Related Experiment Video
Updated: Aug 19, 2025
Multicellular Human Alveolar Model Composed of Epithelial Cells and Primary Immune Cells for Hazard Assessment
Published on: May 6, 2020
Establishment and characterization of the immortalized porcine lung-derived mononuclear phagocyte cell line
Takato Takenouchi1, Kentaro Masujin2, Shunichi Suzuki1
1Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Tsukuba, Japan.
Abstract:
Mononuclear phagocytes (MNP), including monocytes, dendritic cells (DC), and macrophages, play critical roles in innate immunity. MNP are abundant in the lungs and contribute to host defense against airborne agents and pulmonary immune homeostasis. In this study, we isolated porcine lung-derived MNP (PLuM) from primary cultures of parenchymal lung cells and then immortalized them by transferring the SV40 large T antigen gene and porcine telomerase reverse transcriptase gene using lentiviral vectors. The established cell line, immortalized PLuM (IPLuM), expressed DC/macrophage markers; i.e., CD163, CD172a, and major histocompatibility complex class II, whereas they did not express a porcine monocyte-specific marker, CD52. The expression patterns of these cell surface markers indicate that IPLuM originate from the DC/macrophage lineage rather than the monocyte lineage. The bacterial cell wall components muramyl dipeptide and lipopolysaccharide induced the production of the interleukin-1 family of pro-inflammatory cytokines in IPLuM. Phagocytotic activity was also detected by time-lapse fluorescence imaging of live cells when IPLuM were cultured in the presence of pHrodo dye-conjugated E. coli BioParticles. It is worth noting that IPLuM are susceptible to African swine fever virus infection and support the virus' efficient replication in vitro. Taken together, the IPLuM cell line may be a useful model for investigating host-agent interactions in the respiratory microenvironments of the porcine lung.
Insights
A new cell line of immortalized porcine lung mononuclear phagocytes (IPLuM) was developed. This IPLuM cell line effectively models respiratory immune responses and African swine fever virus infection in pigs.
Area of Science:
- Immunology
- Cell Biology
- Virology
Background:
- Mononuclear phagocytes (MNP), including monocytes, dendritic cells (DC), and macrophages, are crucial for lung immunity and homeostasis.
- Porcine lung-derived MNP (PLuM) are vital for host defense against airborne pathogens.
Purpose of the Study:
- To establish and characterize an immortalized cell line of porcine lung mononuclear phagocytes (IPLuM).
- To evaluate the utility of IPLuM as a model for studying host-agent interactions in the porcine respiratory system.
Main Methods:
- Isolation of porcine lung-derived MNP (PLuM) from primary lung cell cultures.
- Immortalization of PLuM using lentiviral vectors carrying SV40 large T antigen and porcine telomerase reverse transcriptase genes.
- Characterization of the IPLuM cell line using cell surface marker expression, cytokine production assays, phagocytosis assays, and African swine fever virus infection studies.
Main Results:
- The established IPLuM cell line expressed DC/macrophage markers (CD163, CD172a, MHC class II) but not a monocyte-specific marker (CD52), indicating a DC/macrophage lineage origin.
- IPLuM produced pro-inflammatory cytokines (interleukin-1 family) in response to bacterial components (muramyl dipeptide, lipopolysaccharide).
- IPLuM exhibited phagocytic activity and supported efficient African swine fever virus replication in vitro.
Conclusions:
- The IPLuM cell line represents a valuable tool for studying innate immunity and host-pathogen interactions in the porcine lung.
- This cell line can be utilized to investigate respiratory immune responses and viral pathogenesis, particularly African swine fever virus.
- IPLuM provides an in vitro model for understanding the complex interplay between porcine lung immune cells and infectious agents.
