f424 rAra h 3 Peanut

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• Latin name: Arachis hypogaea
• Common names: Glycinin
• Source material: An E. coli strain carrying a cloned cDNA encoding Arachis hypogaea allergen Ara h 3.

Recombinant allergens:

• rAra h 1
• rAra h 2
• rAra h 3
• rAra h 8

Allergen
rAra h 3 (1-6).

Biological function
Trypsin inhibitor.

Mw
Approximately 57 kDa

Other allergens isolated
Peanut contains at least 32 different proteins, of which around 18 have been identified as capable of binding specific IgE (7).
Allergens characterised to date include:

• Ara h 1, a 7S vicilin-like globulin (8).
   
• Ara h 2, a 2S albumin, a conglutin seed storage protein, a trypsin inhibitor (9).
   
• Ara h 3, an 11S globulin, a glycinin, a trypsin inhibitor (2).
   
• Ara h 4, a glycinin (10).
   
• Ara h 5, a profilin (10).
   
• Ara h 6, a conglutin, 2S albumin (11).
   
• Ara h 7, a conglutin (10).
   
• Ara h 8, a Bet v 1-homologous allergen (12).
   
• Ara h Agglutinin (13).
   
• Ara h LTP, a lipid transfer protein (14).
   
• Ara h Oleosin (15).

Ara h 3 and Ara h 4 have a 91% identity and are regarded as isoforms of each, i.e., Ara h 4 and Ara h 3 are considered to be the same allergen (2, 16-17).

Allergen description

Ara h 3 is a glycinin, a member of the 11S globulin family, and may also function as a trypsin inhibitor (2, 16, 18-24). Ara h 3 was in first identified as a 14 kDa protein (25), but cloning revealed a 57 kDa protein that appears to be posttranslationally cleaved to smaller subunits (1).

Ara h 3 consists of a series of polypeptides ranging from approximately 14 to 45 kDa that can be classified as acidic and basic subunits, similar to the subunit organization of soy glycinin. Ara h 4 and Ara h 3 are considered to be the same allergen (2). Iso-allergens may be as a result of medication by post-translational cleavage (26).

Between 20% -55% of Peanut-allergic individuals are sensitized to Ara h 3 (1-2, 27-28). However, mono-sensitization to a single Peanut allergen is relatively rare; instead the recognition of a high degree of patient heterogeneity has been described (27). However, this may not be true for all population groups (6).

A recent study also concluded, that Peanut-derived Ara h 3, in contrast to earlier reported recombinant Ara h 3, resembles, to a large extent, the molecular organization typical for proteins from the glycinin family. Posttranslational processing of Ara h 3 was shown to affect the IgE-binding properties and have impact on the allergenicity of Ara h 3 (2).

A comparison of the Peanut allergen sequences of Ara h 3/4, Ara h 3, Ara h 4 and Peanut trypsin inhibitor and the proteins Gly 1 and iso-Ara h 3 (not yet described as allergens), concluded that these proteins are isoallergens of each other, and that these isoallergens are post-translationally cleaved and held together by disulfide bonds in accordance to the 11S plant seed storage proteins signature (16).

The 11S globulins, also known as legumins, are classified into the Cupin superfamily, and are composed of 2 polypeptide chains of different molecular masses and amino acid sequences (heterodimeric form composed of a 20- to 40-kDa chain plus a 20- to 25-kDa chain), which are linked together by one disulfide bridge (29).

Roasting of Peanut uses higher temperatures (150-170 degrees C) than boiling (100 degrees C) or frying (120 degrees C) and roasting has been shown to increase the allergenic property of Peanut proteins. In an investigation of how roasting affects Peanut allergens, it was shown that compared with roasted Peanuts, the relative amount of Ara h 1 was reduced in the fried and boiled preparations, resulting in a significant reduction of IgE-binding intensity. There was significantly less IgE binding to Ara h 2 and Ara h 3 in fried and boiled Peanuts compared with that in roasted Peanuts, even though the protein amounts were similar in all 3 preparations (30).

Also, allergen content may vary depending on the Peanut variety (breeding lines) and may explain the differences in the prevalence of sensitization between different population studies. For example, an accession from India had the lowest level of Ara h 1 (7.0%). An accession from Nigeria had the highest level of Ara h 1 (18.5%), but the lowest level of Ara h 2 (6.2%). An accession from Zambia had the highest level of Ara h 2 (13.2%), but the lowest level of Ara h 3 (21.8%). Two accessions, 20 lines, and two Peanut cultivars (Florunner and Georgia Red) contained no or little of a 36 kDa Ara h 3 isoform, Ara h 3-im (31).

