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Interaction between Leptospiral Lipopolysaccharide and Toll-like Receptor 2 in Pig Fibroblast Cell Line, and Inhibitory Effect of Antibody against Leptospiral Lipopolysaccharide on Interaction.

Guo Y, Fukuda T, Nakamura S, Bai L, Xu J, Kuroda K, Tomioka R, Yoneyama H, Isogai E - Asian-australas. J. Anim. Sci. (2015)

Bottom Line: Leptospiral lipopolysaccharide (L-LPS) has shown potency in activating toll-like receptor 2 (TLR2) in pig fibroblasts (PEFs_NCC1), and causes the expression of proinflammatory cytokines.In this study, the immune response of pig embryonic fibroblast cell line (PEFs_SV40) was investigated and was found to be the high immune response, thus TLR2 is the predominate receptor of L-LPS in pig cells.Further, we found a strategy using the antibody against L-LPS, to prevent L-LPS interaction with TLR2 in pig cells which could impact on immune activation.

View Article: PubMed Central - PubMed

Affiliation: Laboratories of Animal Microbiology, Graduate School of Agricultural Science, Tohoku University, Sendai 981-8555, Japan ; Department of Immunobiology and Pathogenic Biology, Medical School of Xi'an Jiaotong University, Xi'an 710061, China.

ABSTRACT
Leptospiral lipopolysaccharide (L-LPS) has shown potency in activating toll-like receptor 2 (TLR2) in pig fibroblasts (PEFs_NCC1), and causes the expression of proinflammatory cytokines. However, the stimulation by L-LPS was weak eliciting the function of TLR2 sufficiently in pig innate immunity responses during Leptospira infection. In this study, the immune response of pig embryonic fibroblast cell line (PEFs_SV40) was investigated and was found to be the high immune response, thus TLR2 is the predominate receptor of L-LPS in pig cells. Further, we found a strategy using the antibody against L-LPS, to prevent L-LPS interaction with TLR2 in pig cells which could impact on immune activation.

No MeSH data available.


Related in: MedlinePlus

Dose and time-dependent proinflammatory cytokines protein production by LPS-stimulated PEFs_SV40 cells. For dose experiment, cells supernatant fluids were collected at 3 h after incubation with 0.1, 1.0, and 10.0 μg/mL concentrations of LPS. (A) IL-6, (B) IL-8. For time experiment, cells supernatant fluids were collected at 3, 12, and 24 h after incubation with concentrations of 0.1 μg/mL LPS. (C) IL-6, (D) IL-8. Data shown is an average of n = 3 biological replicates±SD. Asterisks indicate significant differences (* p<0.05, ** p<0.01). LPS, lipopolysaccharide; IL, interleukin; SD, standard deviation.
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f5-ajas-28-2-273: Dose and time-dependent proinflammatory cytokines protein production by LPS-stimulated PEFs_SV40 cells. For dose experiment, cells supernatant fluids were collected at 3 h after incubation with 0.1, 1.0, and 10.0 μg/mL concentrations of LPS. (A) IL-6, (B) IL-8. For time experiment, cells supernatant fluids were collected at 3, 12, and 24 h after incubation with concentrations of 0.1 μg/mL LPS. (C) IL-6, (D) IL-8. Data shown is an average of n = 3 biological replicates±SD. Asterisks indicate significant differences (* p<0.05, ** p<0.01). LPS, lipopolysaccharide; IL, interleukin; SD, standard deviation.

Mentions: The amount of IL-6 and IL-8 secreted in culture supernatant were quantitatively determined, showing considerable up-regulation of these cytokines by E-LPS and L-LPS stimulations (p<0.05; Figure 5). In L-LPS stimulations, the secretion level of IL-6 protein was increased until dose of 1.0 μg/mL L-LPS and was slightly decreased at 10 μg/mL L-LPS stimulation (Figure 5A). In contrast, IL-8 secretion was monotonically enhanced with dose concentration of L-LPS (Figure 5B). The secretion levels of IL-6 and IL-8 reached the highest level at 3 h after L-LPS or E-LPS stimulation and retained the level throughout the experiment. Thus, we conclude that L-LPS can rapidly induce production of a large amount of cytokines.


