Chemistry: analytical and immunological testing – Involving iga – igd – ige – or igm
Patent
1994-03-30
1995-10-24
Warden, Jill
Chemistry: analytical and immunological testing
Involving iga, igd, ige, or igm
436 63, 436811, 530324, 530326, 435 29, A61K 3800, C07K 500, C07K 700, C07K 1700
Patent
active
054609771
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to a peptide which causes the blast formation of lymphocytes of allergic patients sensitized with mite allergen. Such a peptide can be used for the diagnosis of mite allergy.
In order to diagnose so-called type I allergy, such as allergic asthma and allergic rhinitis, a method has heretofore been used which employs a causative antigen (i.e. allergen).
That is, there has been widely used a method wherein an allergen is injected intradermally or dropped onto slightly injured skin and the resulting skin reaction is observed to identify the allergen, or such a method as RAST, wherein allergen specific IgE in blood serum is detected in vitro.
In these methods of diagnosis, allergens extracted from naturally occurring substances have hitherto been used. That is, crude allergens extracted from allergen-containing substances, as pollen, house dust and mites, with, for example, the Coca solution (Practice Allergy, third edition, written by W. T. Vaughhan & J. H. Black) have been used. Consequently, these methods often cause side reactions and hence tend to lead to diagnosis of insufficient accuracy. Recently, for the purpose of obtaining pure allergens, a number of attempts have been made to prepare major allergens of pollen or mites by means of genetic recombination (R. Valena et al., Int. Arch. Allergy & Applied Immunol., 97, 287 (1992), Japanese Patent Application Nos. H-3-262538 and H-3-310069).
Mites are one of the most important allergens causing type I allergy. For preparing a mite allergen, there are known, as described above, a method which extracts it from mite bodies and a method which prepare it by genetic recombination. By the method of extraction from mite bodies, however, it is difficult to prepare the purified allergen in large quantity. By the method of preparing a mite allergen through genetic recombination, on the other hand, it is impossible to prepare solely the allergenic activity-bearing low molecular weight portion which constitutes a part of the major allergen protein of mites, unlike in the process of the present invention.
Accordingly, the present inventors have attempted to prepare a substance having allergenic activity easily and in a pure form by chemically synthesizing solely the allergenic activity-bearing portion of the major allergen protein of mites. Thus, the inventors have synthesized various parts of the amino acid sequence of Der f II, which is one of the major allergens of Dermatophagoides farinae, and resultantly found a peptide which has allergenic activity.
The cell which mainly participates in the humoral immunity of mammalia, including human beings, is a lymphocyte.
When the lymphocyte of a patient sensitized by an allergen is reacted with the allergen, the cell is stimulated to cause lively nucleic acid synthesis; that is, so-called blast formation takes place. As the result, the incorporation of thymidine, which is a constituent of nucleic acid, into the cell increases.
Making use of the above-mentioned phenomenon, the present inventors have made a thymidine labelled with radioactivity and the peptide of the present invention to react with the lymphocyte of a mite-allergic patient or a non-allergic subject. As the result, virtually no incorporation of thymidine to the lymphocyte of non-allergic subject was recognized, while a marked incorporation of thymidine into the lymphocyte of allergic patient was observed, revealing that blast formation took place.
This demonstrates that the peptide of the present invention was clearly recognized by the lymphocyte of a mite-allergic patient. In other words, by using the peptide of the present invention, the diagnosis of allergy becomes possible based on the fact that the lymphocyte of a patient sensitized by mites undergoes blast formation, whereas the lymphocyte of a non-allergic subject not sensitized by mites undergoes no blast formation.
According to the present invention, there is provided a peptide which causes the blast formation of lymphocytes represented by mites. In more particular, peptide
REFERENCES:
Hof, Wim Van't, et al., "Epitope mapping of the Dermatophagoides pteronyssinus house dust mite major allergen Der p II using overlapping syntheitc peptides", Molecular Immunology (1991), vol. 28, pp. 1225-1232.
Ando Tohru
Ikeda Shigeru
Okumura Yasushi
Asahi Breweries Ltd.
Huff Sheela J.
Torii & Co.
Warden Jill
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