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R&D Systems anti mouse s1p1 apc
Anti Mouse S1p1 Apc, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti mouse s1p1 apc/product/R&D Systems
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti mouse s1p1 apc - by Bioz Stars, 2022-09
93/100 stars

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    R&D Systems s1pr1 pe
    Increased Fr. D and loss of Fr. F cells in bone marrow of Grk2 −/− mb1 -cre mice. ( A ) Flow cytometry results from analysis bone marrow cells from Grk2 fl/fl and Grk2 fl/fl mb1-cre mice. ( B ) Flow cytometry results from bone marrow showing CD43 expression and the % Fr.A - Fr.F cells. ( C ) Flow cytometry of bone marrow cells gated on B220 + CD19 + CD43 − examining IgD versus IgM. ( D ) Flow cytometry results showing receptors expression and representative <t>S1PR1</t> histograms for each fraction. ( E ) Spontaneous and directed migration of Grk2 fl/fl and Grk2 fl/fl mb1-cre bone marrow B cell fractions to CXCL12 and S1P. All results from B cells purified from a minimum of 5 Grk2 fl/fl and Grk2 fl/fl mb1-cre mice. Data shown as mean +/− standard error of the mean (p
    S1pr1 Pe, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/s1pr1 pe/product/R&D Systems
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    s1pr1 pe - by Bioz Stars, 2022-09
    92/100 stars
      Buy from Supplier

    95
    R&D Systems rat monoclonal anti mouse s1p1 antibody
    B cells are capable of degrading S1P (a) <t>S1P1</t> surface abundance on follicular B cells from the indicated tissues of control (CTL), S1pr1 -deficient (S1P1 KO), or Mx1-cre + Sphk1 f/− or f/f Sphk2 −/− (S1P-deficient) mice. Data are representative of more than 3 mice of each type. (b) Transcript abundance of S1P lyase ( Sgpl ), sphingosine-1-phosphate phosphatase-1 ( Sgpp1 ), and lipid phosphate phosphatases ( Lpp ) 1–3 in B cells and T cells, shown relative to HPRT . (c) Relative amount of S1P remaining in culture supernatants after incubation with B or T cells for the indicated number of minutes, determined by the extent of down-modulation of Flag-S1P1 on a reporter cell line. Data are plotted as MFI of Flag-S1P1 staining relative to reporter cells not exposed to S1P (data are from 3 experiments).
    Rat Monoclonal Anti Mouse S1p1 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat monoclonal anti mouse s1p1 antibody/product/R&D Systems
    Average 95 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    rat monoclonal anti mouse s1p1 antibody - by Bioz Stars, 2022-09
    95/100 stars
      Buy from Supplier

    93
    R&D Systems murine s1p1
    Hindering <t>S1P1</t> internalization abrogates T-cell sequestration in bone marrow a , Relative sequestration of adoptively transferred CFSE-labeled T-cells within the bone marrow of CT2A IC recipient mice at 2 hours (left) or 24 hours (right) after transfer. As indicated, transferred cells were splenocytes from either control C57BL/6 donors (control) or from S1P1 stabilized “knock-in” (S1P1 KI) donors (n=5 recipient mice per group). Data in a are representative findings from one of a minimum of two independently repeated experiments with similar results. b , T-cell counts in the bone marrow of n=10 IC CT2A-bearing wild type (WT) C57BL/6 or n=11 S1P1 KI mice. Counts were assessed 18 days following tumor implantation and are shown relative to baseline counts in n=6 tumor-naïve control WT or n=6 tumor-naïve S1P1 KI mice. Cumulative data from three experiments are depicted in b. c , IC CT2A tumors were harvested from n=6 WT C57BL/6 (WT) or n=6 S1P1 KI mice 18 days following tumor implantation. TIL were assessed by flow cytometry and the number of total T-cells per gram of tumor quantified. d , The frequency of activated effector (CD44 hi CD62L lo ) T-cells in IC CT2A tumors from the same n=6 WT C57BL/6 (WT) or n=6 S1P1 KI mice in c was also quantified. Data in c, d are representative findings from one of a minimum of three independently repeated experiments with similar results. e , C57BL/6 (WT) or S1P1 KI mice were implanted with IC CT2A tumors and treated with a 4-1BB agonist antibody or isotype control (n=8 per group). All data in a–d are shown as mean ± s.e.m. P values in a–d were determined by two-tailed, unpaired Student’s t-test. Survival in e was assessed by two-tailed generalized Wilcoxon test. P value for overall comparison is depicted.
    Murine S1p1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/murine s1p1/product/R&D Systems
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    murine s1p1 - by Bioz Stars, 2022-09
    93/100 stars
      Buy from Supplier

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    Increased Fr. D and loss of Fr. F cells in bone marrow of Grk2 −/− mb1 -cre mice. ( A ) Flow cytometry results from analysis bone marrow cells from Grk2 fl/fl and Grk2 fl/fl mb1-cre mice. ( B ) Flow cytometry results from bone marrow showing CD43 expression and the % Fr.A - Fr.F cells. ( C ) Flow cytometry of bone marrow cells gated on B220 + CD19 + CD43 − examining IgD versus IgM. ( D ) Flow cytometry results showing receptors expression and representative S1PR1 histograms for each fraction. ( E ) Spontaneous and directed migration of Grk2 fl/fl and Grk2 fl/fl mb1-cre bone marrow B cell fractions to CXCL12 and S1P. All results from B cells purified from a minimum of 5 Grk2 fl/fl and Grk2 fl/fl mb1-cre mice. Data shown as mean +/− standard error of the mean (p

