RNA sequencing analyses of gene expressions in a canine macrophages cell line DH82 infected with canine distemper virus

Zheng, Xiaodong; Zhu, Yanyan; Zhao, Zhiguang; Yan, Lei; Xu, Tong; Wang, Xianwei; He, Hongbin; Xia, Xianzhu; Zheng, Wenwen; Xue, Xiaolin

doi:10.1016/j.meegid.2020.104206

Cited by 5 publications

(4 citation statements)

References 57 publications

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“…When compared to larger human in vitro datasets also obtained using mRNA sequencing, we found that canine polarized macrophages also matched transcriptomic patterns seen in human M1 and M2 macrophages (11). Compared to published datasets of single cell sequencing in human M1 or M2 macrophages, STAT3 and STAT1 upregulation in M1 macrophages (7,15,36). Future studies are underway in our lab, which use single cell RNA sequencing to better define the heterogeneity of canine macrophages in normal tissues as well as tumors.…”

Section: Discussionmentioning

confidence: 99%

“…When macrophages are exposed to IFN-γ, a series of stereotypic changes occur which result in an inflammatory macrophage phenotype. These changes include: changes to surface protein expression, upregulation of anti-tumor and anti-microbial activity, secretion of inflammatory cytokines, and enhanced antigen presentation processes (2, 3, [7][8][9][10][11]. In contrast, exposure to IL-4 and IL-13 promote the differentiation of macrophages toward anti-inflammatory phenotype.…”

Section: Introductionmentioning

confidence: 99%

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Canine polarized macrophages express distinct functional and transcriptomic profiles

Chow

Soontararak²,

Wheat³

et al. 2022

Front. Vet. Sci.

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Macrophage differentiation and function in disease states is highly regulated by the local microenvironment. For example, macrophage exposure to IFN-γ (interferon gamma) initiates the development of inflammatory (M1) macrophages, which acquire anti-tumoral and antimicrobial activity, while exposure to IL-4 (interleukin-4) and IL-13 (interleukin-13) drives an anti-inflammatory (M2) macrophage phenotype, which promotes healing and suppression of inflammatory responses. Previous studies of canine polarized macrophages have identified several surface markers that distinguished GM-CSF (granulocyte macrophage colony stimulating factor), IFN-γ and LPS (lipopolysaccharide) derived M1 macrophages or M2 macrophages; and reported a subset of genes that can be used to differentiate between polarization states. However, the need remains to understand the underlying biological mechanisms governing canine macrophage polarization states. Therefore, in the present study we used transcriptome sequencing, a larger panel of flow cytometry markers, and the addition of antimicrobial functional assays to further characterize canine macrophage polarization. Transcriptome analysis revealed unique, previously unreported signatures and pathways for polarized canine M1 and M2 macrophages. New flow cytometric markers were also identified, along with new characterization of how macrophage polarization impacted antimicrobial functions. Taken together, the findings reported here provide new insights into canine macrophage biology and identify new tools for the evaluation of polarized macrophages in dogs.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Canine polarized macrophages express distinct functional and transcriptomic profiles

Chow

Soontararak²,

Wheat³

et al. 2022

Front. Vet. Sci.

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“…Nevertheless, tumor cell lines have previously been used in RNA-seq studies. More specifically, DH82 cells have been employed to explore their interaction with canine distemper virus, finding important variations in gene expression patterns after infection [26], although their use with Leishmania has not been described to date. In the case of L. infantum, another assay performed in 2020 used the human monocyte cell line THP-1 to analyze inflammasome activation and in agreement with our results, the authors described a poor response of cell line to the infection [27].…”

Section: Infection With L Infantum Bcn150 and Bos1fl1 Induces Poor Regulation Of The Dh82 Canine Macrophage Gene Expression Profilementioning

confidence: 99%

Transcriptomic Profile of Canine DH82 Macrophages Infected by Leishmania infantum Promastigotes with Different Virulence Behavior

