Allergy information for: Apple (Malus domestica)

Clinical History

• Number of Studies:6-10
• Number of Patients:>50
• Symptoms:

  When apple allergy is associated with birch pollinosis symptoms generally appears within 5-15 min and comprise local reactions of the mucosa of the upper aero-digestive tract with itching, inflammation and angioedema (called oral allergy syndrome, OAS). (Pastorello et al. 1999) [157]; (Anhøj et al. 2001) [591].

  When apple allergy is not linked to birch pollinosis the most frequent clinical presentation is not OAS, but systemic symptoms such as generalised urticaria and anaphylaxis are more commonly observed, specially in those patients without an associated pollen allergy. (Fernandez-Rivas et al. 1997) [1060]. In these patients, clinical data also suggests that adverse reactions appear more frequently and are more severe when fruits are eaten with the peel (Fernandez-Rivas and Cuevas 1999) [833].

Skin Prick Test

• Number of Studies:6-10
• Food/Type of allergen:Apple flesh (prick-prick technique)
  • Whole fruit (cvs. Golden Delicious, and/or Gloster): Rodriguez et al. (2000) [491]; Skamstrup Hansen et al. (2001) [589] and (2001) [590], Asero et al. 2001 [596], Crespo et al. (2002) [834]
  • Pulp (cv. Golden Delicious): Fernandez-Rivas and Cuevas (1999) [157], Anhøj et al. (2001) [591], Osterballe et al. (2003) [581]
  • Peel (cv. Golden Delicious): Fernandez-Rivas and Cuevas (1999) [157], Osterballe et al. (2003) [581]

  Whole-apple extract:

  • French cv. Golden Delicious and German cv. Gloster apples (Skamstrup Hansen et al. (2001) [590]
  • cv. Golden delicious apple peel and pulp (Asero et al. 2001) [596]

  Commercial apple extract: Anhøj et al. (2001) [591], Osterballe et al. (2003) [581].

  Low temperature acetone powder extract of apple (A72): Osterballe et al. (2003) [581].

  Recombinant Mal d 1: Osterballe et al. (2003) [581]

  Purified Mal d 1: Skamstrup Hansen et al. (2001) [590]

  Purified Mal d 3: Garcia-Selles et al. (2002) [594]

  Authors agreed that although the diagnostic value of the prick-prick technique with fresh apple is useful, it is still very problematic that the expression of the major allergen Mal d1 varies between different apple strains and maturation stages (Osterballe et al. 2003) [581]. It is also of concern that fruit allergen extracts are often readily degradable or contain clinically irrelevant cross-reacting epitopes, resulting in diagnostic discrepancies (Anhøj et al. 2001) [591].

• Protocol: (controls, definition of positive etc)

  In most of the studies skin prick tests (SPTs) were considered positive when the wheal area was ≥7 mm² (diameter ≥3 mm).

  Asero et al. (2001) [596] described negative SPTs wheals <2 (with diameters <50% of the diameter of the positive control which was 10 mg/ml histamine).

  Osterballe et al. (2003) [581] also peformed scratch-chamber test (SCT) which was regarded as positive with a wheal-and-flare reaction.

• Number of Patients:

  33 patients showing clinical symptoms after the ingestion of apple, peach and/or pear (at least one of them) and a positive SPT response to the involved fruit/s. 22 patients had an associated grass pollinosis (fruit and pollinosis group) and 11 did not present any type of inhalant allergy (fruit group). Adverse reactions to apple were reported by 25 subjects, 15 of the fruit and pollinosis group and 10 patients of the fruit group. (Fernandez-Rivas and Cuevas, 1999) [833].

  22 patients with adverse reactions to Rosaceae fruits is studied; 9 patients reported adverse reactions to apple which were confirmed by a DBPCFC in 6. Rodriguez et al. (2000) [491]

  47 peach allergic patients. 24 of them with allergic symptoms to apple (Garcia-Selles et al. 2001) [594]

  36 grass- and or birch-allergic patients based on a clinical history followed by an oral food challenge test and SPT with fresh fruits. The control group comprised five nonatopics and 12 birch and grass allergics with no symptoms of OAS and frequently eating fresh fruits. All controls had negative SPT to fresh fruits. Anhøj et al. (2001) [591]

  36 patients with OAS after ingestion of fruits. 18 patients did not have birch pollen sensitivity and the other 18 had bich pollinosis (Asero et al. 2001) [596].

  65 birch pollen-allergic patients with a history of rhinitis in the birch-pollen season, positive specific IgE to birch and a positive open oral challenge with apple. Skamstrup Hansen et al. (2001) [589].

  26 patients based on a clinical history of rhinoconjunctivitis in the birch pollen season and OAS or other allergic manifestations upon ingestion of apple, together with positive specific IgE and a positive SPT to birch pollen. Skamstrup Hansen et al. (2001) [590]

  10 adults diagnosed with IgE-mediated clinical allergy to apple from history and by skin tests (SPT), and/or food-specific IgE antibodies, and oral challenge. Crespo et al. (2002) [834].

  10 patients with clinical allergy to birch pollen, a positive SPT to birch, and a positive case history of OAS to apple and 10 nonatopics without birch pollinosis, no symptoms of OAS and a negative SPT to birch as control subjects. Osterballe et al. (2003) [581]

• Summary of Results:

  Fernandez-Rivas and Cuevas (1999) [833] found that more than 40% of patients allergic to apple tolerated the ingestion of the pulp, and reactions were only elicited by the intake of the whole fruit. SPTs with peels of apple induced significantly higher responses than the pulps, both in fruit and pollinosis and in fruit allergic patients.

  Overall, 28 of 34 patients reporting adverse reactions had positive skin testing or serum-specific IgE antibodies for the evaluated Rosaceae fruits. 18 patients showed a positive SPT for apple. Rodriguez et al. (2000) [491]
  77% of the 47 peach allergic patients and 92% of the 24 apple allergic patients had positive SPT with purified Mal d 3. Garcia-Selles et al. (2001) [594]

  Anhøj et al. (2001) [591] found that the diagnostic sensitivity of the SPT with fresh apple fruit was 92% and specificity of 72%. Good concordance (test efficiency of >80%) was found between case history, SPT, and oral challenge tests. The negative predictive value of the SPT was >90%. A combination of case history of Oral Allergy syndrome with a SPT or basophil histamine release with fresh apple resulted in >90% positive predictive value.

  In both patients groups with and without birch pollinosis, the SPTs with fresh apple and apple peel extract were positive and negative with apple pulp. In non birch pollen-allergic patients, the SPTs with fresh apple gave inconsistent results, therefore Asero et al. (2001) [596] suggests diagnosis of vegetable food hypersensitivity in birch pollen-allergic patients using fresh foods but fruit peel extracts for patients not allergic to birch pollen.