Similarly, in a study of Peanut allergen expression during seed development and upon germination and seedling growth, patterns were heterogeneous depending on the specific Peanut allergen gene and the cultivars tested. Ara h 3 expression patterns among the cultivars were more variable than Ara h 1 and Ara h 2. Transcripts were tissue specific, seen in seeds, but not in leaves, flowers, or roots and were undetectable during seed germination (32).

See Peanut, f13, for further clinical information and details on Peanut allergy.

Sensitization to Peanut occurs with a high degree of heterogeneity to a number of Peanut allergens. Mono-sensitization to a single Peanut allergen is relatively rare (27).

For example, in a British study, evaluating sera of 40 Peanut-allergic individuals, of 18 allergens identified, 8 were bound by >50% of patients and the total number of bands per patient correlated significantly with challenge score and serum-IgE. Ara h 2 was recognized by 71% and Ara h 3 by 68-73% of the sera. The study concluded that promiscuity of IgE binding appears more important than the recognition of individual proteins (33).

Between 20% -55% of Peanut-allergic individuals are sensitized to Ara h 3 (1-2, 27-28). Ara h 3 was regarded as a minor allergen, but it was found that a group of Peanut-allergic Italian children were specifically sensitized to the basic subunit of Ara h 3. The authors stated their surprise that the dominant immunoreactivity in these patients was in a basic subunit of Ara h 3 because previous studies had indicated that Ara h 3 was only a minor Peanut allergen and that the identified allergenic epitopes occurred mainly in the acidic Ara h 3 subunit (23). It is therefore evident that sensitization to Ara h 3 depends on the population group studied and the methodology of the study, but there is a suggestion that the frequency of Ara h 3 sensitization may indeed vary between population groups. In another study, recombinant Ara h 3 was recognized by serum IgE from approximately 45% of a Peanut-allergic patient population (1).

In a study that evaluated the pattern of IgE binding to specific Peanut allergens with the severity of clinical symptoms, 40 Peanut-allergic patients underwent a double-blind placebo-controlled low-dose Peanut challenge, during which the severity of the patients' Peanut allergy was scored. Seventeen IgE binding bands were identified between 5 and 100 kDa with eight bound by >50% of patients and the total number of bands per patient correlated significantly with challenge score and serum IgE. However, two protein bands, identified as subunits of Ara h 3/4, had peak intensities that correlated positively with challenge score and a third band (Ara h 1) that correlated negatively. The study concluded that promiscuity of IgE binding appears more important than the recognition of individual proteins (33).

A Dutch study investigated the relevance of Peanut allergens in 32 adult Peanut-allergic patients by in vitro, ex vivo and in vivo assays. Ara h 2 was reported as the most frequently recognized allergen (81%) (26/32) in skin-specific IgE evaluation and induced basophil degranulation at low concentrations, followed by Ara h 1 (44%) then Ara h 3 (37.5%). Ara h 2 was also deemed more potent eliciting reactions at 100-fold lower concentrations than Ara h 1 and 3 as analyzed by skin specific IgE testing and basophil histamine release. Besides these three allergens evaluated for, proteins smaller than 15 kDa were also identified as binding IgE in the majority of the patients (20/32) (28).

The availability of recombinant Peanut allergens has resulted in a greater ability to assess the sensitization and clinical profiles of individual Peanut allergens in different population groups. This is illustrated by a number of studies.

In an evaluation of recombinant allergens, Ara h 1, Ara h 2, and Ara h 3, using sera of 77 Peanut-allergic patients, seven different patterns of sensitization were identified to these allergens. The majority of patients (97%) had specific IgE to at least one of the recombinant allergens (Ara h 1, Ara h 2, and Ara h 3), and 77%, 75% and 77% recognized rAra h1, rAra h 2 and rAra h 3 respectively. High epitope diversity was found in patients with a history of more severe allergic reactions (27).

An European study of sensitization to six recombinant Peanut allergens with 40 patients sera, showed 14 individual recognition patterns with the following frequency of specific IgE binding: Ara h 1 was recognized by 65%, Ara h 2 by 85%, Ara h 4 by 53%, Ara h 5 by 13%, Ara h 6 by 38% and Ara h 7 by 43% of the selected sera (10).