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

    Article Title: Biased S1PR1 Signaling in B cells Subverts Responses to Homeostatic Chemokines Severely Disorganizing Lymphoid Organ Architecture

    doi: 10.4049/jimmunol.1900678

    Figure Lengend Snippet: Increased Fr. D and loss of Fr. F cells in bone marrow of Grk2 −/− mb1 -cre mice. ( A ) Flow cytometry results from analysis bone marrow cells from Grk2 fl/fl and Grk2 fl/fl mb1-cre mice. ( B ) Flow cytometry results from bone marrow showing CD43 expression and the % Fr.A - Fr.F cells. ( C ) Flow cytometry of bone marrow cells gated on B220 + CD19 + CD43 − examining IgD versus IgM. ( D ) Flow cytometry results showing receptors expression and representative S1PR1 histograms for each fraction. ( E ) Spontaneous and directed migration of Grk2 fl/fl and Grk2 fl/fl mb1-cre bone marrow B cell fractions to CXCL12 and S1P. All results from B cells purified from a minimum of 5 Grk2 fl/fl and Grk2 fl/fl mb1-cre mice. Data shown as mean +/− standard error of the mean (p

    Article Snippet: Cells were washed stained with S1PR1-PE prior to incubation and at the indicated times.

    Techniques: Mouse Assay, Flow Cytometry, Expressing, Migration, Purification

    Increased non-specific migration, decreased CXCL13 but enhanced S1P triggered migration of Grk2 fl/fl mb1-cre B cells. ( A ) Chemotaxis assays to CXCL12 (ng/ml), CCL19 (ng/ml), CXCL13 (ng/ml), and S1P (nM). Shown on left, is non-specific migration, and on the right, specific migration of T1, FO, and MZ B cells. ( B ) Flow cytometry examining S1PR1, CXCR4, CXCR5, and CCR7 expression on B cell subsets from Grk2 fl/fl and Grk2 fl/fl mb1-cre mice. FACS profile shown only for S1PR1. ( C ) Time dependent S1PR1 internalization following exposure to S1P (1 μM) and recovery of S1PR1 expression following placement of splenic B cells in media lacking S1P. N=3 vs. 3 ( Grk2 fl/fl , black lines; Grk2 fl/fl mb1-cre, grey lines). Experiments repeated 3–4 times. Data shown as mean +/− standard error of the mean (p

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

    Article Title: Biased S1PR1 Signaling in B cells Subverts Responses to Homeostatic Chemokines Severely Disorganizing Lymphoid Organ Architecture

    doi: 10.4049/jimmunol.1900678

    Figure Lengend Snippet: Increased non-specific migration, decreased CXCL13 but enhanced S1P triggered migration of Grk2 fl/fl mb1-cre B cells. ( A ) Chemotaxis assays to CXCL12 (ng/ml), CCL19 (ng/ml), CXCL13 (ng/ml), and S1P (nM). Shown on left, is non-specific migration, and on the right, specific migration of T1, FO, and MZ B cells. ( B ) Flow cytometry examining S1PR1, CXCR4, CXCR5, and CCR7 expression on B cell subsets from Grk2 fl/fl and Grk2 fl/fl mb1-cre mice. FACS profile shown only for S1PR1. ( C ) Time dependent S1PR1 internalization following exposure to S1P (1 μM) and recovery of S1PR1 expression following placement of splenic B cells in media lacking S1P. N=3 vs. 3 ( Grk2 fl/fl , black lines; Grk2 fl/fl mb1-cre, grey lines). Experiments repeated 3–4 times. Data shown as mean +/− standard error of the mean (p

    Article Snippet: Cells were washed stained with S1PR1-PE prior to incubation and at the indicated times.

    Techniques: Migration, Chemotaxis Assay, Flow Cytometry, Expressing, Mouse Assay, FACS

    Leukocytosis, but a dearth of B cells in LNs and Peyer’s patches from Grk 2 fl/fl mb1-cre mice. ( A ) Flow cytometry analysis of the blood from control and Grk 2 fl/fl mb1-cre mice. Shown are total cell count and composition of blood leukocytes on a % basis. ( B ) Percentage of B cells in the indicated subsets from blood of Grk 2 fl/fl and Grk 2 fl/fl mb1-cre mice. ( C ) Flow cytometry profiles of S1PR1 expression on indicated blood cells. (D) Flow cytometry results analyzing expression of S1PR1 and homing receptors on blood B cells. (E) Chemotaxis assays to CXCL12 (ng/ml), CCL19 (ng/ml), and CXCL13 (ng/ml). Shown on left, is non-specific migration, and on the right, specific migration of blood B, and CD4 T cells. ( F ) Cell recovery and subset distribution in LNs and Peyer’s patches (PP) from Grk 2 fl/fl and Grk 2 fl/fl mb1-cre mice. ( G ) Spontaneous migration of LN B and CD4 T cells. ( H ) Specific migration of LN B and CD4 T cells to indicated chemokines (ng/ml) or S1P (nM), cells prepared from Grk 2 fl/fl and Grk 2 fl/fl mb1-cre mice. ( I ) Flow cytometry results assessing S1PR1, CXCR4, CXCR5, and CCR7 expression on LN B cells from control and Grk 2 fl/fl mb1-cre mice. All results from B cells purified from a minimum of 5 Grk2 fl/fl and Grk2 fl/fl mb1-cre mice. Data shown as mean +/− standard error of the mean (p

