BACKGROUND: Grass pollen allergens belong to the potent elicitors of type I allergy. Approximately 40% of allergic individuals display IgE reactivity with grass pollen allergens. In previous studies we have reported the complementary DNA cloning and expression in Escherichia coli of three of the most relevant timothy grass pollen allergens: Phl p 1, Phl p 2, and Phl p 5. OBJECTIVE: To achieve high level expression of immunologically active timothy grass pollen allergens in E. coli, the cDNAs were inserted into expression plasmids. METHODS: The three recombinant grass pollen allergens were expressed at high levels in E. coli as recombinant nonfusion proteins, purified by conventional protein chemical methods and tested for their IgE-binding capacity by immunoblot and ELISA, as well as in histamine release assays. RESULTS: Milligram amounts of pure recombinant allergens were obtained from cultured E. coli. IgE binding to purified recombinant Phl p 1, Phl p 2, and Phl p 5 could be demonstrated by immunoblot and ELISA. With ELISAs the percentage of grass pollen-specific IgE directed against the individual recombinant allergens could be estimated. In addition, the purified recombinant timothy grass pollen allergens induced dose-dependent and specific histamine release from patients' blood basophils. CONCLUSION: Purified recombinant timothy grass pollen allergens represent useful tools for diagnosis and therapy of grass pollen allergy.
BACKGROUND: Grass pollen allergens belong to the potent elicitors of type I allergy. Approximately 40% of allergic individuals display IgE reactivity with grass pollen allergens. In previous studies we have reported the complementary DNA cloning and expression in Escherichia coli of three of the most relevant timothy grass pollen allergens: Phl p 1, Phl p 2, and Phl p 5. OBJECTIVE: To achieve high level expression of immunologically active timothy grass pollen allergens in E. coli, the cDNAs were inserted into expression plasmids. METHODS: The three recombinant grass pollen allergens were expressed at high levels in E. coli as recombinant nonfusion proteins, purified by conventional protein chemical methods and tested for their IgE-binding capacity by immunoblot and ELISA, as well as in histamine release assays. RESULTS: Milligram amounts of pure recombinant allergens were obtained from cultured E. coli. IgE binding to purified recombinant Phl p 1, Phl p 2, and Phl p 5 could be demonstrated by immunoblot and ELISA. With ELISAs the percentage of grass pollen-specific IgE directed against the individual recombinant allergens could be estimated. In addition, the purified recombinant timothy grass pollen allergens induced dose-dependent and specific histamine release from patients' blood basophils. CONCLUSION: Purified recombinant timothy grass pollen allergens represent useful tools for diagnosis and therapy of grass pollen allergy.
Authors: Hans Grönlund; Susanne Vrtala; Ursula Wiedermann; Gerhard Dekan; Dietrich Kraft; Rudolf Valenta; Marianne Van Hage-Hamsten Journal: Immunology Date: 2002-12 Impact factor: 7.397
Authors: S Vrtala; K Hirtenlehner; L Vangelista; A Pastore; H G Eichler; W R Sperr; P Valent; C Ebner; D Kraft; R Valenta Journal: J Clin Invest Date: 1997-04-01 Impact factor: 14.808
Authors: V Niederberger; F Horak; S Vrtala; S Spitzauer; M-T Krauth; P Valent; J Reisinger; M Pelzmann; B Hayek; M Kronqvist; G Gafvelin; H Grönlund; A Purohit; R Suck; H Fiebig; O Cromwell; G Pauli; M van Hage-Hamsten; R Valenta Journal: Proc Natl Acad Sci U S A Date: 2004-08-13 Impact factor: 11.205
Authors: Katarzyna Niespodziana; Margarete Focke-Tejkl; Birgit Linhart; Vera Civaj; Katharina Blatt; Peter Valent; Marianne van Hage; Hans Grönlund; Rudolf Valenta Journal: J Allergy Clin Immunol Date: 2011-03-16 Impact factor: 10.793