title: Cloning of three new allergens from the dust mite Lepidoglyphus destructor using phage surface display technology.
abstract: The dust mite Lepidoglyphus destructor is a common species in Europe and a major cause of dust mite allergy in rural surroundings, but it also contributes to dust mite allergy in urban areas. One major allergen, Lep d 2, has been expressed as a recombinant protein and evaluated both in vivo and in vitro and shown to detect 60% or more of L. destructor-sensitized subjects. Additional recombinant allergens are needed to obtain a reliable diagnostic tool for L. destructor allergy. The aim of this study was to clone and express new allergens from L. destructor and determine their recognition frequency among sensitized individuals. A phage display cDNA expression library was constructed and screened with sera from L. destructor-sensitized individuals. The cDNAs encoding the allergens were cloned into the pET17b vector and subsequently expressed in Escherichia coli as C-terminal His6-tagged proteins. Immunoblotting of the recombinant proteins was performed using sera from 45 subjects allergic to L. destructor. Three new allergens from L. destructor, Ld 5 (originating from a partial Lep d 5 clone), Lep d 7 and Lep d 13, were identified and recognized by 4/45 (9%), 28/45 (62%) and 6/45 (13%) sera from L. destructor-sensitized subjects, respectively.
authors.affiliation: Unit of Clinical Immunology and Allergy, Karolinska Institute and Hospital, S-171 76 Stockholm, Sweden. tove.eriksson@ks.se
authors.forename: T L
authors.initials: TL
authors.lastname: Eriksson
authors.forename: O
authors.initials: O
authors.lastname: Rasool
authors.forename: S
authors.initials: S
authors.lastname: Huecas
authors.forename: P
authors.initials: P
authors.lastname: Whitley
authors.forename: R
authors.initials: R
authors.lastname: Crameri
authors.forename: U
authors.initials: U
authors.lastname: Appenzeller
authors.forename: G
authors.initials: G
authors.lastname: Gafvelin
authors.forename: M
authors.initials: M
authors.lastname: van Hage-Hamsten
mesh_terms: D000485:Allergens / Q000235:genetics* / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D015723:Gene Library; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood* / Q000276:immunology; D008925:Mites / Q000235:genetics* / Q000276:immunology*; D008969:Molecular Sequence Data; D019151:Peptide Library; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D017386:Sequence Homology, Amino Acid
chemical_list: D000485:Allergens; C422451:Ld 5 allergen; C422453:Lep d 13 allergen; C422452:Lep d 7 allergen; D019151:Peptide Library; D011994:Recombinant Proteins; D007073:Immunoglobulin E