Journal: Immunity

Article Title: Engagement of the type I interferon receptor on dendritic cells inhibits T helper 17 cell development: role of intracellular osteopontin.

doi: 10.1016/j.immuni.2008.05.008

Figure Lengend Snippet: Figure 7. Microglia Require Opn-i Expression to Induce Th17 Cells Opn-deficient microglia were infected with lenti-Opn-i or lenti-GFP and incubated 2D2+CD4 T cells and MOG peptide. (A and B) Reconstitution of Opn-deficient microglia with Opn-i rescues IL-17 production by CD4+ 2D2 T cells. (A) IL-17 concentrations in culture supernatants 24 hr after T cell restimulation on day 4 were determined by ELISA in triplicate wells. (B) Concentrations of IL-27p28 in day 4 cultures before T cell restimulation. The amounts of IL-27p28 were determined by ELISA in triplicate wells; empty and filled bars denote supernatants from Opn-i-transfected and GFP-transfected DC, respectively, and data are representative of two experiments. (C) EAE development is inhibited in irradiated Opn-deficient hosts. Total BM cells (1.7 3 107 cells/mouse) from 2D2+ Tg Opn-deficient mice were i.v. trans- ferred to irradiated (800 rads) hosts. Ten weeks later, EAE was induced. Disease development (mean clinical score) and incidence (%) is shown for each group (n = 5). Error bars in disease score denote mean ± SEM and t test values on the last day of observation are shown. Mice displayed >95% engraftment of donor bone marrow according to immunofluorescence with Ly5.2 Ab (donor BM cells were obtained from B6.Ly5.2+ Opn-deficient mice expressing the 2D2 TCR transgene) 10 weeks after reconstitution and before disease induction, as judged by immunofluorescence of peripheral blood cells.

Article Snippet: Antibodies to IL-17 (TC1118H10) and IFN-g (XMG1.2) for flow cytometry were from BD PharMingen; Foxp3 (FJK-16 s) and IL-10 (JES5-16E3) Abs were from eBioscience; IL-27p28 Ab (BAF1834), recombinant proteins for mouse IL-23, IL-6, Flt-3L, and human TGF-b1 were from R&D systems; and recombinant mouse IFN-a was from HyCult.

Techniques: Expressing, Infection, Incubation, Enzyme-linked Immunosorbent Assay, Transfection, Irradiation

Journal: Virology Journal

Article Title: Interleukin-27 ameliorates coxsackievirus-B3-induced viral myocarditis by inhibiting Th17 cells

doi: 10.1186/s12985-015-0418-x

Figure Lengend Snippet: Expression kinetics of IL-27 and IL-27R in VMC. Male BALB/c mice were infected with 10 3 TCID 50 of CVB3 at day 0. a Body weight was monitored daily until day 10 post-infection. b The severity of myocarditis was scored every 2 days until day 10 post-infection using a scale based on foci of mononuclear infiltration and myocardial necrosis. c The myocarditis viral replication (positive strand of CVB3) and the viral loads (negative-strand of CVB3) were detected using qRT-PCR. d The survival rate of CVB3-infected mice and control mice after CVB3 infection were recorded ( n = 20 per group). The expression of IL-27 in serum ( e ) and cardiac tissue ( f ) were monitored every two days until day 10 post-infection by ELISA assay. The expression of IL-27R mRNA ( g ) and protein ( h ) levels in cardiac tissue on day 7 post infection. Data are showed as the means ± SEM, Each time point contained three mice. *** P <0.001

Article Snippet: For in vivo IL-27 neutralization, mice were injected i.p. with Mouse Anti-Mouse IL-27 p28Ab (500 μg per mouse, MM27.7B1, BD Biosciences, San Diego, CA, USA) or control Ab (500 μg per mouse, Mouse IgG2a κ Isotype Control, BD Biosciences, San Diego, CA, USA) on day 0 and 5 after CVB3 infection.

