Kc. Thompson et al., Primary rat and mouse hepatic stellate cells express the macrophage inhibitor cytokine interleukin-10 during the course of activation in vitro, HEPATOLOGY, 28(6), 1998, pp. 1518-1524
Activation of local tissue macrophages (Kupffer cells) and of quiescent hep
atic stellate cells (HSCs) to a myofibroblast phenotype are two key events
in liver inflammation and fibrosis. It is known that products of activated
macrophages may activate stellate cells. We have hypothesized that the prod
ucts of activated HSCs may also modulate the activity of Kupffer cells. The
cytokine interleukin-10 (IL-10), produced by lymphocytes and macrophages,
has profound inhibitory actions on macrophages. Normal rat and mouse HSCs t
hat differentiate in vivo and in vitro to activated myofibroblasts were iso
lated by enzyme perfusion and density centrifugation with or without centri
fugal elutriation, confirmed by vitamin A autofluorescence and positive imm
unostaining for the myofibroblast markers desmin and smooth muscle actin (S
MA). Conditioned media and lysates from these cells were found to downregul
ate lipopolysaccharide (LPS)-induced tumor necrosis factor-alpha (TNF-alpha
) secretion by the mouse macrophage line RAW 267.4. In highly purified prep
arations of rat HSCs, messenger RNA (mRNA) for IL-10 was detected by revers
e-transcription polymerase chain reaction (RT-PCR), from the time of isolat
ion to up to 120 days of culture on plastic. Long-term cultures of unstimul
ated mouse HSCs secreted IL-10 protein as detected by immunoblotting and sp
ecific enzyme-linked immunosorbent assay (ELISA). IL-10 protein was undetec
table by immunohistochemistry in mouse HSCs for the first 3 days in culture
. After this, the percentage of IL-10-positive cells increased to 45% at da
y 7 and 100% by day 14, and expression of IL-10 continued in long-term cult
ures of up to 120 days. The expression of IL-10 by the stromal cells that g
overn the fibrotic process in the liver may have important implications for
the regulation of inflammation and fibrosis in the liver.