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

    Article Title: Biased S1PR1 Signaling in B cells Subverts Responses to Homeostatic Chemokines Severely Disorganizing Lymphoid Organ Architecture

    doi: 10.4049/jimmunol.1900678

    Figure Lengend Snippet: Leukocytosis, but a dearth of B cells in LNs and Peyer’s patches from Grk 2 fl/fl mb1-cre mice. ( A ) Flow cytometry analysis of the blood from control and Grk 2 fl/fl mb1-cre mice. Shown are total cell count and composition of blood leukocytes on a % basis. ( B ) Percentage of B cells in the indicated subsets from blood of Grk 2 fl/fl and Grk 2 fl/fl mb1-cre mice. ( C ) Flow cytometry profiles of S1PR1 expression on indicated blood cells. (D) Flow cytometry results analyzing expression of S1PR1 and homing receptors on blood B cells. (E) Chemotaxis assays to CXCL12 (ng/ml), CCL19 (ng/ml), and CXCL13 (ng/ml). Shown on left, is non-specific migration, and on the right, specific migration of blood B, and CD4 T cells. ( F ) Cell recovery and subset distribution in LNs and Peyer’s patches (PP) from Grk 2 fl/fl and Grk 2 fl/fl mb1-cre mice. ( G ) Spontaneous migration of LN B and CD4 T cells. ( H ) Specific migration of LN B and CD4 T cells to indicated chemokines (ng/ml) or S1P (nM), cells prepared from Grk 2 fl/fl and Grk 2 fl/fl mb1-cre mice. ( I ) Flow cytometry results assessing S1PR1, CXCR4, CXCR5, and CCR7 expression on LN B cells from control and Grk 2 fl/fl mb1-cre mice. All results from B cells purified from a minimum of 5 Grk2 fl/fl and Grk2 fl/fl mb1-cre mice. Data shown as mean +/− standard error of the mean (p

    Article Snippet: Cells were washed stained with S1PR1-PE prior to incubation and at the indicated times.

    Techniques: Mouse Assay, Flow Cytometry, Cell Counting, Expressing, Chemotaxis Assay, Migration, Purification

    The loss of GRK2 in B cells enhances S1P signaling but decreases CXCL13 signaling except for the intracellular Ca 2+ response. ( A ) Chemoattractant induced pAKT and pERK in B cell subsets. Grk2 fl/fl or Grk2 fl/fl mb1-cre B cells were exposed to CXCL13 (1000 ng/ml), or S1P (1μM) for indicated times and level of pAKT or pERK determined by flow cytometry. Data expressed as % of basal. ( B ) pERM levels in blood and splenic B cells from Grk2 fl/fl or Grk2 fl/fl mb1-cre mice. Level of pERM in blood B cell subsets and CD4 T cells determined by flow cytometry (left). Basal pERM levels in splenic B cell subsets and following exposure S1P (1μM). ( C ) Peak intracellular Ca 2+ levels following chemoattractant exposure and impact of pertussis toxin (200nM). Grk2 fl/fl or Grk2 fl/fl mb1-cre splenic B cells were exposed to varying concentration of CXCL13 (ng/ml) or S1P (nM). ( D ) Effect of a S1PR1 antagonist on S1P and CXCL13 induced increases in intracellular Ca 2+ . Plot of intracellular Ca 2+ levels over time in splenic B cells from Grk2 fl/fl or Grk2 fl/fl mb1-cre mice and exposed to S1P or CXCL13. Where indicated the B cells were pre-incubated with Ex 26 for 1 hours prior to the addition of S1P or CXCL13. Results are from 2–10 mice per group. All experiments repeated a minimum of three times. Data shown as mean +/− standard error of the mean (p

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

    Article Title: Biased S1PR1 Signaling in B cells Subverts Responses to Homeostatic Chemokines Severely Disorganizing Lymphoid Organ Architecture

    doi: 10.4049/jimmunol.1900678

    Figure Lengend Snippet: The loss of GRK2 in B cells enhances S1P signaling but decreases CXCL13 signaling except for the intracellular Ca 2+ response. ( A ) Chemoattractant induced pAKT and pERK in B cell subsets. Grk2 fl/fl or Grk2 fl/fl mb1-cre B cells were exposed to CXCL13 (1000 ng/ml), or S1P (1μM) for indicated times and level of pAKT or pERK determined by flow cytometry. Data expressed as % of basal. ( B ) pERM levels in blood and splenic B cells from Grk2 fl/fl or Grk2 fl/fl mb1-cre mice. Level of pERM in blood B cell subsets and CD4 T cells determined by flow cytometry (left). Basal pERM levels in splenic B cell subsets and following exposure S1P (1μM). ( C ) Peak intracellular Ca 2+ levels following chemoattractant exposure and impact of pertussis toxin (200nM). Grk2 fl/fl or Grk2 fl/fl mb1-cre splenic B cells were exposed to varying concentration of CXCL13 (ng/ml) or S1P (nM). ( D ) Effect of a S1PR1 antagonist on S1P and CXCL13 induced increases in intracellular Ca 2+ . Plot of intracellular Ca 2+ levels over time in splenic B cells from Grk2 fl/fl or Grk2 fl/fl mb1-cre mice and exposed to S1P or CXCL13. Where indicated the B cells were pre-incubated with Ex 26 for 1 hours prior to the addition of S1P or CXCL13. Results are from 2–10 mice per group. All experiments repeated a minimum of three times. Data shown as mean +/− standard error of the mean (p

    Article Snippet: Cells were washed stained with S1PR1-PE prior to incubation and at the indicated times.