Similarly, a French and American study aimed at evaluating the diagnostic value of the 3 major recombinant Peanut allergens utilizing skin and serum specific IgE determination in 30 Peanut-allergic patients. All patients with Peanut allergy demonstrated skin specific IgE to rAra h 2; 40% reacted with rAra h 1 and 27% with rAra h 3. Monosensitization to rAra h 2 was observed in 53% of patients. Levels of specific IgE did not correlate with the disease severity. However, patients with monosensitization to rAra h 2 had a significantly lower severity score than polysensitized subjects and a lower level of specific IgE against Peanut extract and rAra h 2. Cosensitization to rAra h 2 and rAra h 1 and/or rAra h 3 appeared to be predictive of more severe reactions (6).

A Dutch study examined the IgE reactivity to major recombinant Peanut allergens in 20 Peanut-allergic children at two subsequent time-points. Before DBPCFC, all 20 Peanut-allergic children were shown to have specific IgE to Ara h 2, 16 to Ara h 6, and 10 to both Ara h 1 and Ara h 3. After 20 months, Peanut-specific IgE levels and the individual recognition of major allergens were comparable with the levels and recognition before challenge. Skin specific IgE was detected to Ara h 2 and Ara h 6 in most children, whereas for Ara h 1 and Ara h 3 in approximately half of the children. None of the parameters were related to the severity of Peanut allergy. In this study, Ara h 2 and Ara h 6 were found to be the most frequently recognized major Peanut allergens in children (34).

A more recent study also investigated whether sensitization to individual allergens Ara h 1, Ara h 2, Ara h 3 and Ara h 6 could be correlated with clinical severity using purified Peanut allergens for skin and serum IgE evaluation in 30 patients. The majority of patients were found to have specific IgE to Ara h 2 (25/30, 83%) and Ara h 6 (26/30, 87%). Sixteen patients (53%) were sensitized to Ara h 1 and 15 patients (50%) to Ara h 3. All patients with a positive SPT to Ara h 1 and/or Ara h 3 were also sensitized to Ara h 2 and/or Ara h 6. Patients with severe symptoms had a higher skin specific IgE response to Ara h 2 and Ara h 6 at low concentrations (0.1 mug/mL) and to Ara h 1 and Ara h 3 at higher concentrations (100 mug/mL) compared with patients with mild symptoms. They also recognized a greater number of allergens and showed a higher cumulative SPT response compared with patients with mild symptoms. Therefore Ara h 2 and Ara h 6 appeared to be more potent than Ara h 1 and Ara h 3. Both skin specific IgE reactivity to low concentrations of Ara h 2 and Ara h 6 and to higher concentrations of Ara h 1 and Ara h 3 were shown to be indicative of severe symptoms (5).

Recombinant Peanut allergens have been evaluated for their ability to predict the outcome of tolerance in Peanut-allergic individuals. An American study was performed using sera from 15 patients with symptomatic Peanut allergy and 16 patients who were sensitized but tolerant (of which 10 of these 16 patients had "outgrown" their allergy) investigated 8 peptides representing the immunodominant sequential epitopes on Ara h 1, 2, and 3. It was found that regardless of their Peanut-specific IgE levels, most patients with symptomatic Peanut allergy showed IgE binding to the 3 immunodominant epitopes on Ara h 2. Sixty to seventy three percent of symptomatic patients recognized Ara h 2 and 87% to Ara h 3. The most striking difference in IgE binding was for Ara h 2 with 60-73% of symptomatic patients recognizing three immunodominant epitopes. For Ara h 3, 87% of symptomatic and 31% of asymptomatic patients recognized the epitope investigated. In contrast, each of these epitopes was recognized by < 10% of the tolerant patients. Tolerant patients did not recognize 2 immunodominant epitopes on Ara h 1. At least 93% of symptomatic, but only 12.5% of tolerant patients, recognized 1 of these "predictive" epitopes on Ara h 1 or Ara h 2. With up to 50% of patients with Peanut-specific IgE levels below suggested diagnostic decision levels still being clinically reactive, oral food challenges could be avoided in approximately 90% of these patients through the determination of peptide-specific IgE. This study analyzed only selected allergen epitopes rather than whole proteins (35).

It has also been argued that in contrast to recombinant Ara h 3, the allergen isolated from its native source is extensively proteolytically processed, and that native Ara h 3 polypeptides are much more complex than the recombinant protein used for epitope mapping experiments. The authors concluded that characterization of the allergenicity of Ara h 3 should therefore also include IgE-binding studies with Peanut-derived Ara h 3, providing the high degree of variation in the Ara h 3 protein structure, as this is what Peanut-allergic individuals are confronted with (3).

