IgE cross-reactivity of peanut with walnut and soybean in children with food allergy

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Echeverría-Zudaire, S. Fernández-Fernández, A. Rayo-Fernández, C. Muñóz-Archidona, R. Checa-Rodriguez" "autores" => array:5 [ 0 => array:2 [ "nombre" => "L.Á." "apellidos" => "Echeverría-Zudaire" ] 1 => array:2 [ "nombre" => "S." "apellidos" => "Fernández-Fernández" ] 2 => array:2 [ "nombre" => "A." "apellidos" => "Rayo-Fernández" ] 3 => array:2 [ "nombre" => "C." "apellidos" => "Muñóz-Archidona" ] 4 => array:2 [ "nombre" => "R." 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Miceli Sopo, M. Greco, S. Monaco, A. Bianchi, B. Cuomo, L. Liotti, I.D. Iacono" "autores" => array:7 [ 0 => array:2 [ "nombre" => "S." "apellidos" => "Miceli Sopo" ] 1 => array:2 [ "nombre" => "M." "apellidos" => "Greco" ] 2 => array:2 [ "nombre" => "S." "apellidos" => "Monaco" ] 3 => array:2 [ "nombre" => "A." "apellidos" => "Bianchi" ] 4 => array:2 [ "nombre" => "B." "apellidos" => "Cuomo" ] 5 => array:2 [ "nombre" => "L." "apellidos" => "Liotti" ] 6 => array:2 [ "nombre" => "I.D." "apellidos" => "Iacono" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0301054616300635?idApp=UINPBA00004N" "url" => "/03010546/0000004400000006/v1_201611090130/S0301054616300635/v1_201611090130/en/main.assets" ] "en" => array:20 [ "idiomaDefecto" => true "cabecera" => "Original Article" "titulo" => "IgE cross-reactivity of peanut with walnut and soybean in children with food allergy" "tieneTextoCompleto" => true "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "524" "paginaFinal" => "530" ] ] "autores" => array:1 [ 0 => array:4 [ "autoresLista" => "M.N. Kim, K.E. Lee, J.Y. Hong, K.W. Kim, K.-E. Kim, M.H. Sohn, J.W. Park" "autores" => array:7 [ 0 => array:3 [ "nombre" => "M.N." "apellidos" => "Kim" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] ] ] 1 => array:3 [ "nombre" => "K.E." "apellidos" => "Lee" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] ] ] 2 => array:3 [ "nombre" => "J.Y." "apellidos" => "Hong" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] ] ] 3 => array:3 [ "nombre" => "K.W." "apellidos" => "Kim" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] ] ] 4 => array:3 [ "nombre" => "K.-E." "apellidos" => "Kim" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] ] ] 5 => array:4 [ "nombre" => "M.H." "apellidos" => "Sohn" "email" => array:1 [ 0 => "mhsohn@yuhs.ac" ] "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "*" "identificador" => "cor0005" ] ] ] 6 => array:3 [ "nombre" => "J.W." "apellidos" => "Park" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "b" "identificador" => "aff0010" ] ] ] ] "afiliaciones" => array:2 [ 0 => array:3 [ "entidad" => "Department of Pediatrics, Severance Hospital, Institute of Allergy, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea" "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:3 [ "entidad" => "Department of Internal Medicine, Severance Hospital, Institute of Allergy, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea" "etiqueta" => "b" "identificador" => "aff0010" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0025" "etiqueta" => "Figure 5" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr5.jpeg" "Alto" => 875 "Ancho" => 2500 "Tamanyo" => 184429 ] ] "descripcion" => array:1 [ "en" => "Immunoblot analysis of walnut (A) and peanut (B) protein extracts using peanut specific IgE positive individual sera (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>. Group 1)." ] ] ] "textoCompleto" => "IntroductionThe prevalence of food allergies has been increasing in recent years and can affect up to 10% of young children and 2–3% of adults globally.<a class="elsevierStyleCrossRefs" href="#bib0130">1,2</a> Food allergies can cause urticaria, angio-oedema, rhinitis, wheezing, and in some cases can be life threatening due to anaphylactic shock. A recent meta-analysis study reported that fatal food anaphylaxis has an incidence rate of 1.81per million person-years.<a class="elsevierStyleCrossRef" href="#bib0140">3</a>Peanut is one of the primary allergenic foods. It contains at least 18 proteins that have been shown to cause specific IgE reactivity and have a higher potential to cause anaphylactic shock compared to other foods.<a class="elsevierStyleCrossRefs" href="#bib0145">4–6</a> The peanut allergens, vicilin (Ara h 1), 2S albumin (Ara h 2), and 11S globulin (Ara h 3), share significant amino acid homology with their respective allergens from other legumes and tree nuts.<a class="elsevierStyleCrossRefs" href="#bib0160">7–9</a>Tree nut, despite its botanical taxonomic distance, is believed to be cross-reactive with peanut, due to the high number of individuals reported to be co-sensitised or co-allergic to peanut and tree nut.<a class="elsevierStyleCrossRef" href="#bib0175">10</a> A study of the prevalence of peanut and tree nut allergies among 1218 newborns in the United Kingdom revealed that 1 in 200 children could have adverse reactions to either peanuts or tree nuts by the age of four.<a class="elsevierStyleCrossRef" href="#bib0180">11</a> In another study among individuals with nut allergies, 65% reacted specifically to peanuts of which 30% also reacted with other tree nuts.<a class="elsevierStyleCrossRef" href="#bib0185">12</a>Soybean, belonging to the same legume family as the peanut, has also been investigated as a cross-reactive food with peanuts. Of 69 legume-sensitive individuals, 43% showed sensitivity to soybean and 87% to peanut.<a class="elsevierStyleCrossRef" href="#bib0190">13</a> It has also been reported that soybean immunotherapy in a peanut allergy mouse model resulted in a significant reduction of clinical symptoms following peanut challenge compared with placebo-treated mice.<a class="elsevierStyleCrossRef" href="#bib0195">14</a>However, despite the above findings, the clinical significance of cross-reactivity between peanut allergens and those of walnut and soybean remains unclear. With this in mind, we sought to determine the cross-reactivity of peanut allergens with those of walnut and soybean in the sera of allergic patients.Materials and methodsSubjectsPooled sera from eight subjects with allergic sensitisation to both peanut and walnut were used to investigate cross-reactivity of peanut and walnut in an inhibition test (Supplemental Table 1). For the peanut and soybean cross-reactivity test, the pooled sera from 18 subjects with both peanut and soybean specific IgE antibodies were evaluated (Supplemental Table 2). Allergic sensitisation, the presence of specific IgE, was evaluated using an immunoCAP test (Phadia AB, Upsala, Sweden). The clinical data of the subjects used for the immunoblot analysis are summarised in <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>. The pooled sera from eight subjects without an allergic sensitisation were used as a normal control. All sera were obtained from Severance Children's Hospital of Yonsei University, Seoul (South Korea). This study was approved by the institutional Review Board of Severance Children's Hospital of Yonsei University.<elsevierMultimedia ident="tbl0005"></elsevierMultimedia>Protein extractionTo obtain protein extracts, peanuts, walnuts and soybeans were purchased and finely ground. The ground meals were defatted with cold (4°C) acetone (1:5, w/v) under constant stirring for one hour and then filtered with filter paper. The defatting procedure was repeated until the filtrate became clear. The defatted meals were completely air dried at ambient temperature and then stirred in phosphate buffered saline (PBS, pH 7.4) (1:4, w:v) for 48h at 4°C. The extracts were centrifuged twice at 13,000×g for 30min at 4°C. The supernatants were filtered and dialysed against several changes of distilled water for 48h at 4°C. The extracts were centrifuged once again, lyophilised, and stored at −20°C until use.Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)The extracts were resuspended in PBS and the protein concentration of each of the extracts was measured by the Bradford assay. Equal concentration of the samples was mixed with loading buffer (60mM Tris–HCl, 25% glycerol, 2% SDS, 14.4mM 2-mercaptoethanol, 0.1% bromophenol blue) and heated for 5min in a heating block at 4°C. The samples were then analysed by SDS-PAGE. Following electrophoresis, the gels were stained with Coomassie brilliant blue or used for immunoblotting.IgE immunoblottingProteins separated by SDS-PAGE were electrotransfered onto a polyvinyl-difluoride membrane. After blocking with 5% skimmed milk in PBS containing 0.1% tween 20, immunodetection of IgE-binding proteins was performed by sequential incubation with subjects’ sera (diluted 1:10) and alkaline phosphatase-conjugated goat anti-human IgE (¿-chain specific) (Sigma Chemical, MO, USA). IgE reactive protein bands were visualised using 5-bromo-4-chloro-3′-indolyphosphate p-toluidine salt/nitro-blue tetrazolium chloride (Promega, USA).Inhibition testPooled sera, which were positive for both peanut and walnut by ImmunoCAP, were incubated with 0, 0.1, 1, 5, 10, 50, 100μg of either peanut or walnut protein extracts (9:1, w:v) over night at 4°C. The IgE-reactivity of the inhibited sera against peanut and walnut was then measured by immunoCAP. Pooled sera of both peanut and soybean ImmunoCAP positive sera were also inhibited with either peanut or soybean protein and measured by a CAP radio allergosorbent test against peanut and soybean. House dust mite extract (Dermatophagoides pteronyssinus; Yonsei University, Seoul, Korea) was used as a negative control inhibitor.ResultsCross-inhibition of peanut with walnut and soybeanTo observe the cross-reactivity between peanut and walnut, pooled sera from eight subjects who were immunoCAP positive for both peanut and walnut (sex: male 6, female 2, age: 3.1±3.0yrs, peanut specific IgE level of uninhibited pooled serum: 8.57[kU/L], walnut specific IgE level of uninhibited pooled serum: 8.32[kU/L]) were incubated with protein extracts from either peanut or walnut. The sera were then evaluated for specific IgE activities using a peanut and walnut immunoCAP test. As shown in <a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>, peanut specific IgE levels were inhibited up to 80% by peanut protein extracts in a dose dependent manner, while only 20% was inhibited by walnut protein extracts. Walnut specific IgE levels showed similar tendencies and were inhibited up to 91% by walnut protein extracts but only 21% by peanut protein extracts.