  Skamstrup Hansen et al. (2001) [589] reported that groups I and II had higher skin wheal area to birch than group III (P=0.02/0.04). The freeze-dried apple powder proved to be useful for SPT but further investigation of the stability and the allergenic profile of the material is needed.

  Skamstrup Hansen et al. (2001) [590] could not confirm an increase in reactivity during season by SPT.

  Crespo et al. (2002) [834] SPT determinations were positive to apple in 23/30 (77%).

  Osterballe et al. (2003) [581] demonstrated that the SPT had a high sensitivity to fresh apple peel (80%), apple pulp (70%), A72 (90%), birch (100%), and low to the commercial apple extract (10%). The authors concluded "In daily practice a detailed case history about symptoms of oral allergy syndrome combined with a SPT with fresh apple peel or low temperature acetone powder extract of apple will be useful"

IgE assay (by RAST, CAP etc)

• Number of Studies:0
• Food/Type of allergen:Apple extracts
  • Whole fruit cv. Golden Delicious: Skamstrup Hansen et al. (2001) [589] and (2001) [590], Osterballe et al. (2003) [581], Rossi et al. (2003) [835].
  • Pulp (cv. Golden Delicious): Fernandez-Rivas and Cuevas (1999) [833], Anhøj et al. (2001) [591], Osterballe et al. (2003) [581]
  • Peel (cv. Golden Delicious): Fernandez-Rivas and Cuevas (1999) [833], Osterballe et al. (2003) [581]
• IgE protocol:CAP: Fernandez-Rivas and Cuevas (1999) [833], Skamstrup Hansen et al. (2001) [589] and (2001) [590], Osterballe et al. (2003) [581], Rossi et al. (2003) [835].
  • Rossi et al. (2003) [835] expressed the results in classes of positivity from 0 to 6, where class 0 corresponds to <0.35 kUA/l, class 1 to 0.35-0.7 kUA/l, class 2 to 0.7-3.5 kUA/l, class 3 to 3.5-17.5 kUA/l, class 4 to 17.5-50 kUA/l, class 5 to 50-100 kUA/l and class 6 to >100 kUA/l.
  • Measurable specific IgE to apple was classified as a positive when CAP = 0.35 kU A /L by Osterballe et al. (2003) [581]
  Magic Lite (ML): Skamstrup Hansen et al. (2001) [590], Osterballe et al. (2003) [581]. Measurable specific IgE to apple was classified as a positive when ML = 1.44 SU/ml.
  RAST and RAST inhibition: Apple peel RAST was inhibited by itself and by its pulp, and vice versa (Fernandez-Rivas and Cuevas 1999) [833].

  A histamine release test (HRT) was performed considered positive:

  • A release greater than 10% (Fernandez-Rivas and Cuevas (1999)) [833].
  • An AUC of >40 ng/ml and a release of >20 ng/ml histamine (Anhøj et al. (2001)) [591].
  • Maximal release is ≥14 ng/ml (Skamstrup Hansen et al. (2001) [589]).
  • A release greater than 10 ng/ml (Osterballe et al. (2003) [581])
• Number of Patients:

  33 patients showing clinical symptoms after the ingestion of apple, peach and/or pear (at least one of them) and a positive SPT response to the involved fruit/s. 22 patients had an associated grass pollinosis (fruit and pollinosis group) and 11 did not present any type of inhalant allergy (fruit group). Adverse reactions to apple were reported by 25 subjects, 15 of the fruit and pollinosis group and 10 patients of the fruit group. (Fernandez-Rivas and Cuevas, 1999) [833].

  36 grass- and or birch-allergic patients based on a clinical history followed by an oral food challenge test and SPT with fresh fruits. The control group comprised five nonatopics and 12 birch and grass allergics with no symptoms of OAS and frequently eating fresh fruits. All controls had negative SPT to fresh fruits. Anhøj et al. (2001) [591]

  65 birch pollen-allergic patients with a history of rhinitis in the birch-pollen season and positive specific IgE to birch. Skamstrup Hansen et al. (2001) [589].

  26 patients based on a clinical history of rhinoconjunctivitis in the birch pollen season and OAS or other allergic manifestations upon ingestion of apple, together with positive specific IgE and a positive SPT to birch pollen. Skamstrup Hansen et al. (2001) [590]

  10 patients with clinical allergy to birch pollen, a positive SPT to birch, and a positive case history of OAS to apple and 10 nonatopics without birch pollinosis, no symptoms of OAS and a negative SPT to birch as control subjects. Osterballe et al. (2003) [581]

  372 Birch pollen allergic patients with a history of seasonal (February-April) pollen allergy with symptoms of rhinitis and/or asthma. A fraction of them suffered from oral allergy syndrome after eating apple, too. Rossi et al. (2003) [835]

• Summary of Results:

  Fernandez-Rivas and Cuevas (1999) [833] found that total IgE was significantly higher in the fruit and pollinosis group (mean 481.42 kU/L) than in fruit group (mean 152.73 kU/L). RAST to apple peel was significantly higher than RAST to pulps in the fruit patients, but no significant differences were observed in the fruit and pollinosis group. Fruit peel RAST was inhibited (from 60 to 97%) by itself and by its pulp, and viceversa. The area under the curve of histamine release was larger for peels of apple.

  Histamine release tests with apple gave sensitivity of 71% and specificity of 93%. Anhøj et al. (2001) [591].

  The maximum histamine release (ng/ml) against fresh Golden delicious apple was higher in group III than in group I (P=0.02). The freeze-dried apple powder proved to be useful for histamine release but further investigation of the stability and the allergenic profile of the material is needed. Skamstrup Hansen et al. (2001) [589]

  Skamstrup Hansen et al. (2001) [590] found that specific IgE against apples cv. Golden Delicious increased during birch pollen season (P<0.05). None of the patients had positive specific IgE against apple measured by the Magic Lite system.

  Similarly, Osterballe et al. (2003) [581] reported that the CAP test showed a higher diagnostic sensitivity (100%) than the Magic lite test (10%) for apple. The Histamine release test showed a high sensitivity to birch (90%), A72 (90%), Mal d1 (70%) and low to the commercial apple extract (25%).

  Rossi et al. (2003) [835] found that 162 sera (43.55%) contained specific IgE to apple extract (75.35% of birch pollen Bet v 1 positive sera). The authors concluded "In this study we observed that three birch pollen recombinant allergens alone, could sufficiently identify 90% of birch pollen-sensitive patients".

