Intra hepatic leukocytes were responsible for HBcAg-induced TNF-α production in an <i>ex vivo</i> culture system.

<p>(A). C57BL/6 mice received pAAV/HBV1.2 or pAAV/core-null plasmids by hydrodynamic injection into a tail vein. At Day 3 post-injection, hepatocytes were isolated and cultured for 24<i> </i>h. The secretory level of HBsAg and intracellular HBcAg expression were determined by enzyme immunoassay (upper left panel) and Western blotting (upper right panel), respectively. After 24<i> </i>h of hepatocyte incubation, the intra hepatic leukocytes were isolated from naïve syngenic mice and co-cultured with hepatocytes for a further 12 or 24<i> </i>h. The TNF-α production in culture medium was measured by ELISA assay. (B) Wild-type C57BL/6 or TNF-α knockout mice were injected hydrodynamically with pAAV/HBV1.2 plasmids. Hepatocytes were isolated and cultured for 24<i> </i>h. Naïve intra hepatic leukocytes were isolated from wild-type C57BL/6 or TNF-α knockout mice and incubated with hepatocytes for 24<i> </i>h. The TNF-α level in culture supernatants was determined by ELISA assay.