Journal: Transplantation

Article Title: Differential Regulation of T Cell Immunity and Tolerance by Stromal Laminin Expressed in the Lymph Node

doi: 10.1097/TP.0000000000002774

Figure Lengend Snippet: Stromal laminin 411 and 511 differentially modulate CD4+ T cell activation in mice and human. Murine CD4 T lymphocytes evaluated by flow cytometry for proliferation and CD69, CD25, CD44 expression (A, B) after 3 days of culture in plates coated with anti-CD3 (1 μg/mL) and the indicated concentrations of laminin 411 or laminin 511 (A), or with laminin 411 (2 μg/mL) and/or laminin 511 (1 μg/mL) (B). Human CD4+ T lymphocytes (C) evaluated by flow cytometry for proliferation and CD25 expression after 3 days of culture in plates coated with laminin 411 (2 μg/mL) and/or laminin 511 (1 μg/mL) with anti-CD3/CD28 beads (T:Beads ratio = 1:1). Data shown as representative profiles (B, C) and mean ± SEM (A-C) of three independent experiments with samples.

Article Snippet: *P < 0.05, **P < 0.01, ***P < 0.001. table ft1 table-wrap mode="anchored" t5 Table 1: caption a7 Target Molecule Source Clone Company α1 integrin Hamster Ha31/8 BD Bioscience α2 integrin Armenian Hamster HMα2 Biolegend α3 integrin Mouse ASC-1 Biolegend α4 integrin Rat 9C10 Biolegend α5 integrin Armenian Hamster HMα5–1 Biolegend α6 integrin Rat GoH3 Biolegend α7 integrin Mouse 6A11 LSBio αM integrin Rat M1/70 Biolegend αV integrin Rat RMV-7 Biolegend β1 integrin Armenian Hamster HMβ1–1 Biolegend β2 integrin Rat M18/2 Biolegend β3 integrin Armenian Hamster 2C9.G2 Biolegend α dystroglycan Mouse IIH6C4 Merck Millipore Open in a separate window Potential anti-laminin receptor antibodies for in vitro use We next determined if the laminin 511 receptor α6 integrin favored CD4+ T cell activation in vivo.

Techniques: Activation Assay, Flow Cytometry, Expressing

Journal: Transplantation

Article Title: Differential Regulation of T Cell Immunity and Tolerance by Stromal Laminin Expressed in the Lymph Node

doi: 10.1097/TP.0000000000002774

Figure Lengend Snippet: Laminin 511 favors CD4 T cell activation via the receptor α6 integrin. CD4 T lymphocytes evaluated by flow cytometry for proliferation and CD69, CD25, CD44 expression (A), for α6 integrin expression after 3 days of culture (B), or phospho-Erk1/2, phospho-Akt and phospho-P38Mapk (C) after 30 minutes of culture in plates coated with anti-CD3 (1 μg/mL) and with laminin 411 (2 μg/mL) and/or laminin 511 (1 μg/mL) in presence of anti-α6 integrin or isotype control mAb (A, D). Data shown as representative profiles (B, C) and mean ± SEM (A-D) of three independent experiments with duplicate samples. (E) DST or (F) CFA/Ea treated mice received 5 × 106 CFSE-labeled CD45.1 TEa TCR transgenic CD4+ T cells. Mice were euthanized two days post transfer, and donor lymphocytes from spleen and skin LNs assessed for proliferation, CD69, CD25, CD44 and percentage of donor cells (CD4+ CD45.1+) in different organs by flow cytometry. Representative profiles shown together with a quantitative analysis of three independent experiments. Data shown as mean ± SEM of at least six mice per group from three independent experiments.

Article Snippet: *P < 0.05, **P < 0.01, ***P < 0.001. table ft1 table-wrap mode="anchored" t5 Table 1: caption a7 Target Molecule Source Clone Company α1 integrin Hamster Ha31/8 BD Bioscience α2 integrin Armenian Hamster HMα2 Biolegend α3 integrin Mouse ASC-1 Biolegend α4 integrin Rat 9C10 Biolegend α5 integrin Armenian Hamster HMα5–1 Biolegend α6 integrin Rat GoH3 Biolegend α7 integrin Mouse 6A11 LSBio αM integrin Rat M1/70 Biolegend αV integrin Rat RMV-7 Biolegend β1 integrin Armenian Hamster HMβ1–1 Biolegend β2 integrin Rat M18/2 Biolegend β3 integrin Armenian Hamster 2C9.G2 Biolegend α dystroglycan Mouse IIH6C4 Merck Millipore Open in a separate window Potential anti-laminin receptor antibodies for in vitro use We next determined if the laminin 511 receptor α6 integrin favored CD4+ T cell activation in vivo.