Ara h 3 is an 11S globulin and shares homology, and therefore varying degrees of cross-reactivity, with other 11S globulins. Sin a 2, a major allergen from Yellow mustard seed, was shown to have a sequence identity with other allergenic 11S globulins ranging between 27% and 38%. Three peptides described as epitopes in Ara h 3 were moderately conserved in Sin a 2 (36). Similarly, IgE-binding epitopes of Ara h 3 exhibited some structural homology among Peanut and tree nut allergens (Jug r 4 of Walnut, Cor a 9 of Hazelnut, Ana o 2 Cashew nut) to account for the IgE-binding cross-reactivity observed. IgE-binding epitopes similar to those found in 11S globulin allergens do not apparently occur in other vicilin allergens with the cupin fold from Peanut (Ara h 1) or tree nuts (Jug r 2 of Walnut, Cor a 1 of Hazel nut, Ana o 3 of Cashew nut) (37).

Cross-reactivity has also been demonstrated between homologous Ara h 3 proteins (but not related) in Lupin (Lupin conglutin gamma) and Soybean (Soybean Bg7S) (38). A sequence similarity between Ara h 3 and the glycinins in Soybean and Pea of 62% to 72% has been reported (39).

Diagnostic methods e.g., skin and serum specific IgE, etc., are based on natural Peanut extracts that contain both allergenic and non-allergenic proteins. A great variability and difficulty of standardization exists because the extract composition depends on the origin of raw material and extraction, purification and storage procedures (40). Recombinant allergens leads to standardized reagents that are biochemically characterized and therefore results that are comparable. Furthermore, recombinant allergens produced in E coli lack cross-reactive carbohydrate determinants (CCDs), which increases diagnostic specificity (41). As the amount of Ara h 1 and Ara h 3 in Peanut can vary or be higher than Ara 2 and Ara h 6 (42), and considering that the potency of allergens vary (5), utilizing recombinant allergens may be useful for diagnostic purposes in particular for more-appropriate diagnoses when used in Component Resolved Diagnosis (CRD), for exploring cross-reactivity, and for immunotherapy (43).

Compiled by Dr Harris Steinman, harris@zingsolutions.com

References:

