6 research outputs found
Human AAT inhibited pDC maturation and cytokine secretion.
<p>BM pDCs from B6 mice were differentiated with or without hAAT using Flt3L for 8 days then stimulated with 10μg/ml CpG for an additional 24 hr prior to FACS analysis. (A) CD80, (B) CD40, and (C) CCR9 expression. (D) TNF-α and (E) IL-6 levels in pDC culture media. P values of One-Way-ANOVA using Tukey’s post-hoc test are indicated as * P<0.05; ** P<0.01; *** P<0.001, n = 3.</p
Human AAT treatment attenuated lupus nephritis in MRL/lpr mice.
<p>(A) Proteinuria scores after 9 weeks after hAAT treatment (Mann-Whitney test). (B) Albuminuria levels after 11 weeks of hAAT treatment. *P = 0.0727. (C) Average area of glomeruli (um<sup>2</sup>). (D) Average number of nuclei per glomerulus. **P<0.01 and***P<0.001 by student’s <i>t</i>-test. (F) Representative PAS-stained kidney sections from control MRL/lpr mice and hAAT treated MRL/lpr mice. All imaged were photographed at the same magnification (20X).</p
<i>In vivo</i> hAAT treatment attenuated BMDCs differentiation and maturation in MRL/lpr mice.
<p>BMDCs from MRL/lpr mice treated with hAAT or PBS for 11 weeks were stimulated with LPS for 24 hrs. (A) Percentages of CD11c<sup>+</sup> and (B) CD11b<sup>+</sup>CD11c<sup>+</sup>. (C-D) Percentages of CD80<sup>+</sup> (C) and I-A (D) expressing BMDCs. P values of Student’s <i>t</i>-test are indicated as * P<0.05; ** P<0.01; *** P<0.001.</p
Detection of endogenous mouse AAT, exogenous human AAT, and anti-human AAT neutralizing antibody in MRL/lpr mice (N = 10) by ELISA.
<p>(A) Mouse AAT levels (relative unit to C57BL/6) in PBS-treated MRL mice. (B) hAAT levels detected in hAAT injected group. Note: at week 2 and 8, animals were bled at 2 days after hAAT injection; at week 4, animals were bled at 1 day after the injection; at week 11, animals were bled 3 days after the injection. Dashed line is the lower limit of quantification (LLOQ). The serum concentration of human AAT from the PBS-treated group was below LLOQ. (C) Relative levels of anti-human AAT neutralizing antibody in MRL/lpr mice following multiple-dose human AAT administrations in MRL/lpr mice.</p
Human AAT inhibited cDC maturation and cytokine secretion.
<p>BM cDCs from B6 mice were generated in vitro in the presence of GM-CSF and IL-4 for 4 days with or without hAAT and then stimulated with 0.5 μg/ml LPS or 10μg/ml CpG for an additional 24 hr prior to FACS analysis. (A) CD80, CD86 and I-A<sup>b</sup> expression (measured as mean fluorescence intensity, MFI) in B6 DCs stimulated with LPS. (B) TNF-α, IFNI, and IL-1β secretions in supernatants of B6 DCs stimulated with LPS. (C) CD80, CD86 and I-Ab expression in B6 DCs stimulated with CpG. (D) Secretion of TNF-α, IFN-I, IL-1β, and IL-6 by B6 DCs stimulated with CpG. P values of One-Way-ANOVA using Tukey’s post-hoc test are indicated as * P<0.05; ** P<0.01; *** P<0.001, n = 3.</p
Human AAT inhibited autoantibodies production in MRL/lpr mice.
<p>Disease development was evaluated in MRL/lpr mice after 11 weeks of treatment with hAAT or PBS (n = 10 per group). (A) Interscapular lesion surface (cm<sup>2</sup>). (B) Lymph node weight (cervical, brachial, and inguinal). (C) Terminal serum IFN-I, (D) BAFF and (E) TNF-α. (F) Terminal serum anti-dsDNA IgG. (G) Serum ANA staining. Representative images are shown on the left, and FITC relative intensities are graphed on the right. * P<0.05, and*** P<0.001 by Student’s <i>t</i>-test.</p