Techniques: Activation Assay, Flow Cytometry, Expressing, Labeling, Transgenic Assay

Journal: Transplantation

Article Title: Differential Regulation of T Cell Immunity and Tolerance by Stromal Laminin Expressed in the Lymph Node

doi: 10.1097/TP.0000000000002774

Figure Lengend Snippet: Laminin 411 and 511 differentially regulate Th1 and Th2 polarization. CD4 T lymphocytes evaluated by flow cytometry for intracellular expression of IFNγ (A) or IL4 (D) after 5 days or Tbet, Gata3 and phospho-Stat6 after 16 hours culture in pro-Th1 (B,C) or pro-Th2 (E,F) environments with laminin 411 (2 μg/mL) and/or laminin 511 (1 μg/mL) in presence of functional blocking anti-α6 integrin or isotype control mAb (C,F). Data shown as representative profiles and mean ± SEM of at least three independent experiments with duplicate samples.

Article Snippet: *P < 0.05, **P < 0.01, ***P < 0.001. table ft1 table-wrap mode="anchored" t5 Table 1: caption a7 Target Molecule Source Clone Company α1 integrin Hamster Ha31/8 BD Bioscience α2 integrin Armenian Hamster HMα2 Biolegend α3 integrin Mouse ASC-1 Biolegend α4 integrin Rat 9C10 Biolegend α5 integrin Armenian Hamster HMα5–1 Biolegend α6 integrin Rat GoH3 Biolegend α7 integrin Mouse 6A11 LSBio αM integrin Rat M1/70 Biolegend αV integrin Rat RMV-7 Biolegend β1 integrin Armenian Hamster HMβ1–1 Biolegend β2 integrin Rat M18/2 Biolegend β3 integrin Armenian Hamster 2C9.G2 Biolegend α dystroglycan Mouse IIH6C4 Merck Millipore Open in a separate window Potential anti-laminin receptor antibodies for in vitro use We next determined if the laminin 511 receptor α6 integrin favored CD4+ T cell activation in vivo.

Techniques: Flow Cytometry, Expressing, Functional Assay, Blocking Assay

Journal: Transplantation

Article Title: Differential Regulation of T Cell Immunity and Tolerance by Stromal Laminin Expressed in the Lymph Node

doi: 10.1097/TP.0000000000002774

Figure Lengend Snippet: Laminin 511 promotes Th17 polarization via the α-dystroglycan receptor. CD4 T lymphocytes evaluated by flow cytometry for IL-17 after 5 days of culture (A, C), Rorγt, phopho-Stat3, Irf4, Batf and CD126 (B, D) expression after 16 hours of culture in pro-Th17 environment with laminin 411 (2 μg/mL) and/or laminin 511 (1 μg/mL) in presence of functional blocking anti-α6 integrin mAb, anti-α-dystroglycan or isotype control (C, D). Data shown as representative profiles (A, B) and mean ± AEM (A-D) of at least three independent experiments with duplicate samples. (E) CFA/Ea treated mice received 5 × 106 CFSE-labeled CD45.1 Ea TCR transgenic CD4+ T cells, with or without anti-αDG mAb, euthanized three days post transfer, and donor lymphocytes from spleen and skin LNs assessed as percentage of IL17+ donor cells (CD4+ CD45.1+) in different organs by flow cytometry. Representative profile shown with quantitative analysis of three independent experiments. Data shown as mean ± SEM of eight mice per group from three independent experiments.

Article Snippet: *P < 0.05, **P < 0.01, ***P < 0.001. table ft1 table-wrap mode="anchored" t5 Table 1: caption a7 Target Molecule Source Clone Company α1 integrin Hamster Ha31/8 BD Bioscience α2 integrin Armenian Hamster HMα2 Biolegend α3 integrin Mouse ASC-1 Biolegend α4 integrin Rat 9C10 Biolegend α5 integrin Armenian Hamster HMα5–1 Biolegend α6 integrin Rat GoH3 Biolegend α7 integrin Mouse 6A11 LSBio αM integrin Rat M1/70 Biolegend αV integrin Rat RMV-7 Biolegend β1 integrin Armenian Hamster HMβ1–1 Biolegend β2 integrin Rat M18/2 Biolegend β3 integrin Armenian Hamster 2C9.G2 Biolegend α dystroglycan Mouse IIH6C4 Merck Millipore Open in a separate window Potential anti-laminin receptor antibodies for in vitro use We next determined if the laminin 511 receptor α6 integrin favored CD4+ T cell activation in vivo.