Immunoblotting

• Immunoblotting separation:

  Sodium dodecylsulfate-polyacrylamide gel electrophoresis was perdformed using 12% polyacrylamide gels (Vanek-Krebitz et al. 1995) [222], (Ebner et al. 1995) [46]

  The apple extract was separated in a discontinuous buffer system in a SDS-PAGE gradient gel with 6% stacking and 7.5% to 20% separation gradient gel. The sample, at the protein concentration of 7 mg/mL, was diluted 1:2 in sample buffer containing TRIS adjusted to pH 6.1 with concentrated sulfuric acid, 10% SDS, 2-mercaptoethanol, 50% glycerol, and 1% bromphenol blue. The samples were denatured at 100°C for 5 minutes (Pastorello et al. 1999) [157]

• Immunoblotting detection method:

  The proteins were electroblotted to a nitrocellulose sheets with pore size 0.2 µm. The sheets were cut into 5 mm strips and blocked with phosphate-buffered saline pH 7.5 with 0.5% (v/v) Tween 20 and 0.5% (w/v) bovine serum albumin for 30 min. Strips were incubated with diluted (1:4) sera overnight. The IgE-binding components were detected with iodine 125–labeled rabbit anti-human IgE antiserum. Bots were visualised by autoradiography (Ebner et al. 1995) [46].

  The proteins were electroblotted to a nitrocellulose membranes. The IgE-binding components were detected with iodine 125–labeled rabbit anti-human IgE antiserum. Blots were visualised by autoradiography (Vanek-Krebitz et al. 1995) [222]

  Allergens separated by SDS-PAGE were electroblotted onto a nitrocellulose membrane (0.45 µm) with use of a transblot cell from Bio-Rad at 0.45 A, 100 V, for 4 hours at 4°C. The unoccupied protein binding sites in the nitrocellulose membrane were blocked by a 30 minute incubation at 37°C with PBS, pH 7.4, with 0.5% Tween 20. The nitrocellulose was then cut into strips, which were incubated overnight at room temperature with the serum of each single patient. IgE binding by specific antibodies was detected by incubation with iodine 125–labeled anti-human-IgE diluted 1:4 in blocking solution for 6 hours at room temperature and exposure on x-ray film at –70°C for 3 days (Pastorello et al. 1999) [157]

• Immunoblotting results:

  20/20 sera displayed IgE binding to apple extract. 20 sera reacted with a protein of 17 kDa, and 4 sera reacted with an polypeptide of 14 kDa. Sera from 4 patients recognized proteins in the higher MW range (Ebner et al. 1995) [46]

  9/9 patients suffering from allergic reactions to birch pollen and apple displayed igE binding to nMal d1, rMal d 1 and rBet v1 (Vanek-Krebitz et al. 1995) [222]

  Birch-positive patients had IgE binding to proteins with MWs of 9 kDa (9 patients, 28%), 15 kDa (16 patients, 50%), 18 kDa (29 patients, 91%), 28 kDa (2 patients, 6%), 31 kDa (13 patients, 41%), 43, 51, and 84 kDa (11 patients, 34%), and 60 kDa (7 patients, 22%). All birch-negative patients had IgE binding only to the 9-kDa polypeptide, and 6/7 patients with mugwort pollinosis (86%) (Pastorello et al. 1999) [157]

Oral provocation

• Number of Studies:6-10
• Food used and oral provocation vehicle:

  Anhøj et al. (2001) [591] and Pastorello et al. (1999) [157] used fresh apple.

  A total of 17 g of dehydrated whole fruit was masked in a mix of orange (200 mL) and pineapple (200 mL) juices, sugar (16 g), wheat meal (13 g), and liquid coloring (McCormick). An open feeding of the whole fresh fruit in identical conditions and quantity as in the initial OFC followed all negative blinded challenges. The time between initial and final OFCs was not more than 10 days. Rodriguez et al. (2000) [491]

  Skamstrup Hansen et al. (2001) [589].

  • Model I: fresh apple juice and additional blackcurrant juice.
  • Model II: freshly grated apple and additional grated cabbage and apple juice.
  • Model III: freeze-dried apple powder and additional apple juice and blackcurrant juice

  Skamstrup Hansen et al. (2001) [590] used grated cabbage, pasteurisated apple juice and grated apple (Golden delicious) for DBPCFC, and whole fresh apple (Golden delicious) for open challenge tests.

  The meals contained a mixture of fresh shredded apple (Golden Delicious), yogurt, orange juice, apple juice, applesauce, oatmeal flakes and dry rasped coconut. The placebo doses consisted of the same ingredients except fresh apple. Apple juice and sauce did not contain any IgE-reactive Mal d 1 allergen due to processing steps and were added for optimal blinding of taste. Bolhaar et al. (2004) [1143]

• Blind:

  Skamstrup Hansen et al. (2001) [589], Bolhaar et al. (2004) [1143]. Yes.

  Skamstrup Hansen et al. (2001) [590]; Rodriguez et al. (2000) [491]. DBPCFC and open challenge.

  Anhøj et al. (2001) [591] and Pastorello et al. (1999) [157]. Open challenge

• Number of Patients:

  43 patients with oral allergy syndrome for apple and positive open food challenge, skin prick test, and serum specific IgE antibodies to apple. 32 had birch pollinosis (documented by specific IgE for birch) and 11 were not allergic to birch. Pastorello et al. (1999) [157]

  A total of 226 food challenges (including open food challenge and DBPCFC) were performed in the 28 patients with positive skin prick tests or CAP System FEIA. Rodriguez et al. (2000) [491]

  65 birch pollen-allergic patients with a history of rhinitis in the birch-pollen season and positive specific IgE to birch, and positive open challenge with apple. For comparison of the DBPCFC models, patients with a positive open oral challenge ith apple were selected. Skamstrup Hansen et al. (2001) [589].

  26 patients based on a clinical history of rhinoconjunctivitis in the birch pollen season and Oral allergy syndrome or other allergic manifestations upon ingestion of apple, together with positive specific IgE and a positive SPT to birch pollen. Skamstrup Hansen et al. (2001) [590]

  36 grass- and or birch-pollen allergic patients based on a clinical history followed by an oral food challenge test and SPT with fresh fruits. The control group comprised five nonatopics and 12 birch and grass allergics with no symptoms of Oral allergy syndrome and frequently eating fresh fruits. All controls had negative SPT to fresh fruits. Anhøj et al. (2001) [591]

  25 patients (nine males and 16 females, mean age 35.6 years) with a history of birch-pollen and apple allergy. Patients were randomized into two groups: one group to receive birch-pollen immunotheraphy (IT) (n=13) and the control group to receive only symptomatic treatment (n=12). Bolhaar et al. (2004) [1143]

• Dose response:

  From 4 to 64 g. Pastorello et al. (1999) [157].

  Subjects were first challenged randomly with either food or placebo. After an interval of at least 24 hours, the second part of the DBPCFC took place with subjects receiving either food or placebo depending on what they had recieved in the first challenge. Confirmation by DBPCFC was accepted if the subject had symptoms after provocation with the active substance and no symptoms after the placebo challenge. Rodriguez et al. (2000) [491]

  Model I: 5-50 ml. Model II: 20 g. Model III: 3 g. Skamstrup Hansen et al. (2001) [589].