Mas

Martínez‐Rodrigo

Carrión

et al. 2022

IJMS

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Zoonotic visceral leishmaniosis caused by Leishmania infantum is an endemic disease in the Mediterranean Basin affecting mainly humans and dogs, the main reservoir. The leishmaniosis outbreak declared in the Community of Madrid (Spain) led to a significant increase in human disease incidence without enhancing canine leishmaniosis prevalence, suggesting a better adaptation of the outbreak’s isolates by other host species. One of the isolates obtained in the focus, IPER/ES/2012/BOS1FL1 (BOS1FL1), has previously demonstrated a different phenotype than the reference strain MCAN/ES/1996/BCN150 (BCN150), characterized by a lower infectivity when interacting with canine macrophages. Nevertheless, not enough changes in the cell defensive response were found to support their different behavior. Thus, we decided to investigate the molecular mechanisms involved in the interaction of both parasites with DH82 canine macrophages by studying their transcriptomic profiles developed after infection using RNA sequencing. The results showed a common regulation induced by both parasites in the phosphoinositide-3-kinase–protein kinase B/Akt and NOD-like receptor signaling pathways. However, other pathways, such as phagocytosis and signal transduction, including tumor necrosis factor, mitogen-activated kinases and nuclear factor-κB, were only regulated after infection with BOS1FL1. These differences could contribute to the reduced infection ability of the outbreak isolates in canine cells. Our results open a new avenue to investigate the true role of adaptation of L. infantum isolates in their interaction with their different hosts.

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“…Firstly, the original study revealed that ATP could alter cytokine expression in LPS-stimulated DH82 cells [ 26 ]. Secondly, infection of DH82 cells with canine distemper virus modulates inflammatory signalling pathways [ 27 ] that are common to P2X receptor-mediated signalling [ 28 ]. Thirdly, despite few studies analysing the expression of P2 receptors in dogs, it has been demonstrated that canine monocytes express P2X7 receptors [ 29 , 30 ].…”

Section: Introductionmentioning

confidence: 99%

P2Y2 and P2X4 Receptors Mediate Ca2+ Mobilization in DH82 Canine Macrophage Cells

Sophocleous

Miles

Ooi

et al. 2020

IJMS

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Purinergic receptors of the P2 subclass are commonly found in human and rodent macrophages where they can be activated by adenosine 5′-triphosphate (ATP) or uridine 5′-triphosphate (UTP) to mediate Ca2+ mobilization, resulting in downstream signalling to promote inflammation and pain. However, little is understood regarding these receptors in canine macrophages. To establish a macrophage model of canine P2 receptor signalling, the expression of these receptors in the DH82 canine macrophage cell line was determined by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry. P2 receptor function in DH82 cells was pharmacologically characterised using nucleotide-induced measurements of Fura-2 AM-bound intracellular Ca2+. RT-PCR revealed predominant expression of P2X4 receptors, while immunocytochemistry confirmed predominant expression of P2Y2 receptors, with low levels of P2X4 receptor expression. ATP and UTP induced robust Ca2+ responses in the absence or presence of extracellular Ca2+. ATP-induced responses were only partially inhibited by the P2X4 receptor antagonists, 2′,3′-O-(2,4,6-trinitrophenyl)-ATP (TNP-ATP), paroxetine and 5-BDBD, but were strongly potentiated by ivermectin. UTP-induced responses were near completely inhibited by the P2Y2 receptor antagonists, suramin and AR-C118925. P2Y2 receptor-mediated Ca2+ mobilization was inhibited by U-73122 and 2-aminoethoxydiphenyl borate (2-APB), indicating P2Y2 receptor coupling to the phospholipase C and inositol triphosphate signal transduction pathway. Together this data demonstrates, for the first time, the expression of functional P2 receptors in DH82 canine macrophage cells and identifies a potential cell model for studying macrophage-mediated purinergic signalling in inflammation and pain in dogs.

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RNA sequencing analyses of gene expressions in a canine macrophages cell line DH82 infected with canine distemper virus

Cited by 5 publications

References 57 publications

Canine polarized macrophages express distinct functional and transcriptomic profiles

Canine polarized macrophages express distinct functional and transcriptomic profiles

Transcriptomic Profile of Canine DH82 Macrophages Infected by Leishmania infantum Promastigotes with Different Virulence Behavior

P2Y2 and P2X4 Receptors Mediate Ca2+ Mobilization in DH82 Canine Macrophage Cells

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