Techniques: Flow Cytometry, Expressing, Functional Assay, Blocking Assay, Labeling, Transgenic Assay

Journal: Transplantation

Article Title: Differential Regulation of T Cell Immunity and Tolerance by Stromal Laminin Expressed in the Lymph Node

doi: 10.1097/TP.0000000000002774

Figure Lengend Snippet: Laminin 411 and 511 differentially regulate Treg polarization. CD4 T lymphocytes evaluated by flow cytometry for CD25, FoxP3 expression after 5 days of culture (A, C), Rorγt, phopho-Stat3, Irf4 and Batf (B, D) expression after 16 hours of culture in pro-Treg environment, with laminin 411 (2 μg/mL) and/or laminin 511 (1 μg/mL) in presence of functional blocking anti-α6 integrin or isotype control mAb (C, D). Data shown as representative profiles (A-C) and mean ± SEM (A-D) of at least three independent experiments with duplicate samples.

Article Snippet: *P < 0.05, **P < 0.01, ***P < 0.001. table ft1 table-wrap mode="anchored" t5 Table 1: caption a7 Target Molecule Source Clone Company α1 integrin Hamster Ha31/8 BD Bioscience α2 integrin Armenian Hamster HMα2 Biolegend α3 integrin Mouse ASC-1 Biolegend α4 integrin Rat 9C10 Biolegend α5 integrin Armenian Hamster HMα5–1 Biolegend α6 integrin Rat GoH3 Biolegend α7 integrin Mouse 6A11 LSBio αM integrin Rat M1/70 Biolegend αV integrin Rat RMV-7 Biolegend β1 integrin Armenian Hamster HMβ1–1 Biolegend β2 integrin Rat M18/2 Biolegend β3 integrin Armenian Hamster 2C9.G2 Biolegend α dystroglycan Mouse IIH6C4 Merck Millipore Open in a separate window Potential anti-laminin receptor antibodies for in vitro use We next determined if the laminin 511 receptor α6 integrin favored CD4+ T cell activation in vivo.

Techniques: Flow Cytometry, Expressing, Functional Assay, Blocking Assay

Journal: Transplantation

Article Title: Differential Regulation of T Cell Immunity and Tolerance by Stromal Laminin Expressed in the Lymph Node

doi: 10.1097/TP.0000000000002774

Figure Lengend Snippet: Blocking laminin 511 receptor α6 integrin and αDG prolongs allogenic cardiac graft survival. C57BL/6 mice received BALB/c heterotopic cardiac allografts on day 0 along with anti-α6 integrin or isotype control mAb alone (10 μg i.v, day 0 and every 3 days) (A), anti- αDG or isotype control mAb alone (B), anti-α6 integrin or anti- αDG (10 μg i.v, day 0 and once a week) with anti-CD40L (250 μg i.v. day 0 only) (C). Data presented as percent survival. n = at least five mice per group.

Article Snippet: *P < 0.05, **P < 0.01, ***P < 0.001. table ft1 table-wrap mode="anchored" t5 Table 1: caption a7 Target Molecule Source Clone Company α1 integrin Hamster Ha31/8 BD Bioscience α2 integrin Armenian Hamster HMα2 Biolegend α3 integrin Mouse ASC-1 Biolegend α4 integrin Rat 9C10 Biolegend α5 integrin Armenian Hamster HMα5–1 Biolegend α6 integrin Rat GoH3 Biolegend α7 integrin Mouse 6A11 LSBio αM integrin Rat M1/70 Biolegend αV integrin Rat RMV-7 Biolegend β1 integrin Armenian Hamster HMβ1–1 Biolegend β2 integrin Rat M18/2 Biolegend β3 integrin Armenian Hamster 2C9.G2 Biolegend α dystroglycan Mouse IIH6C4 Merck Millipore Open in a separate window Potential anti-laminin receptor antibodies for in vitro use We next determined if the laminin 511 receptor α6 integrin favored CD4+ T cell activation in vivo.

Techniques: Blocking Assay