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Enhanced Allergic Inflammation of Der p 2 Affected by Polymorphisms of MD-2 Promoter.

Liao EC, Hsieh CW, Chang CY, Yu SJ, Sheu ML, Wu SM, Tsai JJ - Allergy Asthma Immunol Res (2015)

Bottom Line: The function of SNPs of MD-2 promoter and the effects of cytokines and immunoglobulin on the secretion and mRNA expression were investigated in 73 allergic subjects with different MD-2 gene promoter variants.Increased secretions of IL-6, IL-8, and IL-10 were found to be up-regulated by Der p 2 stimulation, and an increased secretion of IFN-γ and decreased secretion of IL-4 were noted after LPS stimulation.The high levels of proinflammatory cytokines secreted by Der p 2 were predetermined by MD-2 promoter SNPs (rs1809441/rs1809442).

View Article: PubMed Central - PubMed

Affiliation: Center for Translational Medicine, Department of Medical Research, Taichung Veterans General Hospital, Taichung, Taiwan.; Department of BioIndustry Technology, Da Yeh University, Changhua, Taiwan.; Department of Medical Technology, Jen Ten College of Medicine, Nursing and Management, Miaoli, Taiwan.

ABSTRACT

Purpose: Myeloid differentiation-2 (MD-2) has been associated with endotoxin and inflammatory disorders because it can recognize lipopolysaccharide (LPS) binding and attenuate Toll-like receptor 4 (TLR4)-mediated signaling. However, its role in allergic inflammation has yet to be clarified. We examined whether single nucleotide polymorphisms (SNPs) in MD-2 promoter can affect MD-2 expression and aimed to clarify the relationship between Der p 2 allergy and SNPs of MD-2 promoter.

Methods: The function of SNPs of MD-2 promoter and the effects of cytokines and immunoglobulin on the secretion and mRNA expression were investigated in 73 allergic subjects with different MD-2 gene promoter variants. Peripheral blood mononuclear cells were cultured with or without LPS in the presence of Dermatophagoides pteronyssinus group 2 allergen (Der p 2), followed by mRNA extraction and cytokine expression analysis. The culture supernatants were collected for cytokine measurement.

Results: Patients with the MD-2 promoter SNPs (rs1809441/rs1809442) had increased mRNA expressions of MD-2, ε heavy chain of IgE (Cε), and interleukin (IL)-8; however, only MD-2 and IL-8 were further up-regulated after Der p 2 stimulation. Patients with SNPs of MD-2 promoter tended to have high levels of IL-1β, IL-6, IL-8, IL-10, and tumor necrosis factor (TNF)-α after Der p 2 and LPS stimulation. Increased secretions of IL-6, IL-8, and IL-10 were found to be up-regulated by Der p 2 stimulation, and an increased secretion of IFN-γ and decreased secretion of IL-4 were noted after LPS stimulation.

Conclusions: The high levels of proinflammatory cytokines secreted by Der p 2 were predetermined by MD-2 promoter SNPs (rs1809441/rs1809442). Through cytokine secretion by Der p 2 and LPS, these SNPs may serve as an indicator of the pathological phenotype of Der p 2-induced allergic inflammation.

No MeSH data available.


Related in: MedlinePlus

The mRNA expressions of ε heavy chain of IgE (Cε) and GATA-3 in PBMCs from subjects with the SNP rs1809441/42 mutant and wild genotypes were evaluated using reverse-transcriptase polymerase chain reaction (RT-PCR). (A) The 0, 24, 48, and 72 represent the time points (hours) after allergen rDer p 2 (10 µg/mL) challenge. The GAPDH expression acted as an internal-control. (B) The Cε expression was evaluated using RT-PCR. Data are expressed as mean of each group. Medium, detected without allergen challenge; Der p 2, detected after rDer p 2 (10 µg/mL) challenge. (C) The expression of the transcription factor GATA-3 was evaluated using RT-PCR; **P<0.05 compared between the MD-2 promoter SNPs (+) and SNPs (-) under medium only; ***P<0.01 compared between MD-2 promoter SNPs (+) and SNPs (-) under Der p 2 (10 µg/mL) challenge. ##P<0.05 compared between medium and rDer p 2 (10 µg/mL) challeges in the group of MD-2 promoter SNPs (+).
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Figure 2: The mRNA expressions of ε heavy chain of IgE (Cε) and GATA-3 in PBMCs from subjects with the SNP rs1809441/42 mutant and wild genotypes were evaluated using reverse-transcriptase polymerase chain reaction (RT-PCR). (A) The 0, 24, 48, and 72 represent the time points (hours) after allergen rDer p 2 (10 µg/mL) challenge. The GAPDH expression acted as an internal-control. (B) The Cε expression was evaluated using RT-PCR. Data are expressed as mean of each group. Medium, detected without allergen challenge; Der p 2, detected after rDer p 2 (10 µg/mL) challenge. (C) The expression of the transcription factor GATA-3 was evaluated using RT-PCR; **P<0.05 compared between the MD-2 promoter SNPs (+) and SNPs (-) under medium only; ***P<0.01 compared between MD-2 promoter SNPs (+) and SNPs (-) under Der p 2 (10 µg/mL) challenge. ##P<0.05 compared between medium and rDer p 2 (10 µg/mL) challeges in the group of MD-2 promoter SNPs (+).

Mentions: To determine whether the MD-2 promoter SNPs (rs1809441/rs1809442) affect Cε and GATA-3 mRNA expressions, PBMCs derived from allergic subjects were cultured with or without 24, 48, and 72 hours of Der p 2 challenge, followed by mRNA measurement, RT-PCR, and gel analysis presented by representative data (Fig. 2A). When mRNA expressions of Cε and GATA-3 were analyzed, both mRNA expressions could be up-regulated by Der p 2 in both groups of subjects. There were significantly increased expressions of Cε mRNA (Fig. 2B) and GATA-3 (Fig. 2C) with the Der p 2 challenge after 48-72 hours. The expressions of Cε and GATA-3 were significantly higher in allergic subjects with MD-2 promoter SNPs (rs1809441-T/rs1809442-G) compared to MD-2 wild-type (rs1809441-G/rs1809442-C) both in the conditions of PBMC cultured with or without Der p 2 stimulation after 48-72 hours (Fig. 2B and C).


Enhanced Allergic Inflammation of Der p 2 Affected by Polymorphisms of MD-2 Promoter.

Liao EC, Hsieh CW, Chang CY, Yu SJ, Sheu ML, Wu SM, Tsai JJ - Allergy Asthma Immunol Res (2015)

The mRNA expressions of ε heavy chain of IgE (Cε) and GATA-3 in PBMCs from subjects with the SNP rs1809441/42 mutant and wild genotypes were evaluated using reverse-transcriptase polymerase chain reaction (RT-PCR). (A) The 0, 24, 48, and 72 represent the time points (hours) after allergen rDer p 2 (10 µg/mL) challenge. The GAPDH expression acted as an internal-control. (B) The Cε expression was evaluated using RT-PCR. Data are expressed as mean of each group. Medium, detected without allergen challenge; Der p 2, detected after rDer p 2 (10 µg/mL) challenge. (C) The expression of the transcription factor GATA-3 was evaluated using RT-PCR; **P<0.05 compared between the MD-2 promoter SNPs (+) and SNPs (-) under medium only; ***P<0.01 compared between MD-2 promoter SNPs (+) and SNPs (-) under Der p 2 (10 µg/mL) challenge. ##P<0.05 compared between medium and rDer p 2 (10 µg/mL) challeges in the group of MD-2 promoter SNPs (+).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4509663&req=5

