Allergy information for: Carrot (Daucus carota )

Clinical History

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

  The symptoms of carrot allergy from more than 50 patients have been reported. They include oral allergy syndrome (OAS), angioedema, urticaria, dyspnea, vertigo, tightness of throat or chest, dysphagia, hoarseness, cough, rhinitis and conjunctivitis (Helbling et al. 1994 [1097]; Ballmer-Weber et al. 2001 [573]). In addition, 50 % of the patients studied by Ballmer-Weber et al. (2001) had a history of systemic reactions.

  A case of anaphylaxis and rhinoconjunctivitis induced by raw carrot in a non pollen allergic patient was reported by Fernandez-Rivas et al. (2004) [1187].

Skin Prick Test

• Number of Studies:1-5
• Food/Type of allergen:

  Ballmer-Weber et al. (2001) used either raw carrot by the prick to prick test or commercial carrot extracts.

  Fernandez-Rivas et al. (2004) used raw and boiled carrot.

  Fresh raw and cooked carrot, and its cooking broth (Agustin Ubide et al. 2004)

• Protocol: (controls, definition of positive etc)

  Histamine dihydrochloride (10 mg/mL) was used as a positive control, and the glycerol-containing diluent of the prick solution was used as a negative control. A 3mm diameter wheal was considered positive (Ballmer-Weber et al. 2001).

  Prick to prick (Agustin Ubide et al. 2004)

• Number of Patients:

  26 patients with histories of allergic reactions to carrot (Ballmer-Weber et al. 2001).

  A 50-year-old woman presenting anaphylaxis and rhinoconjunctivitis (Fernandez-Rivas et al. 2004)

  A 38-year-old woman (Agustin Ubide et al. 2004)

• Summary of Results:

  All patients with positive DBPCFC results were positive with raw native carrot (SE = 100%) whereas SPT with commercial carrot extracts were positive in just 5 of 19 subjects tested (SE = 26%). In all patients with negative DBPCFC results, however, SPT with carrot extract was negative (SP = 100%) whereas prick-to-prick testing with raw carrot was positive (SP = 0%) (Ballmer-Weber et al. 2001)

  Prick - prick test with raw carrot was positive (10 mm), but negative with boiled carrot (Fernandez-Rivas et al. 2004)

  Test was positive with carrot, celery and horsetail (Agustin Ubide et al. 2004)
  

IgE assay (by RAST, CAP etc)

• Number of Studies:0
• Food/Type of allergen:

  Commercial carrot extract (Ballmer-Weber et al. 2001) (Agustin Ubide et al. 2004)

  Carrot extract, recombinant Bet v 1, Bet v 2, and carrot LTP, and carrot profilin (Fernandez-Rivas et al. 2004)

• IgE protocol:CAP (Ballmer-Weber et al. 2001) (Fernandez-Rivas et al. 2004) (Agustin Ubide et al. 2004)
• Number of Patients:

  26 patients with histories of allergic reactions to carrot (Ballmer-Weber et al. 2001).

  A 50-year-old woman presenting anaphylaxis and rhinoconjunctivitis (Fernandez-Rivas et al. 2004)

  A 38-year-old woman (Agustin Ubide et al. 2004)

• Summary of Results:

  18/20 patients with positive DBPCFC results and 2/4 patients with negative DBPCFC results were CAP test–positive for carrot (≥0.7 kU/L). Thus, determination of specific IgE to carrot by the CAP method showed an SE of 90% and an SP of 50% (Ballmer-Webner et al. 2001).

  Specific IgE to carrot was 5.97 kU/l and negative to birch and mugwort pollens, recombinant Bet v 1, Bet v 2, and carrot LTP, and carrot profilin (Fernandez-Rivas et al. 2004)

  Specific IgE to carrot was 15.1 kU/l (Agustin Ubide et al. 2004)

Immunoblotting

• Immunoblotting separation:

  Protein separation was performed using 12.5% polyacrylamide 1D SDS-PAGE gels. Samples were reduced with beta-mercaptoethanol. In addition, two-dimensional PAGE was performed at pH 5.3-10.0 and reducing conditions (Fujita et al. 2001).

  Carrot extract was separated with Tricine SDS-PAGE using 14% T, 2.6% C separating gel and a 4% T, 2.6% C stacking gel. Proteins were reduced by 1,4-dithiothreitol (Ballmer-Weber et al. 2001).

  Total protein extract from carrot roots were electrophoresed on 12% SDS polyacrylamide gels (Hoffmann-Sommergruber et al. 1999)

• Immunoblotting detection method:

  Proteins were transferred to nitrocellulose membranes using a semidry device. Membranes were blocked with 2% (w/v) skim milk , incubated with sera (1:6 dilution) and bound IgE detected by horse radish peroxidase conjugated goat anti-human IgE. Blots were stained by Coomassie brilliant blue R-350 (Fujita et al. 2001).

