Gene expression measurements in the context of epidemiological studies

Bieli, Christian; Frei, Remo; Schickinger, V.; Steinle, Johanna; Bommer, Claudine; Loeliger, Susanne; Braun‐Fahrländer, Charlotte; Pershagen, Göran; Lauener, Roger

doi:10.1111/j.1398-9995.2008.01744.x

Cited by 9 publications

(6 citation statements)

References 4 publications

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“…We analyzed the data with the comparative Ct method according to the manufacturer’s instructions (Applied Biosystem). Tests regarding RNA stability were performed [44].…”

Section: Methodsmentioning

confidence: 99%

Expression of Genes Related to Anti-Inflammatory Pathways Are Modified Among Farmers’ Children

Frei

Roduit²,

Bieli³

et al. 2014

PLoS ONE

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BackgroundThe hygiene hypothesis states that children exposed to higher loads of microbes such as farmers’ children suffer less from allergies later in life. Several immunological mechanisms underpinning the hygiene hypothesis have been proposed such as a shift in T helper cell balance, T regulatory cell activity, or immune regulatory mechanisms induced by the innate immunity.ObjectiveTo investigate whether the proposed immunological mechanisms for the hygiene hypotheses are found in farmers’ children.MethodsWe assessed gene expression levels of 64 essential markers of the innate and adaptive immunity by quantitative real-time PCR in white blood cells in 316 Swiss children of the PARSIFAL study to compare farmers’ to non-farmers’ expressions and to associate them to the prevalence of asthma and rhinoconjunctivitis, total and allergen-specific IgE in serum, and expression of Cε germ-line transcripts.ResultsWe found enhanced expression of genes of the innate immunity such as IRAK-4 and RIPK1 and enhanced expression of regulatory molecules such as IL-10, TGF-β, SOCS4, and IRAK-2 in farmers’ children. Furthermore, farmers’ children expressed less of the TH1 associated cytokine IFN-γ while TH2 associated transcription factor GATA3 was enhanced. No significant associations between the assessed immunological markers and allergic diseases or sensitization to allergens were observed.ConclusionFarmers’ children express multiple increased innate immune response and immune regulatory molecules, which may contribute to the mechanisms of action of the hygiene hypothesis.

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“…We analyzed the data with the comparative Ct method according to the manufacturer’s instructions (Applied Biosystem). Tests regarding RNA stability were performed [44].…”

Section: Methodsmentioning

confidence: 99%

Expression of Genes Related to Anti-Inflammatory Pathways Are Modified Among Farmers’ Children

Frei

Roduit²,

Bieli³

et al. 2014

PLoS ONE

View full text Add to dashboard Cite

show abstract

“…For the assessment of mRNA, the blood was collected in a PAXgene Blood RNA tube containing an RNA-stabilizing solution (PreAnalytiX/Qiagen, Hilden, Germany) and then frozen to 2808C within 24 hours. 19 In a central laboratory (Zurich, Switzerland), the RNA was isolated by using the PAXgene 96 Blood RNA Kit (PreAnalytiX/Qiagen) supplemented with RNase-free DNase (Qiagen). The mRNA was reverse-transcribed into cDNA by using the TaqMan Reverse Transcription Reagents (Applied Biosystems, Foster City, Calif).…”

Section: Expression Of Toll-like Receptors and Cd14mentioning

confidence: 99%

Prenatal animal contact and gene expression of innate immunity receptors at birth are associated with atopic dermatitis

Roduit

Wohlgensinger

Frei

et al. 2011

Journal of Allergy and Clinical Immunology

153

128

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“…For the assessment of mRNA, the blood was collected in a PAXgene Blood RNA tube containing an RNA-stabilizing solution (PreAnalytiX/Qiagen, Hilden, Germany) and then frozen to 2808C within 24 hours. 22 At the central laboratory of the Children's Hospital of Zurich, the RNA was isolated with the PAXgene 96 Blood RNA Kit (PreAnalytiX/Qiagen) supplemented with RNase-free DNase (Qiagen). The mRNA was reverse transcribed into cDNA by using the TaqMan Reverse Transcription Reagents (Applied Biosystems, Foster City, Calif).…”

Section: Measurement Of Mrna Expression In Cord Blood and At Yearmentioning

confidence: 99%

Prenatal and early-life exposures alter expression of innate immunity genes: The PASTURE cohort study

Loss

Bitter

Wohlgensinger

et al. 2012

Journal of Allergy and Clinical Immunology

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Gene expression measurements in the context of epidemiological studies

Abstract: Collecting blood samples into tubes containing an RNA-stabilizing solution increases RNA yield and reduces its variability. Long-term storage of samples may lead to RNA degradation, requiring special attention in longitudinal studies.

Cited by 9 publications

References 4 publications

Expression of Genes Related to Anti-Inflammatory Pathways Are Modified Among Farmers’ Children

Expression of Genes Related to Anti-Inflammatory Pathways Are Modified Among Farmers’ Children

Prenatal animal contact and gene expression of innate immunity receptors at birth are associated with atopic dermatitis

Prenatal and early-life exposures alter expression of innate immunity genes: The PASTURE cohort study

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