Techniques: Expressing, Infection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Virology Journal

Article Title: Interleukin-27 ameliorates coxsackievirus-B3-induced viral myocarditis by inhibiting Th17 cells

doi: 10.1186/s12985-015-0418-x

Figure Lengend Snippet: Association between IL-27 expression and disease severity in VMC. Male BALB/c mice ( n = 10) were infected with 10 3 TCID 50 of CVB3 at day 0. IL-27 expression levels in cardiac tissues were negatively correlated with body weight loss ( a ) and myocarditis pathological score ( b ) on day 7 post-infection. IL-37 expression levels in serum were negatively correlated with body weight loss ( c ) and myocarditis pathological score ( d ) on day 7 post-infection

Article Snippet: For in vivo IL-27 neutralization, mice were injected i.p. with Mouse Anti-Mouse IL-27 p28Ab (500 μg per mouse, MM27.7B1, BD Biosciences, San Diego, CA, USA) or control Ab (500 μg per mouse, Mouse IgG2a κ Isotype Control, BD Biosciences, San Diego, CA, USA) on day 0 and 5 after CVB3 infection.

Techniques: Expressing, Infection

Journal: Virology Journal

Article Title: Interleukin-27 ameliorates coxsackievirus-B3-induced viral myocarditis by inhibiting Th17 cells

doi: 10.1186/s12985-015-0418-x

Figure Lengend Snippet: Neutralization of IL-27 exacerbated while administration of rmIL-27 alleviated CVB3-induced myocarditis. Male BALB/c mice were infected with 10 3 TCID 50 of CVB3 at day 0; mice were injected i.p. with IL-27 p28Ab or rmIL-27 on day 0 and 5 after CVB3 infection. The survival rates ( a ), body weight ( b ) and the severity of myocarditis ( c and d ) were monitored. Each group contained ten mice. Results are presented as the mean ± SEM of three independent experiments. * P <0.05, ** P <0.01, *** P <0.001, compared with PBS group; # P <0.05, ## P <0.01, compared with IgG group

Article Snippet: For in vivo IL-27 neutralization, mice were injected i.p. with Mouse Anti-Mouse IL-27 p28Ab (500 μg per mouse, MM27.7B1, BD Biosciences, San Diego, CA, USA) or control Ab (500 μg per mouse, Mouse IgG2a κ Isotype Control, BD Biosciences, San Diego, CA, USA) on day 0 and 5 after CVB3 infection.

Techniques: Neutralization, Infection, Injection

Journal: Virology Journal

Article Title: Interleukin-27 ameliorates coxsackievirus-B3-induced viral myocarditis by inhibiting Th17 cells

doi: 10.1186/s12985-015-0418-x

Figure Lengend Snippet: IL-27 ameliorates CVB3-induced myocarditis by inhibiting Th17 cells. a Representative FACS profile of Th17 cells in the spleen of CVB3 infection mice treated with rmIL-27 or Anti-IL-27 on day 7 post infection. Cells were gated on CD3 + CD4 + T cells. b Statistical analysis of the percentage of Th17 cells. c Expression of Th17 cells characteristic cytokines, IL-17A, IL-22, and key transcription factor ROR-γt in cardiac tissues on day 7 after CVB3 infection were detected by qRT-PCR. d The serum levels of proinflammatory cytokines IL-17A, IFN-γ, IL-6 and TNF-α in CVB3 infection mice were determined by ELISA. Normal, mice not-infected with the virus. Data are showed as the means ± SEM. * P <0.05, ** P <0.01, *** P <0.001

Article Snippet: For in vivo IL-27 neutralization, mice were injected i.p. with Mouse Anti-Mouse IL-27 p28Ab (500 μg per mouse, MM27.7B1, BD Biosciences, San Diego, CA, USA) or control Ab (500 μg per mouse, Mouse IgG2a κ Isotype Control, BD Biosciences, San Diego, CA, USA) on day 0 and 5 after CVB3 infection.

Techniques: Infection, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay