2001 Fiscal Year Final Research Report Summary
Development of the high sensitivity measurements for drug-allergy
| Project/Area Number |
11672310
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Toho university |
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Principal Investigator |
ITO Masatoshi Toho university, professor, 医学部, 教授 (10057698)
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| Co-Investigator(Kenkyū-buntansha) |
OKADA Yayoi Toho university, assistant, 医学部, 助手 (60256758)
ISHIKAWA Fumio Toho university, assistant professor, 医学部, 講師 (10130345)
TAKEUCHI Yoshio Toho university, assistant professor, 医学部, 講師 (40130372)
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| Project Period (FY) |
1999 – 2001
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| Keywords | Drug allergy / Anti-drug antibody / DLST / Interleukin-2 / RT-PCR / RAD / Affinity content / Competitive ELISA |
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| Research Abstract |
Drug allergy was examined on the supersensitive detection method for the determinant of drug antigen and drag-lymphocyte blastogenesis test. In ELISA, the antibody detection was carried out using 3 kinds of cefaclor/OVA complex of which the cross-link position differed. First, the side-chain(R 1) of cefaclor was recognized by anti-cefaclor IgG antibodies obtained from cefaclor-allergy patient. In contrast, the IgM antibody reacted at 7-aminocephalosporanic acid (7-AMA) of cefaclor. Competitive ELISA shown that anti-cefaclor side-chain antibodies were competitive inhibitied by ampicillin, becampicillin, cefaclor and cephalexin, which were composed from α-phenylglycine side-chain. Anti-cefaclor IgG antibodies reacted to the α-phenylglycine (R1) side- chain in a dose response, but not p-amino phenyl acetic acid and 7-AMA. Furthermore, BIAcore analysis indicated that the affinity of anti-cefaclor antibody was Ka=2. 19 x 10-3M-1and Kd=2.56 x 10-4 M-l , which were almost equal to other the immune antibody. On the other hand, DLST was positive in 3 out of 25 patients, and false positives were 3 patients. In contrast, IL-2 was detected from 4 to 8 hrs after the stimulation by drugs. Moreover,IL-2 mRNA was detected in all patients with the IL-2 production.
These results suggested that the detection method of IL-2 protein and mRNA are more super-sensitive than the DLST, and this method is effective diagnosed test for drug-allergy.
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