Reducing environment protects sinusoidal lymphocytes isolated from normal human liver from apoptosis.
OBJECTIVE
We previously reported that the populations of lymphocytes and the expression of activated antigens in human sinusoidal mononuclear cells were different from those in peripheral blood mononuclear cells. Attempts to culture these cells for further study failed because they died rapidly under standard culture conditions in vitro after isolation from the liver. In this study, we evaluated the characteristics of cell death and the effects of various culture conditions on the viability of these cells.


METHODS
Sinusoidal mononuclear cells were isolated from University of Wisconsin solution that had been perfused through the portal veins of normal healthy human livers harvested for transplantation into living related recipients.


RESULTS
70% of sinusoidal mononuclear cells cultured in vitro were nonviable within 48 h after isolation, while only 10% of peripheral blood mononuclear cells died under the same conditions. Sinusoidal mononuclear cells showed DNA ladder formation of DNA on electrophoresis and characteristic morphological pattern on electron microscopic examination that suggested they had died in an apoptotic manner. The addition of human liver extracts or 2-mercaptoethanol and reduced glutathione to the cultures rescued the sinusoidal mononuclear cells from apoptosis. Furthermore, diamide, a sulfhydryl group specific oxidant, negated the effect of the liver extract.


CONCLUSIONS
In comparison with peripheral blood mononuclear cells, human sinusoidal mononuclear cells were more subject to death by apoptosis ex vivo, which was reversed by exogenous agents producing reducing conditions. These results suggested that hepatic sinusoidal mononuclear cells might express a different sensitivity to redox environment than peripheral blood mononuclear cells.
Kinoshita
K
K
Second Department of Surgery, College of Medical Technology, Kyoto University, Japan.
Kanai
M
M
Awane
M
M
Furuke
K
K
Shirakata
Y
Y
Gomi
T
T
Nakamura
Y
Y
Ikai
I
I
Yamauchi
A
A
Inamoto
T
T
Ohmori
K
K
Yamaoka
Y
Y
D017209:Apoptosis / Q000502:physiology*; D002470:Cell Survival; D002478:Cells, Cultured; D003958:Diamide / Q000494:pharmacology; D004587:Electrophoresis, Agar Gel; D005978:Glutathione / Q000494:pharmacology; D006801:Humans; D008110:Liver Extracts / Q000276:immunology*; D008214:Lymphocytes / Q000187:drug effects* / Q000473:pathology; D008623:Mercaptoethanol / Q000494:pharmacology; D010084:Oxidation-Reduction; D012016:Reference Values; D013438:Sulfhydryl Compounds / Q000494:pharmacology*; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D008110:Liver Extracts; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; D003958:Diamide; D008623:Mercaptoethanol; D005978:Glutathione