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Vol. 131, No. 4, 2003
Issue release date: August 2003
Int Arch Allergy Immunol 2003;131:245–255

Cloning and Molecular and Immunological Characterisation of Two New Food Allergens, Cap a 2 and Lyc e 1, Profilins from Bell Pepper (Capsicum annuum) and Tomato (Lycopersicon esculentum)

Willerroider M. · Fuchs H. · Ballmer-Weber B.K. · Focke M. · Susani M. · Thalhamer J. · Ferreira F. · Wüthrich B. · Scheiner O. · Breiteneder H. · Hoffmann-Sommergruber K.
aDepartment of Chemistry and Biochemistry, and bInstitute of Genetics and General Biology, University of Salzburg, Salzburg, cDepartment of Pathophysiology, University of Vienna, Vienna, Austria; dAllergy Unit, Department of Dermatology, University Hospital of Zurich, Zurich, Switzerland; eBiomay, Vienna, Austria

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Background: Profilins are recognised by IgE of about 20% of patients allergic to birch pollen and plant foods. They are ubiquitous intracellular proteins highly cross-reactive among plant species. Therefore, they were called panallergens and are made responsible for cross-sensitisation between plant pollen and food. Objectives: The aim of the present study was to clone the cDNAs encoding profilins from bell pepper and tomato, to produce and purify the recombinant proteins and to compare their IgE-binding capacities to those of the natural proteins. Methods: cDNA clones coding for profilin were obtained by RT-PCR from total RNA of tomato and bell pepper fruits, sequenced and expressed as non-fusion proteins in Escherichia coli. The recombinant profilins were subsequently purified and tested for IgE-binding and inhibition capacity with sera from 34 food-allergic patients. Possible oligomerisation of recombinant profilins was investigated by HPLC analysis and its influence on IgE binding assayed by ELISA. Results: The open reading frame from both profilins encompasses 393 bp with a predicted molecular mass of 14,184 kD and a pI of 4.44 for bell pepper profilin (Cap a 2) and 14,257 kD and a pI of 4.46 for the profilin from tomato (Lyc e 1). The two protein sequences display 91% identity, whereas tomato profilin from pollen shares only 75% identity with tomato fruit profilin. Eleven out of 34 food-allergic patients (32%) display IgE binding to both purified profilins. Preincubation of a serum pool with either purified rCap a 2 or rLyc e 1 nearly abolished IgE binding to natural Cap a 2 and Lyc e 1, respectively. In addition, purified recombinant Cap a 2 was able to inhibit IgE-binding to rLyc e 1 by approximately 50%, whereas rLyc e 1 completely blocked IgE-binding to rCap a 2 in cross-inhibition assays. HPLC analysis showed that in solution Cap a 2 and Lyc e 1 can be found predominantly as dimers, which can be partially reduced to monomers by addition of dithiothreitol (DTT). In ELISA DTT-treated Lyc e 1 displayed a clearly lower IgE-binding capacity than untreated profilin. Conclusions: Purified rCap a 2 and rLyc e 1 proved to be valuable tools for studying cross-reactivity to profilins in patients allergic to pollen and food.