Interaction between Leptospiral Lipopolysaccharide and Toll-like Receptor 2 in Pig Fibroblast Cell Line, and Inhibitory Effect of Antibody against Leptospiral Lipopolysaccharide on Interaction.

Guo Y, Fukuda T, Nakamura S, Bai L, Xu J, Kuroda K, Tomioka R, Yoneyama H, Isogai E - Asian-australas. J. Anim. Sci. (2015)

Dose and time-dependent proinflammatory cytokines protein production by LPS-stimulated PEFs_SV40 cells. For dose experiment, cells supernatant fluids were collected at 3 h after incubation with 0.1, 1.0, and 10.0 μg/mL concentrations of LPS. (A) IL-6, (B) IL-8. For time experiment, cells supernatant fluids were collected at 3, 12, and 24 h after incubation with concentrations of 0.1 μg/mL LPS. (C) IL-6, (D) IL-8. Data shown is an average of n = 3 biological replicates±SD. Asterisks indicate significant differences (* p<0.05, ** p<0.01). LPS, lipopolysaccharide; IL, interleukin; SD, standard deviation.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4283174&req=5

f5-ajas-28-2-273: Dose and time-dependent proinflammatory cytokines protein production by LPS-stimulated PEFs_SV40 cells. For dose experiment, cells supernatant fluids were collected at 3 h after incubation with 0.1, 1.0, and 10.0 μg/mL concentrations of LPS. (A) IL-6, (B) IL-8. For time experiment, cells supernatant fluids were collected at 3, 12, and 24 h after incubation with concentrations of 0.1 μg/mL LPS. (C) IL-6, (D) IL-8. Data shown is an average of n = 3 biological replicates±SD. Asterisks indicate significant differences (* p<0.05, ** p<0.01). LPS, lipopolysaccharide; IL, interleukin; SD, standard deviation.
Mentions: The amount of IL-6 and IL-8 secreted in culture supernatant were quantitatively determined, showing considerable up-regulation of these cytokines by E-LPS and L-LPS stimulations (p<0.05; Figure 5). In L-LPS stimulations, the secretion level of IL-6 protein was increased until dose of 1.0 μg/mL L-LPS and was slightly decreased at 10 μg/mL L-LPS stimulation (Figure 5A). In contrast, IL-8 secretion was monotonically enhanced with dose concentration of L-LPS (Figure 5B). The secretion levels of IL-6 and IL-8 reached the highest level at 3 h after L-LPS or E-LPS stimulation and retained the level throughout the experiment. Thus, we conclude that L-LPS can rapidly induce production of a large amount of cytokines.

Bottom Line: Leptospiral lipopolysaccharide (L-LPS) has shown potency in activating toll-like receptor 2 (TLR2) in pig fibroblasts (PEFs_NCC1), and causes the expression of proinflammatory cytokines.In this study, the immune response of pig embryonic fibroblast cell line (PEFs_SV40) was investigated and was found to be the high immune response, thus TLR2 is the predominate receptor of L-LPS in pig cells.Further, we found a strategy using the antibody against L-LPS, to prevent L-LPS interaction with TLR2 in pig cells which could impact on immune activation.

View Article: PubMed Central - PubMed

Affiliation: Laboratories of Animal Microbiology, Graduate School of Agricultural Science, Tohoku University, Sendai 981-8555, Japan ; Department of Immunobiology and Pathogenic Biology, Medical School of Xi'an Jiaotong University, Xi'an 710061, China.

ABSTRACT
Leptospiral lipopolysaccharide (L-LPS) has shown potency in activating toll-like receptor 2 (TLR2) in pig fibroblasts (PEFs_NCC1), and causes the expression of proinflammatory cytokines. However, the stimulation by L-LPS was weak eliciting the function of TLR2 sufficiently in pig innate immunity responses during Leptospira infection. In this study, the immune response of pig embryonic fibroblast cell line (PEFs_SV40) was investigated and was found to be the high immune response, thus TLR2 is the predominate receptor of L-LPS in pig cells. Further, we found a strategy using the antibody against L-LPS, to prevent L-LPS interaction with TLR2 in pig cells which could impact on immune activation.

No MeSH data available.


Related in: MedlinePlus