中国大陆日本血吸虫品系的研究 XI.酶联免疫电转移印迹的分析

本文报告安徽、湖北、广西、云南和四川5地日本血吸虫成虫抗原与安徽、湖北、云南、四川4地钉螺的抗血清进行酶联免疫电转移印迹(EITB)分析的结果。安徽与湖北两地日本血吸虫EITB图谱基本相似。云南和四川的日本血吸虫EITB图谱虽相近,但在84kDa处云南雌虫呈现明显条带而四川雌虫则无此反应带。广西的日本血吸虫与云南的虫体一样,雌虫在84kDa处亦呈现明显的条带。广西的雄虫在与安徽的钉螺抗血清反应后在>130kDa处有两条主要条带,这与安徽地区雄虫的一致且较之更为浓染致密,而云南、四川两地雄虫的则未见此条带。根据上述结果并结合以前有关大陆各地钉螺对日本血吸虫的易感性结果,进行了初步的分析和讨论。     

日本血吸虫未成熟虫卵26／28kDa抗原诱导抗雌虫生…

目的：探讨日本血吸虫未成熟虫的２６／２８ｋＤａ抗原（ＳＩＥＡ２６－２８ｋＤａ）诱导小鼠产生抗雌虫生殖免疫的效果。方法：采用纯化的ＳＩＥＡ２６／２８ｋＤａ抗原以及ＳＩＥＡ－Ｉ抗原，分别免疫ＢＡＬＢ／ｃ小鼠，于攻击感染后４６ｄ进行粪卵，组织内虫卵定量。结果：证明ＳＩＥＡ２６／２８ｋＤａ抗原能诱导小鼠产生抑制雌虫生殖的免疫力。与对照组比较，ＳＩＥ２６／２８ｋＤａ抗原免疫鼠减虫虽不明显，但肝组织内总卵数、     

日本血吸虫与华支睾吸虫和姜片虫之间的交叉抗原及其血清学反应性

应用ＳＤＳ－ＰＡＧＥ分离ＡＷＡ、ＣＳＡ和ＦＳＡ三种抗原的组分蛋白后，再以酶联免疫印迹技术（ＥＬＩＢ）进一步鉴定其组分蛋白的特异性蛋白带，分别显示１２条、１９条和２１条，其分子量（ＭＷ）范围分别为１３～６４ｋＤｅ、１６～１９８．５ｋＤａ和１４～１５０ｋＤａ。ＡＷＡ组分蛋白与兔抗ＳＥＡ免疫血清出现３条交叉反应带．ＭＷ为１７ｋＤａ、１４ｋＤａ和１３ｋＤａ，与抗ＣＳＡ的兔血清及华支睾吸虫病人血清显示３条（３８ｋＤａ、１７．５ｋＤａ及１７ｋＤｅ）交叉反应带．与抗ＦＳＡ的兔血清及姜片虫病人血清呈现５条（６４ｋＤａ、５９ｋＤｅ、５３ｋＤｅ、１７．５ｋＤａ和１７ｋＤａ）交叉反应带。实验结果表明，日本血吸虫成虫不仅与其虫卵之间存有交叉抗原，而且与华支睾吸虫和姜片虫成虫之间也存有交叉抗原及血清学交叉反应性。本研究为今后制备特异的血吸虫病血清学诊断抗原，提供了科学依据。     

抗日本血吸虫生殖及卵胚发育免疫的研究

抗日本血吸虫生殖及卵胚发育免疫的研究是血吸虫疫苗研究的重要策略之一。本研究探讨了未成熟卵等不同抗原免疫诱导宿主生产抗日本血吸虫生殖及卵胚发育免疫的效果。研究证明ＳＩＥＡ２６－２８ｋＤａ抗原能诱导小鼠产生抑制雌虫生殖和卵胚发育成熟的免疫力。与对照组比较，ＳＩＥＡ２６－２８ｋＤａ抗原免疫鼠减虫虽不明显，但肝组织内每雌产卵数、成熟卵数和粪卵数（ＥＰＧ）分别减少４８．１％、８３．６％、８７．３％，死亡卵数     

日本血吸虫未成熟卵可溶性抗原的初步分析

采用ＳＤＳ－ＰＡＧＥ和ＥＬＩＢ技术分析了日本血吸虫未成熟卵可溶性抗原（ＳＩＥＡ），并与可溶性成熟卵抗原（ＳＥＡ）进行比较，二者主要蛋白区带存在明显差异，同时，ＥＬＩＢ结果表明，ＳＩＥＡ中感染血清和免疫血清所识别的６５ｋＤａ、６２ｋＤａ、５０ｋＤａ、２８ｋＤａ和和２６ｋＤａ等抗原组分不出现于ＳＥＡ，推测其在ＳＩＥＡ诱导宿主产生抗雌虫生殖及抗卵胚发育免疫应答方面起作用。     