    Techniques: Flow Cytometry, Mouse Assay, Concentration Assay, Incubation

    B cells are capable of degrading S1P (a) S1P1 surface abundance on follicular B cells from the indicated tissues of control (CTL), S1pr1 -deficient (S1P1 KO), or Mx1-cre + Sphk1 f/− or f/f Sphk2 −/− (S1P-deficient) mice. Data are representative of more than 3 mice of each type. (b) Transcript abundance of S1P lyase ( Sgpl ), sphingosine-1-phosphate phosphatase-1 ( Sgpp1 ), and lipid phosphate phosphatases ( Lpp ) 1–3 in B cells and T cells, shown relative to HPRT . (c) Relative amount of S1P remaining in culture supernatants after incubation with B or T cells for the indicated number of minutes, determined by the extent of down-modulation of Flag-S1P1 on a reporter cell line. Data are plotted as MFI of Flag-S1P1 staining relative to reporter cells not exposed to S1P (data are from 3 experiments).

    Journal: Nature immunology

    Article Title: The sphingosine 1-phosphate receptor S1P2 maintains germinal center B cell homeostasis and promotes niche confinement

    doi: 10.1038/ni.2047

    Figure Lengend Snippet: B cells are capable of degrading S1P (a) S1P1 surface abundance on follicular B cells from the indicated tissues of control (CTL), S1pr1 -deficient (S1P1 KO), or Mx1-cre + Sphk1 f/− or f/f Sphk2 −/− (S1P-deficient) mice. Data are representative of more than 3 mice of each type. (b) Transcript abundance of S1P lyase ( Sgpl ), sphingosine-1-phosphate phosphatase-1 ( Sgpp1 ), and lipid phosphate phosphatases ( Lpp ) 1–3 in B cells and T cells, shown relative to HPRT . (c) Relative amount of S1P remaining in culture supernatants after incubation with B or T cells for the indicated number of minutes, determined by the extent of down-modulation of Flag-S1P1 on a reporter cell line. Data are plotted as MFI of Flag-S1P1 staining relative to reporter cells not exposed to S1P (data are from 3 experiments).

    Article Snippet: For detection of surface S1P1, we used a rat monoclonal anti-mouse S1P1 antibody that is under development at R & D Systems.

    Techniques: CTL Assay, Mouse Assay, Incubation, Staining

    Most of thymic stromal cells and thymocytes belongs to the Lyve1 lineage . (A) tdTomato expression in the thymocytes of R26 -tdTomato (Cre − ) or Lyve1-CRE/R26 -tdTomato (Cre + ) mice. (B) Histological sections of the thymus from Lyve1-CRE/R26 -tdTomato reporter mice stained with anti-CD31 (green) and anti-Lyve1 (cyan) antibodies, and the Hoechst 33342 nuclear stain (blue). Cre recombinase-mediated tdTomato expression is shown in magenta. Bars: 500 μm. (C,D) Expression of CD62L, CD69, and S1PR1 in tdTomato − or tdTomato + CD4 + SP cells of Lyve1-CRE/R26 -tdTomato mice. S1PR1 expression was examined on mature CD4 + SP cells. (E) tdTomato expression in the T and B cells in the LNs and spleens of R26 -tdTomato (Cre − ) and Lyve1-CRE/R26 -tdTomato (Cre + ) mice. n = 3 (A,E) and n = 2 (B–D) per group.

    Journal: Frontiers in Immunology

    Article Title: Thymocytes in Lyve1-CRE/S1pr1f/f Mice Accumulate in the Thymus due to Cell-Intrinsic Loss of Sphingosine-1-Phosphate Receptor Expression

    doi: 10.3389/fimmu.2016.00489

    Figure Lengend Snippet: Most of thymic stromal cells and thymocytes belongs to the Lyve1 lineage . (A) tdTomato expression in the thymocytes of R26 -tdTomato (Cre − ) or Lyve1-CRE/R26 -tdTomato (Cre + ) mice. (B) Histological sections of the thymus from Lyve1-CRE/R26 -tdTomato reporter mice stained with anti-CD31 (green) and anti-Lyve1 (cyan) antibodies, and the Hoechst 33342 nuclear stain (blue). Cre recombinase-mediated tdTomato expression is shown in magenta. Bars: 500 μm. (C,D) Expression of CD62L, CD69, and S1PR1 in tdTomato − or tdTomato + CD4 + SP cells of Lyve1-CRE/R26 -tdTomato mice. S1PR1 expression was examined on mature CD4 + SP cells. (E) tdTomato expression in the T and B cells in the LNs and spleens of R26 -tdTomato (Cre − ) and Lyve1-CRE/R26 -tdTomato (Cre + ) mice. n = 3 (A,E) and n = 2 (B–D) per group.

    Article Snippet: Briefly, single-cell suspensions were incubated with rat anti-S1PR1 monoclonal antibody (40 μg/ml, 713412; R & D Systems) followed by incubation with phycoerythrin-conjugated goat anti-rat IgG antibody (Southern Biotech).

