Quantification of angiogenic and inflammatory mRNA signals under various experimental conditions

type 1 infected livers demonstrated significantly higher expression of both SOCS3 and AXL, with no effect of liver fibrosis grade on expression of either SOCS3 of AXL in HCV infected livers. AXL expression also demonstrated a strong correlation with viral load, supporting the direct role of HCV in AXL induction. The rs12979860 IFNL3 CC SNP correlated with lower baseline hepatic AXL expression in genotype 1 HCV infected livers and JFH1 infected PHHs. PBMC expression of AXL was up-regulated in HCV infected individuals, and similar to hepatic expression, was lower in patients possessing the CC genotype. Stronger AXL up-regulation in PBMCs 12 h after the first IFN injection was also observed . doi:10.1371/journal.pone.0136227.g006 observed between hepatic AXL expression and HCV viral load, suggesting that increased viral replication and subsequent activation of immune signalling pathways may drive AXL expression in the liver. We next examined AXL expression in liver and blood from HCV infected patients, stratified by the IFNL3 rs12979860 SNP. Hepatic AXL expression was higher in patients with the nonresponder rs12979860 CT/TT genotype, consistent with published data showing elevated hepatic ISGs in patients possessing the IFNL3 non-responder haplotype. To examine the effect of IFNL3 genotype on AXL in a PHH model, we utilized publicly available microarray data examining expression of primary human hepatocytes infected with HCV. Laser capture microdissection was utilized to compare infected and infection adjacent cells, and was further stratified by IFNL3 rs12979860 genotype. By analysing AXL expression in both infected and adjacent PHHs we found similar results as in vivo HCV infection, with elevated AXL expression in HCV infected PHH isolated from patients with the “non-responder” rs12979860 CT/TT genotype. To determine whether AXL induction is mediated directly by viral infection or by subsequent cytokine expression, we examined AXL expression in infected, adjacent and mock infected PHHs from the same dataset. AXL expression increased in infected cells from day 1 to day 7, and was consistently higher in infected cells than either adjacent cells PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19729663 or mock 11 / 16 HCV Induced AXL Suppresses the Hepatic Type I Interferon Response infected controls. This suggests that although cytokines may contribute to AXL expression in vivo, AXL up-regulation occurs primarily in infected PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19728767 cells. AXL expression was also significantly elevated in peripheral blood mononuclear cells from HCV infected individuals compared to healthy controls . Furthermore, patients possessing the responder rs12979860 CC genotype displayed lower AXL expression than CT/TT carriers, in agreement with hepatic AXL expression. Finally, induction of AXL 12 h after the first dose of pegylated IFN was stronger in PBMCs from patients possessing the responder CC genotype, compared with CT/TT nonresponder patients. Discussion We previously demonstrated that AXL is up-regulated following HCV infection in vitro, and may contribute to SOCS3 expression and interferon refractoriness. We now demonstrate that AXL is induced in a genotype dependent manner in vitro as well as in vivo, and is likely induced by IFNs and other inflammatory 487-52-5 site mediators. AXL knockdown showed little effect on IFN signalling, however AXL overexpression reduced Huh-7 responsiveness to IFN, as well as the antiviral response against HCV. Our current findings build on our previous report that HCV induces AXL expression in vitro. W