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Multi-Approach Analysis for the Identification of Proteases within Birch Pollen.

ABSTRACT: Birch pollen allergy is highly prevalent, with up to 100 million reported cases worldwide. Proteases in such allergen sources have been suggested to contribute to primary sensitisation and exacerbation of allergic disorders. Until now the protease content of Betula verrucosa, a birch species endemic to the northern hemisphere has not been studied in detail. Hence, we aim to identify and characterise pollen and bacteria-derived proteases found within birch pollen. The pollen transcriptome was constructed via de novo transcriptome sequencing and analysis of the proteome was achieved via mass spectrometry; a cross-comparison of the two databases was then performed. A total of 42 individual proteases were identified at the proteomic level. Further clustering of proteases into their distinct catalytic classes revealed serine, cysteine, aspartic, threonine, and metallo-proteases. Further to this, protease activity of the pollen was quantified using a fluorescently-labelled casein substrate protease assay, as 0.61 ng/mg of pollen. A large number of bacterial strains were isolated from freshly collected birch pollen and zymographic gels with gelatinase and casein, enabled visualisation of proteolytic activity of the pollen and the collected bacterial strains. We report the successful discovery of pollen and bacteria-derived proteases of Betula verrucosa.

SUBMITTER: McKenna OE

PROVIDER: S-EPMC5535924 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature

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Multi-Approach Analysis for the Identification of Proteases within Birch Pollen.

McKenna Olivia E OE Posselt Gernot G Briza Peter P Lackner Peter P Schmitt Armin O AO Gadermaier Gabriele G Wessler Silja S Ferreira Fatima F

International journal of molecular sciences 20170704 7

Birch pollen allergy is highly prevalent, with up to 100 million reported cases worldwide. Proteases in such allergen sources have been suggested to contribute to primary sensitisation and exacerbation of allergic disorders. Until now the protease content of <i>Betula verrucosa</i>, a birch species endemic to the northern hemisphere has not been studied in detail. Hence, we aim to identify and characterise pollen and bacteria-derived proteases found within birch pollen. The pollen transcriptome ...[more]

PMID: 28677627

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Project description:AimTo investigate whether birch pollen allergy symptoms are linked with gut microbiota changes and whether probiotics have an effect on these.MethodsForty seven children with confirmed birch pollen allergy were randomized to receive either a probiotic combination of Lactobacillus acidophilus (L. acidophilus) NCFM (ATCC 700396) and Bifidobacterium lactis (B. lactis) Bl-04 (ATCC SD5219) or placebo in a double-blind manner for 4 mo, starting prior to onset of the birch pollen season. Symptoms were recorded in a diary. Blood samples were taken for analysis of cytokines and eosinophils. Fecal samples were analysed for microbiota components, calprotectin and IgA. Nasal swabs were taken for analysis of eosinophils.ResultsThe pollen season induced a reduction in Bifidobacterium, Clostridium and Bacteroides which could not be prevented by the probiotic intervention. During the intervention, significantly higher numbers of B. lactis 11.2 x 10(7) +/- 4.2 x 10(7) vs 0.1 x 10(7) +/- 0.1 x 10(7) bacteria/g feces (P < 0.0001) and L. acidophilus NCFM 3.5 x 10(6) +/- 1.3 x 10(6) vs 0.2 x 10(6) +/- 0.1 x 10(6) bacteria/g feces (P < 0.0001) were observed in the probiotic group compared to the placebo group. During May, there was a tendency for fewer subjects, (76.2% vs 95.2%, P = 0.078) to report runny nose, while during June, fewer subjects, 11.1% vs 33.3%, reported nasal blocking in the probiotics group (P = 0.101). Concomitantly, fewer subjects in the probiotic group had infiltration of eosinophils in the nasal mucosa compared to the placebo group, 57.1% vs 95% (P = 0.013). Eye symptoms tended to be slightly more frequent in the probiotic group, 12.5 d [interquartile range (IQR) 6-18] vs 7.5 d (IQR 0-11.5) (P = 0.066) during May. Fecal IgA was increased in the placebo group during the pollen season; this increase was prevented by the probiotics (P = 0.028).ConclusionBirch pollen allergy was shown to be associated with changes in fecal microbiota composition. The specific combination of probiotics used was shown to prevent the pollen-induced infiltration of eosinophils into the nasal mucosa, and indicated a trend for reduced nasal symptoms.

Dataset Information

Multi-Approach Analysis for the Identification of Proteases within Birch Pollen.

Publications

Multi-Approach Analysis for the Identification of Proteases within Birch Pollen.

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