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NAOSITE : Nagasaki University's Academic Output SITE > Institute of Tropical Medicine > Bulletin > Tropical medicine > Volume 29, No. 3 >

イムノブロット法を用いた住血吸虫抗原の解析 : stage-specific抗原及びspecies-specific抗原の検索


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Title: イムノブロット法を用いた住血吸虫抗原の解析 : stage-specific抗原及びspecies-specific抗原の検索
Other Titles: Immunoblot Analysis of Schistosome Antigens : Trial of Identification of Stage-specific and Species-specific Antigens
Authors: 佐藤, 克之
Authors (alternative): Sato, Katsuyuki
Issue Date: 30-Sep-1987
Publisher: 長崎大学熱帯医学研究所 / Institute of Tropical Medicine, Nagasaki University
Citation: 熱帯医学 Tropical medicine 29(3). p139-152, 1987
Abstract: イムノブロット法を用いて,マンソン住血吸虫感染BALE/Cマウスより経時的に採集した血清(感染後1,2,4,8,12週)と,マンソン住血吸虫各種抗原(セルカリア分泌物,セルカリア及び成虫粗抗原)との反応を調べマンソン住血吸虫のstage-specific抗原について検討した。各種抗原をSDS-ポリアクリルアミドゲル電気泳動で分離した後,ニトロセルロース膜に転写し,被検血清との反応後,酵素抗体法で各種抗原に反応した抗体を検出した。その結果,マンソン住血吸虫感染BALE/Cマウスでは,各種抗原に対する感染血清の反応が,各感染時期(感染初期,産卵開始期及び慢性期)で特徴的なパターンを示すことがわかった。しかし,ある感染時期に特異的な抗原は成虫由来の分子量26.5kDaの抗原を除いて見つけることはできなかった。また,本研究においては同じくイムノブロット法により,マンソン及びビルハルツ住血吸虫感染血清(GNハムスター,ヒト)とマンソン,ビルハルツ成虫粗抗原とを用いて,住血吸虫のspecies-specific抗原についても調べてみた。マンソン住血吸虫成虫粗抗原を使って,ハムスター感染血清との反応を行なったところ,分子量45kDaと31kDaの2つの抗原に対しては,マンソン住血吸虫感染血清のみが反応し,分子量33kDaの抗原に対しては,ビルハルッ住血吸虫感染血清のみが反応した。ビルハルツ住血吸虫粗抗原を用いた場合でも同じ結果が得られた。次に,マンソン住血吸虫成虫粗抗原を使って,ヒト患者血清との反応を調べてみると,マンソン感染患者12例中11例が,45kDa抗原に反応したのに対し,ビルハルツ感染患者9例のいずれもこの抗原に対しては反応しなかった。31kDa抗原に対しては,交叉反応があり,33kDaの成分は,どのヒト患者血清とも反応しなかった。以上の結果より,分子量45kDaの抗原は,マンソン・ビルハルツ両住血吸虫症の混合流行地において,マンソン住血吸虫症の特異的な免疫診断に応用できるかもしれない。 / The experiment was designed to identify the stage-specific and species-specific immunoreactive components in Schistosomiasis mansoni. Cercarial secretion materials (CSM), whole cercarial and adult worm antigens of S. mansoni were separated by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis and assayed in immunoblot for reaction with mouse sera obtained at given Intervals after infection. The striking difference in the appearance of antibodies reacting with CSM, cercanal antigens and adult worm antigens was observed during the course of infection. When the cercarial antigens were used, as early as 1 week postinfection, IgG antibodies reacted prominently with the components in the region of 34, 28.5 and 26 kDa of molecular weight. The immunoreactivity of these components to the sera remained unchanged up to 4 weeks postinfection. CSM reacted with IgM antibodies as well, but the molecular weights of immunoreactive components differed from those of cercarial antigens during the first 4 weeks postinfection. From week 1 to week 4 postinfection, none of the adult worm antigens reacted with the sera. At week 8, 2 components of 31 and 26.5 kDa of adult worm antigens reacted strongly with IgG antibodies. Other additional major immunoreactive components also reacted with the sera at this stage were 40 kDa of cercarial antigens and 90 kDa of CSM. At week 12, the adult worm antigens of 34 and 60 kDa molecular weights were recognized by the sera. These results suggest that comparison of antibodies reacting with different developmental stages of schistosome antigens clearly distinguish the stage of schistosome infection. S. mansoni species-specific antigens were examined by comparing the immunoreactivity of S. mansoni adult worm antigens with S. mansoni and S. haematobium infected human and hamster sera. The immunogens of 45 and 31 kDa of S. mansoni adult worm antigens reacted strongly with sera of hamsters infected with S. mansoni, but did not with sera of animals infected with S. haematobium. S. haematobium adult worm antigens, however, contained two components of 45 and 31 kDa which gave slight reaction with S. mansoni infected sera. The 45 kDa component of S. mansoni adult worm antigens reacted strongly with 11 out of 12 sera collected from patients infected with S. mansoni. This antigen also reacted with none of the 9 sera from patients infected with S. haematobium. The 31 kDa component of S. mansoni adult worm antigens, however, reacted with 7 out of 12 sera collected from the patients infected with S. mansoni and with 4 sera from the 9 patients infected with S. haematobium. The use of this 45 kDa antigen will possibly improve the diagnosis of S. mansoni infection in the areas where both S. mansoni and S. haematobium coexist.
URI: http://hdl.handle.net/10069/4498
ISSN: 03855643
Type: Departmental Bulletin Paper
Appears in Collections:Volume 29, No. 3

Citable URI : http://hdl.handle.net/10069/4498

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