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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

A relative mRNA expression ratio of proinflammatory cytokines were quantified by qRT-PCR in LPS-induced PEFs_SV40 cells. (A) (B) IL-6 and IL-8 mRNA expression in PEFs_SV40 cells after incubation with 0.1, 1.0, and 10.0 μg/mL concentrations of LPS at 3 h. (C) (D) IL-6 and IL-8 mRNA expression in SV40 PEFs after incubation with 0.1 μg/mL LPS at 3, 12, and 24 h. Data shown is an average of n = 3 biological replicate±SD. Levels of expression and significance are relative to unstimulated cells (at each time point for time-dependent experiments). Asterisks indicate significant differences (* p<0.05, ** p<0.01). qRT-PCR, quantitative real-time polymerase chain reaction; LPS, lipopolysaccharide; IL, interleukin; SD, standard deviation.
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f4-ajas-28-2-273: A relative mRNA expression ratio of proinflammatory cytokines were quantified by qRT-PCR in LPS-induced PEFs_SV40 cells. (A) (B) IL-6 and IL-8 mRNA expression in PEFs_SV40 cells after incubation with 0.1, 1.0, and 10.0 μg/mL concentrations of LPS at 3 h. (C) (D) IL-6 and IL-8 mRNA expression in SV40 PEFs after incubation with 0.1 μg/mL LPS at 3, 12, and 24 h. Data shown is an average of n = 3 biological replicate±SD. Levels of expression and significance are relative to unstimulated cells (at each time point for time-dependent experiments). Asterisks indicate significant differences (* p<0.05, ** p<0.01). qRT-PCR, quantitative real-time polymerase chain reaction; LPS, lipopolysaccharide; IL, interleukin; SD, standard deviation.

Mentions: To understand the inflammatory response triggered by L-LPS, we examined the expression level of cytokines mRNA in the PEFs_SV40 cells by qRT-PCR. The cells were treated with various concentrations of L-LPS, showing significant increase of the mRNA expression of IL-6 (p<0.05) and IL-8 (p<0.05) up to more than 30 folds and 40 folds of unstimulated cells, respectively (Figure 4A and B). Also marked enhancement of IL-6 and IL-8 mRNA expression were caused at 3 h after L-LPS stimulation and then rapidly decreased but the expression level was still higher than that of unstimulated cells (p<0.05; Figure 4C and D). As a positive control, the cells were treated with E-LPS, shows significantly high expression of IL-6 and IL-8 as compared with unstimulated cells (p<0.05; Figure 4A and B). These expressions were gradually decreased over time (Figure 4C and D).


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)

A relative mRNA expression ratio of proinflammatory cytokines were quantified by qRT-PCR in LPS-induced PEFs_SV40 cells. (A) (B) IL-6 and IL-8 mRNA expression in PEFs_SV40 cells after incubation with 0.1, 1.0, and 10.0 μg/mL concentrations of LPS at 3 h. (C) (D) IL-6 and IL-8 mRNA expression in SV40 PEFs after incubation with 0.1 μg/mL LPS at 3, 12, and 24 h. Data shown is an average of n = 3 biological replicate±SD. Levels of expression and significance are relative to unstimulated cells (at each time point for time-dependent experiments). Asterisks indicate significant differences (* p<0.05, ** p<0.01). qRT-PCR, quantitative real-time polymerase chain reaction; LPS, lipopolysaccharide; IL, interleukin; SD, standard deviation.
© Copyright Policy
Related In: Results  -  Collection

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

f4-ajas-28-2-273: A relative mRNA expression ratio of proinflammatory cytokines were quantified by qRT-PCR in LPS-induced PEFs_SV40 cells. (A) (B) IL-6 and IL-8 mRNA expression in PEFs_SV40 cells after incubation with 0.1, 1.0, and 10.0 μg/mL concentrations of LPS at 3 h. (C) (D) IL-6 and IL-8 mRNA expression in SV40 PEFs after incubation with 0.1 μg/mL LPS at 3, 12, and 24 h. Data shown is an average of n = 3 biological replicate±SD. Levels of expression and significance are relative to unstimulated cells (at each time point for time-dependent experiments). Asterisks indicate significant differences (* p<0.05, ** p<0.01). qRT-PCR, quantitative real-time polymerase chain reaction; LPS, lipopolysaccharide; IL, interleukin; SD, standard deviation.
Mentions: To understand the inflammatory response triggered by L-LPS, we examined the expression level of cytokines mRNA in the PEFs_SV40 cells by qRT-PCR. The cells were treated with various concentrations of L-LPS, showing significant increase of the mRNA expression of IL-6 (p<0.05) and IL-8 (p<0.05) up to more than 30 folds and 40 folds of unstimulated cells, respectively (Figure 4A and B). Also marked enhancement of IL-6 and IL-8 mRNA expression were caused at 3 h after L-LPS stimulation and then rapidly decreased but the expression level was still higher than that of unstimulated cells (p<0.05; Figure 4C and D). As a positive control, the cells were treated with E-LPS, shows significantly high expression of IL-6 and IL-8 as compared with unstimulated cells (p<0.05; Figure 4A and B). These expressions were gradually decreased over time (Figure 4C and D).

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