscience, San Diego, CA, USA), anti-Foxp3-FITC, anti-IL-4-PerCP, anti-PDL1-PE (eBioscience, San Diego, CA, USA), anti-CD4-PE (Sysmex, Kobe, Hyogo, Japan), IgG1 negative control-PE, IgG1 negative control-FITC, IgG1 negative control-APC, IgG1 negative control PerCP, (Thermo Fisher, Waltham, MA, USA), IgG2 negative control APCCy7 (Millipore, Burlington, MA, USA). .. Surface staining with primary Abs was conducted in PBS/0.1% NaN3/0.5% FBS prior to the intracellular staining that was carried out using a flow cytometry fixation and permeabilization kit (Biolegend, San Diego, CA, USA).

    Article Title: Asthma Increases Susceptibility to Heterologous but Not Homologous Secondary Influenza
    Article Snippet: Dead cells were identified using 7-aminoactinomycin D (eBioscience). .. For B- and T-cell analysis, cells were stained with PE-Cy7-conjugated anti-B220 (BD Pharmingen), FITC-conjugated anti-CD3 (BD Pharmingen), FITC-conjugated anti-CD4 (BD Pharmingen), PE-Cy7-conjugated anti-CD8 (BD Pharmingen), FITC-conjugated anti-CD62L (Biolegend), and PE-conjugated anti-CD127 (Biolegend) MAbs. .. For analysis of influenza virus nucleoprotein (NP)-specific CD8+ T lymphocytes, cells were stained with APC-conjugated major histocompatibility complex (MHC) class I H-2Kd tetramers loaded with the NP peptide (amino acids [aa] 147 to 155; TYQRTRALV).

    Article Title: A Combination of Polybacterial MV140 and Candida albicans V132 as a Potential Novel Trained Immunity-Based Vaccine for Genitourinary Tract Infections
    Article Snippet: Cells were washed with PBS/EDTA 2 mM/0.5% BSA and stained for 15 min at room temperature in the darkness. .. For analysis of FOXP3 expression in human T cells primed with hmoDCs, cells were first subjected to surface staining with anti-human CD127-PE, CD4-PerCP, and CD25-APC antibodies. .. After fixation and permeabilization, cells were stained with anti-human FOXP3-Alexa Fluor 488, according to manufacturer’s recommendations.

    Article Title: Immune recovery in HIV-infected patients after Candida esophagitis is impaired despite long-term antiretroviral therapy
    Article Snippet: Generation of heat-inactivated C. albicans A mixture of Candida albicans strain SC5314 yeast and hyphae was cultured and heat-inactivated as previously described [ , ]. .. Phenotypic characterization T-cell activation/exhaustion was analyzed by staining with anti-CD3-PerCP, anti-CD4-PacificBlue, anti-CD8-APC, anti-CD25-PE/Cy7 and anti-CD279-PE (PD-1); ILCs by staining with anti-Lineage-Cocktail-APC (anti-CD3/CD14/CD16/CD19/CD20/CD56) and anti-CD127-PE; NK-cell subsets by staining with anti-CD3-PerCP, anti-CD14-FITC, anti-CD19-PE/Cy7, anti-CD56-APC/Cy7, anti-CD16-PacificBlue (all BioLegend, Fell, Germany), anti-NKG2A-APC (clone 131411) and anti-NKG2C-PE (both R & D Systems, Abingdon, UK) [ ]. .. Samples were acquired on a CyAn ADP Analyzer (Beckman Coulter, Nyon, Switzerland) and data analyzed with FlowJo software vX.0.7 (FlowJo, Ashland, Oregon, USA).