    Techniques: Expressing, Mouse Assay, Staining

    T cells are remarkably decreased in the peripheral lymphoid tissues of Lyve1-CRE/S1pr1 f/f mice . (A) T and B cell numbers in the inguinal LNs (ILNs), spleen, blood, thymus, and bone marrow (BM) of S1pr1 f/f and Lyve1-CRE/S1pr1 f/f mice. DP, CD4 + CD8 + double positive cells; CD4 or CD8 SP, CD4 + or CD8 + single-positive cells. (B) Numbers of CD4 + T cell subsets in the inguinal LNs of S1pr1 f/f and Lyve1-CRE/S1pr1 f/f mice. Naïve, memory, and effector CD4 + T cells were defined as CD62L + CD44 − , CD62L + CD44 + , and CD62L − CD44 + , respectively. (C) Migration of WT lymphocytes into the LNs of S1pr1 f/f and Lyve1-CRE/S1pr1 f/f mice. Lymphocytes from beta-actin-eGFP mice were injected intravenously into recipient mice. After 12 h, the inguinal LNs were collected, and donor-derived lymphocytes were quantified by flow cytometry. These data are pooled from three independent experiments ( n > 4) (A) and are representative of two (B,C) independent experiments with three mice per group. These data were evaluated by Student’s t -test and represent the mean ± SD (* P

    Journal: Frontiers in Immunology

    Article Title: Thymocytes in Lyve1-CRE/S1pr1f/f Mice Accumulate in the Thymus due to Cell-Intrinsic Loss of Sphingosine-1-Phosphate Receptor Expression

    doi: 10.3389/fimmu.2016.00489

    Figure Lengend Snippet: T cells are remarkably decreased in the peripheral lymphoid tissues of Lyve1-CRE/S1pr1 f/f mice . (A) T and B cell numbers in the inguinal LNs (ILNs), spleen, blood, thymus, and bone marrow (BM) of S1pr1 f/f and Lyve1-CRE/S1pr1 f/f mice. DP, CD4 + CD8 + double positive cells; CD4 or CD8 SP, CD4 + or CD8 + single-positive cells. (B) Numbers of CD4 + T cell subsets in the inguinal LNs of S1pr1 f/f and Lyve1-CRE/S1pr1 f/f mice. Naïve, memory, and effector CD4 + T cells were defined as CD62L + CD44 − , CD62L + CD44 + , and CD62L − CD44 + , respectively. (C) Migration of WT lymphocytes into the LNs of S1pr1 f/f and Lyve1-CRE/S1pr1 f/f mice. Lymphocytes from beta-actin-eGFP mice were injected intravenously into recipient mice. After 12 h, the inguinal LNs were collected, and donor-derived lymphocytes were quantified by flow cytometry. These data are pooled from three independent experiments ( n > 4) (A) and are representative of two (B,C) independent experiments with three mice per group. These data were evaluated by Student’s t -test and represent the mean ± SD (* P

    Article Snippet: Briefly, single-cell suspensions were incubated with rat anti-S1PR1 monoclonal antibody (40 μg/ml, 713412; R & D Systems) followed by incubation with phycoerythrin-conjugated goat anti-rat IgG antibody (Southern Biotech).

    Techniques: Mouse Assay, Migration, Injection, Derivative Assay, Flow Cytometry, Cytometry

    Mature CD4 + and CD8 + SP cells are accumulated in the Lyve1-CRE/S1pr1 f/f thymus . (A) Frequencies of CD4 + and CD8 + SP cells in the thymus of Lyve1-CRE/S1pr1 f/f and S1pr1 f/f mice ( n = 5). (B,C) Frequencies of immature (CD62L low CD69 high ), mature (CD62L high CD69 low ), and CD62 high CD69 high cells in the CD4 + or CD8 + SP subset of S1pr1 f/f and Lyve1-CRE/S1pr1 f/f mice. Data were pooled from nine mice (C) . The data were evaluated by Student’s t -test and represent the mean ± SD (** P

    Journal: Frontiers in Immunology

    Article Title: Thymocytes in Lyve1-CRE/S1pr1f/f Mice Accumulate in the Thymus due to Cell-Intrinsic Loss of Sphingosine-1-Phosphate Receptor Expression

    doi: 10.3389/fimmu.2016.00489

    Figure Lengend Snippet: Mature CD4 + and CD8 + SP cells are accumulated in the Lyve1-CRE/S1pr1 f/f thymus . (A) Frequencies of CD4 + and CD8 + SP cells in the thymus of Lyve1-CRE/S1pr1 f/f and S1pr1 f/f mice ( n = 5). (B,C) Frequencies of immature (CD62L low CD69 high ), mature (CD62L high CD69 low ), and CD62 high CD69 high cells in the CD4 + or CD8 + SP subset of S1pr1 f/f and Lyve1-CRE/S1pr1 f/f mice. Data were pooled from nine mice (C) . The data were evaluated by Student’s t -test and represent the mean ± SD (** P

    Article Snippet: Briefly, single-cell suspensions were incubated with rat anti-S1PR1 monoclonal antibody (40 μg/ml, 713412; R & D Systems) followed by incubation with phycoerythrin-conjugated goat anti-rat IgG antibody (Southern Biotech).

