In the fenugreek model (Fig. 3C,D) only peanut displayed a partial inhibition of fenugreek positive sera at this concentration. In general, all antibody reactions, total and specific IgE as well as specific IgG1, were elevated in immunized Adriamycin animals compared to control groups, regardless of challenge (Figs 2 and 3). Fenugreek had an inhibitory effect on the levels of all cytokines in both models both in vivo, after challenge, and ex vivo, after spleen cell stimulation (Fig. 4, IL-4 and IL-13; and supplementary figure (Fig. S1), IL-2, IL-5, IL-10 and IFN-γ). This is reflected by lower cytokine levels in spleen cells from fenugreek immunized mice when stimulated with fenugreek compared to cells stimulated

with lupin. In both models, stimulation with the primary allergen yielded strong responses with a mixed Th1/Th2 profile, but with an emphasis on Th2 responses, as reported earlier [25, 26]. A positive cytokine response was defined as a response significantly higher than the cytokine release from unstimulated cells and significantly higher than cytokine release from cells of control animals stimulated with the same allergen.

When looking at the responses after stimulation with cross-allergens in the model of lupin allergy, stimulation with selleck kinase inhibitor soy extract yielded higher IL-4 and IL-13 responses compared to unstimulated cells and control cells stimulated with soy (Fig. 4A,B). Peanut stimulated Carteolol HCl cells from mice challenged with lupin also released higher levels of the same cytokines, however only significantly higher than unstimulated cells and not to peanut stimulated control cells. In the model of fenugreek allergy, the inhibitory

effect of fenugreek on the spleen cells both in vivo and ex vivo makes it difficult to evaluate possible cross-reactions. There is, however, a tendency to increased responses after lupin stimulation regarding IL-2, IL-4 and IL-10 when compared to unstimulated cells, but no differences could be seen between the different groups of mice (Fig. 4C,D). In two mouse models of legume allergy, we have shown clinically relevant cross-allergy to other legumes. The proportion of cross-allergy in sensitized mice varied from 12.5% up to 75% with a clinical score of 2 or higher. The majority of the legumes displayed a cross-allergy of 30% or more. This is in contrast to Lifrani et al. [28] who demonstrated cross-reactivity in vitro between peanut and lupin, but could not find any cross-allergy to lupin in peanut sensitized mice. Our finding is, however, in concordance with findings from the Norwegian Food Allergy Register [24] and other publications on cross-allergy to lupin [15, 19–22, 29] and fenugreek in peanut-sensitized individuals [10]. This illustrates the potential for cross-allergy in legume allergic patients, even though this has earlier been regarded as relatively rare [30, 31].