日本因吸虫各期虫体的酶联免疫电转移印迹分析及抗原定位研究

本文采用酶联免疫电转移印迹技术（ＥＩＴＢ）分析、比较日本血吸虫不同发育时期虫体特定的组分蛋白分子、雌虫、雄虫和虫卵抗原分别与相应的雌、雄虫和虫卵免疫血清反应呈现１７、２０和８条蛋白带，这三种抗原与其它不同时期虫体免疫血清反应，三者间及彼此间均出现交叉反应，而雌、雄虫和虫卵抗原与其它寄生虫感染血清作用没发现有叉及反应。应用间接荧光抗体试验（ＩＦＡ）和免疫酶染色试验（ＩＥＳ）对日本血吸虫抗的进行定位研     

日本血吸虫31／32kDa抗原纯化及诊断应用的研究

采用超凝胶柱层析结合超滤、透析、沉淀等方法分离纯化了日本血吸虫成虫３１／３２ｋＤａ抗原。经ＳＤＳ－ＰＡＧＥ、银染和免疫印迹分析，证明免疫及生化纯度。银染及ＰＡＳ证实此抗原是一种不含糖的多肽类组分。用于ＥＬＩＳＡ和ＩＨＡ诊断日本血吸虫病，与ＳＥＡ同样敏感，而特异性明显较优。     

日本血吸虫不同发育阶段抗原与抗LSA兔血清的交叉反应分析

目的为进一步阐明水蛭与日本血吸虫抗原交叉性机理提供依据。方法制备日本血吸虫子胞蚴抗原、尾蚴抗原、肝期童虫抗原、雌(雄)成虫抗原、虫卵抗原，采用Western blot方法对制备的日本血吸虫不同发育阶段抗原与水蛭可溶性抗原(LSA)免疫兔血清分别进行免疫印迹反应，分析其交叉性。结果LSA免疫兔血清能识别日本血吸虫不同发育阶段抗原，且所识别的抗原成分有明显差异，雌性成虫抗原和雄性成虫抗原分别有3条和2条蛋白组分可被LSA免疫血清识别，而不含抗虫卵和抗尾蚴抗体。结论在LSA成分中与血吸虫的共同成分主要存在于成虫中，LSA免疫血清中含有较多的抗雌性成虫抗体，而雄虫次之。     

钩虫抗原组份分析及其免疫反应性的研究

应用ＳＤＳ－ＰＡＧＥ和ＥＬＩＢ对十二指肠钩虫第三期幼虫和成虫可溶性抗原白组份及免疫反应性进行了比较研究。ＳＤＳ－ＰＡＧＥ电泳结果显示,钩虫幼虫和成虫蛋白区带数分别为２１条和１９条。ＥＬＩＢ反应表明,钩虫幼虫和成虫特异性抗原组份为４０～４１ｋＤａ和５４～５６ｋＤａ。该结果为钩虫病免疫学及其疫苗的研究提供了科学资料。     

日本血吸虫不同时期虫体蛋白质的SDS-聚丙烯酰胺凝胶电泳分析

用聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）分析，比较日本血吸虫不同发育时期虫体的蛋白质组成。尾蚴、雌虫、雄虫和虫卵各分离出２６、５１、５６和６０条蛋白带，其主带也不相同。雌、雄虫比较有９处蛋白带呈明显差异。日本血吸虫蛋白质组成有期和性的特异性，尾蚴、雌虫、雄虫和虫卵不仅有各自的特异蛋白，而且还有大量相同蛋白带。     

中国大陆日本血吸虫品系的研究Ⅺ.酶联免疫...

Antigens prepared from S. japonicum adult worms of different isolates i. e. Anhui, Hubei, Guangxi, Yunnan and Sichuan by origin were analyzed by enzyme linked immunoelectrotransfer blot (EITB) probed with rabbit anti-snail antibody (Figs 1,2). Anti-sera against Oncomelania h. hupensis from Anhui, Hubei, Yunnan and Sichuan localities were prepared separately and used in the tests. The EITB patterns were similar in S. j. isolates of Anhui and Hubei, and it was also the case among S. j. isolates of Yunnan and Sichuan except Yunnan female worms with a marked band of 84 kDa but it was almost invisible in EITB pattern of Sichuan female worms. Like Yunnan isolate, female worms of Guangxi isolate also showed marked 84 kDa bands. The EITB pattern of male worms from Guangxi isolate showed 2 main bands of mw > 130 kDa against anti-Anhui snail anti-serum, which corresponded with the male worms of Anhui isolate whereas the color of the bands was darker and denser in the former isolates, and these bands can not be seen in the male worms from isolates of Yunnan and Sichuan. Based on the above mentioned results in connection with the information about the susceptibility between different isolates of schistosomes and their snail hosts which we have reported before, some preliminary analysis and discussion were made.     

