Journal: Molecular medicine reports

Article Title: IL-22 exacerbates the severity of CVB3-induced acute viral myocarditis in IL-17A-deficient mice.

doi: 10.3892/mmr.2013.1323

Figure Lengend Snippet: Figure 2. Neutralization of IL-22 in IL-17A-/- mice promoted viral replication and decreased the production of TNF-α, IL-6 and IFN-γ. (A) The levels of cardiac CVB3 titers on day 14. Data show the mean values of CVB3 PFU/g of heart. (B) The relative expression levels of cardiac CVB3 RNA in different groups. (C-E) The result of statistical analysis for the alterations in cardiac TNF-α, IL-6 and IFN-γ mRNA, as measured by RT-PCR. (F) The correlation analysis of cardiac CVB3 RNA, cardiac IL-22 mRNA and IFN-γ mRNA. Each point represents an individual mouse. *P<0.01 and #P<0.05 versus the normal, AVMC and IgG groups. **P<0.01 and ##P<0.05 versus the AVMC, anti-IL-22 and IgG groups. Values are presented as the mean ± SD. IL, interleukin; CVB3, coxsackie virus B3; RT-PCR, real-time polymerase chain reaction; AVMC, acute viral myocarditis; SD, standard deviation.

Article Snippet: For in vivo IL-22 neutralization, a total of 32 IL-17A-/- mice were randomly divided into four groups: Mice in the AVMC group were injected with CVB3 and PBS (50 μg per mouse, n=8); in the anti-IL-22 Ab group, mice were administered with CVB3 and anti-IL-22 Ab (AF582; 50 μg per mouse; R&D Systems, Inc., Minneapolis, MN, USA; n=8); IgG control group, mice were injected with CVB3 and normal IgG control (AB-108-C; 50 μg per mouse; R&D Systems; n=8); and in the normal group, IL-17A-/- mice received no treatments (n=8).

Techniques: Neutralization, Expressing, Reverse Transcription Polymerase Chain Reaction, Virus, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Molecular medicine reports

Article Title: IL-22 exacerbates the severity of CVB3-induced acute viral myocarditis in IL-17A-deficient mice.

doi: 10.3892/mmr.2013.1323

Figure Lengend Snippet: Figure 3. Neutralization of IL-22 in IL-17A-/- mice decreased the levels of IL-22 and STAT3. (A) Results of the statistical analysis for the alteration of serum IL-22 protein levels, as investigated by ELISA. (B) The relative cardiac expression levels of IL-22, as analyzed by RT-PCR. (C) The correlation analysis of cardiac CVB3 RNA and cardiac IL-22 mRNA. (D and E) Results of the statistical analysis for the levels of cardiac STAT3 mRNA and STAT3 proteins, as mea sured by RT-PCR and western blot analysis, respectively. (F) Representative images for the levels of cardiac STAT3 proteins from each treated group. #P<0.05 versus the normal, AVMC and IgG groups. *P<0.01 versus the AVMC, anti-IL-22 and IgG groups. Values are presented as the mean ± SD. IL, interleukin; STAT3, activator of transcription 3; RT-PCR, real-time polymerase chain reaction; CVB3, coxsackievirus B3; SD, standard deviation; GAPDH, glyceralde hyde phosphate dehydrogenase.

Article Snippet: For in vivo IL-22 neutralization, a total of 32 IL-17A-/- mice were randomly divided into four groups: Mice in the AVMC group were injected with CVB3 and PBS (50 μg per mouse, n=8); in the anti-IL-22 Ab group, mice were administered with CVB3 and anti-IL-22 Ab (AF582; 50 μg per mouse; R&D Systems, Inc., Minneapolis, MN, USA; n=8); IgG control group, mice were injected with CVB3 and normal IgG control (AB-108-C; 50 μg per mouse; R&D Systems; n=8); and in the normal group, IL-17A-/- mice received no treatments (n=8).

Techniques: Neutralization, Enzyme-linked Immunosorbent Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Real-time Polymerase Chain Reaction, Standard Deviation

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

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