Generation and functional characterization of a clonal murine periportal Kupffer cell line from H-2Kb -tsA58 mice.
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-2K(b) promoter. Thirty-three degrees Celsius and 37 degrees C but not 39 degrees 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 tryptophan aspartate-containing 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.
Dory
Daniel
D
Division of Infectious Diseases, Department of Research, University Hospital, Basel, Switzerland.
Echchannaoui
Hakim
H
Letiembre
Maryse
M
Ferracin
Fabrizia
F
Pieters
Jean
J
Adachi
Yoshiyuki
Y
Akashi
Sachiko
S
Zimmerli
Werner
W
Landmann
Regine
R
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods; D002461:Cell Line, Transformed; D002999:Clone Cells; D016207:Cytokines / Q000096:biosynthesis; D004705:Endocytosis; D004798:Enzymes / Q000378:metabolism; D007728:Kupffer Cells / Q000737:chemistry / Q000166:cytology* / Q000276:immunology; D008070:Lipopolysaccharides / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D009569:Nitric Oxide / Q000096:biosynthesis; D010587:Phagocytosis; D010641:Phenotype; D011401:Promoter Regions, Genetic
D000952:Antigens, Polyomavirus Transforming; D016207:Cytokines; D004798:Enzymes; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D009569:Nitric Oxide