我国与日本山梨不同地域品系日本血吸虫DNA的限制性片段长度多态性分析

目的 :为了进一步了解中国各地日本血吸虫的特性 ,将其与日本山梨的日本血吸虫进行了虫体 DNA限制性片段长度多态性 ( RFLP)的分析和比较。方法 :以 p SM889为探针 ,与我国云南、四川、广西、湖北、湖南、江西、安徽、台湾和日本山梨的日本血吸虫雄虫 DNA进行杂交。结果 :经限制性内切酶 Eco RI酶切的我国 7地日本血吸虫 DNA与探针杂交的图谱显示弱杂交带数目呈现差异 ,而其 4条强杂交带数目则相同。但是 ,台湾及日本山梨的日本血吸虫 DNA杂交图谱不仅弱杂交带数目呈现差异 ,而且强杂交带数目亦有不同 ;前者的强杂交带数目为 3条 ,后者则为 5条。结论 :我国与日本山梨不同地区的日本血吸虫核基因组存在着遗传变异 ,台湾省的日本血吸虫的遗传变异程度远较其他省份的更为显著。     

我国与日本山梨不同地域品系日本血吸虫DNA的限制性片段长度多态性分析

目的 :为了进一步了解中国各地日本血吸虫的特性 ,将其与日本山梨的日本血吸虫进行了虫体 DNA限制性片段长度多态性 ( RFLP)的分析和比较。方法 :以 p SM889为探针 ,与我国云南、四川、广西、湖北、湖南、江西、安徽、台湾和日本山梨的日本血吸虫雄虫 DNA进行杂交。结果 :经限制性内切酶 Eco RI酶切的我国 7地日本血吸虫 DNA与探针杂交的图谱显示弱杂交带数目呈现差异 ,而其 4条强杂交带数目则相同。但是,台湾及日本山梨的日本血吸虫 DNA杂交图谱不仅弱杂交带数目呈现差异 ,而且强杂交带数目亦有不同 ;前者的强杂交带数目为 3条 ,后者则为 5条。结论 :我国与日本山梨不同地区的日本血吸虫核基因组存在着遗传变异, 台湾省的日本血吸虫的遗传变异程度远较其他省份的更为显著。     

Preliminary observations on the biology of the Indonesian strain of Schistosoma japonicum: animals and oncomelanid snails.

Laboratory animals and subspecies of Oncomelania hupensis were exposed to experimental infections with a strain of Schistosoma japonicum from the Lindu Valley, Central Sulawesi, Indonesia. All laboratory animals given 50 cercariae were suspecptible to infection with the greatest number of worms being found in mice followed by Wistar rats, Long Evans rats, Mongolian gerbils, wild rats, hamsters and guinea pigs. Rabbits and the Taiwan monkeys given larger numbers of cercariae were also susceptible to infections. Adult worms were found to reach the largest size in the rabbit and Taiwan monkeys but many worms from rats were immature. Eggs from the Taiwan monkey were larger than those from a dog and from mice. Six subspecies or geographic strains of Oncomelania hupensis were exposed to 3 to 5 miracidia of the Indonesia strain of Schistosoma japonicum and only Oncomelania hupensis lindoensis and Oncomelania upensis chiui from Taiwan were susceptibile and produced cercariae while Oncomelania hupesis formosana from Changhua, Ilan and Kaohsium countries of Taiwan as well as Oncomalania hupensis quadrasi from the Philippines were considered refractory to infection.     

