A major IgE epitope-containing grass pollen allergen domain from Phl p 5 folds as a four-helix bundle

Phl p 5, a 29 kDa major allergen from timothy grass pollen, is one of the most reactive members of group 5 allergens. Its sequence comprises two repeats of a novel alanine-rich motif (AR) whose structure and allergenic response are still mostly unknown. We report here a structural characterization of an immunodominant fragment of Phl p 5, Phl p 5(56-165) which comprises the first AR repeat. Recombinant (r)Phl p 5(56-165) was expressed in Escherichia coli, purified to homogeneity and shown to be sufficient to react with serum IgE from 90% of grass pollen allergic patients. Using NMR spectroscopy, we show conclusively that the fragment forms a compact globular domain which is, however, prone to degradation with time. The rPhl p 5(56-165) fold consists of a four-helix bundle held together by hydrophobic interactions between the aromatic rings and aliphatic side chains. This evidence gives clear indications about the structure of the full-length Phl p 5 and provides a rational basis for finding ways to stabilize the fold and designing therapeutic vaccines against grass pollen allergy

A major IgE epitope-containing grass pollen allergen domain from Phl p 5 folds as a four-helix bundle

Phl p 5, a 29 kDa major allergen from timothy grass pollen, is one of the most reactive members of group 5 allergens. Its sequence comprises two repeats of a novel alanine-rich motif (AR) whose structure and allergenic response are still mostly unknown. We report here a structural characterization of an immunodominant fragment of Phl p 5, Phl p 5(56-165) which comprises the first AR repeat. Recombinant (r)Phl p 5(56-165) was expressed in Escherichia coli, purified to homogeneity and shown to be sufficient to react with serum IgE from 90% of grass pollen allergic patients. Using NMR spectroscopy, we show conclusively that the fragment forms a compact globular domain which is, however, prone to degradation with time. The rPhl p 5(56-165) fold consists of a four-helix bundle held together by hydrophobic interactions between the aromatic rings and aliphatic side chains. This evidence gives clear indications about the structure of the full-length Phl p 5 and provides a rational basis for finding ways to stabilize the fold and designing therapeutic vaccines against grass pollen allergy

Evaluation of laboratory data by conventional statistics and by three types of neural networks

Abstract Sixty-three patients with lung (34 small-cell, 18 squamous, 11 adeno-) carcinomas and 43 patients with benign lung diseases were characterized with seven tumor markers: neuron-specific enolase (NSE); cancer antigens CA 19-9, CA 125, CA 15-3, and CA 50; carcinoembryonic antigen (CEA); and tissue polypeptide antigen. Diagnosis had been established by histological examination after surgery and used for classification. After vector transformation, the tumor marker data were fed into neural networks (NNs) based on three different types of learning algorithms: backpropagation (BP), competitive learning (CL), and Hopfield (H). For comparison, the data were evaluated with multivariate stepwise discriminant analysis (MVSDA). BP-NNs are equal to (NSE, CA 19-9, CEA) or better than (100% correct classification when using all seven markers) MVSDA in assigning the correct diagnosis to the patients. Cross-validation data yielded shrinkage effects ranging from 0% to 12.5%. Quality-control (QC) data were evaluated by traditional QC algorithms and compared with the results obtained by a BP-NN. The results show that the BP-NNs could only partly fulfill the tasks of three QC algorithms regarding the violation of static borders but gave good results with respect to dynamic changes.</jats:p

In situ localization of latex allergens in 3 different brands of latex gloves by means of immunogold field emission scanning and transmission electron microscopy

Background: Latex proteins represent relevant allergens, particularly fur those persons who are frequently exposed to latex products (eg, health care workers and patients with chronic disorders), Although several latex allergens have been characterized by biochemical and molecular biologic techniques, little information is available concerning the in situ localization of allergenic proteins in latex products. Objective: The objective of the present study was the in situ localization of latex allergens. Methods: Serum IgE from patients with latex allergy reacting with a broad range (5-200 kd) of latex allergens was used for the in situ localization of latex allergens. One surgical and 2 examination latex slave brands were investigated by using immunogold field emission scanning and transmission electron microscopy, Results: Allergens were detected on the inner and outer surface of the gloves, particularly near the edges, crests, or folds of the bleb-like structures visible on the surface of the latex material at high magnifications. In ultrathin cross-sections, latex allergens were Pound throughout the sections. Conclusions: Latex allergens were localized on the outer and inner surface hut also in the interior of latex gloves. The occurrence of latex allergens on the surface of latex products may be related to their potential to induce local reactions and, perhaps, to sensitize individuals by means of contact