<elsevierMultimedia ident="fig0005"></elsevierMultimedia>To investigate peanut and soybean cross-reactivity, pooled sera from 18 subjects who were immunoCAP positive for both peanut and soybean (sex: male 16, female 2, age: 3.3±3.4yrs, peanut specific IgE level of uninhibited pooled serum: 31.8[kU/L], soybean specific IgE level of uninhibited pooled serum: 20.3[kU/L]) were incubated with either peanut or soybean protein extracts as described above. Then, an immunoCAP test for peanut and walnut specific IgE was conducted with the inhibited sera. Peanut specific IgE activity was inhibited up to 86% at an inhibitor concentration of 50μg/ml by peanut protein extracts, while only 26% was inhibited by soybean protein extracts (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>A). Soybean specific IgE also showed similar pattern inhibited up to 83% by soybean protein extracts but only 23% by peanut protein extracts (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>B).<elsevierMultimedia ident="fig0010"></elsevierMultimedia>Constitutive protein distribution and IgE reactive peptidesSDS-PAGE was conducted with protein extracts from peanuts, walnuts, and soybeans to observe the distribution of constitutive proteins (<a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>). This was followed by immunoblotting with various sera to ascertain which specific peptides were responsible for IgE reactivity and to determine whether the pooled sera positive for either peanut, walnut, or soybean reacted with corresponding allergens from each other (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>, <a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>).<elsevierMultimedia ident="fig0015"></elsevierMultimedia><elsevierMultimedia ident="fig0020"></elsevierMultimedia>As shown in <a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>A, lane 2, pooled serum of the peanut immunoCAP positive (>0.35kU/L) sera reacted strongly with peanut protein extracts as we expected. Pooled serum of each walnut and soybean immunoCAP positive sera also showed weak IgE reactive bands to peanut protein extracts. However, this binding was probably non-specific as a similar pattern was seen in the membrane with the normal control serum which is from the subjects without an allergic sensitisation (<a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>A).Again, as we expected, pooled serum of walnut imunoCAP positive (>0.35kU/L) sera reacted with walnut protein extracts (<a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>B, lane 2). Interestingly, peanut immunoCAP positive serum also showed a significant reactive band with walnut protein extracts (<a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>B, lane 3). Even though several bands were visible on the membrane using the normal control serum, the peanut specific IgE positive serum produced a stronger and different pattern of reactive bands especially between 20 and 60kDa with walnut protein (<a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>B).Pooled sera of soybean immunoCAP test positive sera also bound well to soybean proteins while peanut immunoCAP test positive serum showed non-specific bands similar to the control sera (<a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>C). Of the soybean proteins, high molecular weight peptides reacted more strongly than low molecular peptides particularly between 60 and 70kDa.Individual sera immunoblot analysisThe sera showed reactive IgE band to walnut protein in <a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>B, lane 3, even though the sera were positive for peanut but not to walnut by the immunoCAP test. To confirm whether the sera could react to walnut protein, an immunoblot analysis was repeated using individual sera. As shown in <a class="elsevierStyleCrossRef" href="#fig0025">Fig. 5</a>, significant IgE reactive bands were seen in three of the eight individual sera (<a class="elsevierStyleCrossRef" href="#fig0025">Fig. 5</a>A. lanes 2, 6 and 8). Each cross-reacting serum revealed a different IgE band pattern binding to walnut proteins of different sizes. However, each of the three sera bound to a peptide of around 30kDa. In addition, significant cross-reactive IgE bands were observed around 15kDa in lanes 2 and 6.<elsevierMultimedia ident="fig0025"></elsevierMultimedia>DiscussionThe cross-reactivity of peanut with walnut has been studied because they share IgE binding epitopes structurally related to the vicilin allergen in addition to having some structural homology to legumin allergens. Indeed, avoidance or restricted consumption of walnut has been recommended for peanut-sensitised individuals.<a class="elsevierStyleCrossRefs" href="#bib0165">8,15</a> In this study, our results from the inhibition test showed only 20% inhibition of peanut specific IgE by walnut protein extracts. This level of inhibition seems low, but when immunoblots of walnut protein extracts were performed with group 1 individual serum that were positive for peanut but negative to walnut by immunoCAP test, three out of the eight subjects showed significantly reactive IgE bands. We were careful to distinguish whether the vicilin and legumin peptides were reactive in both the peanut and walnut protein extracts, but this was difficult to confirm based on identification of the peptides by molecular weight alone. However, IgE reactive bands around 30kDa were seen in all of the three reactive subjects, and one of them was particularly strong (<a class="elsevierStyleCrossRef" href="#fig0025">Fig. 5</a>A, lane 8). This peptide is not known as a major allergen of walnut; therefore, further investigation is required for its identification and characterisation. Furthermore, IgE reactive bands were also seen around 15kDa, a similar molecular weight to Jug r 1, in two of the three reactive sera (<a class="elsevierStyleCrossRef" href="#fig0025">Fig. 5</a>A, lane 2, 6). As Jug r 1 (14–16kDa) and Jug r 3 (9kDa) are known to be associated with systemic reactions, it is recommended that even though the result of the immunoCAP test for walnut is negative, individuals with peanut allergies should be cautious about ingesting walnuts.<a class="elsevierStyleCrossRefs" href="#bib0205">16–20</a>Soybeans are used widely in various foods for both humans and animals. Peanut Ara h 3 has been reported to share IgE epitopes with the glycinin G1 acidic chain of soybean. A study with the sera of individuals allergic to both soybean and peanut showed that the absorption of soybean-binding antibodies by antigen-affinity chromatography resulted in a 73% reduction of IgE binding to peanut allergens.<a class="elsevierStyleCrossRefs" href="#bib0230">21,22</a> In our results, although the inhibition of peanut specific IgE by soybean protein extracts was higher than by walnut protein extracts, it was still only an inhibition of 26.7%. In previous studies, it has been demonstrated that individuals allergic to both peanut and soybean have IgE antibodies that bind preferentially to larger proteins, while the antibodies of those reacting only to soybean bind strongly to proteins with lower molecular weight.<a class="elsevierStyleCrossRefs" href="#bib0240">23,24</a> In our study, we could not see a significant difference between the IgE binding pattern of pooled sera of both peanut and soybean positive subjects and soybean only positive subjects. Peanut only positive sera did not show significant cross-reactive IgE bands to soybean protein extracts either. Extensive cross-reactivity studies have been undertaken on the different legume species. And the majority of studies do not provide the clinical cross-reactions among the legume species.<a class="elsevierStyleCrossRefs" href="#bib0190">13,22</a> Nevertheless, as has been reported in a previous study, soy could be a cause of food anaphylaxis, and, as such, individuals with peanut allergies are at risk of a cross-reactive response.