DNA杂交鉴定血吸虫种株的研究

曼氏血吸虫及日本血吸虫成虫基因组DNA经内切酶BelII、BamHI、XbaI、及EcoRI消化后，与^32P标记的pSM889探针杂交，或经EcoRI消化后与pSM389探针杂交。上述两种情况下的杂交带型在两种血吸虫之间均有明显区别。日本血吸虫湖北、湖南、江西、浙江及云南隔离群成虫基因组DNA经EcoRI消化后与探针pSM389的杂交带型显示各隔离群间主带相同，而次要带则有不同程度的区别。其中湖南、湖北及浙江隔离群的次要带型互相接近，而江西及云南隔离群的次要带型与前3个隔离群有明显区别，它们二者之间亦显著不同。本工作表明非主要杂交带型可作为日本血吸虫种下分类的依据。     

间接免疫酶染法检测日本血吸虫与湖北钉螺相同抗原的研究

用日本血吸虫成虫及湖北钉螺冰冻切片作抗原,采用间接免疫酶染法分别检测兔抗钉螺免疫血清和感染血清。结果表明日本血吸虫雌雄成虫、钉螺头足部、肝脏分别出现阳性反应。提示:日本血吸虫与其中间宿主湖北钉螺之间确实存在相同的抗原成分,     

Studies on the strain differences of Schistosoma japonicum in the mainland of China. II. Susceptibility of mammalian hosts

Six species of animals were percutaneously exposed to cercariae obtained from pools of naturally infected snails from different isolates of S. japonicum in the mainland of China as shown in Tables 1 and 2. Our results indicated that with the exception of rat, all animals under study were permissive host though their worm recovery rates varied with different isolates. The mean prepatent period in different host species were 35.0 +/- 0.8 to 36.4 +/- 1.0 days for Anhui isolate; 34.5 +/- 1.2 to 36.4 +/- 1.2 days for Hubei isolate; 34.5 +/- 1.3 to 35.8 +/- 0.6 days for Sichuan isolate; 35.1 +/- 1.0 to 37.3 +/- 1.9 days for Guangxi isolate and 36.1 +/- 1.9 to 37.8 +/- 0.8 days for Yunnan isolate. In general, the prepatent period was longer in the C57 BL inbred mice, hamsters and rhesus monkeys infected with Yunnan and Guangxi isolates, than that with Sichuan isolate. This result also indicates that the prepatent period of the strain of S. japonicum defined by Dr. Vogel as a Chinese strain which had been originated from the cercariae in infected Oncomelania hupensis hupensis snails from Jiaxing, Zhejiang (Kashing, Chekiang), China, and established in Tropeninstitut in Hamburg since 1937 by repeated passages in dogs and laboratory-bred O. h. hupensis snails, was 5-8 days longer than that of all the isolates under our study, probably due to some behavioral change. We suggest, therefore, that the Vogel's stock of S. japonicum should be termed as "Chinese Vogel strain".     

Susceptibility of Oncomelania hupensis subspecies to infection with geographic strains of Schistosoma japonicum

Subspecies of Oncomelania hupensis were exposed to infection with zoophilic and anthropophilic strains of Schistosoma japonicum. The snails were Philippine O. h. quadrasi from the islands of Bohol, Leyte and Mindanao, O. h. formosana from Changhua and Ilan, and O. h. chiui from Taiwan, O. h. hupensis from China, O. h. nosophora from Japan and O. h. lindoensis from Indonesia. Anthropophilic strains of S. japonicum were from Bohol and Leyte in the Philippines, China, Japan and Indonesia. The zoophilic strains were from Changhua and Ilan, Taiwan. All geographic strains of the parasite developed exceptionally well and produced cercariae in 32-50% of O. h. chiui and 5-43% of the natural snail hosts. Development in other subspecies of snails with geographic strains of the parasite were variable. Only a few O. h. formosana (Changhua) could be infected with the parasites from other geographic areas and only one O. h. quadrasi each from Mindanao became infected with the Bohol and Changhua strains of S. japonicum.     

中国大陆日本血吸虫品系的研究：Ⅰ,幼虫—钉螺的相容性

本文进行了安徽贵池、湖北监利、广西桂平、四川天全、云南洱源及福建福清6地日本血吸虫和钉螺的人工交互感染实验。结果表明各地日本血吸虫与钉螺呈现不同的相容性。湖北、安徽两地的血吸虫幼虫与钉螺的相容性无差别。湖北、安徽两地血吸虫幼虫很难在云南、四川两地钉螺体内发育成熟逸出尾蚴:反之,云南、四川两地血吸虫幼虫与湖北、安徽两地钉螺却可相容。广西的血吸虫幼虫或钉螺与安徽的钉螺或幼虫亦可交互感染而相容,但尾蚴逸出前期均较感染本地钉螺的为长;广西的血吸虫幼虫与四川、云南、福建的钉螺并不相容。福建的钉螺与安徽、云南两地血吸虫幼虫均可相容。本文还提出了在研究日本血吸虫与钉螺的相容性时,必须包括逸出尾蚴的钉螺感染率和尾蚴逸出前期的数据,才能正确评价两者的相互关系。