Figure 2: The mRNA expressions of ε heavy chain of IgE (Cε) and GATA-3 in PBMCs from subjects with the SNP rs1809441/42 mutant and wild genotypes were evaluated using reverse-transcriptase polymerase chain reaction (RT-PCR). (A) The 0, 24, 48, and 72 represent the time points (hours) after allergen rDer p 2 (10 µg/mL) challenge. The GAPDH expression acted as an internal-control. (B) The Cε expression was evaluated using RT-PCR. Data are expressed as mean of each group. Medium, detected without allergen challenge; Der p 2, detected after rDer p 2 (10 µg/mL) challenge. (C) The expression of the transcription factor GATA-3 was evaluated using RT-PCR; **P<0.05 compared between the MD-2 promoter SNPs (+) and SNPs (-) under medium only; ***P<0.01 compared between MD-2 promoter SNPs (+) and SNPs (-) under Der p 2 (10 µg/mL) challenge. ##P<0.05 compared between medium and rDer p 2 (10 µg/mL) challeges in the group of MD-2 promoter SNPs (+).
Mentions: To determine whether the MD-2 promoter SNPs (rs1809441/rs1809442) affect Cε and GATA-3 mRNA expressions, PBMCs derived from allergic subjects were cultured with or without 24, 48, and 72 hours of Der p 2 challenge, followed by mRNA measurement, RT-PCR, and gel analysis presented by representative data (Fig. 2A). When mRNA expressions of Cε and GATA-3 were analyzed, both mRNA expressions could be up-regulated by Der p 2 in both groups of subjects. There were significantly increased expressions of Cε mRNA (Fig. 2B) and GATA-3 (Fig. 2C) with the Der p 2 challenge after 48-72 hours. The expressions of Cε and GATA-3 were significantly higher in allergic subjects with MD-2 promoter SNPs (rs1809441-T/rs1809442-G) compared to MD-2 wild-type (rs1809441-G/rs1809442-C) both in the conditions of PBMC cultured with or without Der p 2 stimulation after 48-72 hours (Fig. 2B and C).

Bottom Line: The function of SNPs of MD-2 promoter and the effects of cytokines and immunoglobulin on the secretion and mRNA expression were investigated in 73 allergic subjects with different MD-2 gene promoter variants.Increased secretions of IL-6, IL-8, and IL-10 were found to be up-regulated by Der p 2 stimulation, and an increased secretion of IFN-γ and decreased secretion of IL-4 were noted after LPS stimulation.The high levels of proinflammatory cytokines secreted by Der p 2 were predetermined by MD-2 promoter SNPs (rs1809441/rs1809442).

View Article: PubMed Central - PubMed

Affiliation: Center for Translational Medicine, Department of Medical Research, Taichung Veterans General Hospital, Taichung, Taiwan.; Department of BioIndustry Technology, Da Yeh University, Changhua, Taiwan.; Department of Medical Technology, Jen Ten College of Medicine, Nursing and Management, Miaoli, Taiwan.

ABSTRACT

Purpose: Myeloid differentiation-2 (MD-2) has been associated with endotoxin and inflammatory disorders because it can recognize lipopolysaccharide (LPS) binding and attenuate Toll-like receptor 4 (TLR4)-mediated signaling. However, its role in allergic inflammation has yet to be clarified. We examined whether single nucleotide polymorphisms (SNPs) in MD-2 promoter can affect MD-2 expression and aimed to clarify the relationship between Der p 2 allergy and SNPs of MD-2 promoter.

Methods: The function of SNPs of MD-2 promoter and the effects of cytokines and immunoglobulin on the secretion and mRNA expression were investigated in 73 allergic subjects with different MD-2 gene promoter variants. Peripheral blood mononuclear cells were cultured with or without LPS in the presence of Dermatophagoides pteronyssinus group 2 allergen (Der p 2), followed by mRNA extraction and cytokine expression analysis. The culture supernatants were collected for cytokine measurement.

Results: Patients with the MD-2 promoter SNPs (rs1809441/rs1809442) had increased mRNA expressions of MD-2, ε heavy chain of IgE (Cε), and interleukin (IL)-8; however, only MD-2 and IL-8 were further up-regulated after Der p 2 stimulation. Patients with SNPs of MD-2 promoter tended to have high levels of IL-1β, IL-6, IL-8, IL-10, and tumor necrosis factor (TNF)-α after Der p 2 and LPS stimulation. Increased secretions of IL-6, IL-8, and IL-10 were found to be up-regulated by Der p 2 stimulation, and an increased secretion of IFN-γ and decreased secretion of IL-4 were noted after LPS stimulation.

Conclusions: The high levels of proinflammatory cytokines secreted by Der p 2 were predetermined by MD-2 promoter SNPs (rs1809441/rs1809442). Through cytokine secretion by Der p 2 and LPS, these SNPs may serve as an indicator of the pathological phenotype of Der p 2-induced allergic inflammation.

No MeSH data available.


Related in: MedlinePlus