<a class="elsevierStyleCrossRef" href="#bib0250">25</a> Individuals with peanut allergies should, therefore, be aware of this cross-reactivity, so that they can take the necessary measures if they have abnormal responses following ingestion of soybean.In our study, three out of the eight sera that were positive for peanut but were negative for walnut in the immunoCAP test showed a significant IgE reactivity to walnut proteins in the immunoblot analysis. This contradictory result between the immunoCAP test and immunoblot analysis should be considered in the diagnosis of other food allergies, especially for individuals who are immunoCAP negative but still have allergic symptoms. However, the sera used for the experiments were collected based on the result of the immunoCAP test. Positivity by the immunoCAP test does not necessarily mean that the subject will show clinical symptoms following ingestion of the allergenic food. Therefore, future work needs to be undertaken with sera collected from symptomatic individuals.Taken together, these results indicate that although the clinical significance of cross-reactivity of peanut with walnut and soybean has not been established, individuals with peanut allergies need to be cautious when consuming walnut and soybean.Ethical disclosuresConfidentiality of dataThe authors declare that they have followed the protocols of their work centre on the publication of patient data and that all the patients included in the study have received sufficient information and have given their informed consent in writing to participate in that study.Right to privacy and informed consentThe authors have obtained the informed consent of the patients and/or subjects mentioned in the article. The author for correspondence is in possession of this document.Protection of human subjects and animals in researchThe authors declare that no experiments were performed on humans or animals for this investigation.Conflicts of interestNone declared." "textoCompletoSecciones" => array:1 [ "secciones" => array:10 [ 0 => array:3 [ "identificador" => "xres753553" "titulo" => "Abstract" "secciones" => array:5 [ 0 => array:2 [ "identificador" => "abst0005" "titulo" => "Background" ] 1 => array:2 [ "identificador" => "abst0010" "titulo" => "Objective" ] 2 => array:2 [ "identificador" => "abst0015" "titulo" => "Methods" ] 3 => array:2 [ "identificador" => "abst0020" "titulo" => "Results" ] 4 => array:2 [ "identificador" => "abst0025" "titulo" => "Conclusion" ] ] ] 1 => array:2 [ "identificador" => "xpalclavsec755870" "titulo" => "Keywords" ] 2 => array:2 [ "identificador" => "sec0005" "titulo" => "Introduction" ] 3 => array:3 [ "identificador" => "sec0010" "titulo" => "Materials and methods" "secciones" => array:5 [ 0 => array:2 [ "identificador" => "sec0015" "titulo" => "Subjects" ] 1 => array:2 [ "identificador" => "sec0020" "titulo" => "Protein extraction" ] 2 => array:2 [ "identificador" => "sec0025" "titulo" => "Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)" ] 3 => array:2 [ "identificador" => "sec0030" "titulo" => "IgE immunoblotting" ] 4 => array:2 [ "identificador" => "sec0035" "titulo" => "Inhibition test" ] ] ] 4 => array:3 [ "identificador" => "sec0040" "titulo" => "Results" "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0045" "titulo" => "Cross-inhibition of peanut with walnut and soybean" ] 1 => array:2 [ "identificador" => "sec0050" "titulo" => "Constitutive protein distribution and IgE reactive peptides" ] 2 => array:2 [ "identificador" => "sec0055" "titulo" => "Individual sera immunoblot analysis" ] ] ] 5 => array:2 [ "identificador" => "sec0060" "titulo" => "Discussion" ] 6 => array:3 [ "identificador" => "sec0065" "titulo" => "Ethical disclosures" "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0070" "titulo" => "Confidentiality of data" ] 1 => array:2 [ "identificador" => "sec0075" "titulo" => "Right to privacy and informed consent" ] 2 => array:2 [ "identificador" => "sec0080" "titulo" => "Protection of human subjects and animals in research" ] ] ] 7 => array:2 [ "identificador" => "sec0085" "titulo" => "Conflicts of interest" ] 8 => array:2 [ "identificador" => "xack250546" "titulo" => "Acknowledgements" ] 9 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2015-12-14" "fechaAceptado" => "2016-04-27" "PalabrasClave" => array:1 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec755870" "palabras" => array:9 [ 0 => "Cross-reactivity" 1 => "Food allergy" 2 => "IgE-hypersensitivity" 3 => "Legume" 4 => "Peanut" 5 => "Soybean" 6 => "Tree nut" 7 => "Walnut" 8 => "Inhibition test" ] ] ] ] "tieneResumen" => true "resumen" => array:1 [ "en" => array:3 [ "titulo" => "Abstract" "resumen" => "BackgroundPeanut allergies are common and can be life-threating for sensitised individuals. Peanut allergens share significant amino acid homology with those of other legumes and tree nuts, but their cross-reactivity still remains unclear. ObjectiveWe sought to determine the clinical significance of the cross-reactivity of peanut allergens with those of walnut and soybean. MethodsPooled sera from eight subjects with both peanut and walnut specific IgE were investigated in an inhibition test. After the sera were incubated with either peanut or walnut protein extracts, the quantity of IgE antibodies against the peanut and walnut was measured using an immunoCAP test. Likewise, pooled sera from 18 subjects with both peanut and soybean specific IgE antibodies were incubated with either peanut or soybean protein extracts and evaluated with a peanut and soybean immunoCAP test. SDS-PAGE and immunoblotting were also performed with peanut, walnut and soybean protein extracts and relevant sera. ResultsPeanut specific IgE was inhibited up to 20% and 26% by walnut and soybean protein extracts, respectively. In reverse, walnut and soybean specific IgE were inhibited up to 21% and 23% by peanut protein extracts, respectively. In the immunoblot analysis, pooled serum from the subjects with peanut specific IgE antibodies reacted with walnut protein extracts significantly. ConclusionAlthough the clinical significance of the cross-reactivity of peanut specific IgE with walnut and soybean protein extracts has not been established, we believe that individuals who are allergic to peanuts need to be cautious about consuming walnuts and soybeans." "secciones" => array:5 [ 0 => array:2 [ "identificador" => "abst0005" "titulo" => "Background" ] 1 => array:2 [ "identificador" => "abst0010" "titulo" => "Objective" ] 2 => array:2 [ "identificador" => "abst0015" "titulo" => "Methods" ] 3 => array:2 [ "identificador" => "abst0020" "titulo" => "Results" ] 4 => array:2 [ "identificador" => "abst0025" "titulo" => "Conclusion" ] ] ] ] "apendice" => array:1 [ 0 => array:1 [ "seccion" => array:1 [ 0 => array:4 [ "apendice" => "The following are the supplementary data to this article:<elsevierMultimedia ident="upi0005"></elsevierMultimedia><elsevierMultimedia ident="upi0010"></elsevierMultimedia>" "etiqueta" => "Appendix A" "titulo" => "Supplementary data" "identificador" => "sec0095" ] ] ] ] "multimedia" => array:8 [ 0 => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 1041 "Ancho" => 2506 "Tamanyo" => 185006 ] ] "descripcion" => array:1 [ "en" => "Cross-inhibition of peanut with walnut. Pooled sera from eight subjects who were immunoCAP positive for both peanut and walnut (>3.66kU/L) were incubated with the indicated concentration of either peanut or walnut protein extracts at 4°C overnight. House dust mite extract was also incubated with the pooled sera as a negative control. The IgE levels of the inhibited sera against peanut (A) and walnut (B) were then measured by the immunoCAP test. The specific IgE level of uninhibited pooled serum was used as a control." ] ] 1 => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 1034 "Ancho" => 2497 "Tamanyo" => 188724 ] ] "descripcion" => array:1 [ "en" => "Cross-inhibition of peanut with soybean. Pooled sera from 18 individuals that were immunoCAP positive for both peanut and walnut (>3.62kU/L) were incubated with the indicated concentration of either peanut or soybean protein extracts at 4°C overnight. House dust mite extract was used as a negative control. Then, the IgE levels of the inhibited sera against peanut (A) and soybean (B) were measured by the immunoCAP test. The specific IgE level of uninhibited pooled sera was used as a control." ] ] 2 => array:7 [ "identificador" => "fig0015" "etiqueta" => "Figure 3" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr3.jpeg" "Alto" => 1434 "Ancho" => 2917 "Tamanyo" => 295414 ] ] "descripcion" => array:1 [ "en" => "SDS-PAGE analysis of protein extract from peanut (A), walnut (B) and soybean (C). M: Molecular weight marker." ] ] 3 => array:7 [ "identificador" => "fig0020" "etiqueta" => "Figure 4" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr4.jpeg" "Alto" => 1371 "Ancho" => 2917 "Tamanyo" => 285923 ] ] "descripcion" => array:1 [ "en" => "Immunoblot analysis of peanut (A), walnut (B) and soybean (C) protein extracts with pooled sera each containing peanut, walnut and soybean specific IgE. M: Molecular weight marker, PN 1: pooled sera with peanut specific IgE without walnut sensitisation (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>. Group1), WN: pooled sera with walnut specific IgE without peanut sensitisation (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>. Group 2), PN 2: pooled sera with peanut specific IgE without soybean sensitisation (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>. Group 3), Soy: pooled sera with soybean specific IgE without peanut sensitisation (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>. Group 4), Normal: pooled sera without allergic sensitisation, Buff Con: buffer control." ] ] 4 => array:7 [ "identificador" => "fig0025" "etiqueta" => "Figure 5" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr5.jpeg" "Alto" => 875 "Ancho" => 2500 "Tamanyo" => 184429 ] ] "descripcion" => array:1 [ "en" => "Immunoblot analysis of walnut (A) and peanut (B) protein extracts using peanut specific IgE positive individual sera (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>. Group 1)." ] ] 5 => array:8 [ "identificador" => "tbl0005" "etiqueta" => "Table 1" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "detalles" => array:1 [ 0 => array:3 [ "identificador" => "at1" "detalle" => "Table " "rol" => "short" ] ] "tabla" => array:1 [ "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td-with-role" title="table-head ; entry_with_role_rowhead " align="left" valign="top" scope="col">No. \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col">Group \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col">Sex \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col">Age \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col">Peanut IgE \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col">Walnut IgE \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col">Soybean IgE \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col">Total IgE \t\t\t\t\t\t\n \t\t\t\t</th></tr><tr title="table-row"><th class="td" title="table-head " align="" valign="top" scope="col" style="border-bottom: 2px solid black"> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="" valign="top" scope="col" style="border-bottom: 2px solid black"> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="" valign="top" scope="col" style="border-bottom: 2px solid black"> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">(Yrs) \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">(kU/L) \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">(kU/L) \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">(kU/L) \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">(kU/L) \t\t\t\t\t\t\n \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " rowspan="8" align="left" valign="middle">Group 1 Peanut positive 1 (Walnut negative)</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">10.2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.28 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1024 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">12 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.03 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">181 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">3 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">4.88 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.08 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">415 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">5.56 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">26.8 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">5.45 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.22 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">488 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">21.5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.12 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">693 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">7 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">23.4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.25 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1024 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">8 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">F \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">>100 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.12 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1203 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " colspan="8" align="left" valign="top"></td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " rowspan="5" align="left" valign="middle">Group 2 Walnut positive</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.03 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.85 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">150 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.32 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">7.54 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">58.4 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">3 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">F \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.03 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">164 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.15 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">10.9 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">398 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">F \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.16 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">37.6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">209 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " colspan="8" align="left" valign="top"></td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " rowspan="6" align="left" valign="middle">Group 3 Peanut positive 2 (Soybean negative)</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.34 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">116 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">9.02 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">21.9 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">3 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.53 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">32.7 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">F \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.85 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.01 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">23.5 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">F \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.35 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.25 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">102 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">F \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.44 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.08 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">271 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " colspan="8" align="left" valign="top"></td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " rowspan="10" align="left" valign="middle">Group 4 Soybean positive</td><td class="td" title="table-entry " align="left" valign="top">F \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">9.06 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">119 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.13 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">4.98 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">116 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">3 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.11 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.77 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">102 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.07 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.71 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">273 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top"><0.35 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">499 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.26 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">3.27 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">386 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">7 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">F \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.84 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">94.1 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">8 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.22 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2.42 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">154 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">9 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.04 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">35.5 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.07 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">– \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1.82 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">418 \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab1247026.png" ] ] ] ] "descripcion" => array:1 [ "en" => "Characteristics of the subjects from whom sera was used for immunoblot analysis." ] ] 6 => array:5 [ "identificador" => "upi0005" "tipo" => "MULTIMEDIAECOMPONENTE" "mostrarFloat" => false "mostrarDisplay" => true "Ecomponente" => array:2 [ "fichero" => "mmc1.docx" "ficheroTamanyo" => 16847 ] ] 7 => array:5 [ "identificador" => "upi0010" "tipo" => "MULTIMEDIAECOMPONENTE" "mostrarFloat" => false "mostrarDisplay" => true "Ecomponente" => array:2 [ "fichero" => "mmc2.docx" "ficheroTamanyo" => 18661 ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0005" "bibliografiaReferencia" => array:25 [ 0 => array:3 [ "identificador" => "bib0130" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:3 [ "comentario" => "668–676.e1–2" "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Prevalence of challenge-proven IgE-mediated food allergy using population-based sampling and predetermined challenge criteria in infants" "autores" => array:1 [ 0 => array:2 [ "etal" => true "autores" => array:6 [ 0 => "N.J. 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"vista" => "all" ] ] ] "idiomaDefecto" => "en" "url" => "/03010546/0000004400000006/v1_201611090130/S030105461630074X/v1_201611090130/en/main.assets" "Apartado" => array:4 [ "identificador" => "6702" "tipo" => "SECCION" "es" => array:2 [ "titulo" => "Original articles" "idiomaDefecto" => true ] "idiomaDefecto" => "es" ] "PDF" => "https://static.elsevier.es/multimedia/03010546/0000004400000006/v1_201611090130/S030105461630074X/v1_201611090130/en/main.pdf?idApp=UINPBA00004N&text.app=https://www.elsevier.es/" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S030105461630074X?idApp=UINPBA00004N"]

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M.N. Kima, K.E. Leea, J.Y. Honga, K.W. Kima, K.-E. Kima, M.H. Sohna,

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mhsohn@yuhs.ac

Corresponding author.