  20 g of grated apple. The authors note that only a low dose of apple could be used because of the flavour and suggest that more positives would occur if a higher dose were used. Skamstrup Hansen et al. (2001) [590].

  A bite of an apple. Anhøj et al. (2001) [591].

  The challenge meals were prepared within 5 min before administration and contained 4, 10, 40 or 120 g of fresh shredded apple (Golden Delicious). The patients of the immunotheraphy (IT) group received a standardized aluminium hydroxide adsorbed birch-pollen extract. All patients were given IT according to a modified cluster schedule, between November 2001 and January 2002, followed by a monthly maintenance injection of 100 000 standard quality units (SQ-U) for 1 year with a dose reduction of 50% during the pollen season. All patients were pre-treated with an antihistamine at least 1 h before injection, since this reduces side-effects and has been reported to possibly enhance efficacy. Bolhaar et al. (2004) [1143]

• Symptoms:

  Oral allergy syndrome symptoms were classified in 4 grades of increasing severity: grade 1, only oral mucosa symptoms (22/32 and 9/11) ; grade 2, oral mucosa and gastrointestinal symptoms (1/11); grade 3, oral mucosa symptoms plus systemic symptoms (urticaria, rhinoconjunctivitis, or asthma) (2/32 and 1/11); grade 4, oral mucosa symptoms plus life-threatening symptoms (laryngeal edema, anaphylactic shock) (7/32). Pastorello et al. (1999) [157].

  6/28 patients were positive after DBPCFC with apple. 2 patients had oral symptoms and 4 had generalised anaphylaxis (Rodriguez et al. 2000) [491]

  Oral allergy syndrome: rhinitis, conjunctivitis, and local reactions at the oral cavity. Skamstrup Hansen et al. (2001) [590]; Anhøj et al. (2001).

  Skamstrup Hansen et al. (2001) [589] found: Model I (17 positive, 13 negative, placebo 10 positive), Model II (14 positive, 5 negative, placebo 0 positive) and Model III (15 positive, 6 negative, placebo 4 positive). The authors concluded "It is highly desirable that the sensitivity of the DBPCFC with apple is increased further. Lyophilization of apples produces material useful for oral challenges and has some practical advantages over fresh apples, but an investigation of the long-time stability and the allergenic profile of the freeze-dried material is needed.

  Skamstrup Hansen et al. (2001) [590] found that before the pollen season, 14/26 blinded challenges were positive, against 19/27 open challenges. During the pollen season, 16/26 blinded and 20/26 open challenges were positive. 1 positive placebo in each case with a different patient. None of the patients reacted to the blinded challenge without a subsequent reaction to the open challenge. The scores of the open challenge were significantly higher than the scores of the DBPCFC both before the season and during the in-season challenges (P<0.05).

  Authors agreed that the diagnosis of oral allergy syndrome to apple has been complicated by unstable apple extracts and the changes of the allergenicity of apples during ripening, the test technique, the disease prevalence and the number of the patients included in studies.

  Symptoms started within 5-10 min after ingestion of the challenge meal, as the oral allergy syndrome (OAS): itching or feeling of tightness in the oral cavity or tingling of the lips. Patient reactivity was expressed using VAS scores. After 1 year of immunotherapy (IT), both groups were tested with DBPCFC. In the IT group VAS scores were significantly decreased at t=12 (P<0.001) compared with those at the start of the study. This resulted in an increase of the amount of apple tolerated of a factor of 24. Nine of the 13 patients treated with IT improved significantly, whereas four patients showed VAS scores similar to t=0. In three of the nine patients who showed improvement, the provocation was completely negative after 1 year of birch-pollen IT, suggesting they had overcome their apple allergy. In the control group, nine out of 10 patients showed unchanged or even increased VAS scores relative to the year before, and no significant differences were found between t=0 and 12. One patient in the control group was not willing to undergo a challenge for the second time. No placebo reactions were observed in both groups. Bolhaar et al. (2004) [1143]

IgE cross-reactivity and Polysensitisation

Other Clinical information

Reviews (3)

• Marion D, Douliez JP, Gutier MF, Elmorjani K.
  Plant Lipid transfer proteins: Relationships between allergenicity and structural, biological and technological properties.
  Plant Food Allergens. Mills ENC and Shewry PR editors. Chapter 4; 57-69. 2004
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  Bet v 1 homologous allergens
  Plant Food Allergens. Mills ENC and Shewry PR editors. Chapter 8; 125-140. 2004
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• Salcedo G, Sanchez-Monge R, Diaz-Perales A, Garcia-Casado G, Barber D
  Plant non-specific lipid transfer proteins as food and pollen allergens
  Clin Exp Allergy. 34(9):1336-41. 2004
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• Diaz-Perales A, Lombardero M, Sanchez-Monge R, Garcia-Selles FJ, Pernas M, Fernandez-Rivas M, Barber D, Salcedo G.
  Lipid-transfer proteins as potential plant panallergens: cross-reactivity among proteins of Artemisia pollen, Castanea nut and Rosaceae, with different IgE-binding capacities.
  Clin Exp Allergy 2000, 30: 1403-1410.. 2000
  PUBMEDID: 10998016
• Bolhaar ST, Tiemessen MM, Zuidmeer L, van Leeuwen A, Hoffmann-Sommergruber K, Bruijnzeel-Koomen CA, Taams LS, Knol EF, van Hoffen E, van Ree R, Knulst AC
  Efficacy of birch-pollen immunotherapy on cross-reactive food allergy confirmed by skin tests and double-blind food challenges
  Clin Exp Allergy. 34(5):761-9. 2004
  PUBMEDID: 15144469
• Ortolani C, Pastorello EA, Farioli L, Ispano M, Pravettoni V, Berti C, Incorvaia C, Zanussi C
  IgE-mediated allergy from vegetable allergens
  Ann Allergy. 71(5):470-6.. 1993
  PUBMEDID: 8250353
• Asero R, Mistrello G, Roncarolo D, Amato S, Zanoni D, Barocci F, Caldironi G.
  Detection of clinical markers of sensitization to profilin in patients allergic to plant-derived foods.
  J Allergy Clin Immunol. 112(2):427-432. 2003
  PUBMEDID: 12897752
• García Sellés FJ, Diaz-Perales A, Sanchez-Monge R, Alcantara M, Lombardero M, Barber D, Salcedo G, Fernandez-Rivas M
  Patterns of reactivity to lipid transfer proteins of plant foods and Artemisia pollen: an in vivo study
  Int Arch Allergy Immunol. 128(2):115-122. 2002
  PUBMEDID: 12065911

Biochemical Information for Mal d 1

• Allergen Name:Mal d 1
• Alternatve Allergen Names:
• Allergen Designation:Major
• Protein Family:Bet v 1 homologue, Pfam PF00407
• Sequence Known?:Yes
• Allergen accession No.s:

