Allergy information for: Melon (Cucumis melo (Muskmelon))

Clinical History

• Number of Studies:1-5
• Number of Patients:>50
• Symptoms:

  Oral allergy syndrome (OAS), rinithis, gastrointestinal symptoms, pruritus inside the mouth, respiratory difficulty, generalized urticaria, and hypotension. Cuesta-Herranz et al. 2003 [876], Garcia Ortiz et al. 1996 [877], Rodriguez et al. 2000 [917]

Skin Prick Test

• Number of Studies:1-5
• Food/Type of allergen:Fresh melon and commercial extracts
• Protocol: (controls, definition of positive etc)

  Prick by prick test with fresh melon pulp and three commercially available melon extracts. (Cuesta-Herranz et al. 2003; [876]; and Figueredo et al 2004). Prick by prick test with fresh melon pulp (Rodriguez et al. 2000) [917]

  Histamine phosphate (10 mg/mL) and saline solution were used as a positive control and negative control respectively. A weal size of 3 mm larger than the negative control was regarded as positive (Rodriguez et al. 2000) [917] (Cuesta-Herranz et al. 2003) [876]

• Number of Patients:

  35 individuals allergic to melon (Cuesta-Herranz et al. 2003) [876]

  53 adult patients were evaluated and 19 patients were allergic to melon (Rodriguez et al. 2000) [917]

  65 melon patients (Figueredo et al 2004)

• Summary of Results:

  All the patients showed positive response to SPT (Cuesta-Herranz et al. 2003) [876].

  68% of the patients had a positive response (Rodriguez et al. 2000) [917]

  All of the patients (65) showed positive skin prick-prick test (inclusion criteria), but it was only positive in 12%, 17% and 91% with three commercial melon extracts (Figueredo et al 2004)

IgE assay (by RAST, CAP etc)

• Number of Studies:0
• Food/Type of allergen:Extracts prepared according to the manufactures (Pharmacia) (Rodriguez et al. 2000) [917]
• IgE protocol:CAP. A value of 0.35 kU_A/L was considered a positive result (Rodriguez et al. 2000) [917].
• Number of Patients:

  53 adult patients (Rodriguez et al. 2000) [917]

• Summary of Results:43% of the patients had specific IgE for melon (Rodriguez et al. 2000) [917]

Immunoblotting

• Immunoblotting separation:

  Polyacrylamide concentrations of 15% and 5% were used for separating and stacking gels, respectively. The samples were mixed with 0.1 m Tris-HCl, pH 6.8 containing 4% (w/v) SDS, 20% (w/v) glycerol, 10% (w/v) 2-beta-mercaptoethanol and 0.005% (w/v) bromophenol blue. The samples were denatured by heating at 100 °C for 5 min (Cuesta-Herranz et al. 2003) [876]

  Fruit extract was fractionated by means of SDS-PAGE following the Laemmli system on Bio-Rad Miniprotean II System gels (15% polyacrylamide) (Rodriguez et al. 2003) [1005]

• Immunoblotting detection method:

  Protein bands were transferred by semidry blotting onto nitrocellulose sheets. Membranes were blocked with 3% (w/v) skimmed milk powder and incubated with the patients' sera diluted 1 : 5. After washing, blots were incubated with anti-human IgE peroxidase-conjugate antibody diluted 1 : 1000 in TBS-T containing 5% (v/v) foetal calf serum. The protein bands were visualized by chemiluminiscence with ECL (Cuesta-Herranz et al. 2003) [876]

  Protein bands were electrotransferred onto polyvinylidene difluoride (PVDF) membranes. After washing and blocking, membranes were incubated with a serum pool or individual sera from patients with melon allergy or with control sera (1:3 dilutions) and then with alkaline phosphatase-conjugated monoclonal anti-human IgE (clon GE-1; 1:500 dilution) and developed by adding a 5-bromo-4-chloro-3 indoyl phosphate/nitro blue tetrazolium solution. (Rodriguez et al. 2003) [1005]

• Immunoblotting results:

  Incubation with serum pool or individual sera revealed six stained bands with an apparent molecular weight of 67, 54, 49, 36, 26 and 14 kDa. Four IgE binding bands were recognized by more than 50% of patient sera: 67 kDa, 54 kDa, 36 kDa and 14 kDa. The 36 kDa was the most frequent IgE-binding band, which was detected by 100% of the patient sera. (Cuesta-Herranz et al. 2003) [876]

  Immunodetection of melon-blotted extract with the serum pool from patients with OAS recognized several IgE-binding components between 13 and 60 kDa and a major reactive band of 13 kDa. At least 15 (71%) sera reacted with the 13-kDa band (profilin), and therefore it was identified as a major melon allergen. This band was the strongest IgE-binding component in 6 of these sera (Rodriguez et al. 2003) [1005]

Oral provocation

• Number of Studies:1-5
• Food used and oral provocation vehicle:

