Immunodiagnostic value and properties of an antigen isolated with affinity chromatography using a monoclonal antibody to Trichinella spiralis
| Project/Area Number |
02670165
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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 |
Experimental pathology
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| Research Institution | Chiba University |
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Principal Investigator |
NIIMURA Munetoshi Chiba Univ. School of Med. A.Prof., 医学部, 助教授 (60059095)
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| Co-Investigator(Kenkyū-buntansha) |
HATA Hidekazu Chiba Univ. School of Med. Assist., 医学部, 助手 (00110304)
KOBAYASHI Masashi Chiba Univ. School of Med. Assist., 医学部, 助手 (80009654)
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| Project Period (FY) |
1990 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1992: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1991: ¥500,000 (Direct Cost: ¥500,000)
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| Keywords | Trichinella spiralis / alpha -Stichosome / Monoclonal antibody / Antigen / Immunodiagnosis / Anthelmintic agent / 治療剤 / 局在 / αースチコソ-ム / モノクロ-ナル抗体 / 防御免疫 |
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| Research Abstract |
Monoclonal antibodies were generated for the isolation of specific antigen from Trichinella spiralis(T.s). A monoclonal antibody (TS32D12) of IgG_1 class was selected according to its reactivity and specificity. We isolated 160kDa molecule (Ts-alpha 160) from alpha -stichocyte of T.s muscle larvae(Ts-L_1) with affinity chromatography using TS32D12 and examined immunodiagnostic value of Ts-alpha 160 against serum samples which had been obtained from 13 patients 33-109 days after infection. Antibodies against Ts-alpha 160 were detected with higher titers than those against Ts-crude antigen in every case. The application of Ts-alpha 160 to the ELISA eliminated false-positive reactions which were observed against Ts-crude antigen in serum samples from patients with proven helminthiasis. On the other hand, the 160kDa molecule was not detected in adult worms, embryos, nor in muscle larvae at day 14 post infection. However, it was demonstrated in alpha -stichosome of the late muscle stage lar
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vae from day 21 at least up to day 180. Antibody responses of infected BALB/c mice of T.s against the Ts- alpha 160 became detectable with low titers at week 4, followed by a rapid increased in titers at week 8. These results suggested that Ts-alpha 160 can be utilized as a unique antigen for the immunodiagnosis of human trichinosis from about 5-6 week after infection. In addition, Ts-alpha 160 was found consist of glycoprotein and E-S product derived from Ts-L_1, the epitopes recognized by the TS32D12 seemed to be lost after treatment at 100ﾟC, 2 min. The Ts-alpha 160 was elucidated by the immunogold method using electron microscope that is located in alpha granules of stichocyte of Ts-L_1. There were no significant decrease in recovery of worms among experimental groups of mice both passively transferred TS32D12 and actively transferred Ts-alpha 160. Freshly isolated T.s L_1 were incubated containing ammonium embelate in PBS, large precipitates accumulated and adhered to the oral end of the worms,and were observed to react to TS32D12 by immuno-fluorescence stain. Less
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Report
(4 results)
Research Products
(9 results)
