Time Course Study of Cytokine mRNA Expression in LPS-Stimulated Porcine Alveolar Macrophages

Choi, In Soo; Shin, Namsoo; Shin, Sung Jae; Lee, Deog‐Yong; Cho, Young Wook; Yoo, Han Sang

doi:10.4142/jvs.2002.3.2.97

Cited by 12 publications

(4 citation statements)

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“…This was similar to other data observed in LPS-stimulated porcine epithelial cells [25]. Our data, however, showed some di¡erences compared to other data observed in LPS-stimulated porcine alveolar macrophages [26]. These di¡erences might be due to di¡erent experimental conditions, including the analyzing method.…”

Section: Discussionsupporting

confidence: 88%

Kinetic study of porcine GM-CSF expression in porcine alveolar macrophages and spleen cells

Cho

Lee

Shin

et al. 2003

FEMS Immunology &Amp; Medical Microbiology

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Granulocyte-macrophage colony-stimulating factor (GM-CSF) is an important regulator in inducing differentiation and proliferation of immune cells. The functional roles of porcine GM-CSF (pGM-CSF) have not yet been revealed. Therefore, expression patterns of pGM-CSF were investigated in immune cells after cloning and sequencing of whole pGM-CSF cDNA. Whole cDNA of pGM-CSF was amplified from porcine alveolar macrophages stimulated by lipopolysaccharide (LPS), using 5'- and 3'-rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR) methods. The products of 5'- and 3'-RACE-PCR were cloned, and the nucleotide sequence of whole pGM-CSF cDNA was determined (GenBank accession number AY116504). The kinetics of pGM-CSF mRNA expression were studied in porcine immune cells such as alveolar macrophages and spleen cells, using a real-time quantitative PCR. The expression of pGM-CSF in LPS-, phytohemagglutinin (PHA)-, or concanavalin A (ConA)-stimulated cells was always higher as compared to the control cells. The expression levels of pGM-CSF in alveolar macrophages were highest at 5 h after LPS stimulation and then continuously decreased in the late phase. In spleen cells, the LPS-stimulated group showed the highest levels after 5 h, but the PHA- and the ConA-stimulated groups showed slightly increased expression levels at the early phase and peaked at 24 h. To our knowledge, this is the first published report describing the nucleotide sequence of whole cDNA and the expression pattern of pGM-CSF using real-time quantitative PCR. These results indicate that pGM-CSF has its own characteristic expression profile in different immune cells.

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Section: Discussionsupporting

confidence: 88%

Kinetic study of porcine GM-CSF expression in porcine alveolar macrophages and spleen cells

Cho

Lee

Shin

et al. 2003

FEMS Immunology &Amp; Medical Microbiology

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“…47 There is contradictory data that IFN-γ was not evident in LPS-induced porcine AMs. 48 This might indicate, in the present study, that AMs contaminated with PMN cells and lymphocytes may lead to the alteration of cytokine secretion. Differences in our study between LPS- and LTA-activated cytokines in vitro concur with previous findings.…”

Section: Discussionmentioning

confidence: 47%

Alveolar macrophage phagocytic activity is enhanced with LPS priming, and combined stimulation of LPS and lipoteichoic acid synergistically induce pro-inflammatory cytokines in pigs

et al. 2013

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The objective of the present study was to investigate LPS and lipoteichoic acid (LTA)-induced TLRs, associated signaling molecules and inflammatory mediators, as well as to compare their combined effect in porcine alveolar macrophages. Macrophages were incubated for 24 h with various concentrations of LPS, LTA, LPS + LTA or control. Multiple concentrations of LPS elicited marked up-regulation in mRNA for TLR2 and TLR4, CD14, MD2, MyD88, IRAK-4 and TRAF6 compared with the control. LTA had no effect on TLR4 and MD2; only higher doses up-regulated TLR2, CD14, MyD88, IRAK-4 and TRAF6 mRNA. LPS-activated cells released IL1-β, IL12-β, TNF-α, IL-6, IL-8, IFN-γ and IL-10 in a dose-dependent manner, while LTA had no effect on IL-1β, IL-6 and IFN-γ. Higher doses of LTA induced IL-12β, TNF-α, IL-8 and IL-10. Combined stimulation augmented TLR2, CD14 and MyD88 mRNA, and subsequently produced elevated levels of IL-6, TNF-α and IL-8 when compared with LPS and LTA alone. Additionally, phagocytosis of macrophages was significantly increased following low concentration of LPS treatment. Only low levels of NO (nitric oxide) were detected in the LPS group. Overall, compared with LPS, LTA was a relatively weak inducer, and co-stimulation accelerated gene and cytokine production associated with pulmonary innate immune function.

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“…We, thus, hypothesized that myeloid cells are the source of disease-triggering pathogenic IL-6, which stimulates the development of the inflammatory responses in the respiratory tract, while the increase in the proportion of lymphoid cells in the pool of IL-6-producing cells may be a secondary event. Additionally, taking into account that alveolar macrophages and dendritic cells may produce IL-6 with different kinetics ( 49 ), we hypothesized that the effects of IL-6 deficiency in macrophages and dendritic cells on allergic airway inflammation may differ.…”

Section: Discussionmentioning

confidence: 99%

Non-redundant Functions of IL-6 Produced by Macrophages and Dendritic Cells in Allergic Airway Inflammation

et al. 2018

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Asthma is a common inflammatory disease of the airway caused by a combination of genetic and environmental factors and characterized by airflow obstruction, wheezing, eosinophilia, and neutrophilia of lungs and sputum. Similar to other proinflammatory cytokines, IL-6 is elevated in asthma and plays an active role in this disease. However, the exact molecular mechanism of IL-6 involvement in the pathogenesis of asthma remains largely unknown and the major cellular source of pathogenic IL-6 has not been defined. In the present study, we used conditional gene targeting to demonstrate that macrophages and dendritic cells are the critical sources of pathogenic IL-6 in acute HDM-induced asthma in mice. Complete genetic inactivation of IL-6 ameliorated the disease with significant decrease in eosinophilia in the lungs. Specific ablation of IL-6 in macrophages reduced key indicators of type 2 allergic inflammation, including eosinophil and Th2 cell accumulation in the lungs, production of IgE and expression of asthma-associated inflammatory mediators. In contrast, mice with deficiency of IL-6 in dendritic cells demonstrated attenuated neutrophilic, but regular eosinophilic response in HDM-induced asthma. Taken together, our results indicate that IL-6 plays a pathogenic role in the HDM-induced asthma model and that lung macrophages and dendritic cells are the predominant sources of pathogenic IL-6 but contribute differently to the disease.

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Time Course Study of Cytokine mRNA Expression in LPS-Stimulated Porcine Alveolar Macrophages

Cited by 12 publications

References 0 publications

Kinetic study of porcine GM-CSF expression in porcine alveolar macrophages and spleen cells

Kinetic study of porcine GM-CSF expression in porcine alveolar macrophages and spleen cells

Alveolar macrophage phagocytic activity is enhanced with LPS priming, and combined stimulation of LPS and lipoteichoic acid synergistically induce pro-inflammatory cytokines in pigs

Non-redundant Functions of IL-6 Produced by Macrophages and Dendritic Cells in Allergic Airway Inflammation

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