  Fresh melon pulp. Patients had to chew and swallow several doses until a positive response was obtained (Cuesta-Herranz et al. 2003) [876], (Figueredo et al 2004)

  Fresh melon (200 g) masked in orange and pineapple juices, sugar, wheat meal and liquid coloring. Subjects were challenged first randomly with either food or placebo (vehicle). The interval before the second part of the double-blind placebo-controlled food challenge (DBPCFC) was at least 24 hours (Rodriguez et al. 2000) [917]

• Blind:

  Open challenge (Cuesta-Herranz et al. 2003) [876] (Figueredo et al 2004)

  Open challenge and double-blind placebo-controlled food challenge (DBPCFC) (Rodriguez et al. 2000) [917]

• Number of Patients:

  35 individuals allergic to melon (Cuesta-Herranz et al. 2003) [876]

  51 patients on an open challenge. Subjects showing a positive reaction on open provocation were subsequently challenged in a DBPCFC (Rodriguez et al. 2000) [917].

  65 melon patients (Figueredo et al 2004)

• Dose response:Maximal cumulative food dose of the challenge was 200 g (Rodriguez et al. 2000) [917]
• Symptoms:

  All the patients showed OAS following oral challenge. One patient displayed rinitis and another gastrointestinal symptoms (Cuesta-Herranz et al. 2003) [876] .

  All patients (65) showed oral symptoms, but 19.7 % of them also experienced extraoral symptoms and none experienced generalized, urticaria or anaphylaxis. (Figueredo et al. 2003)

  25/51 patients were positive on open food challenges with melon and only 17 (68%) of 25 had reactions with DBPCFC. Most of the patients suffered from OAS whilst 11% of the patients had severe reactions that began quickly, within a few minutes after ingestion of melon, with pruritus inside the mouth. This progressed rapidly with feelings of respiratory difficulty, generalized urticaria, and hypotension (Rodriguez et al. 2000) [917]

IgE cross-reactivity and Polysensitisation

Other Clinical information

Reviews (0)

References (5)

• Cuesta-Herranz J, Pastor C, Figueredo E, Vidarte L, De las Heras M, Duran C, Fernandez-Caldas E, de Miguel J, Vivanco F
  Identification of Cucumisin (Cuc m 1), a subtilisin-like endopeptidase, as the major allergen of melon fruit
  Clin Exp Allergy. 33(6):827-33.. 2003
  PUBMEDID: 12801320
• Garcia Ortiz JC, Ventas P, Cosmes P, Lopez-Asunsolo A.
  An immunoblotting analysis of cross-reactivity between melon, and plantago and grass pollens
  J Investig Allergol Clin Immunol. 6(6):378-82.. 1996
  PUBMEDID: 9015782
• Rodriguez J, Crespo JF, Burks W, Rivas-Plata C, Fernandez-Anaya S, Vives R, Daroca P
  Randomized, double-blind, crossover challenge study in 53 subjects reporting adverse reactions to melon (Cucumis melo).
  J Allergy Clin Immunol. 106(5):968-72.. 2000
  PUBMEDID: 11080722
• Rodriguez-Perez R, Crespo JF, Rodriguez J, Salcedo G
  Profilin is a relevant melon allergen susceptible to pepsin digestion in patients with oral allergy syndrome
  J Allergy Clin Immunol. 111(3):634-9. 2003
  PUBMEDID: 12642849
• Figueredo E, Cuesta-Herranz J, De-Miguel J, Lázaro M, Sastre J, Quirce S, Lluch Bernal M, De las Heras M
  Clinical characteristics of melon (Cucumis melo) allergy
  Ann Allergy asthma Immunol 91(3):303-8. 2003
  PUBMEDID: 14533664

Biochemical Information for Cuc m 1

• Allergen Name:Cuc m 1
• Alternatve Allergen Names:Cucumisin
• Allergen Designation:Major
• Protein Family:Subtilisin, Pfam, PF05922
• Sequence Known?:

  Yes

• Allergen accession No.s:

  Q39547: Swissprot: http://us.expasy.org/cgi-bin/niceprot.pl?Q39547

  D32206; BAA06905.1; -.

  EMBL

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

  78819 Da

• Experimental Masses:67 kDa
• Oligomeric Masses:

  None

• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:Cucumisin is a thermostable protein
• Nature of main cross-reacting proteins:Not known
• Allergen properties & biological function:Sequence comparisons reveal that cucumisin has several features in common with the microbial proteases of the subtilisin family.
• Allergen purification:Not purified
• Other biochemical information:Examination of the primary structure of cucumisin revealed that it is synthesized as a precursor, consisting of four functional domains: a possible signal peptide (22 amino acid residues), an NH2-terminal pro-sequence (88 residues), a 54-kDa protease domain (505 residues), which is the active enzyme domain of the 67-kDa native cucumisin, and a 14-kDa COOH-terminal polypeptide (116 residues), which arises by limited autolysis of the 67-kDa native cucumisin. (Yamagata et al 1994 [1004]). Three of them, 67-kDa native cucumisin, 54-kDa mature cucumisin and 36 kDa NH2-terminal cucumisin fragment, were identified of major allergen corresponding to different molecular forms of cucumisin produced during the processing or degradation of the enzyme.

