[PubMed] [Google Scholar]Tovey ER, Chapman MD, Platts-Mills TA

[PubMed] [Google Scholar]Tovey ER, Chapman MD, Platts-Mills TA. atopic dermatitis (AD), whereas it is only a minor allergen for patients suffering from respiratory forms of HDM allergy. Rabbit Polyclonal to ZC3H8 Thus, rDer p 11 might be a useful serological marker allergen for the identification of a subgroup of HDM-allergic patients suffering from HDM-associated AD. INTRODUCTION House dust mites (HDMs) belong 6-Thioguanine to the most important allergen sources worldwide (Platts-Mills and Chapman, 1987; Thomas, 2011). Approximately 50% of allergic patients in Central Europe below an altitude of 1500 m are sensitized to allergens from HDM, which causes numerous allergic symptoms such as 6-Thioguanine allergic rhinoconjunctivitis, allergic asthma, and skin manifestations, in particular atopic dermatitis (AD; Platts-Mills and Chapman, 1987; Platts-Mills is usually shown in the Supplementary Physique S1 online. Der p 11 is usually a protein of 874 amino acids with a deduced molecular mass of ~103 kDa, and it shows high sequence identities (>85% sequence identity) with paramyosins from HDM, itchy mites, and tropical mites (Supplementary Physique S1 online). All four predicted (“type”:”entrez-protein”,”attrs”:”text”:”AAO73464.1″,”term_id”:”37778944″,”term_text”:”AAO73464.1″AAO73464.1), (“type”:”entrez-protein”,”attrs”:”text”:”Q967Z0″,”term_id”:”42559514″,”term_text”:”Q967Z0″Q967Z0), (“type”:”entrez-protein”,”attrs”:”text”:”ACC65584.1″,”term_id”:”183987114″,”term_text”:”ACC65584.1″ACC65584.1), (“type”:”entrez-protein”,”attrs”:”text”:”Q8MUF6″,”term_id”:”42559485″,”term_text”:”Q8MUF6″Q8MUF6), (“type”:”entrez-protein”,”attrs”:”text”:”XP_002407289.1″,”term_id”:”241048541″,”term_text”:”XP_002407289.1″XP_002407289.1), (“type”:”entrez-protein”,”attrs”:”text”:”XP_002432355.1″,”term_id”:”242023873″,”term_text”:”XP_002432355.1″XP_002432355.1), (“type”:”entrez-protein”,”attrs”:”text”:”XP_393281.2″,”term_id”:”66510482″,”term_text”:”XP_393281.2″XP_393281.2), (“type”:”entrez-protein”,”attrs”:”text”:”NP_001124374.1″,”term_id”:”195963325″,”term_text”:”NP_001124374.1″NP_001124374.1), (“type”:”entrez-protein”,”attrs”:”text”:”XP_314309.4″,”term_id”:”158293004″,”term_text”:”XP_314309.4″XP_314309.4), (“type”:”entrez-protein”,”attrs”:”text”:”CAA41557.1″,”term_id”:”10959″,”term_text”:”CAA41557.1″CAA41557.1), (“type”:”entrez-protein”,”attrs”:”text”:”XP_003371652.1″,”term_id”:”339252858″,”term_text”:”XP_003371652.1″XP_003371652.1), (“type”:”entrez-protein”,”attrs”:”text”:”Q9NJA9″,”term_id”:”42559536″,”term_text”:”Q9NJA9″Q9NJA9.1), (“type”:”entrez-protein”,”attrs”:”text”:”ADY40789.1″,”term_id”:”324501780″,”term_text”:”ADY40789.1″ADY40789.1), and (“type”:”entrez-protein”,”attrs”:”text”:”AAX26229.2″,”term_id”:”76154815″,”term_text”:”AAX26229.2″AAX26229.2). Expression and purification of folded rDer p 11 rDer p 11 was purified to homogeneity by affinity chromatography and compared with purified natural Der p 11 (nDer p 11) by SDS-PAGE under reducing and nonreducing conditions and subsequent metallic staining (Physique 1a). Under nonreducing conditions, natural as well as recombinant Der p 11 occurred mainly as high-molecular-weight aggregates of more than 250 kDa. However, under reducing conditions, recombinant and nDer p 11 migrate as bands of ~100 kDa and several smaller fragments in SDS-PAGE, which is in agreement with the predicted molecular excess weight of total monomeric Der p 11 (i.e., ~103 kDa) and degradation products (Physique 1a) (Tsai body (left panel) and feces (right panel) were incubated with anti-rDer p 11, anti-rDer p 2 antibodies, or preimmune Ig. Molecular weights (kDa) are indicated. (b) Immunogold electron microscopy. Ultrathin sections from muscle mass and feces were stained with anti-Der p 11 antiserum (right panel) 6-Thioguanine or preimmune antiserum (left panel). Arrows show two of the platinum particles localizing Der p 11. Black bars denote the level. IgE reactivity of rDer p 11 In a first series of experiments, we compared recombinant Der p 11 with nDer p 11 regarding IgE reactivity, which showed that both proteins exhibited comparable IgE binding in dot-blot and western blot assays (data not shown). Therefore, we used rDer p 11 for IgE reactivity studies and compared the IgE 6-Thioguanine reactivity of a random sample of HDM-allergic patients from Zimbabwe with dot-blotted rDer p 11 and rDer p 2 (Physique 3a). rDer p 11 reacted with fewer patients, and the IgE reactivity was less pronounced than that of rDer p 2. None of the HDM-allergic patients sera reacted with BSA, and serum from a nonallergic person or buffer gave no reaction with any of the dotted proteins. Open in a separate window Physique 3 IgE reactivity of rDer p 11(a) Comparison of the intensity of IgE acknowledgement of rDer p 11, rDer p 2, and BSA (unfavorable control) as tested by dot blot with sera from mite-allergic patients (1C20), a nonallergic individual (NC), and with buffer alone (BC). (b) Prevalence of IgE reactivity to rDer p 11 and rDer p 2 (y axis: percentages of reactive patients) in different populations of house dust mite-allergic patients (x axis: Austria: = 47; France: = 38; Italy: = 67; Sweden: = 52; Zimbabwe: = 34). Higher prevalence of IgE reactivity to Der p 11 in populations comprising AD patients We then analyzed the frequency of IgE acknowledgement of Der p 11 using sera from HDM-allergic patients from several European countries (Austria: = 47, France: = 38, Italy: = 67, Sweden: = 52) and from southern Africa (i.e., Zimbabwe: = 34; Physique 3b). In the European HDM-allergic patients, the frequency of IgE reactivity to Der p 11 was relatively low (Austria 13%, France 16%, Italy 7%, and Sweden 10%) when compared with the major HDM allergen Der p 2.