1989 Fiscal Year Final Research Report Summary
Immunochemical assay of mite infestation toward devising a "house-hold diagnostic kit"
| Project/Area Number |
62860011
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
応用生物化学・栄養化学
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
OKA Satoru Hiroshima University Fac. of Eng. Professor, 工学部, 教授 (80034320)
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| Co-Investigator(Kenkyū-buntansha) |
WADA Takeshi Fumakilla Limited, Dept. Development, Chief, 開発管理部部長
OGAWA Kengo Fumakilla Limited, Dept. Development, Director, 本部長
JYO Toshihiko Hiroshima Prefectural Hospital Director, 院長
SHIGETA Seiko Hiroshima University Fac. of Eng. Research Associate, 工学部, 助手 (10034381)
ONO Kazuhisa Hiroshima University Fac. of Eng. Research Associate, 工学部, 助手 (10144883)
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| Project Period (FY) |
1987 – 1989
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| Keywords | Allergen / Mite Allergen / Mite Asthma / Mite Infestation / Histamine / Hyposensitization Therapy / cDNA / Genetic Engineering |
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| Research Abstract |
1. Diagnoses of mite allergic disease by determination of histamine. Micro-assay of histamine using HPLC was established. The patient's allergen sensitivity or the titer of blocking antibody estimated by the assay made it possible to evaluate which component was major allergen responsible for the patient or balance of specific IgE and IgG titers in the patient, respectively. The two measured values are expected to improve the accuracy of diagnosis of the patient with mite allergy.
2. Immunochemical assay of mite infestation. Rabbit, mouse, hen, and guinea pig antisera and mouse monoclonal antibody against mite body and feces antigens were prepared in addition to the patient's sera. Proper use of the antibodies in addition to the results on the allergen sensitivity and blocking antibody titer of the patient enabled us to evaluate the actual condition of mite infestation under living environment.
3. Purification of major allergen. By monitoring the ability of fractions obtained from gel-filtration of crude mite feces and body antigens to release histamine from patient's leukocyte, unknown diverse major allergens were found to occur over a wide range of molecular weight. Of them new mite fecal allergens with MW 74k and 4k were isolated. The high molecular mite allergen was a glycoprotein (saccharide content 70%) and possessed high immunogenicity but induced no anaphylactic reaction. Therefore, the allergen is expected to be a valuable antigen for hyposensitization therapy. The low molecular mite allergen was also a glycoprotein (saccharide content 45%) and cross-reactive to the high molecular mite allergen, but did not possess immunogenicity and ability to induce anaphylactic reaction.
4. Production of mite allergen by genetic engineering. cDNA-inserted phages coding for mite allergen were isolated in recombinant phages prepared using mRNA in mite. The nucleotide sequence coding for a mite allergen is now under investigation by dideooxy-mediated chain-termination method.
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