, J.W. Parkb

a Department of Pediatrics, Severance Hospital, Institute of Allergy, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea

b Department of Internal Medicine, Severance Hospital, Institute of Allergy, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea

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    "textoCompleto" => "<span class="elsevierStyleSections"><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0040">Introduction</span><p id="par0005" class="elsevierStylePara elsevierViewall">The prevalence of food allergies has been increasing in recent years and can affect up to 10&#37; of young children and 2&#8211;3&#37; of adults globally&#46;<a class="elsevierStyleCrossRefs" href="#bib0130"><span class="elsevierStyleSup">1&#44;2</span></a> Food allergies can cause urticaria&#44; angio-oedema&#44; rhinitis&#44; wheezing&#44; and in some cases can be life threatening due to anaphylactic shock&#46; A recent meta-analysis study reported that fatal food anaphylaxis has an incidence rate of 1&#46;81<span class="elsevierStyleHsp" style=""></span>per million person-years&#46;<a class="elsevierStyleCrossRef" href="#bib0140"><span class="elsevierStyleSup">3</span></a></p><p id="par0010" class="elsevierStylePara elsevierViewall">Peanut is one of the primary allergenic foods&#46; It contains at least 18 proteins that have been shown to cause specific IgE reactivity and have a higher potential to cause anaphylactic shock compared to other foods&#46;<a class="elsevierStyleCrossRefs" href="#bib0145"><span class="elsevierStyleSup">4&#8211;6</span></a> The peanut allergens&#44; vicilin &#40;Ara h 1&#41;&#44; 2S albumin &#40;Ara h 2&#41;&#44; and 11S globulin &#40;Ara h 3&#41;&#44; share significant amino acid homology with their respective allergens from other legumes and tree nuts&#46;<a class="elsevierStyleCrossRefs" href="#bib0160"><span class="elsevierStyleSup">7&#8211;9</span></a></p><p id="par0015" class="elsevierStylePara elsevierViewall">Tree nut&#44; despite its botanical taxonomic distance&#44; is believed to be cross-reactive with peanut&#44; due to the high number of individuals reported to be co-sensitised or co-allergic to peanut and tree nut&#46;<a class="elsevierStyleCrossRef" href="#bib0175"><span class="elsevierStyleSup">10</span></a> A study of the prevalence of peanut and tree nut allergies among 1218 newborns in the United Kingdom revealed that 1 in 200 children could have adverse reactions to either peanuts or tree nuts by the age of four&#46;<a class="elsevierStyleCrossRef" href="#bib0180"><span class="elsevierStyleSup">11</span></a> In another study among individuals with nut allergies&#44; 65&#37; reacted specifically to peanuts of which 30&#37; also reacted with other tree nuts&#46;<a class="elsevierStyleCrossRef" href="#bib0185"><span class="elsevierStyleSup">12</span></a></p><p id="par0020" class="elsevierStylePara elsevierViewall">Soybean&#44; belonging to the same legume family as the peanut&#44; has also been investigated as a cross-reactive food with peanuts&#46; Of 69 legume-sensitive individuals&#44; 43&#37; showed sensitivity to soybean and 87&#37; to peanut&#46;<a class="elsevierStyleCrossRef" href="#bib0190"><span class="elsevierStyleSup">13</span></a> It has also been reported that soybean immunotherapy in a peanut allergy mouse model resulted in a significant reduction of clinical symptoms following peanut challenge compared with placebo-treated mice&#46;<a class="elsevierStyleCrossRef" href="#bib0195"><span class="elsevierStyleSup">14</span></a></p><p id="par0025" class="elsevierStylePara elsevierViewall">However&#44; despite the above findings&#44; the clinical significance of cross-reactivity between peanut allergens and those of walnut and soybean remains unclear&#46; With this in mind&#44; we sought to determine the cross-reactivity of peanut allergens with those of walnut and soybean in the sera of allergic patients&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Materials and methods</span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Subjects</span><p id="par0030" class="elsevierStylePara elsevierViewall">Pooled sera from eight subjects with allergic sensitisation to both peanut and walnut were used to investigate cross-reactivity of peanut and walnut in an inhibition test &#40;Supplemental Table 1&#41;&#46; For the peanut and soybean cross-reactivity test&#44; the pooled sera from 18 subjects with both peanut and soybean specific IgE antibodies were evaluated &#40;Supplemental Table 2&#41;&#46; Allergic sensitisation&#44; the presence of specific IgE&#44; was evaluated using an immunoCAP test &#40;Phadia AB&#44; Upsala&#44; Sweden&#41;&#46; The clinical data of the subjects used for the immunoblot analysis are summarised in <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#46; The pooled sera from eight subjects without an allergic sensitisation were used as a normal control&#46; All sera were obtained from Severance Children&#39;s Hospital of Yonsei University&#44; Seoul &#40;South Korea&#41;&#46; This study was approved by the institutional Review Board of Severance Children&#39;s Hospital of Yonsei University&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Protein extraction</span><p id="par0035" class="elsevierStylePara elsevierViewall">To obtain protein extracts&#44; peanuts&#44; walnuts and soybeans were purchased and finely ground&#46; The ground meals were defatted with cold &#40;4<span class="elsevierStyleHsp" style=""></span>&#176;C&#41; acetone &#40;1&#58;5&#44; w&#47;v&#41; under constant stirring for one hour and then filtered with filter paper&#46; The defatting procedure was repeated until the filtrate became clear&#46; The defatted meals were completely air dried at ambient temperature and then stirred in phosphate buffered saline &#40;PBS&#44; pH 7&#46;4&#41; &#40;1&#58;4&#44; w&#58;v&#41; for 48<span class="elsevierStyleHsp" style=""></span>h at 4<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; The extracts were centrifuged twice at 13&#44;000<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">g</span> for 30<span class="elsevierStyleHsp" style=""></span>min at 4<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; The supernatants were filtered and dialysed against several changes of distilled water for 48<span class="elsevierStyleHsp" style=""></span>h at 4<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; 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Walnut specific IgE levels showed similar tendencies and were inhibited up to 91&#37; by walnut protein extracts but only 21&#37; by peanut protein extracts&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><p id="par0060" class="elsevierStylePara elsevierViewall">To investigate peanut and soybean cross-reactivity&#44; pooled sera from 18 subjects who were immunoCAP positive for both peanut and soybean &#40;sex&#58; male 16&#44; female 2&#44; age&#58; 3&#46;3<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>3&#46;4<span class="elsevierStyleHsp" style=""></span>yrs&#44; peanut specific IgE level of uninhibited pooled serum&#58; 31&#46;8<span class="elsevierStyleHsp" style=""></span>&#91;kU&#47;L&#93;&#44; soybean specific IgE level of uninhibited pooled serum&#58; 20&#46;3<span class="elsevierStyleHsp" style=""></span>&#91;kU&#47;L&#93;&#41; were incubated with either peanut or soybean protein extracts as described above&#46; Then&#44; an immunoCAP test for peanut and walnut specific IgE was conducted with the inhibited sera&#46; Peanut specific IgE activity was inhibited up to 86&#37; at an inhibitor concentration of 50<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml by peanut protein extracts&#44; while only 26&#37; was inhibited by soybean protein extracts &#40;<a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>A&#41;&#46; Soybean specific IgE also showed similar pattern inhibited up to 83&#37; by soybean protein extracts but only 23&#37; by peanut protein extracts &#40;<a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>B&#41;&#46;</p><elsevierMultimedia ident="fig0010"></elsevierMultimedia></span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Constitutive protein distribution and IgE reactive peptides</span><p id="par0065" class="elsevierStylePara elsevierViewall">SDS-PAGE was conducted with protein extracts from peanuts&#44; walnuts&#44; and soybeans to observe the distribution of constitutive proteins &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>&#41;&#46; This was followed by immunoblotting with various sera to ascertain which specific peptides were responsible for IgE reactivity and to determine whether the pooled sera positive for either peanut&#44; walnut&#44; or soybean reacted with corresponding allergens from each other &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#44; <a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>&#41;&#46;</p><elsevierMultimedia ident="fig0015"></elsevierMultimedia><elsevierMultimedia ident="fig0020"></elsevierMultimedia><p id="par0070" class="elsevierStylePara elsevierViewall">As shown in <a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>A&#44; lane 2&#44; pooled serum of the peanut immunoCAP positive &#40;&#62;0&#46;35<span class="elsevierStyleHsp" style=""></span>kU&#47;L&#41; sera reacted strongly with peanut protein extracts as we expected&#46; Pooled