Lentil (Lens culinaris) Lipid Transfer Protein Len c 3: A Novel Legume Allergen

Background: Lentils are increasingly consumed in many parts of the world. Two allergens, Len c 1 and 2, have been reported previously. Recently, peanut and green bean lipid transfer proteins (LTPs) have been identified as the first two members of an important group of allergens that might be associated with severe food allergies. Objective:To investigate lentil LIP as a potential new allergen. Methods: Efficacy of LIP extraction was monitored at different acidic pH values, using immunoblotting with cross-reactive anti-peach LIP antiserum. Natural LIP was purified from lentil extract and expressed as recombinant allergen in Escherichia Sera from 10 lentil-allergic and/or-sensitized patients (Spain: 6, Italy: 1 and the Netherlands: 3) were used to further characterize lentil LIP. Results: Natural lentil LIP, purified from the homogenized germinated seeds and optimally extracted at pH 3, was identified and designated as allergen Len c 3. By CAP, 9/10 sera showed specific IgE to Len c 3. Recombinant (r) Len c 3 was successfully purified. The natural (n) Len c 3 CAP was completely inhibited by rLen c 3/rPru p 3. IgE binding to lentil pH 3 extract blot was completely inhibited by rLen c 3. Conclusion: The availability of immunochemically active nLen/rLen c 3 as a novel legume allergen facilitates further development and implementation of a third (next to peanut and green bean) legume LIP in component-resolved diagnosis strategies and contributes to evaluate the clinical importance of legume LTPs. Preferential extraction of Len c 3 (pH 3) will affect the production of sensitive extract-based diagnostic tests. Copyright (C) 2011 S. Karger AG, Base

Characterization of Monoclonal Antibodies to turbot (Scophthalmus maximus L.)

Monoclonal antibodies (mAbs) can be used in several applications, v.g in the aquaculture field. In attempt to study the immune system of the turbot fish, we wanted to develop mAbsdirected against turbot’s lymphocytes, mainly T cells. BALB/c mice were immunized with turbot thymic cells and mouse splenocytes were fused with mouse myeloma cells to genera-te hybridomas. The screening of secretory hybridomas was first performed by ELISA. Then, specificity of different monoclonal antibodies was determined by using immunohistoche-mical and immunofluorescence methods and, on the basis on their reactivity with cells from lymphoid tissues of turbot, three antibodies called RoA3, RoA4 and RoD4 were selected.RoA3 positively reacted with the membrane components of cells resembling to thymocytes, RoA4 stained cells with extended cytoplasmic processes, compatible with reticular cells.In the case of RoD4 mAb, it stains large cells mainly in the medullar zone of the thymus, compatible with macrophages and reticular cells. Flow cytometry studies showed its specifici-ty for thymic cells (no reaction with spleen or pronephros cells), further confirmed by western blot, indicating that RoD4 mAb recognizes a specific protein of 130-135 kD only onthymic tissue