    Article Title: Cytochalasin B-Induced Membrane Vesicles from Human Mesenchymal Stem Cells Overexpressing IL2 Are Able to Stimulate CD8+ T-Killers to Kill Human Triple Negative Breast Cancer Cells
    Article Snippet: Cells were incubated with CIMVs for 72 h. The activation of PBMC populations was determined by flow cytometry using staining with antibodies specific to surface markers of various human immune cell populations. .. After 72 h, PBMCs treated with hADSCs or CIMVs were washed twice with PBS and stained with the following antibodies for 30 min in the absence of light at room temperature: PE anti-human HLA-DR antibody (#307605, BioLegend, San Diego, CA, USA), APC anti-human CD25 antibody (#302610, BioLegend, San Diego, CA, USA), FITC anti-human CD8a antibody (#300906, BioLegend, San Diego, CA, USA), PE anti-human CD3 antibody (#300308, BioLegend, San Diego, CA, USA), Pacific Blue anti-human CD4 antibody (#317429, BioLegend, San Diego, CA, USA), PerCP/Cy5.5 anti-human CD38 antibody (#356614, BioLegend, San Diego, CA, USA), PE anti-human CD127 (IL-7Rα) antibody (#351304, BioLegend, San Diego, CA, USA), APC anti-human CD183 (CXCR3) antibody (#353708, BioLegend, San Diego, CA, USA), PE anti-human CD196 (CCR6) antibody (#353410, BioLegend, San Diego, CA, USA), APC anti-human CD4 antibody (#300514, BioLegend, San Diego, CA, USA), FITC anti-human CD3 antibody (#317306, BioLegend, San Diego, CA, USA), PerCP/Cyanine5.5 anti-human CD56 (NCAM) antibody (#362506, BioLegend, San Diego, CA, USA). .. Then, the cells were washed twice with PBS and analyzed on the FACSAria III flow cytometer (BD Biosciences, San Jose, CA, USA) and data were analyzed using BD FACSDiva™ software version 7.0.

    Negative Control:

    Article Title: Plasma-Activated Medium Potentiates the Immunogenicity of Tumor Cell Lysates for Dendritic Cell-Based Cancer Vaccines
    Article Snippet: For all mixed cell cultures, T cells cultivated without DCs and A375 cells cultivated without T cells were used as controls. .. Flow CytometryThe flow cytometry analysis of DCs and T cells was performed on the flow cytometers CyFlow Cube 6 (Sysmex, Kobe, Hyogo, Japan) and LSR II (BD) by staining the cells with the following directly conjugated antibodies: anti-CD83-FITC, anti-CD86-PE, CD86-PerCPCy5.5, anti-CD40-APC, anti-CCR7-FITC, anti-IL17A-Alexa Fluor 488, anti-CD25-PerCPcy5.5, anti-CD127-PE, anti-IL10-APC, anti-HLA-DR-APCCy7 (Biolegend Inc., San Diego, CA, USA), anti-HLA-DR-PerCP (Miltenyi Biotec, Bergisch Gladbach, Germany), anti-ILT4-PE, anti-IFNγ-FITC (R & D Systems Minneapolis, MN, USA), anti-IL-12p40/p70-PE, anti-IL-10-FITC (BioRad, Hercules, CA, USA), anti-CD4-APC, anti-CD8-PerCPCy5.5, anti-Granzyme A-PE (eBioscience, San Diego, CA, USA), anti-Foxp3-FITC, anti-IL-4-PerCP, anti-PDL1-PE (eBioscience, San Diego, CA, USA), anti-CD4-PE (Sysmex, Kobe, Hyogo, Japan), IgG1 negative control-PE, IgG1 negative control-FITC, IgG1 negative control-APC, IgG1 negative control PerCP, (Thermo Fisher, Waltham, MA, USA), IgG2 negative control APCCy7 (Millipore, Burlington, MA, USA). .. Surface staining with primary Abs was conducted in PBS/0.1% NaN3/0.5% FBS prior to the intracellular staining that was carried out using a flow cytometry fixation and permeabilization kit (Biolegend, San Diego, CA, USA).

    Expressing:

    Article Title: A Combination of Polybacterial MV140 and Candida albicans V132 as a Potential Novel Trained Immunity-Based Vaccine for Genitourinary Tract Infections
    Article Snippet: Cells were washed with PBS/EDTA 2 mM/0.5% BSA and stained for 15 min at room temperature in the darkness. .. For analysis of FOXP3 expression in human T cells primed with hmoDCs, cells were first subjected to surface staining with anti-human CD127-PE, CD4-PerCP, and CD25-APC antibodies. .. After fixation and permeabilization, cells were stained with anti-human FOXP3-Alexa Fluor 488, according to manufacturer’s recommendations.