serum of each walnut and soybean immunoCAP positive sera also showed weak IgE reactive bands to peanut protein extracts&#46; However&#44; this binding was probably non-specific as a similar pattern was seen in the membrane with the normal control serum which is from the subjects without an allergic sensitisation &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>A&#41;&#46;</p><p id="par0075" class="elsevierStylePara elsevierViewall">Again&#44; as we expected&#44; pooled serum of walnut imunoCAP positive &#40;&#62;0&#46;35<span class="elsevierStyleHsp" style=""></span>kU&#47;L&#41; sera reacted with walnut protein extracts &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>B&#44; lane 2&#41;&#46; Interestingly&#44; peanut immunoCAP positive serum also showed a significant reactive band with walnut protein extracts &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>B&#44; lane 3&#41;&#46; Even though several bands were visible on the membrane using the normal control serum&#44; the peanut specific IgE positive serum produced a stronger and different pattern of reactive bands especially between 20 and 60<span class="elsevierStyleHsp" style=""></span>kDa with walnut protein &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>B&#41;&#46;</p><p id="par0080" class="elsevierStylePara elsevierViewall">Pooled sera of soybean immunoCAP test positive sera also bound well to soybean proteins while peanut immunoCAP test positive serum showed non-specific bands similar to the control sera &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>C&#41;&#46; Of the soybean proteins&#44; high molecular weight peptides reacted more strongly than low molecular peptides particularly between 60 and 70<span class="elsevierStyleHsp" style=""></span>kDa&#46;</p></span><span id="sec0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Individual sera immunoblot analysis</span><p id="par0085" class="elsevierStylePara elsevierViewall">The sera showed reactive IgE band to walnut protein in <a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>B&#44; lane 3&#44; even though the sera were positive for peanut but not to walnut by the immunoCAP test&#46; To confirm whether the sera could react to walnut protein&#44; an immunoblot analysis was repeated using individual sera&#46; As shown in <a class="elsevierStyleCrossRef" href="#fig0025">Fig&#46; 5</a>&#44; significant IgE reactive bands were seen in three of the eight individual sera &#40;<a class="elsevierStyleCrossRef" href="#fig0025">Fig&#46; 5</a>A&#46; lanes 2&#44; 6 and 8&#41;&#46; Each cross-reacting serum revealed a different IgE band pattern binding to walnut proteins of different sizes&#46; However&#44; each of the three sera bound to a peptide of around 30<span class="elsevierStyleHsp" style=""></span>kDa&#46; In addition&#44; significant cross-reactive IgE bands were observed around 15<span class="elsevierStyleHsp" style=""></span>kDa in lanes 2 and 6&#46;</p><elsevierMultimedia ident="fig0025"></elsevierMultimedia></span></span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">Discussion</span><p id="par0090" class="elsevierStylePara elsevierViewall">The cross-reactivity of peanut with walnut has been studied because they share IgE binding epitopes structurally related to the vicilin allergen in addition to having some structural homology to legumin allergens&#46; Indeed&#44; avoidance or restricted consumption of walnut has been recommended for peanut-sensitised individuals&#46;<a class="elsevierStyleCrossRefs" href="#bib0165"><span class="elsevierStyleSup">8&#44;15</span></a> In this study&#44; our results from the inhibition test showed only 20&#37; inhibition of peanut specific IgE by walnut protein extracts&#46; This level of inhibition seems low&#44; but when immunoblots of walnut protein extracts were performed with group 1 individual serum that were positive for peanut but negative to walnut by immunoCAP test&#44; three out of the eight subjects showed significantly reactive IgE bands&#46; We were careful to distinguish whether the vicilin and legumin peptides were reactive in both the peanut and walnut protein extracts&#44; but this was difficult to confirm based on identification of the peptides by molecular weight alone&#46; However&#44; IgE reactive bands around 30<span class="elsevierStyleHsp" style=""></span>kDa were seen in all of the three reactive subjects&#44; and one of them was particularly strong &#40;<a class="elsevierStyleCrossRef" href="#fig0025">Fig&#46; 5</a>A&#44; lane 8&#41;&#46; This peptide is not known as a major allergen of walnut&#59; therefore&#44; further investigation is required for its identification and characterisation&#46; Furthermore&#44; IgE reactive bands were also seen around 15<span class="elsevierStyleHsp" style=""></span>kDa&#44; a similar molecular weight to Jug r 1&#44; in two of the three reactive sera &#40;<a class="elsevierStyleCrossRef" href="#fig0025">Fig&#46; 5</a>A&#44; lane 2&#44; 6&#41;&#46; As Jug r 1 &#40;14&#8211;16<span class="elsevierStyleHsp" style=""></span>kDa&#41; and Jug r 3 &#40;9<span class="elsevierStyleHsp" style=""></span>kDa&#41; are known to be associated with systemic reactions&#44; it is recommended that even though the result of the immunoCAP test for walnut is negative&#44; individuals with peanut allergies should be cautious about ingesting walnuts&#46;<a class="elsevierStyleCrossRefs" href="#bib0205"><span class="elsevierStyleSup">16&#8211;20</span></a></p><p id="par0095" class="elsevierStylePara elsevierViewall">Soybeans are used widely in various foods for both humans and animals&#46; Peanut Ara h 3 has been reported to share IgE epitopes with the glycinin G1 acidic chain of soybean&#46; A study with the sera of individuals allergic to both soybean and peanut showed that the absorption of soybean-binding antibodies by antigen-affinity chromatography resulted in a 73&#37; reduction of IgE binding to peanut allergens&#46;<a class="elsevierStyleCrossRefs" href="#bib0230"><span class="elsevierStyleSup">21&#44;22</span></a> In our results&#44; although the inhibition of peanut specific IgE by soybean protein extracts was higher than by walnut protein extracts&#44; it was still only an inhibition of 26&#46;7&#37;&#46; In previous studies&#44; it has been demonstrated that individuals allergic to both peanut and soybean have IgE antibodies that bind preferentially to larger proteins&#44; while the antibodies of those reacting only to soybean bind strongly to proteins with lower molecular weight&#46;<a class="elsevierStyleCrossRefs" href="#bib0240"><span class="elsevierStyleSup">23&#44;24</span></a> In our study&#44; we could not see a significant difference between the IgE binding pattern of pooled sera of both peanut and soybean positive subjects and soybean only positive subjects&#46; Peanut only positive sera did not show significant cross-reactive IgE bands to soybean protein extracts either&#46; Extensive cross-reactivity studies have been undertaken on the different legume species&#46; And the majority of studies do not provide the clinical cross-reactions among the legume species&#46;<a class="elsevierStyleCrossRefs" href="#bib0190"><span class="elsevierStyleSup">13&#44;22</span></a> Nevertheless&#44; as has been reported in a previous study&#44; soy could be a cause of food anaphylaxis&#44; and&#44; as such&#44; individuals with peanut allergies are at risk of a cross-reactive response&#46;<a class="elsevierStyleCrossRef" href="#bib0250"><span class="elsevierStyleSup">25</span></a> Individuals with peanut allergies should&#44; therefore&#44; be aware of this cross-reactivity&#44; so that they can take the necessary measures if they have abnormal responses following ingestion of soybean&#46;</p><p id="par0100" class="elsevierStylePara elsevierViewall">In our study&#44; three out of the eight sera that were positive for peanut but were negative for walnut in the immunoCAP test showed a significant IgE reactivity to walnut proteins in the immunoblot analysis&#46; This contradictory result between the immunoCAP test and immunoblot analysis should be considered in the diagnosis of other food allergies&#44; especially for individuals who are immunoCAP negative but still have allergic symptoms&#46; However&#44; the sera used for the experiments were collected based on the result of the immunoCAP test&#46; Positivity by the immunoCAP test does not necessarily mean that the subject will show clinical symptoms following ingestion of the allergenic food&#46; Therefore&#44; future work needs to be undertaken with sera collected from symptomatic individuals&#46;</p><p id="par0105" class="elsevierStylePara elsevierViewall">Taken together&#44; these results indicate that although the clinical significance of cross-reactivity of peanut with walnut and soybean has not been established&#44; individuals with peanut allergies need to be cautious when consuming walnut and soybean&#46;</p></span><span id="sec0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0100">Ethical disclosures</span><span id="sec0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0105">Confidentiality of data</span><p id="par0110" class="elsevierStylePara elsevierViewall">The authors declare that they have followed the protocols of their work centre on the publication of patient data and that all the patients included in the study have received sufficient information and have given their informed consent in writing to participate in that study&#46;</p></span><span id="sec0075" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0110">Right to privacy and informed consent</span><p id="par0115" class="elsevierStylePara elsevierViewall">The authors have obtained the informed consent of the patients and&#47;or subjects mentioned in the article&#46; The author for correspondence is in possession of this document&#46;</p></span><span id="sec0080" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0115">Protection of human subjects and animals in research</span><p id="par0120" class="elsevierStylePara elsevierViewall">The authors declare that no experiments were performed on humans or animals for this investigation&#46;</p></span></span><span id="sec0085" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0120">Conflicts of interest</span><p id="par0125" class="elsevierStylePara elsevierViewall">None declared&#46;</p></span></span>"