  Proteins were transferred onto nitrocellulose membranes by semidry blotting and blocked twice in 50 mmol/L Tris(hydroxy-methyl)aminomethane/HCl buffer (pH 7.4) containing 0.15 mol/L sodium chloride and 0.3% (v/v) Tween 20 (TBST). Blots were incubated with patients’ sera and bound IgE detected with alkaline phosphatase-conjugated mouse antihuman IgE (Ballmer-Weber et al. 2001).

  The IgE-binding capacity of aqueous protein extracts from carrot roots was investigated by using 6 allergic patients' sera followed by incubation with ¹²⁵I rabbit antihuman IgE (Hoffmann-Sommergruber et al. 1999)

• Immunoblotting results:

  14% of the patients recognised the 20 kDa protein (Cyclophilin), and proteins with masses of aproximately 27 and 12-14 kDa (Fujita et al. 2001).

  Patients recognised the Bet v 1–related major carrot allergen Dau c 1 (85%) and 20% the carrot profilin. Other sera recognised proteins between 34 and 70 kDa (Ballmer-Weber et al. 2001).

  All patients displayed IgE-binding to a protein of ~16 kDa molecular mass. In addition, patients 1, 2 and 4 revealed IgE-antibodies directed against several proteins in the range of ~40-66 kDa. Patient six possessed specific IgE-antibodies against a protein of 14 kDa, possibly profilin (Hoffmann-Sommergruber et al. 1999)

  Immunoblot revealed IgE binding to a band at 17 kDa in celery, carrot, horsetail and Bet v 1 (Agustin Ubide et al. 2004)

Oral provocation

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

  Carrot was given raw or blind. Two different drinks, identical in color, texture, and taste, were prepared. The active drink contained 70 g of carrot, 100 g of cooked pumpkin, 20 g of cream, 60 g of yogurt without flavoring, 35 g of water, 2 pinches of saffron, and salt and pepper, which were mixed in a blender. With the exception of carrot, the placebo drink and the active drink contained the same ingredients. (Ballmer-Weber et al. 2001)

• Blind:

  Open challenge and DBPCFC (Ballmer-Weber et al. 2001)

  Open challenge (Fernandez-Rivas et al. 2004)

• Number of Patients:

  26 patients for DBPCFC. Open challenge was performed in patients with a negative DBPCFC (4 patients). (Ballmer-Weber et al. 2001)

  A 50-year-old woman presenting anaphylaxis and rhinoconjunctivitis (Fernandez-Rivas et al. 2004)

• Dose response:

  For the open challenges patients had to chew 5 g of raw carrot and spit it out; if no reaction was experienced, they chewed and swallowed 5 g, then 10 g, and then 20 g. OAS occurs at 1.9 ± 1.0 g of carrot and symptoms not restricted to the oral cavity occurred at a mean provocation dose of 1.7 ± 0.9 g of carrot. DBPCFC was performed through use of the 2-step spit (local mucosal challenge) and swallow procedure. (Ballmer-Weber et al. 2001)

  100 g (Fernandez-Rivas et al. 2004)

• Symptoms:

  Sixteen patients complained of oral allergy syndrome (OAS). OAS appeared in 14 patients during the local mucosal challenge (“spit” phase) and in 2 patients after the swallowing of 13 mL of the active drink. In 4 patients, symptoms were not restricted to the oral cavity and occurred during the “spit” phase in 3 patients and after the swallowing of 13 mL of the active drink in 1 patient. Only one patient of 4 reported symptoms of an OAS after swallowing 5 g of raw carrot during the open challenge. The reactions under provocation were clearly weaker as indicated by history (Ballmer-Weber et al. 2001)

  Cooked carrot was tolerate by the patient (Fernandez-Rivas et al. 2004)

IgE cross-reactivity and Polysensitisation

Other Clinical information

Reviews (0)

References (8)

