1988 Fiscal Year Final Research Report Summary
Preparation of site-specific antibodies by a novel immunization procedure involving induction of tolerance to cross-reacting determinants and its utilization
| Project/Area Number |
61570148
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pathological medical chemistry
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| Research Institution | Fukuoka University |
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Principal Investigator |
TATEISHI Kayoko Fukuoka University, School of Medicine, Instructor, 医学部, 助手 (60179728)
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| Co-Investigator(Kenkyū-buntansha) |
KUROKI Masahide Fukuoka University, School of Medicine, Associate professor, 医学部, 助教授 (40122692)
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| Project Period (FY) |
1986 – 1988
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| Keywords | -GL / Specific anti-neurokinin A antiserum / Specific anti-neurokinin B antiserum / Production of site-specific antibody / Distribution of neurokinin A and neurokinin B / Nucleus tractus solitarii / 高血圧とニューロキニンB |
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| Research Abstract |
Neurokinin A (NKA) and neurokinin B (NKB) have the common C-terminal amino acid sequence. By virtue of such homologies among tachykinins, most antisera raised against NKA markedly cross-react with NKB and other tackykinins. Difficulty in producing highly specific antisera discriminating NKA and NKB has prevented detailed studies of these peptides. Therefore, preparations of highly specific antisera for NKA and NKB were tried by the novel immunization procedure which involved immunization with NKA or NKB, both conjugated with keyhole limpet hemocyanin, and treatments with a tolerogenic conjugate of kassinin and a copolymer of D-glutamic acid and D-lysine (D-GL) to inhibit the production of cross-reactive antibodies against common C-terminal region of tachykinins. Cross-reactivities of anti-NKA antiserum, thus obtained, with NKB and kassinin were 12.6% and 10.6%, respectively. This was in sharp contrast with those (129.0% and 42.5%, respectively) of antiserum obtained from the rabbit not
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treated with kassinin-D-GL. Anti-NKB antiserum by the above tolerizing regimen was highly specific for NKB and the cross-reactivities with NKA and other tachykinins were all less than 0.001%. RIAs using these specific antisera allowed us to measure directly NKA and NKB in tissue extracts without their fractionation by chromatography prior to RIAs. Measurements of immunoreactive NKA and NKB in rat brain regions and spinal cord revealed that they are present with various ratios (NKA/NKB : 1.1 - 9.9) depending on the region. Microinjection of senktide (NKB receptor peptide) into the nucleus tractus solitrarii (NTS) of rats caused long-lasting hypertension. NKB content in the nts was not affected by nodose ganglionectomy. NKB levels in supraoptic nucleus of spontaneously bypertensive rats (SHR) were significantly higher than those in control rats (WKY). These findings suggest that NKB may be a neuromodulator on cardiovascular responses in the NTS and stimulate the ascending pathway from the NTS to the hypothalamus. Less
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