    Techniques: Mouse Assay

    Thymocytes accumulating in the thymus of Lyve1-CRE/S1pr1 f/f mice resemble egress-competent cells, but lack S1PR1 expression . (A,B) Expression of 6C10 and heat stable antigen (HSA) (A) or S1PR1 (B) in immature (CD62L low CD69 high ), mature (CD62L high CD69 low ), and CD62L high CD69 high cells of the CD4 + SP subset in S1pr1 f/f and Lyve1-CRE/S1pr1 f/f thymuses. The mean fluorescence intensity was normalized against CD62L low CD69 high cells. 6C10 ( n = 3), HAS ( n = 3), and S1PR1 ( n = 3). Data are representative of two independent experiments. These data were evaluated by Student’s t -test and represent the mean ± SD (* P

    Journal: Frontiers in Immunology

    Article Title: Thymocytes in Lyve1-CRE/S1pr1f/f Mice Accumulate in the Thymus due to Cell-Intrinsic Loss of Sphingosine-1-Phosphate Receptor Expression

    doi: 10.3389/fimmu.2016.00489

    Figure Lengend Snippet: Thymocytes accumulating in the thymus of Lyve1-CRE/S1pr1 f/f mice resemble egress-competent cells, but lack S1PR1 expression . (A,B) Expression of 6C10 and heat stable antigen (HSA) (A) or S1PR1 (B) in immature (CD62L low CD69 high ), mature (CD62L high CD69 low ), and CD62L high CD69 high cells of the CD4 + SP subset in S1pr1 f/f and Lyve1-CRE/S1pr1 f/f thymuses. The mean fluorescence intensity was normalized against CD62L low CD69 high cells. 6C10 ( n = 3), HAS ( n = 3), and S1PR1 ( n = 3). Data are representative of two independent experiments. These data were evaluated by Student’s t -test and represent the mean ± SD (* P

    Article Snippet: Briefly, single-cell suspensions were incubated with rat anti-S1PR1 monoclonal antibody (40 μg/ml, 713412; R & D Systems) followed by incubation with phycoerythrin-conjugated goat anti-rat IgG antibody (Southern Biotech).

    Techniques: Mouse Assay, Expressing, Fluorescence

    CD4 + SP cells in the Lyve1-CRE/S1pr1 f/f thymus are egress-incompetent . (A) Egress of S1pr1 f/f or Lyve1-CRE/S1pr1 f/f thymocytes from the WT thymus. CD62L + CD4 + SP cells of the S1pr1 f/f or Lyve1-CRE/S1pr1 f/f thymus were labeled with CFSE or CMTMR, respectively, and cotransferred into the WT thymus in equal numbers (1 × 10 6 cells per mouse) by intrathymic (i.t.) injection. Two days after the injection, the numbers of donor-derived cells in the thymus, blood, and LNs of recipient mice were analyzed by flow cytometry. (B) Ratio of Lyve1-CRE/S1pr1 f/f CD4 + SP cells (CMTMR) to S1pr1 f/f CD4 + SP cells (CFSE) that migrated into the WT recipient’s LNs. PLN, peripheral LNs; MLN, mesenteric LNs. Data are representative of two independent experiments with three mice per group. (C) Egress of WT thymocytes from the S1pr1 f/f or Lyve1-CRE/S1pr1 f/f thymus. CD62L + CD4 + SP cells from beta-actin-DsRed mice were injected into the S1pr1 f/f or Lyve1-CRE/S1pr1 f/f thymus. Two days later, the frequencies of DsRed + cells in the thymus, blood, and LNs were analyzed by flow cytometry. Data are pooled from two independent experiments ( n = 3 per group) and represent the mean ± SD. Differences between the groups were evaluated by one-way ANOVA (B) or Student’s t -test (C) (*** P

    Journal: Frontiers in Immunology

    Article Title: Thymocytes in Lyve1-CRE/S1pr1f/f Mice Accumulate in the Thymus due to Cell-Intrinsic Loss of Sphingosine-1-Phosphate Receptor Expression

    doi: 10.3389/fimmu.2016.00489

    Figure Lengend Snippet: CD4 + SP cells in the Lyve1-CRE/S1pr1 f/f thymus are egress-incompetent . (A) Egress of S1pr1 f/f or Lyve1-CRE/S1pr1 f/f thymocytes from the WT thymus. CD62L + CD4 + SP cells of the S1pr1 f/f or Lyve1-CRE/S1pr1 f/f thymus were labeled with CFSE or CMTMR, respectively, and cotransferred into the WT thymus in equal numbers (1 × 10 6 cells per mouse) by intrathymic (i.t.) injection. Two days after the injection, the numbers of donor-derived cells in the thymus, blood, and LNs of recipient mice were analyzed by flow cytometry. (B) Ratio of Lyve1-CRE/S1pr1 f/f CD4 + SP cells (CMTMR) to S1pr1 f/f CD4 + SP cells (CFSE) that migrated into the WT recipient’s LNs. PLN, peripheral LNs; MLN, mesenteric LNs. Data are representative of two independent experiments with three mice per group. (C) Egress of WT thymocytes from the S1pr1 f/f or Lyve1-CRE/S1pr1 f/f thymus. CD62L + CD4 + SP cells from beta-actin-DsRed mice were injected into the S1pr1 f/f or Lyve1-CRE/S1pr1 f/f thymus. Two days later, the frequencies of DsRed + cells in the thymus, blood, and LNs were analyzed by flow cytometry. Data are pooled from two independent experiments ( n = 3 per group) and represent the mean ± SD. Differences between the groups were evaluated by one-way ANOVA (B) or Student’s t -test (C) (*** P

    Article Snippet: Briefly, single-cell suspensions were incubated with rat anti-S1PR1 monoclonal antibody (40 μg/ml, 713412; R & D Systems) followed by incubation with phycoerythrin-conjugated goat anti-rat IgG antibody (Southern Biotech).