    Activation Assay:

    Article Title: Immune recovery in HIV-infected patients after Candida esophagitis is impaired despite long-term antiretroviral therapy
    Article Snippet: Generation of heat-inactivated C. albicans A mixture of Candida albicans strain SC5314 yeast and hyphae was cultured and heat-inactivated as previously described [ , ]. .. Phenotypic characterization T-cell activation/exhaustion was analyzed by staining with anti-CD3-PerCP, anti-CD4-PacificBlue, anti-CD8-APC, anti-CD25-PE/Cy7 and anti-CD279-PE (PD-1); ILCs by staining with anti-Lineage-Cocktail-APC (anti-CD3/CD14/CD16/CD19/CD20/CD56) and anti-CD127-PE; NK-cell subsets by staining with anti-CD3-PerCP, anti-CD14-FITC, anti-CD19-PE/Cy7, anti-CD56-APC/Cy7, anti-CD16-PacificBlue (all BioLegend, Fell, Germany), anti-NKG2A-APC (clone 131411) and anti-NKG2C-PE (both R & D Systems, Abingdon, UK) [ ]. .. Samples were acquired on a CyAn ADP Analyzer (Beckman Coulter, Nyon, Switzerland) and data analyzed with FlowJo software vX.0.7 (FlowJo, Ashland, Oregon, USA).

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    BioLegend anti cd127 pe
    Gene expression context does not define the quality of CD8 responses to MHC-I restricted epitopes of MCMV. 129/Sv mice were infected intraperitoneally (i.p.) with 2x10 5 PFU of MCMV M45SL . Blood leukocytes were stimulated with the SSIEFARL or the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. Cells were surface-stained for CD3, CD4, CD8, CD11a, CD44, KLRG1, <t>CD127</t> and intracellularly for IFNγ expression and analyzed by flow cytometry. ( A ) Cells responding to the SSIEFARL (gB K b response) or the HGIRNASFI (M45D b response) peptide. ( B ) Upper graph—epitope specific cells with the EM phenotype (CD127 - KLRG1 + ). Lower graph—epitope specific cells with the CM phenotype (CD127 + KLRG1 - ). The experiment was performed three times independently, at 5 mice per group in each experiment, and grouped averages +/- SEM from all three experiments are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. **** p
    Anti Cd127 Pe, supplied by BioLegend, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Characterization of macrophages and T cells. (A, B, C) Representative plots for the identification of the macrophage population (A) and of their two subtypes, M1-like (B) and M2-like (C) macrophages versus days after implantation. (D, E) Percentage of macrophages (D) and macrophage subtypes (E) versus days after implantation. (F, G) T cells (CD3) identification strategy (F) for the CD4+ and CD8+ subpopulations (G). (H, I) Percentage of T cells among viable cells (H) and T cells subpopulations (I) among all T cells, both versus days after implantation. (J, K, L) Representative plots for identifying T regulatory cells <t>(CD4+CD127-,</t> J, CD25+, K) among T cells, versus days after implantation. (L). Data shown as mean ± SEM. *: p
    Pe Cy7 Anti Mouse Cd127 Il 7rα, supplied by BioLegend, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MV140/V132-activated hmoDCs generate IFN-γ-, IL-17A-, and IL-10-producing T cell production as well as FOXP3 + Treg cells. (A) Percentage of CD3 + CD4 + T cells producing IFN-γ, IL-10 and IL-17A generated after 3 days of co-culture with control excipients-, V132-, MV140-, and MV140/V132-stimulated hmoDCs as determined by intracellular staining and flow cytometry analysis. Representative dot plots are shown for the intracellular staining after flow cytometry analysis. (B) Percentage of induced CD4 + CD25 high <t>CD127</t> - FOXP3 + Treg cells gated on CD4 + T cells after 3 days of co-culture with allogeneic control excipients-, V132-, MV140-, and MV140/V132-activated hmoDCs. Results are mean ± s.e.m. of 7-8 (A) , and 5 (B) independent experiments. Paired Student t test, * P
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    BioLegend pe conjugated anti mouse cd127
    Surface molecule expression on pulmonary γδ T cells. Single lung cell suspensions were separated from naïve and infected mice, and cells were stained with monoclonal antibodies against mice: CD3, γδTCR, CD4, CD8, Vγ2, CD25, CD69, <t>CD127,</t> CD62L, MHC II, CD80, PDL1, PDL2, CXCR3, CXCR4, CXCR6, and CX3CR1. (A) Representative FACS analysis of three independent experiments is shown. (B) Percentages of surface molecules calculated from FACS analysis, a representative experiment from five independent experiments was shown. The error bars are SD, ** P
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    Gene expression context does not define the quality of CD8 responses to MHC-I restricted epitopes of MCMV. 129/Sv mice were infected intraperitoneally (i.p.) with 2x10 5 PFU of MCMV M45SL . Blood leukocytes were stimulated with the SSIEFARL or the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. Cells were surface-stained for CD3, CD4, CD8, CD11a, CD44, KLRG1, CD127 and intracellularly for IFNγ expression and analyzed by flow cytometry. ( A ) Cells responding to the SSIEFARL (gB K b response) or the HGIRNASFI (M45D b response) peptide. ( B ) Upper graph—epitope specific cells with the EM phenotype (CD127 - KLRG1 + ). Lower graph—epitope specific cells with the CM phenotype (CD127 + KLRG1 - ). The experiment was performed three times independently, at 5 mice per group in each experiment, and grouped averages +/- SEM from all three experiments are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. **** p

