Journal: European journal of immunology

Article Title: Comparison of stable human Treg and Th clones by transcriptional profiling.

doi: 10.1002/eji.200838807

Figure Lengend Snippet: Figure 4. Human Treg clones, but not Th clones, produce the mature and active form of TGF-b. (A) Schematic representation of TGF-b processing. Double lines represent cell membrane. Sites of proteolytic cleavages are indicated by arrow heads. Small bars indicate disulfide bonds. LAP: latency associated peptide. LTBP: latent TGF-b binding protein; TGFBR: TGF-b receptors. (B) Three Treg clones (Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated in X-VIVO-10 serum-free medium for 24 h with anti-CD3 and anti-CD28 Ab. Active and total TGF-b concentrations in the supernatants were measured by ELISA, before or after treatment with acid, respectively. Values represent means of duplicate1SD. (C) Four Treg clones (Treg C2, Treg D1, Treg B2, and Treg B3) and four Th clones (Th C2, Th D1, Th B1, and Th A2) were activated for 24 h with anti-CD3 and anti-CD28 Ab in the presence of IL-2, prior to staining with biotinylated polyclonal anti-LAP antibody (filled gray histograms) or isotype control (empty histograms) and Streptavidin-PE. (D) Western blot analysis of cell lysates from Treg and Th clones collected at rest (T 5 0), or 6 or 24 h after activation with anti-CD3 and anti-CD28 Ab. Samples in lanes 4 and 8 were treated with 5 ng/mL of recombinant human TGF-b1 (rhTGF-b1) during the last 15 min of the activation period and serve as positive controls. Samples in lanes 17 and 18 were prepared as detailed in Fig. 3D. Briefly, suppressed (‘‘S’’) or control (‘‘Ctrl’’) clone Th A2 was sorted by FACS out of a co-culture with clone Treg A1 or clone Th A2, respectively. Blots were hybridized with an anti-phosphorylated SMAD2 (P-SMAD2) or anti-b-actin antibody.

Article Snippet: For surface LAP expression, cells activated for 24 h with anti-CD3 and anti-CD28 antibodies in the presence of IL-2 (160 IU/mL) were labeled with a biotinylated polyclonal anti-LAP antibody or an isotype control (R&D Systems, BAF246 and BAF108, respectively), followed by incubation with Streptavidin coupled to PE (BD Pharmingen).

Techniques: Clone Assay, Membrane, Binding Assay, Enzyme-linked Immunosorbent Assay, Staining, Control, Western Blot, Activation Assay, Recombinant, Co-Culture Assay

Journal: JVS-Vascular Science

Article Title: EphB4 monomer inhibits chronic graft vasculopathy in an aortic transplant model

doi: 10.1016/j.jvssci.2023.100109

Figure Lengend Snippet: EphB4 monomer reduces CD68 + cell infiltration in allografts. (A) Representative immunofluorescence of isografts and allografts (day 28) treated with either control IgG or EphB4 monomer; top row, 40× (scale bar, 250 μm); bottom row, 400× (scale bar, 25 μm). (B) Mean number of CD68 + cells per high-power field ( HPF ) in isografts; P = .66 (Mann-Whitney U test; n = 5-6 rats per group). (C) Mean number of CD68 + cells per HPF in allografts; P ≤ .01 (Mann-Whitney U test; n = 5-6 rats per group).

Article Snippet: Rats then had subdermal injections with either EphB4 monomer (20 μg/kg; Sino Biological Inc., Beijing, China) or IgG control (Sino Biological Inc.).

Techniques: Immunofluorescence, MANN-WHITNEY

Journal: JVS-Vascular Science

Article Title: EphB4 monomer inhibits chronic graft vasculopathy in an aortic transplant model

doi: 10.1016/j.jvssci.2023.100109

Figure Lengend Snippet: EphB4 monomer reduces CD3 + cell infiltration in allografts. (A) Representative immunofluorescence of isografts and allografts (day 28) treated with either control IgG or EphB4 monomer; top row, 40× (scale bar, 250 μm); bottom row, 400× (scale bar, 25 μm). (B) Mean number of CD3 + cells per high-power field ( HPF ) in isografts ( P = .17, Mann-Whitney U test ; n = 5-6 rats per group). (C) Mean number of CD3 + cells per HPF in allografts; P = .02 (Mann-Whitney U test; n = 5-6 rats per group).

Article Snippet: Rats then had subdermal injections with either EphB4 monomer (20 μg/kg; Sino Biological Inc., Beijing, China) or IgG control (Sino Biological Inc.).

Techniques: Immunofluorescence, MANN-WHITNEY

Journal: JVS-Vascular Science

Article Title: EphB4 monomer inhibits chronic graft vasculopathy in an aortic transplant model

doi: 10.1016/j.jvssci.2023.100109

Figure Lengend Snippet: EphB4 monomer reduces neointimal thickness in allografts. (A) Representative EVG staining of isografts and allografts (day 28) treated with either control IgG or EphB4 monomer; top row, 40× (scale bar, 250 μm); bottom row, 400× (scale, bar 25 μm). (B) Mean neointimal thickness in isografts ( P = .08, Mann-Whitney U test; n = 5-6 rats per group). (C) Mean medial thickness in isografts ( P = .08, Mann-Whitney U test; n = 5-6 rats per group). (D) Ratio of neointima:media thickness in isografts ( P = .66, Mann-Whitney U test; n = 5-6 rats per group). (E) Mean neointimal thickness in allografts ( P = .05, Mann-Whitney U test; n = 5-6 rats per group). (F) Mean medial thickness in allografts ( P = .12, Mann-Whitney U test; n = 5-6 rats per group). (G) Ratio of neointima:media thickness in allografts ( P = .05, Mann-Whitney U test; n = 5-6 rats per group).

Article Snippet: Rats then had subdermal injections with either EphB4 monomer (20 μg/kg; Sino Biological Inc., Beijing, China) or IgG control (Sino Biological Inc.).

Techniques: Staining, MANN-WHITNEY