Decreased mast cell protease induction by heated-OVA.

<p>Heated OVA (h-OVA, black bar) induced significantly lower amounts of mast cell protease (MMCP-1), the marker of mast cell activation, compared to mice fed with native OVA (white bar). Data are represented as mean ± SEM (n = 10 mice/group), representative data from one out of three independent experiments. *P≤0.05, **P≤0.01, ***P≤0.001.</p

Numbers of Tregs in splenocytes and mesenteric lymph nodes of OVA treated mice.

<p>Typical plots depicting numbers of Tregs in mouse splenocytes (a) and mesenteric lymph node (b) in gated CD3+CD4+CD8– T helper cells after feeding with OVA, h-OVA or PBS, respectively. Numbers in upper quadrants shows proportions (mean ± SD) of either CD25–Foxp3+ or CD25+Foxp3+ Th cells out of all cells. Representative data from one out of three independent experiments. *P≤0.05.</p

Number of Tregs in spleen cell suspensions co-cultured <i>in vitro</i> with OVA digests.

<p>The percentage of Tregs in cell suspension isolated from spleens of non-stimulated (naïve) BALB/c mice cultured <i>in vitro</i> for 48 hours with undigested (0′) and after 20 (20′) and 40 minutes (40′) peptic digest of OVA (white bars), h-OVA (black bars) or b-OVA (grey bars). The data represent the percentage of CD4+Foxp3+ cells out of all live cells as measured by FACS. Representative data from one out of three independent experiments are shown. Data are represented as mean ± SEM.</p

Cytokine production after <i>in vitro</i> restimulation with OVA.

<p>The cytokine production from mesenteric lymph nodes (a) and splenocytes (b) of BALB/c mice fed with OVA (white bars) or h-OVA (black bars) and stimulated <i>in vitro</i> with appropriate allergen. Cytokine levels are expressed after subtraction of base line levels of unstimulated lymph node cells or splenocytes. Data shown are mean values ± SEM (n = 4–7 mice/group), representative data from one out of three independent experiments. *P≤0.05, **P≤0.01, ***P≤0.001, n.d. =  not detectable.</p

RP-HPLC separation profile of native-OVA and heated-OVA peptic digests.

<p>RP-HPLC separation profile monitored at 280 nm corresponds to OVA and OVA heated at 70°C (h)-OVA or boiled at 95°C (b)-OVA undigested (0′) and after 20 (20′) and 40 minutes (40′) of digestion by pepsin. RT – retention time.</p