    Techniques: Labeling, Injection, Derivative Assay, Mouse Assay, Flow Cytometry, Cytometry

    S1PR1 is expressed in lymphatic endothelial cells of the thymus and LNs . (A) S1PR1 expression was examined immunohistologically in the thymus and LNs. Lyve1-positive lymphatics were observed in the vicinity of the cortico-medullary junction (dotted line). C, cortex; M, medulla. Bars, 100 μm. (B,C) Flow cytometric analysis of S1PR1 expression in thymic and LN stromal cells of S1pr1 f/f and Lyve1-CRE/S1pr1 f/f mice. (B) CD45-negative stromal cell populations from the thymus and LNs. Lymphatic and blood vessel endothelial cells (LECs and BECs) were identified by their expression of Gp38 and CD31. (C) S1PR1 expression in LECs and BECs in the thymus and LNs of S1pr1 f/f (blue line) and Lyve1-CRE/S1pr1 f/f mice (red line). Data are representative of two (A) or three (B,C) independent experiments.

    Journal: Frontiers in Immunology

    Article Title: Thymocytes in Lyve1-CRE/S1pr1f/f Mice Accumulate in the Thymus due to Cell-Intrinsic Loss of Sphingosine-1-Phosphate Receptor Expression

    doi: 10.3389/fimmu.2016.00489

    Figure Lengend Snippet: S1PR1 is expressed in lymphatic endothelial cells of the thymus and LNs . (A) S1PR1 expression was examined immunohistologically in the thymus and LNs. Lyve1-positive lymphatics were observed in the vicinity of the cortico-medullary junction (dotted line). C, cortex; M, medulla. Bars, 100 μm. (B,C) Flow cytometric analysis of S1PR1 expression in thymic and LN stromal cells of S1pr1 f/f and Lyve1-CRE/S1pr1 f/f mice. (B) CD45-negative stromal cell populations from the thymus and LNs. Lymphatic and blood vessel endothelial cells (LECs and BECs) were identified by their expression of Gp38 and CD31. (C) S1PR1 expression in LECs and BECs in the thymus and LNs of S1pr1 f/f (blue line) and Lyve1-CRE/S1pr1 f/f mice (red line). Data are representative of two (A) or three (B,C) independent experiments.

    Article Snippet: Briefly, single-cell suspensions were incubated with rat anti-S1PR1 monoclonal antibody (40 μg/ml, 713412; R & D Systems) followed by incubation with phycoerythrin-conjugated goat anti-rat IgG antibody (Southern Biotech).

    Techniques: Expressing, Flow Cytometry, Mouse Assay

    Hindering S1P1 internalization abrogates T-cell sequestration in bone marrow a , Relative sequestration of adoptively transferred CFSE-labeled T-cells within the bone marrow of CT2A IC recipient mice at 2 hours (left) or 24 hours (right) after transfer. As indicated, transferred cells were splenocytes from either control C57BL/6 donors (control) or from S1P1 stabilized “knock-in” (S1P1 KI) donors (n=5 recipient mice per group). Data in a are representative findings from one of a minimum of two independently repeated experiments with similar results. b , T-cell counts in the bone marrow of n=10 IC CT2A-bearing wild type (WT) C57BL/6 or n=11 S1P1 KI mice. Counts were assessed 18 days following tumor implantation and are shown relative to baseline counts in n=6 tumor-naïve control WT or n=6 tumor-naïve S1P1 KI mice. Cumulative data from three experiments are depicted in b. c , IC CT2A tumors were harvested from n=6 WT C57BL/6 (WT) or n=6 S1P1 KI mice 18 days following tumor implantation. TIL were assessed by flow cytometry and the number of total T-cells per gram of tumor quantified. d , The frequency of activated effector (CD44 hi CD62L lo ) T-cells in IC CT2A tumors from the same n=6 WT C57BL/6 (WT) or n=6 S1P1 KI mice in c was also quantified. Data in c, d are representative findings from one of a minimum of three independently repeated experiments with similar results. e , C57BL/6 (WT) or S1P1 KI mice were implanted with IC CT2A tumors and treated with a 4-1BB agonist antibody or isotype control (n=8 per group). All data in a–d are shown as mean ± s.e.m. P values in a–d were determined by two-tailed, unpaired Student’s t-test. Survival in e was assessed by two-tailed generalized Wilcoxon test. P value for overall comparison is depicted.

    Journal: Nature medicine

    Article Title: Sequestration of T-cells in bone marrow in the setting of glioblastoma and other intracranial tumors

    doi: 10.1038/s41591-018-0135-2

    Figure Lengend Snippet: Hindering S1P1 internalization abrogates T-cell sequestration in bone marrow a , Relative sequestration of adoptively transferred CFSE-labeled T-cells within the bone marrow of CT2A IC recipient mice at 2 hours (left) or 24 hours (right) after transfer. As indicated, transferred cells were splenocytes from either control C57BL/6 donors (control) or from S1P1 stabilized “knock-in” (S1P1 KI) donors (n=5 recipient mice per group). Data in a are representative findings from one of a minimum of two independently repeated experiments with similar results. b , T-cell counts in the bone marrow of n=10 IC CT2A-bearing wild type (WT) C57BL/6 or n=11 S1P1 KI mice. Counts were assessed 18 days following tumor implantation and are shown relative to baseline counts in n=6 tumor-naïve control WT or n=6 tumor-naïve S1P1 KI mice. Cumulative data from three experiments are depicted in b. c , IC CT2A tumors were harvested from n=6 WT C57BL/6 (WT) or n=6 S1P1 KI mice 18 days following tumor implantation. TIL were assessed by flow cytometry and the number of total T-cells per gram of tumor quantified. d , The frequency of activated effector (CD44 hi CD62L lo ) T-cells in IC CT2A tumors from the same n=6 WT C57BL/6 (WT) or n=6 S1P1 KI mice in c was also quantified. Data in c, d are representative findings from one of a minimum of three independently repeated experiments with similar results. e , C57BL/6 (WT) or S1P1 KI mice were implanted with IC CT2A tumors and treated with a 4-1BB agonist antibody or isotype control (n=8 per group). All data in a–d are shown as mean ± s.e.m. P values in a–d were determined by two-tailed, unpaired Student’s t-test. Survival in e was assessed by two-tailed generalized Wilcoxon test. P value for overall comparison is depicted.