• Fujita C, Moriyama T, Ogawa T
  Identification of Cyclophilin as an IgE-Binding Protein from Carrots.
  International Archives of Allergy and Immunology 125(1): 44-50. 2001
  PUBMEDID: 11385287
• Hoffmann-Sommergruber K, O'Riordain G, Ahorn H, Ebner C, Laimer da Camara Machado M, Pühringer H, Scheiner O, Breiteneder H.
  Molecular characterization of Dau c 1, the Bet v 1-homologous protein from carrot and its crossreactivity with Bet v 1 and Api g 1.
  Clin Exp Allergy 29, 840-847.. 1999
  PUBMEDID: 10336602
• Ballmer-Weber BK, Wuthrich B, Wangorsch A, Fotisch K, Altmann F, Vieths S
  Carrot allergy: double-blinded, placebo-controlled food challenge and identification of allergens
  J Allergy Clin Immunol. 108(2):301-307. 2001
  PUBMEDID: 11496252
• Helbling A, Schwartz HJ, Lopez M, Lehrer SB
  Lettuce and carrot allergy: are they related?
  Allergy Proc. 15(1):33-8.. 1994
  PUBMEDID: 8005454
• Moneo I, Gomez M, Sanchez-Monge R, Alday E, de las Heras M, Esteban I, Bootello A, Salcedo G
  Lack of crossreaction with Bet v 1 in patients sensitized to Dau c 1, a carrot allergen
  Ann Allergy Asthma Immunol. 83(1):71-5. 1999
  PUBMEDID: 10437820
• Helbling A, Lopez M, Schwartz HJ, Lehrer SB
  Reactivity of carrot-specific IgE antibodies with celery, apiaceous spices, and birch pollen
  Ann Allergy. 70(6):495-9. 1993
  PUBMEDID: 8507047
• Agustin-Ubide MP, Martinez-Cocera C, Alonso-Llamazares A, Robledo T, Lombardero M, Dominguez J, Carballo MA
  Diagnostic approach to anaphylaxis by carrot, related vegetables and horsetail (Equisetum arvense) in a homemaker.
  Allergy 59 (7), 786-787. 2004
  PUBMEDID: 15180768
• Fernandez-Rivas M, Gonzalez-Mancebo E, van Leeuwen WA, Leon F, van Ree R.
  Anaphylaxis to raw carrot not linked to pollen allergy
  Allergy. 59(11):1239-40. 2004
  PUBMEDID: 15461613

Biochemical Information for Cyclophilin

• Allergen Name:Cyclophilin
• Alternatve Allergen Names:None
• Allergen Designation:Minor
• Protein Family:Cyclophilin type peptidyl-prolyl cis-trans isomerase family, Pfam, PF00160
• Sequence Known?:No
• Allergen accession No.s:None
• 3D Structure Accession No.:Not determined
• Calculated Masses:
• Experimental Masses:20 kDa
• Oligomeric Masses:
• Allergen epitopes:Not Known
• Allergen stability:
  Process, chemical, enzymatic:Not known
• Nature of main cross-reacting proteins:Cyclophilins are ubiquitous but sera reactive to carrot cyclophilin does not cross-react with the allergenic cyclophilin from birch pollen (Bet v 7).
• Allergen properties & biological function:Cyclophilins catalyse cis-trans isomerization and thus may assist in protein folding.
• Allergen purification:Fresh carrots were homogenized in 30 mM Tris-HCl (pH 8), filtered through cheesecloth and centrifuged. Proteins were precipitated by 70% ammonium sulfate. The precipitate was dialysed against 30 mM Tris-HCl with 1 mM EDTA (pH 8) and passed through a TSK gel Super Q-Toyopearl 650 column and again precipitated, disolved in 20 mM MOPS with 1 mM EDTA (pH 6), desalted with a PD-10 column and applied to a Hi-trap SP column. Fractions were eluted with 20 mM MOPS with 1 mM EDTA (pH 6) and a gradient of 0-1.0 M NaCl. The IgE binding fraction was concentrated and chromatograhed on a TSK gel G3000SW HPLC column in 20 mM Tris-HCl, 1 mM EDTA and 0.15 M NaCl (pH 7.5).
• Other biochemical information:

  The allergen was identified using pooled sera from patients with atopic dermatitis and was found to be an active cyclophilin after the 15 N-terminal residues were found to be closely homologous to other plant cyclophilins.

  The protein has a pI of 8.3-9.1

References (1)

• Fujita C, Moriyama T, Ogawa T
  Identification of Cyclophilin as an IgE-Binding Protein from Carrots.
  International Archives of Allergy and Immunology 125(1): 44-50. 2001
  PUBMEDID: 11385287

Biochemical Information for Dau c 1

• Allergen Name:Dau c 1
• Alternatve Allergen Names:Dau c 1.0101, Dau c 1.0102 (CR16), Dau c 1.0103, Dau c 1.0104, Dau c 1.0105, Dau c 1.0201
  
• Allergen Designation:Major
• Protein Family:Pathogenesis-related protein Bet v I family, Pfam, PF00407
• Sequence Known?:Yes
• Allergen accession No.s:

  O04298:Swissprot:http://ca.expasy.org/cgi-bin/niceprot.pl?O04298

  Z84376; CAB06416.1; EMBL
  Z81361; CAB03715.1; EMBL
  Z81362; CAB03716.1; EMBL
  D88388; BAA13604.1; EMBL
  U47087; AAB01092.1; EMBL

  Q8SAE7: Swissprot: http://us.expasy.org/cgi-bin/niceprot.pl?Q8SAE7

  AF456481; AAL76932.1;

  EMBL

• 3D Structure Accession No.:Not determined
• Calculated Masses:17799 Da for Dau c 1.0101, 16125 for Dau c 1.0102, 16071 for Dau c 1.0103, 1603 for Dau c 1.0104, 16049 for Dau c 1.0105 and 16518 for Dau c 1.0201.
• Experimental Masses:16 kDa
• Oligomeric Masses:
• Allergen epitopes:Not Known
• Allergen stability:
  Process, chemical, enzymatic:

  Not known but likely to be thermolabile as other members of the Bet v 1 family.