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  1. Bruijnzeel-Koomen C, Ortolani C, Aas K, Bindslev-Jensen C, Bjorksten B, Moneret-Vautrin D, Wuthrich B: Adverse reactions to food. European Academy of Allergology and Clinical Immunology Subcommittee. Allergy 1995;50:623–635.
  2. Baatout S: Profilin: An update (letter). Eur J Clin Chem Clin Biochem 1996;34:575–577.
  3. Gibbon BC, Zonia LE, Kovar DR, Hussey PJ, Staiger CJ: Pollen profilin function depends on interaction with proline-rich motifs. Plant Cell 1998;10:981–993.
  4. van Ree R, Viotenko V, van Leeuwen WA, Aalberse RC: Profilin is a cross-reactive allergen in pollen and vegetable foods. Int Arch Allergy Immunol 1992;98:97–104.
  5. Valenta R, Duchene M, Ebner C, Valent P, Sillaber C, Deviller P, Ferreira F, Tejkl M, Edelmann H, Kraft D, et al: Profilins constitute a novel family of functional plant pan-allergens. J Exp Med 1992;175:377–385.
  6. Diez-Gomez ML, Quirce S, Cuevas M, Sanchez-Fernandez C, Baz G, Moradiellos FJ, Martinez A: Fruit-pollen-latex cross-reactivity: Implication of profilin (Bet v 2). Allergy 1999;54:951–961.
  7. Bauer L, Ebner C, Hirschwehr R, Wuthrich B, Pichler C, Fritsch R, Scheiner O, Kraft D: IgE cross-reactivity between birch pollen, mugwort pollen and celery is due to at least three distinct cross-reacting allergens: Immunoblot investigation of the birch-mugwort-celery syndrome. Clin Exp Allergy 1996;26:1161–1170.
  8. Ballmer-Weber BK, Hoffmann A, Wuthrich B, Luttkopf D, Pompei C, Wangorsch A, Kastner M, Vieths S: Influence of food processing on the allergenicity of celery: DBPCFC with celery spice and cooked celery in patients with celery allergy. Allergy 2002;57:228–235.
  9. Luttkopf D, Ballmer-Weber BK, Wuthrich B, Vieths S: Celery allergens in patients with positive double-blind placebo-controlled food challenge. J Allergy Clin Immunol 2000;106:390–399.
  10. Ebner C, Jensen-Jarolim E, Leitner A, Breiteneder H: Characterization of allergens in plant-derived spices: Apiaceae spices, pepper (Piperaceae), and paprika (bell peppers, Solanaceae). Allergy 1998;53:52–54.
  11. Petersen A, Vieths S, Aulepp H, Schlaak M, Becker WM: Ubiquitous structures responsible for IgE cross-reactivity between tomato fruit and grass pollen allergens. J Allergy Clin Immunol 1996;98:805–815.
  12. van Toorenenbergen AW, Waanders J, van Wijk RG, Vermeulen AM: Immunoblot analysis of IgE-binding antigens in paprika and tomato pollen. Int Arch Allergy Immunol 2000;122:246–250.
  13. Groenewoud GC, de Jong NW, van Oorschot-van Nes AJ, Vermeulen AM, van Toorenenbergen AW, Mulder PG, Burdorf A, de Groot H, van Wijk RG: Prevalence of occupational allergy to bell pepper pollen in greenhouses in the Netherlands. Clin Exp Allergy 2002;32:434–440.
  14. Jensen-Jarolim E, Santner B, Leitner A, Grimm R, Scheiner O, Ebner C, Breiteneder H: Bell peppers (Capsicum annuum) express allergens (profilin, pathogenesis-related protein P23 and Bet v 1) depending on the horticultural strain. Int Arch Allergy Immunol 1998;116:103–109.
  15. Bleumink E, Berrens L, Young E: Studies on the atopic allergen in ripe tomato fruits. I. Isolation and identification of the allergen. Int Arch Allergy Appl Immunol 1966;30:132–145.
  16. Kondo Y, Urisu A, Tokuda R: Identification and characterization of the allergens in the tomato fruit by immunoblotting. Int Arch Allergy Immunol 2001;126:294–299.
  17. Foetisch K, Son DY, Altmann F, Aulepp H, Conti A, Haistein D, Vieths S: Tomato (Lycopersicon esculentum) allergens in pollen-allergic patients. Eur Food Res Technol 2001;213:259–266.
  18. Yu LX, Nasrallah J, Valenta R, Parthasarathy MV: Molecular cloning and mRNA localization of tomato pollen profilin. Plant Mol Biol 1998;36:699–707.

