Skin test diagnosis of grass pollen allergy with a recombinant hybrid molecule

• Published in: Journal of allergy and clinical immunology 2007-08, Vol.120 (2), p.315-321
• Main Authors: Metz-Favre, Carine, MD, Linhart, Birgit, PhD, Focke-Tejkl, Margarete, PhD, Purohit, Ashok, MD, de Blay, Frédéric, MD, Valenta, Rudolf, MD, Pauli, Gabrielle, MD
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; Allergens - administration & dosage; Allergens - genetics; Allergies; Allergy; Allergy and Immunology; Basophils - drug effects; Basophils - metabolism; Biological and medical sciences; diagnosis; Dose-Response Relationship, Drug; Epitopes; Escherichia coli; Female; Fundamental and applied biological sciences. Psychology; Fundamental immunology; grass pollen; Histamine Release; Humans; Hypersensitivity - diagnosis; Hypersensitivity - immunology; Immunoglobulin E - immunology; Male; Medical sciences; Middle Aged; Phleum - immunology; Plant Extracts - immunology; Poaceae - chemistry; Poaceae - immunology; Pollen; Pollen - chemistry; Pollen - immunology; Product introduction; Quality of life; recombinant hybrid allergen; Recombination, Genetic; Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis; Sensitivity and Specificity; Serologic Tests; skin testing; Skin Tests - methods
• ISSN: 0091-6749; 1097-6825
• DOI: 10.1016/j.jaci.2007.03.046

Background A recombinant hybrid molecule (HM) consisting of 4 major allergens from timothy grass (Phl p 1, 2, 5, and 6) was expressed in Escherichia coli , purified, and characterized regarding its immunologic properties. Objective We sought to determine whether the recombinant HM can be used for the diagnosis of grass pollen allergy by means of skin testing. Methods Skin prick testing was performed in 32 patients with grass pollen allergy and in 9 control individuals by using increasing concentrations (4, 12, 36, and 108 μg/mL) of the HM and using commercial grass pollen extract. Specific IgE reactivities against the HM, grass pollen extract, and a panel of purified grass pollen allergens (recombinant Phl p 1, 2, 5, 6, 7, 12, and 13 and natural Phl p 4) were measured by means of ELISA, and timothy grass pollen–specific IgE levels were determined by using ImmunoCAP. Results Grass pollen allergy was diagnosed in all patients by means of skin testing with the HM. No false-positive skin test responses were obtained in the control individuals. There was an excellent correlation between IgE levels obtained with the HM and natural grass pollen extract measured by means of ELISA ( r = 0.98, P < .0001) and by means of ImmunoCAP ( r = 0.98, P < .0001). Conclusions The recombinant HM permitted accurate and specific in vivo diagnosis of grass pollen allergy in all tested patients. It can be considered a well-defined tool for the diagnosis and perhaps for immunotherapy of grass pollen allergy. Clinical implications A recombinant HM can replace traditional allergen extracts for skin test–based diagnosis of grass pollen allergy.