Allergenic activity of purified proteins of obeche wood dust.

<p><b>4A</b>. IgE-binding (ELISA) to purified obeche proteins (24 and 12 kDa). Solid line shows the positivity cut-off point, dotted lines show mean OD for each group. <b>4B</b>. Basophil activation tests using the <i>in-house</i> obeche wood dust extract and purified proteins (24 and 12 kDa). The solid line shows the threshold of positivity.</p

Demographic and clinical characteristics of subjects with confirmed occupational rhinitis/asthma (ORA+) <i>vs.</i> asymptomatic exposed (ORA−) and controls (CG). SPT: skin prick test; SD: standard deviation; *: p<0.05.

<p>Demographic and clinical characteristics of subjects with confirmed occupational rhinitis/asthma (ORA+) <i>vs.</i> asymptomatic exposed (ORA−) and controls (CG). SPT: skin prick test; SD: standard deviation; *: p<0.05.</p

Results of <i>in vivo</i> and <i>in vitro</i> tests with obeche extract and purified proteins in subjects with confirmed occupational rhinitis and asthma (ORA+; R = rhinitis; R+A = rhinitis and asthma).

<p>Results regarding food allergy assessment and CCD recognition are also included.</p

ELISA inhibition studies in ORA+ subjects.

<p>Bromelain (CCD, solid line) and obeche (dotted line) are inhibitors and obeche extract is the solid phase.</p

Measurement of specific antibodies to obeche wood dust.

<p>Means are represented by dotted lines and a horizontal line indicates cut-off value. *p<0.05 symptomatic vs asymptomatics; **p<0.05 symptomatic vs controls. <b>1A</b>. Levels of specific IgE in ORA+, ORA− and CG subjects; <b>1B</b>. Levels of specific IgG in ORA+, ORA− and CG subjects.</p

Calculated affinities energies and dissociation constants of ligands to Bos d 5.

<p>Calculated affinities energies and dissociation constants of ligands to Bos d 5.</p

Annexin V staining of PBMCs.

<p>PBMCs were left unstimulated or treated with iron-catechol alone or with PMA alone or in the presence of <i>apo</i>- or <i>holo</i>-Bos d 5. Living cells were gated on forward and side scatter and then plotted for CD4 and Annexin V. (A) Representative pictograms of PBMCs plotted for CD4 and Annexin V. (B) Summary of percentage of CD4+Annexin V+ cells. (C) Summary of percentage of CD4-Annexin V+ cells. Statistical analyses were conducted with repeated measures ANOVA following Newman-Keuls Multiple Comparison test. ***P<0.001, *P<0.05.</p

<i>Apo</i>-, but not <i>holo</i>-Bos d 5 promotes CD4 cells.

<p>PBMCs were treated with PMA alone or in the presence of <i>apo</i>- or <i>holo-</i>Bos d 5. (A) Living cells were gated on forward and side scatter, before gating on CD3 positive cells were performed. CD3 gated cells were then analyzed for their CD4 and CD8 expression.Representative pictograms of CD3 gated PBMCs plotted for CD4 and CD8 (B) Percentage of CD3+CD4+ cells. (C) Percentage of CD3+CD8+ cells. Statistical analyses were conducted with repeated measures ANOVA following Newman-Keuls Multiple Comparison test. **<i>P</i><0.01, ****<i>P</i><0.0001, ns not significant.</p

Bos d 5 is able to bind iron via siderophores.

<p>(A) Bos d 5 (1GX9) with Fe(DHBA)₂ (B) chemical structure of siderophores (C) UV-VIS spectra of Fe(DHBA) complex in the presence or absence of Bos d 5 or with Bos d 5 alone. (D) Iron-staining of <i>apo</i>- or <i>holo</i>-Bos d 5 as well as controls (buffer and iron-siderophore-complex) dotted on nitrocellulose-membrane.</p

Bos d 5 devoid of iron promotes IL13 and IFNγ secretion.

<p>(A) IL13 levels and (B<b>)</b> IFNγ levels of stimulated PBMCs. Statistical analyses were conducted with repeated measures ANOVA following Newman-Keuls Multiple Comparison test. **<i>P</i><0.01, *** P<0.001, ns not significant.</p