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      Evaluating the Allergic Risk of Genetically Modified Soybean

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          Abstract

          Genetically modified (GM) soybean (carrying the EPSPS transgene) is the most common GM food in Korea. In order to assess whether genetic modification increases the allergenic risk of soybeans, the allergenicity and IgE-reactive components of wild-type and GM soybean extracts were compared in allergic adults who had been sensitized to soybeans. We enrolled 1,716 adult allergy patients and 40 healthy, non-atopic controls. Skin prick tests and IgE enzyme linked immunosorbent assays (ELISAs) were performed using wild-type and GM soybean extracts, along with other common inhaled allergens. The specificities of serum IgE antibodies from allergic patients and the identities of the IgE-reactive components of the soybean extracts were compared using ELISA inhibition testing, 2-dimensional gel electrophoresis, and IgE immunoblotting. To evaluate the effects of digestive enzymes and heat treatment, the soybean extracts were heated or pre- incubated with or without simulated gastric and intestinal fluids. The IgE sensitization rates to wild-type and GM soybeans were identical (3.8% of allergic adults), and circulating IgE antibodies specific for the two extracts were comparable. The results of the ELISA inhibition test, SDS-PAGE, and IgE immunoblotting showed a similar composition of IgE-binding components within the wild-type and GM extracts, which was confirmed using two-dimensional gel electrophoresis, IgE immunoblotting, and amino acid sequencing. None of the subjects had a positive response to purified EPSPS protein in the skin prick test, ELISA, or IgE immunoblot analysis. These findings suggest that the IgE sensitization rate to GM soybean extracts is identical to that of wild-type soybean extracts in adult allergy patients. In addition, based on both in vivo and in vitro methods, the allergenicity of wild type and GM soybean extracts was identical.

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          Lack of detectable allergenicity of transgenic maize and soya samples.

          The safety issues regarding foods derived from genetically modified (GM) plants are central to their acceptance into the food supply. The potential allergenicity of proteins newly introduced in GM foods is a major safety concern. We sought to monitor, in potentially sensitive human populations, the allergenicity effects of 5 GM materials obtained from sources with no allergenic potential and already under commercialization in the European Union. We have performed skin prick tests with protein extracts prepared from transgenic maize (MON810, Bt11, T25, Bt176) and soya (Roundup Ready) samples and from nontransgenic control samples in 2 sensitive groups: children with food and inhalant allergy and individuals with asthma-rhinitis. We have also tested IgE immunoblot reactivity of sera from patients with food allergy to soya (Roundup Ready) and maize (MON810, Bt11, Bt176) samples, as well as to the pure transgenic proteins (CryIA[b] and CP4 5-enolpyruvylshikimate-3-phosphate synthase). None of the individuals undergoing tests reacted differentially to the transgenic and nontransgenic samples under study. None of the volunteers tested presented detectable IgE antibodies against pure transgenic proteins. The transgenic products under testing seem to be safe in terms of allergenic potential. We propose postmarket testing as an important screening strategy for putative allergic sensitization to proteins introduced in transgenic plants.
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            Two-spotted spider mite allergy: immunoglobulin E sensitization and characterization of allergenic components.

            Previous investigations demonstrated that two-spotted spider mite (TSM) inhalation causes allergic asthma in agricultural workers. This work investigates whether TSM causes similar problems in the urban population. We determined the sensitization rate to TSM. We also identified immunoglobulin (Ig)E-binding components and evaluated their relationship with house-dust mite (HDM) allergens. We carried out skin prick test (SPT) with TSM in 1,806 respiratory allergy patients over 1 year. TSM-IgE was detected by enzyme-linked immunoadsorbent assay (ELISA). TSM-sensitized patients were classified into two groups: patients who were skin test-positive to both TSM and HDM were included in group A and patients who were skin test-positive to TSM only were included in group B. ELISA inhibition test using sera from group A and B were conducted. IgE-immunoblotting was used to identify major allergens. These were purified by 2-dimensional electrophoresis and blotted onto polyvinylidene difluoride, and N-terminal sequences were identified. SPT (> or = 2+ of allergen/histamine) was positive in 358 (19.8%) patients. Twelve (6.6%) showed positive response to TSM only, and 54.5% had positive specific IgE. ELISA inhibition test using sera from two groups showed significant inhibition by TSM with minimal inhibition by HDM. Amino acid sequence of three major allergens was not homologous with any previously characterized allergens. IgE-sensitization rate to TSM was 19.8% in respiratory allergy patients. Eleven IgE-binding components and three major allergens were identified. The pIs and amino acid sequences of the major allergens were determined.
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              Molecular cloning of a protein associated with soybean seed oil bodies that is similar to thiol proteases of the papain family.

              A 34,000-Da protein (P34) is one of the four major soybean oil body proteins observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of isolated organic solvent-extracted oil bodies from mature seeds. P34 is processed during seedling growth to a 32,000-Da polypeptide (P32) by the removal of an amino-terminal decapeptide (Herman, E.M., Melroy, D.L., and Buckhout, T.J. (1990) Plant Physiol, in press). A soybean lambda ZAP II cDNA library constructed from RNA isolated from midmaturation seeds was screened with monoclonal antibodies directed against two different epitopes of P34. The isolated cDNA clone encoding P34 contains 1,350 base pairs terminating in a poly(A)+ tail and an open reading frame 1,137 base pairs in length. The open reading frame includes a deduced amino acid sequence which matches 23 of 25 amino-terminal amino acids determined by automated Edman degradation of P34 and P32. The cDNA predicts a mature protein of 257 amino acids and of 28,641 Da. The open reading frame extends 5' from the known amino terminus of P34 encoding a possible precursor and signal sequence segments with a combined additional 122 amino acids. Prepro-P34 is deduced to be a polypeptide of 42,714 Da, indicating that the cDNA clone apparently encodes a polypeptide of 379 amino acids. A comparison of the nucleotide and deduced amino acid sequences in the GenBank Data Bank with the sequence of P34 has shown considerable sequence similarity to the thiol proteases of the papain family. Southern blot analysis of genomic DNA indicated that the P34 gene has a low copy number.
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                Author and article information

                Journal
                Yonsei Med J
                YMJ
                Yonsei Medical Journal
                Yonsei University College of Medicine
                0513-5796
                1976-2437
                31 August 2006
                31 August 2006
                : 47
                : 4
                : 505-512
                Affiliations
                [1 ]Department of Allergy and Rheumatology, Ajou University School of Medicine, Suwon, Korea.
                [2 ]Department of Food science and Technology, Seoul National University, Seoul, Korea.
                [3 ]College of Agriculture, Chonbuk National University, Chonju, Korea.
                Author notes
                Reprint address: requests to Dr. Hae-Sim Park, Department of Internal Medicine, Yonsei University Wonju College of Medicnie, 162 Ilsan-dong, Wonju 220-701, Korea. Tel: 82-31-219-5196, Fax: 82-31-219-5154, hspark@ 123456ajou.ac.kr

                *Current address: Department of Internal Medicine, Yonsei University Wonju College of Medicine, 162 Ilsan-dong, Wonju 220-701, Korea.

                Article
                10.3349/ymj.2006.47.4.505
                2687731
                16941740
                c7f6c9df-3ad4-45b4-ae49-2ed03b6d604b
                Copyright © 2006 The Yonsei University College of Medicine

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 01 November 2005
                : 20 March 2006
                Categories
                Original Article

                Medicine
                genetically modified,soybean,allergic risk
                Medicine
                genetically modified, soybean, allergic risk

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