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Bacterial DNA in house and farm barn dust

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1 Bacterial DNA in house and farm barn dust
Sitesh R Roy, MD, Allison M Schiltz, BA, Alex Marotta, MD, Yiqin Shen, BA, Andrew H Liu, MD  Journal of Allergy and Clinical Immunology  Volume 112, Issue 3, Pages (September 2003) DOI: /S (03)

2 FIG 1 Amplification of conserved bacterial segment in ribosomal operons of E coli DNA by qRT PCR. This exemplifies how standard curves were generated in each PCR run to quantify bacterial DNA in dust samples. Duplicate samples of 4-fold serial dilutions of E coli genomic DNA were used as templates for a 2-step nested PCR. Starting amount of E coli DNA is shown next to each amplification curve. Relative fluorescence was measured; cycle threshold at which threshold fluorescence was reached is shown. Journal of Allergy and Clinical Immunology  , DOI: ( /S (03) )

3 FIG 2 Bacterial DNA and endotoxin content in dust samples from 4 different locales: (1) urban homes in Denver, Colo (n = 8); (2) rural homes in India (n = 8); (3) farm homes in the United States (n = 8); and (4) US farm barns (n = 8). Bacterial DNA (left side, blue) was measured with qRT PCR assay. Endotoxin (right side, black) was measured with LAL assay. Bacterial DNA and endotoxin levels in dust samples differed significantly by locale, with highest levels in farm barn samples and lowest levels in urban homes (P ≤ .001). Journal of Allergy and Clinical Immunology  , DOI: ( /S (03) )

4 FIG 3 Correlation of bacterial DNA and endotoxin in dust samples. Log-transformed levels of bacterial DNA and endotoxin were positively correlated (pairwise correlation). Journal of Allergy and Clinical Immunology  , DOI: ( /S (03) )

5 FIG 4 Cytokine release from in vitro stimulations of PBMC from 5 human subjects. Mean IL-10 (A), IL-12p40 (B), and TNF-α (C) release stimulated by DNA (3 μg/mL) from dust of 2 farm barns and 6 urban homes, with and without 300 pg/mL LPS, is shown; 300 pg/mL LPS alone and 10 μg/mL CpG ODN ,29 with and without 300 pg/mL LPS are included as controls. Farm barn DNA showed significant potentiation of IL-10 (13-fold) and IL-12 (3-fold) release when combined with low levels of LPS, above that stimulated by LPS alone and urban home DNA plus LPS (∗P ≤ .05, Wilcoxon). Journal of Allergy and Clinical Immunology  , DOI: ( /S (03) )

6 FIG 4 Cytokine release from in vitro stimulations of PBMC from 5 human subjects. Mean IL-10 (A), IL-12p40 (B), and TNF-α (C) release stimulated by DNA (3 μg/mL) from dust of 2 farm barns and 6 urban homes, with and without 300 pg/mL LPS, is shown; 300 pg/mL LPS alone and 10 μg/mL CpG ODN ,29 with and without 300 pg/mL LPS are included as controls. Farm barn DNA showed significant potentiation of IL-10 (13-fold) and IL-12 (3-fold) release when combined with low levels of LPS, above that stimulated by LPS alone and urban home DNA plus LPS (∗P ≤ .05, Wilcoxon). Journal of Allergy and Clinical Immunology  , DOI: ( /S (03) )

7 FIG 4 Cytokine release from in vitro stimulations of PBMC from 5 human subjects. Mean IL-10 (A), IL-12p40 (B), and TNF-α (C) release stimulated by DNA (3 μg/mL) from dust of 2 farm barns and 6 urban homes, with and without 300 pg/mL LPS, is shown; 300 pg/mL LPS alone and 10 μg/mL CpG ODN ,29 with and without 300 pg/mL LPS are included as controls. Farm barn DNA showed significant potentiation of IL-10 (13-fold) and IL-12 (3-fold) release when combined with low levels of LPS, above that stimulated by LPS alone and urban home DNA plus LPS (∗P ≤ .05, Wilcoxon). Journal of Allergy and Clinical Immunology  , DOI: ( /S (03) )

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