    Journal: PLoS Pathogens

    Article Title: Peptide Processing Is Critical for T-Cell Memory Inflation and May Be Optimized to Improve Immune Protection by CMV-Based Vaccine Vectors

    doi: 10.1371/journal.ppat.1006072

    Figure Lengend Snippet: Gene expression context does not define the quality of CD8 responses to MHC-I restricted epitopes of MCMV. 129/Sv mice were infected intraperitoneally (i.p.) with 2x10 5 PFU of MCMV M45SL . Blood leukocytes were stimulated with the SSIEFARL or the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. Cells were surface-stained for CD3, CD4, CD8, CD11a, CD44, KLRG1, CD127 and intracellularly for IFNγ expression and analyzed by flow cytometry. ( A ) Cells responding to the SSIEFARL (gB K b response) or the HGIRNASFI (M45D b response) peptide. ( B ) Upper graph—epitope specific cells with the EM phenotype (CD127 - KLRG1 + ). Lower graph—epitope specific cells with the CM phenotype (CD127 + KLRG1 - ). The experiment was performed three times independently, at 5 mice per group in each experiment, and grouped averages +/- SEM from all three experiments are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. **** p

    Article Snippet: The antibody panels used for the samples stimulated with the SSIEFARL peptide or stained with SSIEFARL-Kb tetramers shown previously [ ] were expanded with anti-CD62L-eFluor605NC (MEL-14, eBioscience) for tetramer staining or with anti-KLRG1-Biotin (Clone 2F1; Biolegend) and anti-CD127-PE (Clone A7R34; Biolegend) for peptide re-stimulation.

    Techniques: Expressing, Mouse Assay, Infection, Staining, Flow Cytometry, Cytometry, MANN-WHITNEY

    Epitope localization defines the quality of CD8 T-cell responses. ( A ) Representative dot plots of intracellular IFNγ expression at 7 and 180 dpi upon HGIRNASFI peptide stimulation. ( B ) Grouped means +/- SEM of cells responding to the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. ( C ) Representative dot plots of the surface expression of CD127 and KLRG1 on HGIRNASFI specific CD8 T cells at 7 and 180 dpi with MCMV WT or MCMV M45Cterm . The staining was used to define the CM (CD127 + KLRG1 - ) and the EM (CD127 - KLRG1 + ) subsets. ( D, E ) Grouped means +/- SEM of the percentage of EM ( D ) or CM ( E ) cells in the HGIRNASFI-responding subset at indicated time points p.i‥ The experiment was performed twice, at 5 mice per group in each experiment, and pooled results are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. *p