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        0 => array:3 [
          "identificador" => "xres753553"
          "titulo" => "Abstract"
          "secciones" => array:5 [
            0 => array:2 [
              "identificador" => "abst0005"
              "titulo" => "Background"
            ]
            1 => array:2 [
              "identificador" => "abst0010"
              "titulo" => "Objective"
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            2 => array:2 [
              "identificador" => "abst0015"
              "titulo" => "Methods"
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            3 => array:2 [
              "identificador" => "abst0020"
              "titulo" => "Results"
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            4 => array:2 [
              "identificador" => "abst0025"
              "titulo" => "Conclusion"
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        1 => array:2 [
          "identificador" => "xpalclavsec755870"
          "titulo" => "Keywords"
        ]
        2 => array:2 [
          "identificador" => "sec0005"
          "titulo" => "Introduction"
        ]
        3 => array:3 [
          "identificador" => "sec0010"
          "titulo" => "Materials and methods"
          "secciones" => array:5 [
            0 => array:2 [
              "identificador" => "sec0015"
              "titulo" => "Subjects"
            ]
            1 => array:2 [
              "identificador" => "sec0020"
              "titulo" => "Protein extraction"
            ]
            2 => array:2 [
              "identificador" => "sec0025"
              "titulo" => "Sodium dodecyl sulphate-polyacrylamide gel electrophoresis &#40;SDS-PAGE&#41;"
            ]
            3 => array:2 [
              "identificador" => "sec0030"
              "titulo" => "IgE immunoblotting"
            ]
            4 => array:2 [
              "identificador" => "sec0035"
              "titulo" => "Inhibition test"
            ]
          ]
        ]
        4 => array:3 [
          "identificador" => "sec0040"
          "titulo" => "Results"
          "secciones" => array:3 [
            0 => array:2 [
              "identificador" => "sec0045"
              "titulo" => "Cross-inhibition of peanut with walnut and soybean"
            ]
            1 => array:2 [
              "identificador" => "sec0050"
              "titulo" => "Constitutive protein distribution and IgE reactive peptides"
            ]
            2 => array:2 [
              "identificador" => "sec0055"
              "titulo" => "Individual sera immunoblot analysis"
            ]
          ]
        ]
        5 => array:2 [
          "identificador" => "sec0060"
          "titulo" => "Discussion"
        ]
        6 => array:3 [
          "identificador" => "sec0065"
          "titulo" => "Ethical disclosures"
          "secciones" => array:3 [
            0 => array:2 [
              "identificador" => "sec0070"
              "titulo" => "Confidentiality of data"
            ]
            1 => array:2 [
              "identificador" => "sec0075"
              "titulo" => "Right to privacy and informed consent"
            ]
            2 => array:2 [
              "identificador" => "sec0080"
              "titulo" => "Protection of human subjects and animals in research"
            ]
          ]
        ]
        7 => array:2 [
          "identificador" => "sec0085"
          "titulo" => "Conflicts of interest"
        ]
        8 => array:2 [
          "identificador" => "xack250546"
          "titulo" => "Acknowledgements"
        ]
        9 => array:1 [
          "titulo" => "References"
        ]
      ]
    ]
    "pdfFichero" => "main.pdf"
    "tienePdf" => true
    "fechaRecibido" => "2015-12-14"
    "fechaAceptado" => "2016-04-27"
    "PalabrasClave" => array:1 [
      "en" => array:1 [
        0 => array:4 [
          "clase" => "keyword"
          "titulo" => "Keywords"
          "identificador" => "xpalclavsec755870"
          "palabras" => array:9 [
            0 => "Cross-reactivity"
            1 => "Food allergy"
            2 => "IgE-hypersensitivity"
            3 => "Legume"
            4 => "Peanut"
            5 => "Soybean"
            6 => "Tree nut"
            7 => "Walnut"
            8 => "Inhibition test"
          ]
        ]
      ]
    ]
    "tieneResumen" => true
    "resumen" => array:1 [
      "en" => array:3 [
        "titulo" => "Abstract"
        "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0010">Background</span><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Peanut allergies are common and can be life-threating for sensitised individuals&#46; Peanut allergens share significant amino acid homology with those of other legumes and tree nuts&#44; but their cross-reactivity still remains unclear&#46;</p></span> <span id="abst0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0015">Objective</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">We sought to determine the clinical significance of the cross-reactivity of peanut allergens with those of walnut and soybean&#46;</p></span> <span id="abst0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0020">Methods</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Pooled sera from eight subjects with both peanut and walnut specific IgE were investigated in an inhibition test&#46; After the sera were incubated with either peanut or walnut protein extracts&#44; the quantity of IgE antibodies against the peanut and walnut was measured using an immunoCAP test&#46; Likewise&#44; pooled sera from 18 subjects with both peanut and soybean specific IgE antibodies were incubated with either peanut or soybean protein extracts and evaluated with a peanut and soybean immunoCAP test&#46; SDS-PAGE and immunoblotting were also performed with peanut&#44; walnut and soybean protein extracts and relevant sera&#46;</p></span> <span id="abst0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Results</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">Peanut specific IgE was inhibited up to 20&#37; and 26&#37; by walnut and soybean protein extracts&#44; respectively&#46; In reverse&#44; walnut and soybean specific IgE were inhibited up to 21&#37; and 23&#37; by peanut protein extracts&#44; respectively&#46; In the immunoblot analysis&#44; pooled serum from the subjects with peanut specific IgE antibodies reacted with walnut protein extracts significantly&#46;</p></span> <span id="abst0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Conclusion</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">Although the clinical significance of the cross-reactivity of peanut specific IgE with walnut and soybean protein extracts has not been established&#44; we believe that individuals who are allergic to peanuts need to be cautious about consuming walnuts and soybeans&#46;</p></span>"
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            "titulo" => "Objective"
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          2 => array:2 [
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            "apendice" => "<p id="par0140" class="elsevierStylePara elsevierViewall">The following are the supplementary data to this article&#58;<elsevierMultimedia ident="upi0005"></elsevierMultimedia><elsevierMultimedia ident="upi0010"></elsevierMultimedia></p>"
            "etiqueta" => "Appendix A"
            "titulo" => "Supplementary data"
            "identificador" => "sec0095"
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          "en" => "<p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">Cross-inhibition of peanut with walnut&#46; Pooled sera from eight subjects who were immunoCAP positive for both peanut and walnut &#40;&#62;3&#46;66<span class="elsevierStyleHsp" style=""></span>kU&#47;L&#41; were incubated with the indicated concentration of either peanut or walnut protein extracts at 4<span class="elsevierStyleHsp" style=""></span>&#176;C overnight&#46; House dust mite extract was also incubated with the pooled sera as a negative control&#46; The IgE levels of the inhibited sera against peanut &#40;A&#41; and walnut &#40;B&#41; were then measured by the immunoCAP test&#46; The specific IgE level of uninhibited pooled serum was used as a control&#46;</p>"
