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A Simple and Efficient Method to Isolate Macrophages from Mixed Primary Cultures of Adult Liver Cells
Published on: May 24, 2011
Antimicrobial properties of Kupffer cells
1Infectious Disease Section, Veterans Administration Medical Center, Minneapolis, Minnesota 55417.
Abstract:
To characterize the antimicrobial activities of Kupffer cells, I harvested macrophages from livers with a technique involving perfusion with collagenase and DNase. Ninety-nine percent of glass-adherent cells had typical macrophage morphology, 99% were esterase positive, and 60% phagocytosed opsonized zymosan when challenged with four particles per macrophage. Toxoplasma gondii multiplied within Kupffer cells from unmanipulated mice, but multiplication was intermediate between that observed in highly permissive peritoneal macrophages and highly activated macrophages. Intravenous injection of heat-killed Propionibacterium acnes, a stimulus known to activate macrophages in other compartments, resulted in a uniform, highly activated population of liver macrophages. Kupffer cells from P. acnes-injected mice were capable of generating reactive oxygen intermediates as shown by reduction of Nitro Blue Tetrazolium during phagocytosis of T. gondii or opsonized zymosan. In contrast, intravenous P. acnes injection did not activate spleen macrophages. Intravenous injection of P. acnes into athymic mice activated Kupffer cells, which suggested that T cells were not essential for this response. Kupffer cells were not activated in mice with latent Toxoplasma infection or during acute Giardia muris infection. Ordinarily, Kupffer cells became highly permissive for T. gondii during 48 h in culture, but inclusion of recombinant murine gamma interferon maintained their moderate inhibitory activity.
Insights
Kupffer cells, liver macrophages, exhibit antimicrobial activity and can be activated by Propionibacterium acnes. Activation enhances their ability to combat pathogens like Toxoplasma gondii.
Area of Science:
- Immunology
- Cell Biology
- Microbiology
Background:
- Kupffer cells are resident liver macrophages crucial for immune surveillance.
- Understanding Kupffer cell antimicrobial functions is vital for liver immunity.
Purpose of the Study:
- To characterize the antimicrobial activities and activation states of Kupffer cells.
- To investigate Kupffer cell responses to microbial stimuli and infections.
Main Methods:
- Isolation of liver macrophages (Kupffer cells) using collagenase and DNase perfusion.
- Assessing Kupffer cell morphology, esterase activity, and phagocytosis of zymosan.
- Evaluating Toxoplasma gondii multiplication within Kupffer cells.
- Stimulating Kupffer cells with heat-killed Propionibacterium acnes (P. acnes) and assessing reactive oxygen intermediate generation via Nitro Blue Tetrazolium reduction.
- Investigating Kupffer cell activation in athymic mice and during parasitic infections (T. gondii, Giardia muris).
- Assessing the effect of gamma interferon on Kupffer cell permissiveness to T. gondii in vitro.
Main Results:
- Isolated Kupffer cells demonstrated typical macrophage characteristics and phagocytic capacity.
- T. gondii multiplied in Kupffer cells, with intermediate replication compared to other macrophage types.
- Intravenous P. acnes injection uniformly activated Kupffer cells, enhancing their capacity to generate reactive oxygen intermediates.
- P. acnes activated Kupffer cells but not spleen macrophages.
- Kupffer cell activation by P. acnes was independent of T cells.
- Kupffer cells were not activated during latent T. gondii or acute Giardia muris infections.
- In vitro, Kupffer cells became permissive to T. gondii, an effect partially inhibited by gamma interferon.
Conclusions:
- Kupffer cells possess inherent antimicrobial functions and can be significantly activated by P. acnes.
- Activated Kupffer cells exhibit enhanced pathogen-killing capabilities, including reactive oxygen intermediate production.
- Kupffer cell activation pathways are distinct from systemic responses and T cell-dependent activation is not required.
- These findings highlight Kupffer cells as key players in liver innate immunity with potential for therapeutic modulation.
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