  P43211:Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?P43211

  X83672	EMBL
  Z48969	EMBL

  Q9SYW3:Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?Q9SYW3

  AF126402	EMBL
  AJ417551	EMBL

  Q9S7M5:Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?Q9S7M5

  AF124836	EMBL
  AF124822	EMBL
  AF124826	EMBL
  AF124827	EMBL
  AF124828	EMBL
  AF124833	EMBL
  AF124834	EMBL
  AY428578	EMBL

  Q9SYV3:Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?Q9SYV3

  AF124824

  EMBL

  Q9SYV4:Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?Q9SYV4

  AF124825

  EMBL

  Q40280:Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?Q40280

  L42952	EMBL
  AF074721	EMBL
  AF020542	EMBL

  Q9SYV2:Swissprot: http://us.expasy.org/cgi-bin/niceprot.pl?Q9SYV2

  AF124823

  EMBL

  Q9SYV5:Swissprot: http://us.expasy.org/cgi-bin/niceprot.pl?Q9SYV5

  AF124829

  EMBL

  Q9SYV6:Swissprot: http://us.expasy.org/cgi-bin/niceprot.pl?Q9SYV6

  AF124830

  EMBL

  Q9SYV7:Swissprot: http://us.expasy.org/cgi-bin/niceprot.pl?Q9SYV7

  AF124831

  EMBL

  Q9SYV8:Swissprot: http://us.expasy.org/cgi-bin/niceprot.pl?Q9SYV8

  AF124832

  EMBL

  Q9SYV9:Swissprot: http://us.expasy.org/cgi-bin/niceprot.pl?Q9SYV9

  AF124835

  EMBL

  Q8L6K9:Swissprot: http://us.expasy.org/cgi-bin/niceprot.pl?Q8L6K9

  AJ488060

  EMBL

• 3D Structure Accession No.:Not determined
• Calculated Masses:

  17651, Mal d 1.0101 (Acc. No: X83672)

  17559, Mal d 1.0102 (Acc. No: Z48969)

  17565, Mal d 1.0103 (Acc. No: AF124823)

  17649, Mal d 1.0104 (Acc. No: AF124829)

  17563, Mal d 1.0105 (Acc. No: AF124830)

  17609, Mal d 1.0106 (Acc. No: AF124831)

  17523, Mal d 1.0107 (Acc. No: AF124832)

  17623, Mal d 1.0108 (Acc. No: AF126402, AJ417551)

  17539, Mal d 1.0201 and Mal d 1.0202 (Acc. No: L42952; AF124822)

  17513, Mal d 1.0203 (Acc. No: AF124824)

  17571, Mal d 1.0204 (Acc. No: AF124825)

  17542, Mal d 1.0205 (Acc. No: AF124835)

  17525, Mal d 1.0206 (Acc. No: AF020542)

  17485, Mal d 1.0208 (Acc. No: AJ488060)

• Experimental Masses:18 kDa
• Oligomeric Masses:
• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:

  Mal d 1 is not only destroyed by cooking. It is also extremly labile when subjected to gastric digestion assays (pepsin digestion and followed by trypsin digestion. After only 30 sec of pepsin treatment at pH 2 no protein was detectable in immunoblots. (Jensen-Jarolim et al. 1999)

• Nature of main cross-reacting proteins:

  As a consequence of sequence identities, IgE to Mal d 1 cross-reacts with the allergenic Bet v 1 homologous from celery (Api g 1), carot (Dau c 1) and cherry (Pru a 1) and with major pollen allergens from the order Fagales , including birch (Bet v 1) (Ebner et al. 1995), alder (Aln g 1), hazelnut (Cor a 1) and Car b 1.

  Sequence comparison of Mal d 1 to Bet v 1 revealed 64.5% amino acid identity and 55.6% nucleic acid identity (Vanek-Krebitz et al. 1995)

• Allergen properties & biological function:Mal d 1 is considered to be a PR 10 protein based on its sequence identity and are stress- and pathogen-inducible (Puehringer et al. 2000). Bet v 1 homologues have no clear function but have recently been found to bind plant steroids (Markovic-Housley et al. 2003)
• Allergen purification:

  Apple fruits (cv. Golden Delicious) were homogenized with diacetone alcohol for 18 h at -20°C, then, filtered, dried and resolubilized with 0.1 M potassium phosphate buffer containing EDTA, DIECA, benzamidine hydrochloride adn phenylmethylsulfonyl fluoride. The solution was centrifuged (40,000 g, 30 min) and lyophilised. The apple extract was dissolved in Tris-HCl, and loaded first into an anion exchange chromatography column (Q Resource) and then into a reversed-phase HPLC column (Vydac, 214TP54). The proteins were eluted with a linear gradient of 0-50% acetonitrile in 0.1% TFA for 30 min and 5 min of 50-80% (Fahlbusch et al. 1995)

  Apple fruits (Malus domestica, cultivar Granny Smith) were obtained from a local grocery store and extracted according to the method of Björksten et al. followed by removal of pectins by addition of 5 mmol/L Ca²⁺, Mg²⁺, and Mn²⁺ (chloride salts in each case), centrifugation, and dialysis of supernatant against PBS. Mal d 1 was subsequently affinity purified with Sepharose-coupled mAb 5H8 directed to Bet v 1 (Helsper et al. 2002)

  Purification of recombinant Mal d 1 has been reported by several authors (Vanek-Krebitz et al. 1995), (Son et al. 1999)

• Other biochemical information:

  A number of Mal d 1 isoforms have been isolated, cloned and sequenced from different cultivars (Son et al. 1999). The authors concluded that the occurrence of the isoforms is not cultivar-related , and that a mixture of isforms is present in the fruit. Helsper et al. (2002) showed that the main isoform of Mal d 1 expressed in apple is the Mal d 1b. The allergenic potency of different cultivars found by Vieths et al. (1994) is related to total level of expressed Mal d 1 isoforms.