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          "en" => "<p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">Cross-inhibition of peanut with soybean&#46; Pooled sera from 18 individuals that were immunoCAP positive for both peanut and walnut &#40;&#62;3&#46;62<span class="elsevierStyleHsp" style=""></span>kU&#47;L&#41; were incubated with the indicated concentration of either peanut or soybean protein extracts at 4<span class="elsevierStyleHsp" style=""></span>&#176;C overnight&#46; House dust mite extract was used as a negative control&#46; Then&#44; the IgE levels of the inhibited sera against peanut &#40;A&#41; and soybean &#40;B&#41; were measured by the immunoCAP test&#46; The specific IgE level of uninhibited pooled sera was used as a control&#46;</p>"
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          "en" => "<p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">SDS-PAGE analysis of protein extract from peanut &#40;A&#41;&#44; walnut &#40;B&#41; and soybean &#40;C&#41;&#46; M&#58; Molecular weight marker&#46;</p>"
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          "en" => "<p id="spar0045" class="elsevierStyleSimplePara elsevierViewall">Immunoblot analysis of peanut &#40;A&#41;&#44; walnut &#40;B&#41; and soybean &#40;C&#41; protein extracts with pooled sera each containing peanut&#44; walnut and soybean specific IgE&#46; M&#58; Molecular weight marker&#44; PN 1&#58; pooled sera with peanut specific IgE without walnut sensitisation &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#46; Group1&#41;&#44; WN&#58; pooled sera with walnut specific IgE without peanut sensitisation &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#46; Group 2&#41;&#44; PN 2&#58; pooled sera with peanut specific IgE without soybean sensitisation &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#46; Group 3&#41;&#44; Soy&#58; pooled sera with soybean specific IgE without peanut sensitisation &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#46; Group 4&#41;&#44; Normal&#58; pooled sera without allergic sensitisation&#44; Buff Con&#58; buffer control&#46;</p>"
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          "en" => "<p id="spar0050" class="elsevierStyleSimplePara elsevierViewall">Immunoblot analysis of walnut &#40;A&#41; and peanut &#40;B&#41; protein extracts using peanut specific IgE positive individual sera &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#46; Group 1&#41;&#46;</p>"
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                  \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td-with-role" title="table-head ; entry_with_role_rowhead " align="left" valign="top" scope="col">No&#46;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col">Group&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col">Sex&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col">Age&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col">Peanut IgE&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col">Walnut IgE&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col">Soybean IgE&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col">Total IgE&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th></tr><tr title="table-row"><th class="td" title="table-head  " align="" valign="top" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="" valign="top" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="" valign="top" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">&#40;Yrs&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">&#40;kU&#47;L&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">&#40;kU&#47;L&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">&#40;kU&#47;L&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">&#40;kU&#47;L&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " rowspan="8" align="left" valign="middle"><span class="elsevierStyleBold">Group 1</span> Peanut positive 1 &#40;Walnut negative&#41;</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">10&#46;2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;28&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1024&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">12&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;03&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">181&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">4&#46;88&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;08&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">415&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">5&#46;56&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">26&#46;8&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">5&#46;45&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;22&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">488&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">21&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;12&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">693&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">7&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">23&#46;4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;25&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1024&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">8&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">10&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#62;100&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;12&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1203&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " colspan="8" align="left" valign="top"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " rowspan="5" align="left" valign="middle"><span class="elsevierStyleBold">Group 2</span> Walnut positive</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;03&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2&#46;85&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">150&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;32&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">7&#46;54&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">58&#46;4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;03&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&#46;1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">164&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;15&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">10&#46;9&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">398&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;16&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">37&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">209&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " colspan="8" align="left" valign="top"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " rowspan="6" align="left" valign="middle"><span class="elsevierStyleBold">Group 3</span> Peanut positive 2 &#40;Soybean negative&#41;</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&#46;34&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">116&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">9&#46;02&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">21&#46;9&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;53&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">32&#46;7&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2&#46;85&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;01&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">23&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="table-entry ; entry_with_role_rowhead " align="left" valign="top">5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">1&#46;35&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">&#8211;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0&#46;25&nbsp;\t\t\t\t\t\t\n
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Article information

ISSN: 03010546
Original language: English

DOI:10.1016/j.aller.2016.04.012

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Year/Month	Html	Pdf	Total

2024 November	7	1	8
2024 October	55	18	73
2024 September	54	14	68
2024 August	36	14	50
2024 July	36	9	45
2024 June	48	6	54
2024 May	44	12	56
2024 April	31	8	39
2024 March	62	7	69
2024 February	57	28	85
2024 January	127	16	143
2023 December	45	9	54
2023 November	51	9	60
2023 October	93	13	106
2023 September	45	8	53
2023 August	60	18	78
2023 July	92	20	112
2023 June	93	9	102
2023 May	90	9	99
2023 April	51	2	53
2023 March	102	8	110
2023 February	60	10	70
2023 January	57	13	70
2022 December	61	14	75
2022 November	61	23	84
2022 October	70	11	81
2022 September	70	11	81
2022 August	53	12	65
2022 July	37	13	50
2022 June	38	13	51
2022 May	46	41	87
2022 April	40	23	63
2022 March	45	22	67
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2022 January	58	19	77
2021 December	75	12	87
2021 November	75	17	92
2021 October	101	18	119
2021 September	59	17	76
2021 August	78	22	100
2021 July	64	13	77
2021 June	41	15	56
2021 May	52	10	62
2021 April	122	21	143
2021 March	51	16	67
2021 February	44	14	58
2021 January	45	21	66
2020 December	1	1	2
2020 October	1	0	1
2020 July	0	1	1
2018 March	1	0	1
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2018 January	4	3	7
2017 December	2	2	4
2017 November	8	4	12
2017 October	8	10	18
2017 September	0	4	4
2016 December	1	0	1
2016 November	7	2	9

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