    Journal: PLoS Pathogens

    Article Title: Peptide Processing Is Critical for T-Cell Memory Inflation and May Be Optimized to Improve Immune Protection by CMV-Based Vaccine Vectors

    doi: 10.1371/journal.ppat.1006072

    Figure Lengend Snippet: Epitope localization defines the quality of CD8 T-cell responses. ( A ) Representative dot plots of intracellular IFNγ expression at 7 and 180 dpi upon HGIRNASFI peptide stimulation. ( B ) Grouped means +/- SEM of cells responding to the HGIRNASFI peptide at 7, 14, 28, 60, 90, 120, 180 dpi. ( C ) Representative dot plots of the surface expression of CD127 and KLRG1 on HGIRNASFI specific CD8 T cells at 7 and 180 dpi with MCMV WT or MCMV M45Cterm . The staining was used to define the CM (CD127 + KLRG1 - ) and the EM (CD127 - KLRG1 + ) subsets. ( D, E ) Grouped means +/- SEM of the percentage of EM ( D ) or CM ( E ) cells in the HGIRNASFI-responding subset at indicated time points p.i‥ The experiment was performed twice, at 5 mice per group in each experiment, and pooled results are shown. Significance on day 180 p.i. was assessed by a Mann—Whitney U test. *p

    Article Snippet: The antibody panels used for the samples stimulated with the SSIEFARL peptide or stained with SSIEFARL-Kb tetramers shown previously [ ] were expanded with anti-CD62L-eFluor605NC (MEL-14, eBioscience) for tetramer staining or with anti-KLRG1-Biotin (Clone 2F1; Biolegend) and anti-CD127-PE (Clone A7R34; Biolegend) for peptide re-stimulation.

    Techniques: Expressing, Staining, Mouse Assay, MANN-WHITNEY

    Characterization of macrophages and T cells. (A, B, C) Representative plots for the identification of the macrophage population (A) and of their two subtypes, M1-like (B) and M2-like (C) macrophages versus days after implantation. (D, E) Percentage of macrophages (D) and macrophage subtypes (E) versus days after implantation. (F, G) T cells (CD3) identification strategy (F) for the CD4+ and CD8+ subpopulations (G). (H, I) Percentage of T cells among viable cells (H) and T cells subpopulations (I) among all T cells, both versus days after implantation. (J, K, L) Representative plots for identifying T regulatory cells (CD4+CD127-, J, CD25+, K) among T cells, versus days after implantation. (L). Data shown as mean ± SEM. *: p

    Journal: PLoS ONE

    Article Title: Characterization of a B16-F10 melanoma model locally implanted into the ear pinnae of C57BL/6 mice

    doi: 10.1371/journal.pone.0206693

    Figure Lengend Snippet: Characterization of macrophages and T cells. (A, B, C) Representative plots for the identification of the macrophage population (A) and of their two subtypes, M1-like (B) and M2-like (C) macrophages versus days after implantation. (D, E) Percentage of macrophages (D) and macrophage subtypes (E) versus days after implantation. (F, G) T cells (CD3) identification strategy (F) for the CD4+ and CD8+ subpopulations (G). (H, I) Percentage of T cells among viable cells (H) and T cells subpopulations (I) among all T cells, both versus days after implantation. (J, K, L) Representative plots for identifying T regulatory cells (CD4+CD127-, J, CD25+, K) among T cells, versus days after implantation. (L). Data shown as mean ± SEM. *: p

    Article Snippet: Natural T regulatory cells (Tregs) within the CD3+ T cells, were identified by their CD4+ CD127-reactivity (Biolegend, 135014).

    Techniques:

    MV140/V132-activated hmoDCs generate IFN-γ-, IL-17A-, and IL-10-producing T cell production as well as FOXP3 + Treg cells. (A) Percentage of CD3 + CD4 + T cells producing IFN-γ, IL-10 and IL-17A generated after 3 days of co-culture with control excipients-, V132-, MV140-, and MV140/V132-stimulated hmoDCs as determined by intracellular staining and flow cytometry analysis. Representative dot plots are shown for the intracellular staining after flow cytometry analysis. (B) Percentage of induced CD4 + CD25 high CD127 - FOXP3 + Treg cells gated on CD4 + T cells after 3 days of co-culture with allogeneic control excipients-, V132-, MV140-, and MV140/V132-activated hmoDCs. Results are mean ± s.e.m. of 7-8 (A) , and 5 (B) independent experiments. Paired Student t test, * P

