Chemistry: molecular biology and microbiology – Vector – per se
Patent
1992-06-06
1996-12-10
Scheiner, Toni R.
Chemistry: molecular biology and microbiology
Vector, per se
536 236, 4351723, C12N 1529
Patent
active
055830463
DESCRIPTION:
BRIEF SUMMARY
1. FIELD OF THE INVENTION
The invention provides recombinant DNA molecules which code for polypeptides, and the polypeptides per se, that have at least one epitope of a P14 pollen allergen of a tree of the order Fagales, particularly birch (Betula verrucosa), or the entire P14 allergen protein, and exhibit the same or similar antigenicity as a P14 allergen. The invention also provides replicable microbial expression vehicles and microorganisms for use in processes for producing such allergenic polypeptides. Methods are provided for purification of P14 allergen as well as for the diagnosis and therapy of allergic diseases using the synthetic polypeptides of the invention.
2. BACKGROUND OF THE INVENTION
In the springtime large parts of the populations of Central, Eastern and Northern Europe, America and Australia suffer from allergic symptoms (rhinitis, conjunctivitis, dermatitis and pollen asthma). Proteins which can be isolated from pollen of trees of the order Fagales, in particular from pollen of birch, alder, hazel, hornbeam and oak, are responsible for most of these allergic symptoms (1).
At least 10% of the population suffers from pollen allergies at various times and to varying extent. These allergies are mediated by IgE antibodies which react with pollen proteins. The possibility exists for a therapy for pollen allergies by hyposensitization, i.e., by the regular and slowly increasing administration of the proteins producing the allergy.
Diagnostic methods for allergic diseases, such as RIA (radioimmunoassay), IRMA (immuno-radiometric assay), RAST (radio-allergosorbent test), ELISA (enzyme-linked immunosorbent assay), magnetic allergoabsorbent test, immunoblots, LIA (luminescence immunosay), histamine release assays and others depend greatly upon the availability of pure allergens. Protein extracts from pollen isolated from natural sources are difficult to standardize because preparations vary from batch to batch. For example, they may contain unwanted constituents, and/or certain proteins may be lost in the extraction procedure and be missing from the final separation (2). Clearly, diagnostic tests which employ well defined allergens that can be reproducibly prepared would be superior to tests which employ raw pollen extracts with an insufficiently defined mixture of allergens and other components. Recombinant DNA production of allergenic polypeptides, or allergenic fragments thereof, would allow more reprodicible preparations of allergens of defined content for standardized diagnostic and therapeutic methods.
Allergens may be purified to homogenity from pollen by known protein/chemical methods, for example, by means of affinity chromatography (3). These methods are relatively costly and require pollen as an ill-defined source which cannot be standardized. It would, therefore, be cheaper and more efficient to use recombinant DNA methods to produce an allergenic protein, or fragments of that protein.
Hyposensitization has proved to be an effective therapy in allergic diseases. This therapy consists of parenteral or oral administration of allergens in increasing doses over a fairly long period of time. Like diagnostic methods, it requires pure and well defined allergens. The use of purified recombinant allergens or synthetic peptides would greatly reduce the risk of sensitizing patients to unwanted components.
3. SUMMARY OF THE INVENTION
This invention concerns pollen allergens, for example, of white birch (Betula verrucosa), called P14. These pollen allergens are immunologically closely related to allergens which occur in pollen of far distantly related plant species, particularly in trees of the Fagales order (birch, alder, hazel, hornbeam and oak), in grasses and weeds. The cross-reactivity of IgE antibodies of patients to these pollen allergens is illustrated in FIGS. 1A and 1B.
The present invention provides recombinant DNA molecules which contain a nucleotide sequence that codes for a polypeptide which exhibits the same or similar antigenic properties as a natural allergen, P14, which oc
REFERENCES:
patent: 4629706 (1986-12-01), Hammond
patent: 4721668 (1988-01-01), Jones, III et al.
patent: 5116731 (1992-05-01), Wilhelms
Florvaag et al. (1988), Abstract Only, "Comparative Studies on tree pollen allergens XIV" Ann. Allergy 61:392-400.
Marc-Series (1990), Abstract Only, "Mapping of Bet VI epitopes" Int. Arch. Allergy Appl. Immunol. 92:226-32.
Berger et al. (eds) (1987) "Molecular Cloning Manual" Methods in Enzymology 152:316-337, 343-349, 359-371, 451-469.
