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Updated: Jun 9, 2026
Kupffer Cell Isolation for Nanoparticle Toxicity Testing
Published on: August 18, 2015
Characterization of two F4/80-positive Kupffer cell subsets by their function and phenotype in mice
Manabu Kinoshita1, Takefumi Uchida, Atsushi Sato
1Department of Immunology and Microbiology, National Defense Medical College, 3-2 Namiki, Tokorozawa, Saitama 359-8513, Japan.
Background & Aims:
Liver Kupffer cells have been suggested to be heterogeneous macrophage lineage cells. We explored this possibility by classifying the mouse Kupffer cells into subpopulations and characterizing them by their phenotype and function.
Methods:
Liver mononuclear cells (MNCs) from C57BL/6 mice were isolated and their phenotypes and functions were analyzed. The effects of clodronate liposomes and gadolinium chloride (GdCl(3)) on Kupffer cells were also investigated.
Results:
Approximately 25% of liver MNCs were F4/80(+) Kupffer cells. Of these, 46% were CD11b(-)CD68(+), 22% were CD11b(+)CD68(-), and 6% were CD11b(+)CD68(+). CD68(+) cells showed potent phagocytic activity and reactive oxygen species (ROS) production capacity after lipopolysaccharide (LPS) stimulation, whereas CD11b(+) cells did not. CD11b(+) cells showed a strong capacity for the production of cytokines (TNF and IL-12), which was much less prominent in CD68(+) cells. At 24h after LPS or Escherichia coli injection into mice, the proportions of CD11b(+)CD68(-) and CD11b(+)CD68(+) cells increased but that of CD11b(-)CD68(+) cells decreased. The increase in CD11b(+)CD68(+) cells appeared to be derived from the CD11b(+)CD68(-) subset. Although the CD11b(+) cells augmented phagocytic activity after LPS injection, they did not increase ROS production, suggesting their weak lytic activity. Injection of clodronate or GdCl(3) into mice depleted the CD68(+) cells but increased CD11b(+) cells proportionally because CD68(+) cells may phagocytose these toxic reagents and undergo apoptosis. GdCl(3)-treated mice also consistently increased serum TNF after LPS challenge.
Conclusions:
Two F4/80(+) Kupffer cell subsets may exist, a CD68(+) subset with phagocytic activity and a CD11b(+) subset with cytokine-producing capacity.
Insights
Two distinct mouse Kupffer cell subsets, CD68(+) and CD11b(+), were identified. CD68(+) cells excel at phagocytosis, while CD11b(+) cells are key for cytokine production, revealing Kupffer cell heterogeneity.
Area of Science:
- Immunology
- Cell Biology
- Macrophage Biology
Background:
- Liver Kupffer cells, resident macrophages in the liver, are crucial for immune responses.
- Previous research suggested potential heterogeneity within the Kupffer cell population.
- Understanding Kupffer cell subpopulations is vital for comprehending liver immunity and disease.
Purpose of the Study:
- To investigate the heterogeneity of mouse liver Kupffer cells.
- To classify Kupffer cells into distinct subpopulations based on phenotype.
- To characterize the functional differences between identified Kupffer cell subsets.
Main Methods:
- Isolation of liver mononuclear cells (MNCs) from C57BL/6 mice.
- Phenotypic analysis of Kupffer cells using markers such as F4/80, CD11b, and CD68.
- Functional assays including phagocytosis, reactive oxygen species (ROS) production, and cytokine (TNF, IL-12) secretion.
- Investigation of the effects of clodronate liposomes and gadolinium chloride (GdCl(3)) on Kupffer cell populations.
Main Results:
- Kupffer cells (F4/80+) comprised approximately 25% of liver MNCs.
- Three main subpopulations were identified: CD11b(-)CD68(+) (46%), CD11b(+)CD68(-) (22%), and CD11b(+)CD68(+) (6%).
- CD68(+) cells exhibited potent phagocytic activity and ROS production, while CD11b(+) cells were characterized by high cytokine production (TNF, IL-12).
- Treatment with clodronate or GdCl(3) depleted CD68(+) cells and proportionally increased CD11b(+) cells.
Conclusions:
- Evidence suggests the existence of at least two distinct F4/80(+) Kupffer cell subsets in the mouse liver.
- A CD68(+) subset demonstrates significant phagocytic capabilities.
- A CD11b(+) subset is characterized by its capacity for cytokine production, highlighting functional specialization within Kupffer cells.