  The grouping of Mald 1 isoforms into subgroups a ,b, c etc according to Son et al. (1999) can be correlated with the suggested nomenclature from the allergen nomenclature database into: Mal d 1.01 Mal d 1.02 etc. Also on cDNA level Puehringer et al. (2003) identified Mal d 1 .01, Mal d 1.02 and Mal d 1.03 in different levels in the apple fruit when performing Real time PCR

  Cold storage (4°C) of apples increase the levels of Mal d 1 (Hsieh et al. 1995)

References (15)

• Bjorksten F, Halmepuro L, Hannuksela M, Lahti A
  Extraction and properties of apple allergens
  Allergy. 35(8):671-7. 1980
  PUBMEDID: 7224105
• Vieths S, Jankiewicz A, Schoning B, Aulepp H
  Apple allergy: the IgE-binding potency of apple strains is related to the occurrence of the 18-kDa allergen
  Allergy. 49(4):262-71.. 1994
  PUBMEDID: 7518656
• Puehringer H, Moll D, Hoffmann-Sommergruber K, Watillon B, Katinger H, Machado MLD
  The promoter of an apple Ypr10 gene, encoding the major allergen Mal d 1, is stress- and pathogen-inducible
  PLANT SCIENCE 152 (1): 35-50. 2000
  PUBMEDID:
• Ebner C, Hirschwehr, R, Bauer L, Breiteneder H, Valenta R, Ebner, H, Kraft D, Scheiner O.
  Identification of allergens in fruits and vegetables: IgE cross-reactivities with the important birch pollen allergens Bet v 1 and Bet v 2 (birch profilin).
  J Allergy Clin Immunol 95: 962-9.. 1995
  PUBMEDID: 7751517
• Son DY, Scheurer S, Hoffmann A, Haustein D, Vieths S.
  Pollen-related food allergy: cloning and immunological analysis of isoforms and mutants of Mal d 1, the major apple allergen, and Bet v 1, the major birch pollen allergen.
  Eur J Nutr. 38(4):201-15.. 1999
  PUBMEDID: 10502033
• Helsper JP, Gilissen LJ, van Ree R, America AH, Cordewener JH, Bosch D.
  Quadrupole time-of-flight mass spectrometry: a method to study the actual expression of allergen isoforms identified by PCR cloning.
  J Allergy Clin Immunol. 110(1):131-138.. 2002
  PUBMEDID: 12110832
• Schoning B, Ziegler WH, Vieths S, Baltes W
  Apple allergy: the cDNA sequence of the major allergen of apple determined by performing PCR with primers based on the N-terminal amino acid sequence is highly homologous to the sequence of the major birch pollern allergen.
  J. Sci. Food Agric 71:475-482.. 1996
  PUBMEDID:
• Fahlbusch B, Rudeschko O, Muller WD, Schlenvoigt G, Vettermann S, Jager L
  Purification and characterisation of the major allergen from apple and its allergenic cross-reactivity with Bet v 1.
  Int. Arch. Allergy Immunol. 108:119-126. 1995
  PUBMEDID: 7549498
• Hsieh L, Moos M, Lin Y
  Characterisation of apple 18 and 13kD allergens by microsequencing and evaluation of their content during storage and ripening.
  J.Allergy Clin Immunol. 96:960-970.. 1995
  PUBMEDID: 8543755
• Vieths S, Jamek K Aulepp H, Petersen A.
  Isolation and characterisation of 18kD major apple allergen and comparison with the major birch pollen allergen (Bet v 1).
  Allergy 50:421-430.. 1995
  PUBMEDID: 7573831
• Vanek-Krebitz M, Hoffmann-Sommergruber K, Laimer da Camara Machado M, Susani M, Ebner C, Kraft D, Scheiner O, Breiteneder H.
  Cloning and sequencing of Mal d 1, the major allergen from apple (Malus domestica), and its immunological relationship to Bet v 1, the major birch pollen allergen.
  Biochem Biophys Res Commun 214: 538-51.. 1995
  PUBMEDID: 7677763
• Markovic-Housley Z, Degano M, Lamba D, von Roepenack-Lahaye E, Clemens S, Susani M, Ferreira F, Scheiner O, Breiteneder H.
  Crystal structure of a hypoallergenic isoform of the major birch pollen allergen Bet v 1 and its likely biological function as a plant steroid carrier.
  J Mol Biol. 325(1):123-133.. 2003
  PUBMEDID: 12473456
• Hoffmann-Sommergruber K, Radauer C
  Bet v 1 homologous allergens
  Plant Food Allergens. Mills ENC and Shewry PR editors. Chapter 8; 125-140. 2004
  PUBMEDID:
• Puehringer HM, Zinoecker I, Marzban G, Katinger H, Laimer M
  MdAP, a novel protein in apple, is associated with the major allergen Mal d 1.
  Gene. 4;321:173-83. 2003
  PUBMEDID: 14637005
• Jensen-Jarolim E, Wiedermann U, Ganglberger E, Zurcher A, Stadler BM, Boltz-Nitulescu G, Scheiner O, Breiteneder H
  Allergen mimotopes in food enhance type I allergic reactions in mice
  FASEB J. 13(12):1586-92. 1999
  PUBMEDID: 10463950

Biochemical Information for Mal d 2

• Allergen Name:Mal d 2
• Alternatve Allergen Names:Previously known as Mdt1 but now renamed
• Allergen Designation:None
• Protein Family:

  Thaumatin-like proteins, Pfam PF00314

• Sequence Known?:Yes
• Allergen accession No.s:

  O82546:Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?O82546

  AF090143; AAC36740.1; EMBL
  AJ243427; CAC10270.1; EMBL
  AF494393; AAM12886.1; EMBL

• 3D Structure Accession No.:Not determined
• Calculated Masses:23,210.9Da (Krebitz et al. 2003) [595]
• Experimental Masses:31 kDa (Oh et al. 2000) [133]
• Oligomeric Masses:
• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:

  The thaumatin-like proteins (TLPs) contain 8 disulfide bridges and so it might be expected to be thermostable. The TLPs are generally resistant to proteases and pH-induced denaturation (Breiteneder, H. 2004) [1001]

• Nature of main cross-reacting proteins:None identified
• Allergen properties & biological function:

  A 31 kDa protein isolated by Hsieh et al. (1995) [75] shared 46% identity with thaumatin-like proteins belonging to the pathogenesis-related proteins of PR-5 group, involved in plant defence against fungi.

  Recombinant Mal d 2 exhibits antifungal activity against Fusarium oxysporum and Penicillium expansum, implying a function in plant defense against fungal pathogens (Krebitz et al. 2003) [595]. Menu-Bouaouiche et al. (2003) [1176] demonstrated that purified apple TLP did not display any antifungal activity against Aspergillus niger, Botrytis cinerea, Candida albicans, Fusarium gramineum, Penicillium commune, Phytophthora infestans, Trichoderma viride and Verticillium albo-atrum.

  Apple thaumatin like protein displays endo-beta-1,3-glucanase activity. However, this activity is not comparable to that of the banana glucanase used as positive control (Menu-Bouaouiche et al. 2003). [1176]

  The protein is expressed in the fruit but rarely in other tissues and both in the skin and the flesh of the fruit (Kim et al. 2003) [778]

• Allergen purification:

  Natural Mal d 2 was purified from apple protein extract using the reversed-phase HPLC column C8 Hypersil WP 300, 10 µm, 8 mm×250 mm and a linear gradient of 2-propanol (solvent A: 0.1% trifluoroacetic acid in water; solvent B: 90% 2-propanol, 0.1% trifluoroacetic acid; gradient: of 0\Z80% of solvent B) (Krebitz et al., 2003) [595].