Itemmens et al. (1988) "A Comparison of the Antigenic and Allergenic Components of Birch & Alder Pollens in Scandinavia and Australia," Int. Arch. Allergy Appl. Immun. 85:27-37.
Lundberg et al (1988) "the use of poly (L-proline Sephrase in the Isolation of Profilin and Profilactin Complexes." Biochem et Biophys Acata 967:391-400.
Oreste et al., 1991, Purification and characterization of Par o l, major antigen of Parietaria officinalis pollen. Int Arch Appl Immunol 96:19-27.
Geraci et al., 1994, EMBL/Gen Bank/DDBJ data bases Parietaria Judaica cDNA of Par 1.
Halpern, GM, "In vitro diagnosis in asthma: the state-of-the-art", Allerg. Immunool. (Paris) 1991, 23(6):255-262.
Harris Hosen, M.D., Clinical Allergy Based on Provocative Testing, Exposition Press, Hicksville, New York, e.g., pp. 11-29, (1978).
Clinical Experimental Allergy, vol. 20, 1990, Suppl. 1, Meeting 8-11 Jul. 1990, S. d'Abusco et al.: "Characterization of cDNA for Parietaria pollen allergens", pp. 48, see abstract OP52.
Biochimica et Biophysics Acta, vol. 967, 1988, Elsevier, U. Lindberg et al.: "The use of poly(L-proline)-sepharose in the isolation of profilin and profilactin complexes", pp. 391-400.
Int. Arch. Appl. Immunol., vol. 98, 1988, H. Breiteneder et al.: "Isolation and characterization of messenger RNA from male inflorescences and pollen of the white birch (Betula verrucosa"), pp. 19-24, see p. 21, left-hand column, Immunoblotting of aqueous BV pollen extracts.
The EMBO Journal, vol. 8, No. 7, Jul. 1989, (Eynsham, Oxford, GB) H. Breiteneder et al.: "The gene coding for the major birch pollen allergen Betvl, is highly homologous to a pea disease resistance response gene", pp. 1935-1938.
Science, vol. 253, 2 Aug. 1991, R. Valenta et al.: "Identification of profilin as a novel pollen allergen; IgE autoreactivity in sensitized individuals", pp. 557-560.
Allergy, vol. 44, No. 6, Aug. 1989, E. Jarolim et al.; "IgE and igG antibodies of patients with allergy to birch pollen as tools to define the allergen profile of Betula verrucosa", pp. 385-395.
Allergy, vol. 45, No. 6, Aug. 1990, T. Birner et al.: "Evaluation of immunotherapy-induced changes in specific IgE, IgG and IgG subclasses in birch pollen allergic patients by means of immunoblotting", pp. 418-426.
Chemical Abstracts, vol. 97:53816e "Comparative studies on tree pollen allergens, IV, Evaluation of two commercially available allergen extracts of alder (Alnus incana) and birch (Betula verrucosa) pollen", 1982, p. 480.
Chemical Abstracts, vol. 104:184481c "Comparative studies on tree pollen allergens. XII. Partial characterization of the alder (Alnus incana) pollen extract by two-dimensional IEF/SDS-PAG electrophoresis combined with electrophoretic transfer and immunoautoradiography", 1986.
Chemical Abstracts, vol. 104:18392e "Immunochemical characterization of reference alder (Alnus glutinosa) and hazel (Corylus avellana) pollen extracts and the partial immunochemical identity between the major allergens of alder, birch and hazel pollens", 1986, p. 380.
Chemical Abstracts, vol. 107:2344124t "Allergologic-immunochemical investigation of various tree pollens. Part I--Characterization of antigen and allergen components in birch, beech, alder, hazel and oak pollens", 1987.
Ipsen, H. and Hansen, O. C. In: Epitopes of Atopic Allergens. Sehon, A. H., Kraft, D., Kunke, G. (eds) (1990) UCB, Brussels, Belgium, pp. 3-8.
Rumpold, et al. In: Epitopes of Atopic Allergens. Sehon, A. H., Kraft, D., Kunkel, G. (eds) (1990) UCB, Brussels, Belgium, pp. 26-28.
Valenta, et al. In: Epito
Breitenbach Michael
Duchene Michael
Kraft Dietrich
Pettenburger Karin
Rumpold Helmut
Biomay Biotechnik Produktions Und Handelsgesellschaft m.b.H.
Green Lora M.
Scheiner Toni R.
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