Journal of Leukocyte Biology
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© 2003 by The Society for Leukocyte Biology
Journal of Leukocyte Biology, doi:10.1189/jlb.0302133

Received for publication January 23, 2003.

Accepted for publication February 19, 2003.

Article

Generation and functional characterization of a clonal murine periportal Kupffer cell line from H-2Kb -tsA58 mice

Daniel Dory *, Hakim Echchannaoui *, Maryse Letiembre *, Fabrizia Ferracin *, Jean Pieters {dagger}, Yoshiyuki Adachi {ddagger}, Sachiko Akashi §, Werner Zimmerli *, and Regine Landmann *@

*Division of Infectious Diseases, Department of Research, University Hospital, Basel, Switzerland; {dagger}Department of Biochemistry, Biozentrum, University of Basel, Switzerland; {ddagger}Laboratory of Immunopharmacology of Microbial Products, Tokyo University of Pharmacy and Life Science, Japan; and §Division of Infectious Genetics, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Japan

@ To whom correspondence should be addressed. E-mail: Regine.Landmann{at}unibas.ch.


  Abstract

Murine Kupffer cells (KCs) are heterogeneous and survive only for a short time in vitro. Here, a clonal, murine KC line was generated from transgenic mice, expressing the thermolabile mutant tsA58 of the Simian virus 40 large T antigen under the control of the H-2Kb promoter. Thirty-three degrees Celcius and 37°C but not 39°C have been permissive for growth of the clone; it required conditioned media from hepatocytes and endothelial cells for proliferation. In contrast to primary cells, the cells of the clone were uniform, survived detachment, and could therefore be analyzed by cytofluorimetry. The clone, as primary KCs, constitutively expressed nonspecific esterase, peroxidase, MOMA-2, BM8, scavenger receptor A, CD14, and Toll-like receptor 4 (TLR4); the antigen-presenting molecules CD40, CD80, and CD1d; and endocytosed dextran-fluorescein isothiocyanate. It lacked complement, Fc receptors, F4/80 marker, and the phagosomal coat protein TACO. The clone exhibited CD14- and TLR4/MD2-independent, plasma-dependent lipopolysaccharide (LPS) binding, Escherichia coli and Streptococcus pneumoniae phagocytosis, and LPS- and interferon-{gamma}-induced NO production but no tumor necrosis factor {alpha}, interleukin (IL)-6, or IL-10 release. The large size, surface-marker expression, and capacity to clear gram-negative and -positive bacteria indicate that the clone was derived from the periportal, large KC subpopulation. The clone allows molecular studies of anti-infective and immune functions of KCs.

Key Words: mouse • lipopolysaccharide • nitric oxide • phagocytosis




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