• Nature of main cross-reacting proteins:

  As a consequence of sequence homology IgE to Dau c 1 cross-reacts with other Bet v 1 homologues including major food allergens from celery (Api g 1), apple (Mal d 1) and cherry (Pru a 1) and major pollen allergens of the order Fagales (Bet v 1, Cor a 1, Aln g 1 and Car b 1) (Ebner et al. 1995) (Schoning et al. 1995)

  IgE cross-inhibition studies between Dau c 1, Bet v 1 and Api g 1 indicate the presence of common epitopes (Hoffmann et al. 1999).

• Allergen properties & biological function:Dau c 1 is a Bet v 1 homologue allocated to the family PR10, proteins expressed upon pathogen attack, stress and abiotic stimuli. However its precise function is not known.
• Allergen purification:

  Natural Dau c 1 was purified as for Bet v 1 from birch by RP-HPLC (C8) (Hypersil WP300, 8x250mm, 10um particle size) using 0.1% (v:v) trifluroacetic acid as the solvent and eluting the protein with a linear gradient of propan-2-ol (0-60%, 1.0ml/min) (Hoffmann et al. 1999)

  Recombinant Dau c 1 has been purified using a hererologous expression systen, E. coli, as a hexahistidine-tagged fusion protein and further purified by Ni-chelate affinity chromatography (Hoffmann et al. 1999).

• Other biochemical information:Hoffmann et al. 1999 have cloned and expressed 3 proteins called Dau c 1.1, 1.2 and 1.3. The predicted pIs are 4.47, for Dau c 1.1 and Dau c 1.2; and 4.39 for Dau c 1.3. Sequence homology of the 3 isoforms was 97-100% and identity of 96-99%.

References (3)

• 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
• Hoffmann-Sommergruber K, O'Riordain G, Ahorn H, Ebner C, Laimer da Camara Machado M, Pühringer H, Scheiner O, Breiteneder H.
  Molecular characterization of Dau c 1, the Bet v 1-homologous protein from carrot and its crossreactivity with Bet v 1 and Api g 1.
  Clin Exp Allergy 29, 840-847.. 1999
  PUBMEDID: 10336602
• Schoning B, Vieths S, Petersen A, Baltes W
  Identification and characterization of allergens related to Bet v 1, the major birch pollen allergen, in apple, cherry, celery and carrot by 2-dimensional immunoblotting and N-terminal microsequencing.
  Journal of the Science of Food and Agriculture 67 (4): 431-440. 1995
  PUBMEDID:

Biochemical Information for LTP

• Allergen Name:LTP
• Alternatve Allergen Names:Extracellular protein 2
• Allergen Designation:Minor
• Protein Family:Protease inhibitor/seed storage/LTP family, Pfam PF00234 non specific lipid transfer protein (nsLTP)
• Sequence Known?:yes
• Allergen accession No.s:

  P27631:Swissprot:http://us.expasy.org/cgi-bin/niceprot.pl?P27631

  M64746

  EMBL

• 3D Structure Accession No.:Not determined
• Calculated Masses:12504 Da
• Experimental Masses:9 kDa
• Oligomeric Masses:
• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:

  Recombinant and native carrot LTP were resistant to proteases (Asero et al. 2000) [10]

• Nature of main cross-reacting proteins:Not known
• Allergen properties & biological function:The purified protein is capable of binding phospholipids and is able to enhance their transfer between artificial membranes. It could be demonstrated that the protein is also capable of binding palmitic and oleic acid as well as oleyl-CoA (Meijer et al. 1993)
• Allergen purification:Carrot LTP was produced as recombinant protein in Pichia pastoris and natural carrot LTP was purified by ion-exchange chromatography and gel filtration.
• Other biochemical information:

References (2)

• 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
• Meijer EA, de Vries SC, Sterk P, Gadella DW Jr, Wirtz KW, Hendriks T
  Characterization of the non-specific lipid transfer protein EP2 from carrot (Daucus carota L.).
  Mol Cell Biochem. 123(1-2):159-66. 1993
  PUBMEDID: 8232259