1. Rabjohn P, Helm EM, Stanley JC, West CM, Sampson HA, Burks AW, et al. Molecular cloning and epitope analysis of the peanut allergen Ara h 3. J Clin Invest 1999;103:535-542
2. Koppelman SJ, Knol EF, Vlooswijk RA, Wensing M, Knulst AC, Hefle SL, Gruppen H, Piersma S. Peanut allergen Ara h 3: isolation from peanuts and biochemical characterization. Allergy 2003;58(11):1144-51
3. Piersma SR, Gaspari M, Hefle SL, Koppelman SJ. Proteolytic processing of the peanut allergen Ara h 3. Mol Nutr Food Res 2005;49(8):744-55.
4. Viquez OM, Konan KN, Dodo HW. Genomic organization of peanut allergen gene, Ara h 3. Mol Immunol 2004;41(12):1235-40
5. Peeters KA, Koppelman SJ, van HE, van der Tas CW, den Hartog Jager CF, Penninks AH, Hefle SL, Bruijnzeel-Koomen CA, Knol EF, Knulst AC. Does skin prick test reactivity to purified allergens correlate with clinical severity of peanut allergy? Clin Exp Allergy 2007;37(1):108-115
6. Astier C, Morisset M, Roitel O, Codreanu F, Jacquenet S, Franck P, Ogier V, et al. Predictive value of skin prick tests using recombinant allergens for diagnosis of peanut allergy. J Allergy Clin Immunol 2006;118(1):250-256
7. Dean TP. Immunological responses in peanut allergy. [Editorial] Clin Exp Allergy 1998;28:7-9
8. Wichers HJ, De Beijer T, Savelkoul HF, Van Amerongen A. The major peanut allergen Ara h 1 and its cleaved-off N-terminal peptide; possible implications for peanut allergen detection. J Agric Food Chem 2004;52(15):4903-7
9. Maleki SJ, Viquez O, Jacks T, Dodo H, Champagne ET, Chung SY, Landry SJ. The major peanut allergen, Ara h 2, functions as a trypsin inhibitor, and roasting enhances this function. J Allergy Clin Immunol 2003;112(1):190-5
10. Kleber-Janke T, Crameri R, Appenzeller U, Schlaak M, Becker WM Selective cloning of peanut allergens, including profilin and 2S albumins, by phage display technology. Int Arch Allergy Immunol 1999;119:265-274
11. Koppelman SJ, de Jong GA, Laaper-Ertmann M, Peeters KA, Knulst AC, Hefle SL, Knol EF. Purification and immunoglobulin E-binding properties of peanut allergen Ara h 6: evidence for cross-reactivity with Ara h 2. Clin Exp Allergy 2005;35(4):490-7.
12. Mittag D, Akkerdaas J, Ballmer-Weber BK, Vogel L, Wensing M, Becker WM, Koppelman SJ, Knulst AC, Helbling A, Hefle SL, Van Ree R, Vieths S. Ara h 8, a Bet v 1-homologous allergen from peanut, is a major allergen in patients with combined birch pollen and peanut allergy. J Allergy Clin Immunol 2004;114(6):1410-7.
13. Burks AW, Cockrell G, Connaughton C, Guin J, Allen W, Helm RM. Identification of peanut agglutinin and soybean trypsin inhibitor as minor legume allergens. Int Arch Allergy Immunol 1994;105(2):143-9
14. Asero R, Mistrello G, Roncarolo D, Amato S, Caldironi G, Barocci F, Van Ree R. Immunological cross-reactivity between lipid transfer proteins from botanically unrelated plant-derived foods: a clinical study. Allergy 2002;57(10):900-6
15. Pons L, Chery C, Romano A, Namour F, Artesani MC, Gueant JL. The 18 kDa peanut oleosin is a candidate allergen for IgE-mediated reactions to peanuts. Allergy 2002;57 Suppl 72:88-93
16. Boldt A, Fortunato D, Conti A, Petersen A, Ballmer-Weber B, Lepp U, Reese G, Becker WM. Analysis of the composition of an immunoglobulin E reactive high molecular weight protein complex of peanut extract containing Ara h 1 and Ara h 3/4. Proteomics 2005;5(3):675-686
17. Becker W-M, Kleber-Janke T, Lepp U. Four Novel Recombinant Peanut Allergens: More Information, More Problems. Int Arch Allergy Immun 2001;124:100-102
18. International Union of Immunological Societies Allergen Nomenclature: IUIS official list http://www.allergen.org/List.htm 2006
19. Dodo HW, Viquez OM, Maleki SJ, Konan KN. cDNA clone of a putative peanut (Arachis hypogaea L.) trypsin inhibitor has homology with peanut allergens Ara h 3 and Ara h 4. J Agric Food Chem 2004;52(5):1404-9
20. Rabjohn P, West CM, Connaughton C, Sampson HA, Helm RM, Burks AW, Bannon GA. Modification of Peanut Allergen Ara h 3: Effects on IgE Binding and T Cell Stimulation. Int Arch Allergy Immunol 2002;128(1):15-23
21. Kang IH, Gallo M Cloning and characterization of a novel peanut allergen Ara h 3 isoform displaying potentially decreased allergenicity. Plant Sci 2006;172(2):345-353
22. Xiang P, Beardslee TA, Zeece MG, Markwell J, Sarath G. Identification and analysis of a conserved immunoglobulin E-binding epitope in soybean G1a and G2a and peanut Ara h 3 glycinins. Arch Biochem Biophys 2002;408(1):51-7