  Apple fruit pieces were homogenized in 50 mM acetic acid. After filtering the pH was adjusted to 1M acetic acid, pH3 and the extract was centrifuged at 9000 g (15 min). The cleared extract was purified using a S Fast Flow column equilibrated with 20 mM acetic acid and the protein eluted with 50 mM Tris-HCl (pH 7.8). The partially purified protein fraction was adjusted to pH 3.0 with 1 M acetic acid, centrifuged (9000 g, 15 min) and loaded onto the same column. The protein was eluted with 20 mM Na-formate buffer (pH 3.8) containing 0.0\Z0.5 M NaCl in formate buffer (Menu-Bouaouiche et al. 2003) [1176].

  Heterologous expression in Nicotiana benthamiana followed by purification (Krebitz et al., 2003) [595]

• Other biochemical information:

  The protein has been termed Mdt1 by Oh et al (2000) [133]. It has a pI of 5.1 and possesses one putative N-glycosylation site. It is homologous with the thaumatin-like proteins of cherry.

  Krebitz et al. (2003) [595] purified recombinant Mal d 2 with two putative glycosylation sites and a calculated pI of 4.55.

References (6)

• Kim SH, Lee JR, Kim SR.
  Molecular characterization of a fruit-preferential thaumatin-like gene from apple (Malus domestica cv. Fuji)
  J Plant Biol. 46(1):52-58.. 2003
  PUBMEDID:
• Krebitz M, Wagner B, Ferreira F, Peterbauer C, Campillo N, Witty M, Kolarich D, Steinkellner H, Scheiner O, Breiteneder H
  Plant-based heterologous expression of Mal d 2, a thaumatin-like protein and allergen of apple (Malus domestica), and its characterization as an antifungal protein
  J Mol Biol. 329(4):721-730.. 2003
  PUBMEDID: 12787673
• Hsieh L, Moos M, Lin Y
  Characterisation of apple 18 and 13kD allergens by microsequencing and evaluation of their content during storage and ripening.
  J.Allergy Clin Immunol. 96:960-970.. 1995
  PUBMEDID: 8543755
• Oh DH, Song KJ, Shin YU, Chung WI
  Isolation of a cDNA encoding a 31-kDa, pathogenesis-related 5/thaumatin-like (PR5/TL) protein abundantly expressed in apple fruit (Nalus domestica cv. Fuji).
  Biosci Biotechnol Biochem 64:355-62. 2000
  PUBMEDID: 10737193
• Breiteneder H.
  Thaumatin-like proteins -- a new family of pollen and fruit allergens
  Allergy. 59:79-81.. 2004
  PUBMEDID: 15080826
• Menu-Bouaouiche L, Vriet C, Peumans WJ, Barre A, Van Damme EJ, Rouge P
  A molecular basis for the endo-beta 1,3-glucanase activity of the thaumatin-like proteins from edible fruits
  Biochimie. 85:123-131. 2003
  PUBMEDID: 12765782

Biochemical Information for Mal d 3

• Allergen Name:Mal d 3
• Alternatve Allergen Names:Non specific lipid transfer protein (nsLTP)
• Allergen Designation:Minor in Cental and North Europe. Major in Mediterranean population (Sanchez-Monge et al. 1999 [172]; Pastorello et al. 1999 [157]; Garcia-Selles et al. 2002 [594])
• Protein Family:

  Protease inhibitor/seed storage/LTP family, Pfam PF00234

  non specific lipid transfer protein (nsLTP)

• Sequence Known?:Yes
• Allergen accession No.s:

  Q9M5X7:Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?Q9M5X7

  AF221502; AAF26450.1; EMBL
  AJ277164; CAB96874.1; EMBL

• 3D Structure Accession No.:Not determined
• Calculated Masses:9084.44 Da
• Experimental Masses:9078 Da by MALDI-MS (Sanchez-Monge et al. 1999 [172]).
• Oligomeric Masses:
• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:

  nsLTPs are alpha-helix bundle proteins with four disulphide bonds. It has been suggested that this cross-linked structure leads to the heat stability of apple LTP, which binds IgE after 30 minutes at 100 °C (Asero et al. 2003 [1064]). Sancho et al. (2005) [1832] similarly showed that Mal d 3 resists heat treatment at 90°C for 20 min but shows reduced IgE reactivity after more severe heat treatment at 100°C for 2 hours. It is also relatively stable to proteolysis (Asero et al. 2000 [10]).

• Nature of main cross-reacting proteins:

  The deduced primary structures for Pru p 3 and Mal d 3 showed 82% identity which explains the cross-reactions between these proteins (Diaz-Perales et al. 2002 [592]). Immunoblot inhibition assays confirmed these cross-reactivitiy (Sanchez-Monge et al. 1999 [172]).

• Allergen properties & biological function:

  Mal d 3 exact function has not been determined but plant nonspecific lipid-transfer proteins are involved in transport of fatty acids both intracellular and extracellularly; and cutin monomers to the cuticular layer of leaves and fruits. There is an expandable cavity between the four alpha-helices which can bind one or two lipids. nsLTPs have also been reported to act as plant defense proteins against bacterial and fungal infections and form the PR14 family of pathogenesis related proteins. It is possible that a lipid-like post-translational modification is involved (Kader 1996 [1135]; Lindorff-Larsen et al. 2001 [903]).

• Allergen purification:

  Ground apple skins are extracted with PBS (1:10, wt:v) for 1h at 4C, After centrifugation at 10,000rpm for 25 min, 4C, the supernatants are dialysed and freeze dried. The protein was then exchanged into 0.1M ammonium acetate, and defatted with cold acetone, followed by ethanol:ether (1:3, v:v) for 1h at 4C. The residues were dried and applied to a C4 reverse phase column (Vydac 22x250mm, 10um particle size) using a two step linear gradient of acetonitrile in 0.1% (v:v) trifluoroacetic acid (0-40%, 120min; 40-85min 30 min using a flow rate of 1ml/min) (Sanchez-Monge et al 1999 [172]).

  In another study (Pastorello et al 1999 [157]) apple LTP was extracted from apple peel (cv Golden Delicious) by homogenisation 3:1 (w:v) in PBS. 2% (w/v) polyvinylpyrrolidone, 2mM EDTA, 10mM diethyldithiocarbamate, 3mM NaN3. Extracts were clarified by centrifugation (16,000xg) at 4C, 30min, dialysed against PBS and concentrated in a centrifugal concentrator (3kd cut-off filter). This extract was loaded onto a Resource-S cation exchange column equilibrated in 30mM sodium citrate buffer pH 6 and the LTP eluted with 1M NaCl (20 column vols, 6ml/min). A final gel-permeation chromatography step on Superdex 75 in citrate buffer (pH 6.0) was then performed.

  Zuidmeer et al. (2005) [1830] purified recombinant Mal d 3 (and Pru p 3) using the yeast Pichia pastoris. Proteins were purified via cation exchange chromatography.
  