    External Resources

  19. Kingston R: Method for plant RNA preparation. Current Protocols in Molecular Biology. Cambridge, Mass., Wiley & Sons, 1995.
  20. Hoffmann-Sommergruber K, Susani M, Ferreira F, Jertschin P, Ahorn H, Steiner R, Kraft D, Scheiner O, Breiteneder H: High-level expression and purification of the major birch pollen allergen, Bet v 1. Protein Expr Purif 1997;9:33–39.
  21. Lindberg U, Schutt CE, Hellsten E, Tjader AC, Hult T: The use of poly(L-proline)-Sepharose in the isolation of profilin and profilactin complexes. Biochim Biophys Acta 1988;967:391–400.
  22. Jarolim E, Rumpold H, Endler AT, Schlerka G, Ebner H, Scheiner O, Kraft D: Specificities of IgE and IgG antibodies in patients with birch pollen allergy. Int Arch Allergy Appl Immunol 1989;88:180–182.
  23. Laemmli UK: Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 1970;227:680–685.
  24. Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 1979;76:4350–4354.
  25. Hirel PH, Schmitter MJ, Dessen P, Fayat G, Blanquet S: Extent of N-terminal methionine excision from Escherichia coli proteins is governed by the side-chain length of the penultimate amino acid. Proc Natl Acad Sci USA 1989;86:8247–8251.
  26. Vallier P, Balland S, Harf R, Valenta R, Deviller P: Identification of profilin as an IgE-binding component in latex from Hevea brasiliensis: Clinical implications. Clin Exp Allergy 1995;25:332–339.
  27. Hirschwehr R, Valenta R, Ebner C, Ferreira F, Sperr WR, Valent P, Rohac M, Rumpold H, Scheiner O, Kraft D: Identification of common allergenic structures in hazel pollen and hazelnuts: A possible explanation for sensitivity to hazelnuts in patients allergic to tree pollen. J Allergy Clin Immunol 1992;90:927–936.
  28. Pastorello EA, Vieths S, Pravettoni V, Farioli L, Trambaioli C, Fortunato D, Luttkopf D, Calamari M, Ansaloni R, Scibilia J, Ballmer-Weber BK, Poulsen LK, Wuthrich B, Hansen KS, Robino AM, Ortolani C, Conti A: Identification of hazelnut major allergens in sensitive patients with positive double-blind, placebo-controlled food challenge results. J Allergy Clin Immunol 2002;109:563–570.
  29. Jankiewicz A, Aulepp H, Baltes W, Bogl KW, Dehne LI, Zuberbier T, Vieths S: Allergic sensitization to native and heated celery root in pollen-sensitive patients investigated by skin test and IgE binding. Int Arch Allergy Immunol 1996;111:268–278.
  30. 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 2001;108:301–307.
  31. Pastorello EA, Ortolani C, Farioli L, Pravettoni V, Ispano M, Borga A, Bengtsson A, Incorvaia C, Berti C, Zanussi C: Allergenic cross-reactivity among peach, apricot, plum, and cherry in patients with oral allergy syndrome: An in vivo and in vitro study. J Allergy Clin Immunol 1994;94:699–707.
  32. Wensing M, Akkerdaas JH, van Leeuwen WA, Stapel SO, Bruijnzeel-Koomen CA, Aalberse RC, Bast BJ, Knulst AC, van Ree R: IgE to Bet v 1 and profilin: Cross-reactivity patterns and clinical relevance. J Allergy Clin Immunol 2002;110:435–442.
  33. Ganglberger E, Radauer C, Wagner S, Riordain G, Beezhold DH, Brehler R, Niggemann B, Scheiner O, Jensen-Jarolim, E, Breiteneder H: Hev b 8, the Hevea brasiliensis latex profilin, is a cross-reactive allergen of latex, plant foods and pollen. Int Arch Allergy Immunol 2001;125:216–227.
  34. Huang S, McDowell JM, Weise MJ, Meagher RB: The Arabidopsis profilin gene family. Evidence for an ancient split between constitutive and pollen-specific profilin genes. Plant Physiol 1996;111:115–126.
  35. Babich M, Foti LR, Sykaluk LL, Clark CR: Profilin forms tetramers that bind to G-actin. Biochem Biophys Res Commun 1996;218:125–131.
  36. Psaradellis T, Kao NL, Babich M: Recombinant Zea mays profilin forms multimers with pan-allergenic potential. Allergol Int 2000;49:27–35.
  37. Wopfner N, Willerroider M, Hebenstreit D, van Ree R, Aalbers M, Briza P, Thalhamer J, Ebner C, Richter K, Ferreira F: Molecular and immunological characterization of profilin from mugwort pollen. Biol Chem 2002;383:1779–1789.

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