References (2)

• Cuesta-Herranz J, Pastor C, Figueredo E, Vidarte L, De las Heras M, Duran C, Fernandez-Caldas E, de Miguel J, Vivanco F
  Identification of Cucumisin (Cuc m 1), a subtilisin-like endopeptidase, as the major allergen of melon fruit
  Clin Exp Allergy. 33(6):827-33.. 2003
  PUBMEDID: 12801320
• Yamagata H, Masuzawa T, Nagaoka Y, Ohnishi T, Iwasaki T.
  Cucumisin, a serine protease from melon fruits, shares structural homology with subtilisin and is generated from a large precursor
  J Biol Chem. 30;269:32725-31. 1994
  PUBMEDID: 7806492

Biochemical Information for Cuc m 2

• Allergen Name:Cuc m 2
• Alternatve Allergen Names:Profilin
• Allergen Designation:Major
• Protein Family:Profilin; Pfam PF00235
• Sequence Known?:

  Yes

• Allergen accession No.s:

  Q7X837, Swissprot: http://us.expasy.org/cgi-bin/niceprot.pl?Q7X837

  AY292386; AAP42150.2; -.	EMBL / GenBank
  AY292387; AAP42151.2; -.	EMBL / GenBank

• 3D Structure Accession No.:None
• Calculated Masses:

  13920 Da

• Experimental Masses:13 kDa
• Oligomeric Masses:None
• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:Immunoblotting with the pooled sera of patients with melon allergy showed that melon allergens were unaffected by crude human saliva. However, the 13-kDa component (profilin) was digested in the SGF (Rodriguez et al. 2003) [1005]
• Nature of main cross-reacting proteins:Not known
• Allergen properties & biological function:

  Profilin is an actin-binding protein of the cytoskeleton

• Allergen purification:

  Rodriguez et al. 2003 [1005] found difficulties to isolate the protein by means of gel-filtration chromatography or affinity chromatography or reverse-phase HPLC. This is explained by the low level of the allergen in the crude melon extract, as well as its tendency to associate with other components.

• Other biochemical information:

References (1)

• Rodriguez-Perez R, Crespo JF, Rodriguez J, Salcedo G
  Profilin is a relevant melon allergen susceptible to pepsin digestion in patients with oral allergy syndrome
  J Allergy Clin Immunol. 111(3):634-9. 2003
  PUBMEDID: 12642849

Biochemical Information for Cuc m 3

• Allergen Name:Cuc m 3
• Alternatve Allergen Names:PR-1 (Pathogenesis-related protein (Fragments))
• Allergen Designation:Minor
• Protein Family:
• Sequence Known?:Yes
• Allergen accession No.s:P83834: Swissprot: http://us.expasy.org/cgi-bin/niceprot.pl?P83834
• 3D Structure Accession No.:None
• Calculated Masses:

  16,097 Da

• Experimental Masses:17 kDa
• Oligomeric Masses:None
• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:Not known
• Nature of main cross-reacting proteins:Cuc m 3 shows 70% and 65% of sequence identity with PR-1 proteins from grape and cucumber respectively
• Allergen properties & biological function:Cuc m 3 belongs to the phatogenesis-related protein family (PR) which are induced by stress conditions, such as pathogen infection
• Allergen purification:Melon (Cucumis melo) fruits were peeled and sliced to obtain juice and pulp, which were separately freeze-dried, and then defatted with acetone (2 × 1:10 (wt/vol) for 1 hour at 4°C). The dried material was then extracted with PBS buffer (0.1 mol/L sodium phosphate (pH 7.0) and 0.15 mol/L NaCl; 1 × 1:5 (wt/vol) for 1 hour at 4°C) and centrifuged (10,000 rpm for 30 minutes at 4°C), and the supernatant dialyzed against H₂O (cut-off point 3.5 kd) and freeze-dried. The melon juice extract (15 mg protein) was fractionated by means of reverse-phase HPLC on a Vydac C4 column (2.2 × 25 cm; particle size 10 µm; Separations Group, Hesperia, Calif), eluting with a 2-step linear gradient of acetonitrile in 0.1% trifluoroacetic acid (10 minutes at 0%, and 0% to 85% in 280 minutes; 1 mL/min).
• Other biochemical information:Cuc m 3 seems to accumulate mainly in the juice of the central part of melons, where cucumisin Cuc m 1 is also located

References (1)

• Asensio T, Crespo JF, Sanchez-Monge R, Lopez-Torrejon G, Somoza ML, Rodriguez J, Salcedo G
  Novel plant pathogenesis-related protein family involved in food allergy
  J Allergy Clin Immunol. 114(4):896-. 2004
  PUBMEDID: 15480331