23. Restani P, Ballabio C, Corsini E, Fiocchi A, Isoardi P, Magni C, Poiesi C, Terracciano L, Duranti M. Identification of the basic subunit of Ara h 3 as the major allergen in a group of children allergic to peanuts. Ann Allergy Asthma Immunol 2005;94(2):262-6.
24. Beyer K, Grishina G, Bardina L, Grishin A, Sampson HA. Identification of an 11S globulin as a major hazelnut food allergen in hazelnut-induced systemic reactions. J Allergy Clin Immunol 2002;110(3):517-23
25. Eigenmann PA, Burks AW, Bannon GA, Sampson HA. Identification of unique peanut and soy allergens in sera adsorbed with cross-reacting antibodies. J Allergy Clin Immunol 1996;98(5 Pt 1):969-78
26. Liang XQ, Luo M, Holbrook CC, Guo BZ Storage protein profiles in Spanish and runner market type peanuts and potential markers. BMC Plant Biol 2006; 6(1):24-
27. Shreffler WG, Beyer K, Chu TH, Burks AW, Sampson HA. Microarray immunoassay: association of clinical history, in vitro IgE function, and heterogeneity of allergenic peanut epitopes. J Allergy Clin Immunol 2004;113(4):776-82.
28. Koppelman SJ, Wensing M, Ertmann M, Knulst AC, Knol EF. Relevance of Ara h1, Ara h2 and Ara h3 in peanut-allergic patients, as determined by immunoglobulin E Western blotting, basophil-histamine release and intracutaneous testing: Ara h2 is the most important peanut allergen. Clin Exp Allergy 2004;34(4):583-90
29. Rodin J, Sjodahl S, Josefsson LG, Rask L. Characterization of a Brassica napus gene encoding a cruciferin subunit: estimation of sizes of cruciferin gene families. Plant Mol Biol 1992;20(3):559-63.
30. Beyer K, Morrow E, Li XM, Bardina L, Bannon GA, Burks AW, Sampson HA. Effects of cooking methods on peanut allergenicity. J Allergy Clin Immunol 2001;107(6):1077-81
31. Kang IH, Gallo M, Tillman BL Distribution of allergen composition in peanut (Arachis hypogaea l.) and wild progenitor (Arachis) species. Crop Science 2007;47(3):997-1003
32. Kang IH, Srivastava P, Ozias-Akins P, Gallo M Temporal and spatial expression of the major allergens in developing and germinating peanut seed. Plant Physiol 2007;144(2):836-845
33. Lewis SA, Grimshaw KE, Warner JO, Hourihane JO. The promiscuity of immunoglobulin E binding to peanut allergens, as determined by Western blotting, correlates with the severity of clinical symptoms. Clin Exp Allergy 2005;35(6):767-73.
34. Flinterman AE, van HE, den Hartog Jager CF, Koppelman S, Pasmans SG, Hoekstra MO, Bruijnzeel-Koomen CA, Knulst AC, Knol EF. Children with peanut allergy recognize predominantly Ara h2 and Ara h6, which remains stable over time. Clin Exp Allergy 2007;37(8):1221-1228
35. Beyer K, Ellman-Grunther L, Jarvinen KM, Wood RA, Hourihane J, Sampson HA. Measurement of peptide-specific IgE as an additional tool in identifying patients with clinical reactivity to peanuts. J Allergy Clin Immunol 2003;112(1):202-207
36. Palomares O, Vereda A, Cuesta-Herranz J, Villalba M, Rodriguez R. Cloning, sequencing, and recombinant production of Sin a 2, an allergenic 11S globulin from yellow mustard seeds. J Allergy Clin Immunol 2007;119(5):1189-96.
37. Barre A, Jacquet G, Sordet C, Culerrier R, Rouge P. Homology modelling and conformational analysis of IgE-binding epitopes of Ara h 3 and other legumin allergens with a cupin fold from tree nuts. Mol Immunol 2007;44(12):3243-3255
38. Magni C, Ballabio C, Restani P, Sironi E, Scarafoni A, Poiesi C, Duranti M. Two-dimensional electrophoresis and western-blotting analyses with anti Ara h 3 basic subunit IgG evidence the cross-reacting polypeptides of Arachis hypogaea, Glycine max, and Lupinus albus seed proteomes. J Agric Food Chem 2005;53(6):2275-2281
39. Beardslee TA, Zeece MG, Sarath G, Markwell JP. Soybean glycinin G1 acidic chain shares IgE epitopes with peanut allergen Ara h 3. Int Arch Allergy Immunol 2000;123(4):299-307
40. Hefle SL, Helm RM, Burks AW, Bush RK. Comparison of commercial peanut skin test extracts. J Allergy Clin Immunol 1995;95(4):837-42
41. Van Ree R. Carbohydrate epitopes and their relevance for the diagnosis and treatment of allergic diseases. Int Arch Allergy Immunol 2002;129(3):189-97
42. Koppelman SJ, Vlooswijk RA, Knippels LM, Hessing M, Knol EF, van Reijsen FC, Bruijnzeel-Koomen CA. Quantification of major peanut allergens Ara h 1 and Ara h 2 in the peanut varieties Runner, Spanish, Virginia, and Valencia, bred in different parts of the world. Allergy 2001;56(2):132-7
43. Kazemi-Shirazi L, Niederberger V, Linhart B, Lidholm J, Kraft D, Valenta R. Recombinant marker allergens: diagnostic gatekeepers for the treatment of allergy. Int Arch Allergy Immunol 2002;127(4):259-68
44.  

2007

• References