    Article Snippet: Antibodies to murine S1P1 (Cat#FAB7089A, Clone: 713412, Lot#ACNG0216051, 1:10) were obtained from R & D systems (Minneapolis, MN).

    Techniques: Labeling, Mouse Assay, Tumor Implantation, Flow Cytometry, Cytometry, Two Tailed Test

    Loss of surface S1P1 on T-cells directs their sequestration in bone marrow in the setting of intracranial tumor a , The percentage of nascent T-cells expressing surface S1P1 was assessed by flow cytometry in the bone marrow of n=6 control C57BL/6 mice or n=6 mice bearing IC CT2A on Day 18 following tumor implantation. b , Representative flow cytometry plot of data depicted in a. c , Negative correlation between bone marrow T-cell counts and S1P1 levels on bone marrow T-cells across IC and SC murine tumor models. Data in c were obtained from n=6 IC CT2A, n=5 IC E0771, n=6 IC B16F10, n=7 IC LLC, n=6 SC CT2A, n=7 SC E0771, n=6 SC B16F10, and n=7 SC LLC tumor-bearing mice. N=21 control C57BL/6 were also included. Data in c are cumulative results from a minimum of two experiments with each tumor type. d , The percentage of nascent T-cells expressing surface S1P1 was assessed by flow cytometry in the bone marrow of n=14 GBM patients or n=12 age-matched controls. e , Representative flow cytometry plot of data depicted in d. f , Negative correlation between bone marrow T-cell counts and surface S1P1 levels on bone marrow T-cells in n=12 GBM patients and n=10 age-matched controls. g , Relative sequestration of adoptively transferred CFSE-labeled T-cells within the bone marrow of IC CT2A recipient mice either 2 or 24 hours after transfer (n=5 mice per group). As indicated, transferred cells were splenocytes from either control C57BL/6 donors (control) or from S1P1 conditional knockout (S1P1 KO) donors. Data in g are representative findings from one of a minimum of two independently repeated experiments with similar results. All data in a, d , and g are shown as mean ± s.e.m. P values in a, d , and g were determined by two-tailed, unpaired Student’s t-test. Two-tailed, p values and Pearson coefficients for c, f are depicted.

    Journal: Nature medicine

    Article Title: Sequestration of T-cells in bone marrow in the setting of glioblastoma and other intracranial tumors

    doi: 10.1038/s41591-018-0135-2

    Figure Lengend Snippet: Loss of surface S1P1 on T-cells directs their sequestration in bone marrow in the setting of intracranial tumor a , The percentage of nascent T-cells expressing surface S1P1 was assessed by flow cytometry in the bone marrow of n=6 control C57BL/6 mice or n=6 mice bearing IC CT2A on Day 18 following tumor implantation. b , Representative flow cytometry plot of data depicted in a. c , Negative correlation between bone marrow T-cell counts and S1P1 levels on bone marrow T-cells across IC and SC murine tumor models. Data in c were obtained from n=6 IC CT2A, n=5 IC E0771, n=6 IC B16F10, n=7 IC LLC, n=6 SC CT2A, n=7 SC E0771, n=6 SC B16F10, and n=7 SC LLC tumor-bearing mice. N=21 control C57BL/6 were also included. Data in c are cumulative results from a minimum of two experiments with each tumor type. d , The percentage of nascent T-cells expressing surface S1P1 was assessed by flow cytometry in the bone marrow of n=14 GBM patients or n=12 age-matched controls. e , Representative flow cytometry plot of data depicted in d. f , Negative correlation between bone marrow T-cell counts and surface S1P1 levels on bone marrow T-cells in n=12 GBM patients and n=10 age-matched controls. g , Relative sequestration of adoptively transferred CFSE-labeled T-cells within the bone marrow of IC CT2A recipient mice either 2 or 24 hours after transfer (n=5 mice per group). As indicated, transferred cells were splenocytes from either control C57BL/6 donors (control) or from S1P1 conditional knockout (S1P1 KO) donors. Data in g are representative findings from one of a minimum of two independently repeated experiments with similar results. All data in a, d , and g are shown as mean ± s.e.m. P values in a, d , and g were determined by two-tailed, unpaired Student’s t-test. Two-tailed, p values and Pearson coefficients for c, f are depicted.

    Article Snippet: Antibodies to murine S1P1 (Cat#FAB7089A, Clone: 713412, Lot#ACNG0216051, 1:10) were obtained from R & D systems (Minneapolis, MN).

    Techniques: Expressing, Flow Cytometry, Cytometry, Mouse Assay, Tumor Implantation, Labeling, Knock-Out, Two Tailed Test