Abstract
The eastern woodchuck, Marmota monax, represents a useful animal model to study hepatitis B virus infection in humans. However, immunological studies in this model have been impeded by a lack of basic information about the components of the immune system such as cytokines and chemokines. To clarify the role(s) of interleukin 8 (IL-8) in chronic hepatitis B and hepatocellular carcinoma (HCC) in the woodchuck model, we cloned and characterized the woodchuck IL-8 cDNA and genomic DNA. Sequence analysis revealed that the organization of the wk-IL-8 gene is similar to that of the human IL-8 gene and consists of four exons and three introns. Woodchuck IL-8 protein exhibits the conserved ELRCXC motif of IL-8 and shows 87, 82, 82 and 79% similarity with rabbit, ovine, bovine and human IL-8 proteins, respectively. The biological activity of wk-IL-8 was demonstrated using neutrophil chemotaxis assays. Wk-IL-8 could be readily detected in both tumor and non-tumor tissues with higher expression in the non-tumor tissues in most cases. The results from this study will facilitate the investigation of IL-8 in the immunopathogenesis of hepadnavirus-related diseases by the woodchuck model.
Highlights
Hepatitis B virus (HBV) causes acute and chronic hepatitis and leads to the development of life-threatening liver diseases, including cirrhosis and hepatocellular carcinoma (HCC).[1]
We describe the cloning of woodchuck IL-8 cDNA and its genomic DNA
A PCR-based strategy with primers designed to areas of high homology observed in multiple alignments of known mammalian IL-8 gene sequences was used to clone the woodchuck IL-8 (wk-IL-8) cDNA
Summary
Hepatitis B virus (HBV) causes acute and chronic hepatitis and leads to the development of life-threatening liver diseases, including cirrhosis and hepatocellular carcinoma (HCC).[1]. The biological activities of to mediate angiogenesis.[22,23] Residues 57–61 (Ser[57], Gly[58], Pro[59], His[60] and Cys61) form an atypical b turn and, together with the disulfide bridges, provide a structural the cloned wk-IL-8 cDNA were demonstrated and the scaffold for the NH2 terminal region[24] and play sigexpression of wk-IL-8 in liver tissues from woodchucks nificant roles in ligand–receptor interactions. ELR and with chronic hepatitis and HCC was characterized These GPH motif (Gly[58], Pro[59], His60) in the 57–61 turn have data provide a basis for further studies of IL-8 in the been suggested to be important for receptor-binding woodchuck model
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