• Other biochemical information:

References (12)

• Diaz-Perales A, Garcia-Casado G, Sanchez-Monge R, Garcia-Selles FJ, Barber D, Salcedo G.
  cDNA cloning and heterologous expression of the major allergens from peach and apple belonging to the lipid-transfer protein family
  Clin Exp Allergy 32(1):87-92. 2002
  PUBMEDID: 12002744
• Sanchez-Monge R, Lombardero M, Garcia-Selles FJ, Barber D, Salcedo G.
  Lipid-transfer proteins are relevant allergens in fruit allergy.
  J Allergy Clin Immunol. 103:514-519.. 1999
  PUBMEDID: 10069888
• Pastorello EA, Pravettoni V, Farioli L, Ispano M, Fortunato D, Monza M, Giuffrida MG, Rivolta F, Scibola E, Ansaloni R, Incorvaia C, Conti A, Ortolani C.
  Clinical role of a lipid transfer protein that acts as a new apple-specific allergen.
  J Allergy Clin Immunol. 104:1099-106.. 1999
  PUBMEDID: 10550759
• Asero R, Mistrello G, Roncarolo D, de Vries SC, Gautier MF, Ciurana CL, Verbeek E, Mohammadi T, Knul-Brettlova V, Akkerdaas JH, Bulder I, Aalberse RC, van Ree R.
  Lipid transfer protein: a pan-allergen in plant-derived foods that is highly resistant to pepsin digestion.
  Int Arch Allergy Immunol. 122:20-32.. 2000
  PUBMEDID: 10859466
• Asero R, Mistrello, G, Roncarolo, D, Amato, S, Falagiani, P
  Analysis of the heat stability of lipid transfer protein from apple
  J Allergy Clin Immunol. 112:1009-1011.. 2003
  PUBMEDID: 14610497
• Marion D, Douliez JP, Gutier MF, Elmorjani K.
  Plant Lipid transfer proteins: Relationships between allergenicity and structural, biological and technological properties.
  Plant Food Allergens. Mills ENC and Shewry PR editors. Chapter 4; 57-69. 2004
  PUBMEDID:
• Lindorff-Larsen K, Lerche MH, Poulsen FM, Roepstorff P, Winther JR.
  Barley lipid transfer protein, LTP1, contains a new type of lipid-like post-translational modification.
  J Biol Chem. 276(36):33547-33553.. 2001
  PUBMEDID: 11435437
• Kader JC
  Lipid-transfer proteins in plants
  Annu Rev Plant Physiol Plant Mol Biol 47:627-654. 1996
  PUBMEDID: 15012303
• Salcedo G, Sanchez-Monge R, Diaz-Perales A, Garcia-Casado G, Barber D
  Plant non-specific lipid transfer proteins as food and pollen allergens
  Clin Exp Allergy. 34(9):1336-41. 2004
  PUBMEDID: 15347364
• García Sellés FJ, Diaz-Perales A, Sanchez-Monge R, Alcantara M, Lombardero M, Barber D, Salcedo G, Fernandez-Rivas M
  Patterns of reactivity to lipid transfer proteins of plant foods and Artemisia pollen: an in vivo study
  Int Arch Allergy Immunol. 128(2):115-122. 2002
  PUBMEDID: 12065911
• Zuidmeer L, van Leeuwen WA, Budde IK, Cornelissen J, Bulder I, Rafalska I, Besoli NT, Akkerdaas JH, Asero R, Rivas MF, Mancebo EG, van Ree R.
  Lipid transfer proteins from fruit: cloning, expression and quantification.
  Int Arch Allergy Immunol. 137(4):273-281.. 2005
  PUBMEDID: 15970634
• Sancho AI, Rigby NM, Zuidmeer L, Asero R, Mistrello G, Amato S, Gonzalez-Mancebo E, Fernandez-Rivas M, van Ree R, Mills EN.
  The effect of thermal processing on the IgE reactivity of the non-specific lipid transfer protein from apple, Mal d 3.
  Allergy 60(10):1262-8.. 2005
  PUBMEDID: 16134992

Biochemical Information for Mal d 4

• Allergen Name:Mal d 4
• Alternatve Allergen Names:
• Allergen Designation:Minor
• Protein Family:Profilin, Pfam PF00235
• Sequence Known?:Yes
• Allergen accession No.s:

  Q9XF40:Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?Q9XF40
  Q9XF41:Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?Q9XF41
  Q9XF42:Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?Q9XF42

  AF129427; AAD29413.1 EMBL
  AF129426; AAD29412.1 EMBL
  AF129428; AAD29414.1 EMBL

  Q84RR7:Swissprot; http://us.expasy.org/cgi-bin/niceprot.pl?Q84RR7
  Q84RR6:Swissprot; http://us.expasy.org/cgi-bin/niceprot.pl?Q84RR6
  Q84RR5:Swissprot; http://us.expasy.org/cgi-bin/niceprot.pl?Q84RR5

  AJ507457 EMBL
  AJ507458 EMBL
  AJ507459 EMBL

• 3D Structure Accession No.:Not determined
• Calculated Masses:13957, 14052, 14062 Da
• Experimental Masses:

  14 kDa

• Oligomeric Masses:
• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:

  There are only few studies on the stability of profilins most of them on celery profilin. Compared to other allergens, profilin is a moderately stable protein, more resistant than Bet v 1 homologues but less stable than lipid transfer proteins or cross-reactive carbohydrate deteminants of glycoporein allergens.

• Nature of main cross-reacting proteins:IgE cross-reacts with profilins from other plant species such as Bet v 2 from birch pollen (Ebner et al. 1995)
• Allergen properties & biological function:

  Apple profilin is thought to be an actin-binding protein.

• Allergen purification:

  Profilins are generally purified by affinity chormatography. Whilst there is no report for apple, profilins have been purified from other plant species (Radauer and Hoffmann-Sommergruber, 2004)

• Other biochemical information:

References (3)

• van Ree R, Fernandez-Rivas M, Cuevas M, Wijngaarden M, Aalberse RC
  Pollen-related allergy to peach and apple: an important role for profilin.
  J.Allergy Clin. Immunol. 95:726-734. 1995
  PUBMEDID: 7897156
• Ebner C, Hirschwehr, R, Bauer L, Breiteneder H, Valenta R, Ebner, H, Kraft D, Scheiner O.
  Identification of allergens in fruits and vegetables: IgE cross-reactivities with the important birch pollen allergens Bet v 1 and Bet v 2 (birch profilin).
  J Allergy Clin Immunol 95: 962-9.. 1995
  PUBMEDID: 7751517
• Radauer C, Hoffmann-Sommergruber K
  Profilins
  Plant Food Allergens. Mills ENC and Shewry PR editors. Chapter 7; 105-124. 2004
  PUBMEDID: