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Establishment and characterization of the immortalized porcine lung-derived mononuclear phagocyte cell line

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...

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Autores principales: Takenouchi, Takato, Masujin, Kentaro, Suzuki, Shunichi, Haraguchi, Seiki, Hiramatsu, Kanae, Kokuho, Takehiro, Uenishi, Hirohide
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9715978/
https://www.ncbi.nlm.nih.gov/pubmed/36467652
http://dx.doi.org/10.3389/fvets.2022.1058124
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author Takenouchi, Takato
Masujin, Kentaro
Suzuki, Shunichi
Haraguchi, Seiki
Hiramatsu, Kanae
Kokuho, Takehiro
Uenishi, Hirohide
author_facet Takenouchi, Takato
Masujin, Kentaro
Suzuki, Shunichi
Haraguchi, Seiki
Hiramatsu, Kanae
Kokuho, Takehiro
Uenishi, Hirohide
author_sort Takenouchi, Takato
collection PubMed
description 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.
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spelling pubmed-97159782022-12-03 Establishment and characterization of the immortalized porcine lung-derived mononuclear phagocyte cell line Takenouchi, Takato Masujin, Kentaro Suzuki, Shunichi Haraguchi, Seiki Hiramatsu, Kanae Kokuho, Takehiro Uenishi, Hirohide Front Vet Sci Veterinary Science 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. Frontiers Media S.A. 2022-11-18 /pmc/articles/PMC9715978/ /pubmed/36467652 http://dx.doi.org/10.3389/fvets.2022.1058124 Text en Copyright © 2022 Takenouchi, Masujin, Suzuki, Haraguchi, Hiramatsu, Kokuho and Uenishi. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Takenouchi, Takato
Masujin, Kentaro
Suzuki, Shunichi
Haraguchi, Seiki
Hiramatsu, Kanae
Kokuho, Takehiro
Uenishi, Hirohide
Establishment and characterization of the immortalized porcine lung-derived mononuclear phagocyte cell line
title Establishment and characterization of the immortalized porcine lung-derived mononuclear phagocyte cell line
title_full Establishment and characterization of the immortalized porcine lung-derived mononuclear phagocyte cell line
title_fullStr Establishment and characterization of the immortalized porcine lung-derived mononuclear phagocyte cell line
title_full_unstemmed Establishment and characterization of the immortalized porcine lung-derived mononuclear phagocyte cell line
title_short Establishment and characterization of the immortalized porcine lung-derived mononuclear phagocyte cell line
title_sort establishment and characterization of the immortalized porcine lung-derived mononuclear phagocyte cell line
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9715978/
https://www.ncbi.nlm.nih.gov/pubmed/36467652
http://dx.doi.org/10.3389/fvets.2022.1058124
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