Potential conflict of interest: Nothing to report.
Invariant NKT cell activation induces neutrophil accumulation and hepatitis: Opposite regulation by IL-4 and IFN-γ
Version of Record online: 6 AUG 2013
Copyright © 2013 by the American Association for the Study of Liver Diseases
Volume 58, Issue 4, pages 1474–1485, October 2013
How to Cite
Wang, H., Feng, D., Park, O., Yin, S. and Gao, B. (2013), Invariant NKT cell activation induces neutrophil accumulation and hepatitis: Opposite regulation by IL-4 and IFN-γ. Hepatology, 58: 1474–1485. doi: 10.1002/hep.26471
This work was supported by the intramural program of NIAAA, NIH (to B.G.).
- Issue online: 1 OCT 2013
- Version of Record online: 6 AUG 2013
- Accepted manuscript online: 17 MAY 2013 08:42AM EST
- Manuscript Accepted: 11 APR 2013
- Manuscript Received: 20 FEB 2013
Additional Supporting Information may be found in the online version of this article.
|hep26471-sup-0001-suppinfo.doc||48K||Supporting Table 1: Primer sequences for mouse genes used in real-time PCR|
|hep26471-sup-0002-suppfig1.tif||4636K||Fig .1. IFN -γ-l- IL-4-l- dOK mice are more susceptible to α Galcer-induced liver injury and hepatitis than WT mice. WT and dKO mice were treated with α -Galcer, liver tissues were collected 16th post injection for H&E staining.|
|hep26471-sup-0003-suppfig2.tif||3950K||Supporting Fig.2. STAT6, a key downstream signaling molecule of IL-4 is required for α -Galcer-induced hepatitis. WT and STAT6-/- mice were treated with α-Galcer and euthanized 72h post injection. Liver tissues were collected for H&E staining. Representative photos of H&E-stained liver tissues are shown.|
|hep26471-sup-0004-suppfig3.tif||481K||Supporting Fig. 3. MNCs from WT and IFN-γ-l-mice have similar cytotoxicity after α-Galcer treatment in vivo and vitro. (A) Liver MNCs were isolated from WT r IFN-γ-l- mice, and stimulated with α-Galcer (50ng/ml) for 3 hours, then incubated with primary hepatocytes for an additional 4 hours. Cytotoxicity was measured. (B) WT and IFN-γ-l- mice were treated with α-Galer for 3 hours, followed by isolation of liver MNCs. Liver MNCs were then incubated with primary mouse hepatocytes for 4 hours. Cytotovicity was measured.|
|hep26471-sup-0005-suppfig4.tif||307K||Supporting Fig. 4 Depletion of NK cells by using anti-ASGM1 Ab does not affect serum ALT and AST levels in α-Galcer-treated IFN-γ-l- mice. IFN-γ-l- mice were treated with anti-ASGM1 Ab for 24 hours, followed by injection of a single dose of α-Galcer. Mice were euthanized 12 hours post α-Galcer injection, and serum ALT and AST levels were measured.|
|hep26471-sup-0006-suppfig5.tif||2911K||Supporting Fig. 5 Depletion of neutrophils absolished α-Galcer-induced liver injury in IFN-γ-l- mice. IFN-γ-l- mice were treated with control or anti-Ly6G antibodies, as described in the Methods, followed by injection. Liver tissues were stained with H&E. The red arrow indicates massive necrosis.|
|hep26471-sup-0007-suppfig6.tif||325K||Supporting Fig. 6 Decreased expression chemokines in the livers from IL-4-l- or STAT6-l- mice compared with those from WT mice. Real-time PCR analyses of liver tissues from WT, lL-4-l-, and mice treated with or without α-Galcer. *P<0.05, **P<0.01.|
|hep26471-sup-0008-suppfig7.tif||385K||Supporting Fig. 7 Hepatic expression of pro-inflammatory cytokines from α-Galcer-treated WT, lL-4-l- mice. Real-time PCR analyses of liver tissues from WT, lL-4-l-, IFN-γ-l- and mice treated with or without α-Galcer. *P<0.01. HD: not determined.|
|hep26471-sup-0009-suppfig8.tif||579K||Supporting Fig. 8 Neutrophil activation from α-Galcer-treaded WT, lL-4-l- or IFN-γ-l- mice. Liver PNMs were isolated from α-Galcer-treated WT, lL-4-l- mice (A) These liver PMNs were subjected to FACS analyses of neutrophil activation markers on neutrophils. Neturophil activation is associated with up-regulation of CD11b and down-regulation of CD62L expression. (B) These liver PMNs were subjected to FACS analyses of neutrophil ROS production. (C) These liver PMNs were stimulated with PMA in vitro, followed by FACS analyses of neutrophil ROS production.|
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