    Journal: Frontiers in Immunology

    Article Title: A Combination of Polybacterial MV140 and Candida albicans V132 as a Potential Novel Trained Immunity-Based Vaccine for Genitourinary Tract Infections

    doi: 10.3389/fimmu.2020.612269

    Figure Lengend Snippet: MV140/V132-activated hmoDCs generate IFN-γ-, IL-17A-, and IL-10-producing T cell production as well as FOXP3 + Treg cells. (A) Percentage of CD3 + CD4 + T cells producing IFN-γ, IL-10 and IL-17A generated after 3 days of co-culture with control excipients-, V132-, MV140-, and MV140/V132-stimulated hmoDCs as determined by intracellular staining and flow cytometry analysis. Representative dot plots are shown for the intracellular staining after flow cytometry analysis. (B) Percentage of induced CD4 + CD25 high CD127 - FOXP3 + Treg cells gated on CD4 + T cells after 3 days of co-culture with allogeneic control excipients-, V132-, MV140-, and MV140/V132-activated hmoDCs. Results are mean ± s.e.m. of 7-8 (A) , and 5 (B) independent experiments. Paired Student t test, * P

    Article Snippet: For analysis of FOXP3 expression in human T cells primed with hmoDCs, cells were first subjected to surface staining with anti-human CD127-PE, CD4-PerCP, and CD25-APC antibodies.

    Techniques: Generated, Co-Culture Assay, Staining, Flow Cytometry

    Surface molecule expression on pulmonary γδ T cells. Single lung cell suspensions were separated from naïve and infected mice, and cells were stained with monoclonal antibodies against mice: CD3, γδTCR, CD4, CD8, Vγ2, CD25, CD69, CD127, CD62L, MHC II, CD80, PDL1, PDL2, CXCR3, CXCR4, CXCR6, and CX3CR1. (A) Representative FACS analysis of three independent experiments is shown. (B) Percentages of surface molecules calculated from FACS analysis, a representative experiment from five independent experiments was shown. The error bars are SD, ** P

    Journal: Frontiers in Immunology

    Article Title: Adjustments of γδ T Cells in the Lung of Schistosoma japonicum-Infected C56BL/6 Mice

    doi: 10.3389/fimmu.2020.01045

    Figure Lengend Snippet: Surface molecule expression on pulmonary γδ T cells. Single lung cell suspensions were separated from naïve and infected mice, and cells were stained with monoclonal antibodies against mice: CD3, γδTCR, CD4, CD8, Vγ2, CD25, CD69, CD127, CD62L, MHC II, CD80, PDL1, PDL2, CXCR3, CXCR4, CXCR6, and CX3CR1. (A) Representative FACS analysis of three independent experiments is shown. (B) Percentages of surface molecules calculated from FACS analysis, a representative experiment from five independent experiments was shown. The error bars are SD, ** P

    Article Snippet: PE-conjugated anti-mouse CD3 (145-2C11), Brilliant Violet 510-conjugated anti-mouse γδ TCR (GL3), APC-conjugated anti-mouse CD4 (RM4-5), PE-cy5-conjugated anti-mouse CD19 (6D5), PE-conjugated anti-mouse Vγ2 (A7R34), APC-conjugated anti-mouse CD69 (H1.2F3), PE-conjugated anti-mouse CD127 (A7R34), APC-conjugated anti-mouse CD62L (MEL-14), FITC-conjugated anti-mouse CD27 (LG.3A10), APC-conjugated anti-mouse IgD (11-26c.2a), FITC-conjugated anti-mouse MHCII (M5/114.15.2), PE-conjugated anti-mouse CD80 (16-10A1), PE-cy7-conjugated anti-mouse ICOS (C398.4A), APC-conjugated anti-mouse CX3CR1 (SA011F11), APC-conjugated anti-mouse IFN-γ (XMG1.2), Alexa Fluor 647-conjugated anti-mouse IL-21 (BL25168), PerCP-cy5.5-conjugated anti-mouse GM-CSF (MP1-22E9), PE-conjugated anti-mouse IL-1 (ALF-161), and APC-conjugated anti-mouse IL-6 (MP5-20F3) were purchased from BioLegend (San Diego, CA, USA).

    Techniques: Expressing, Infection, Mouse Assay, Staining, FACS