东方田鼠抗日本血吸虫抗体水平检测与分析

东方田鼠具有抗日本血吸虫病的特性，观察东方田鼠感染日本血吸虫尾蚴后的特异性抗体水平变化，可为阐明其抗病机制提供依据。本试验用间接ELISA方法检测东方田鼠抗成虫可溶性抗原（SWAP）和几种日本血吸虫基因重组抗原（Sj28-GST、LHD-SjTP1、C-SjParamyosion）的特异性抗体水平动态变化，并检测东方田鼠感染日本血吸虫尾蚴前、后的IgG3亚类特异性抗体水平。结果东方田鼠在攻击日本血吸虫尾蚴后，抗SWAP的特异性抗体水平有所升高，而抗几体基因重组抗原的特异性抗体水平低，没有显著变化。东方田鼠正常血清中抗SWAP和IgG3亚类抗体的OD值是牛血清白蛋白（BSA）对照的31倍。结果表明，在东方田鼠血清中有较高水平的SWAP的IgG3亚类抗体，值得进一步深入探讨。     

东方田鼠和小鼠感染日本血吸虫后组织与血清蛋白组分差异比较研究

探讨东方田鼠和小鼠感染日本血吸虫前后不同时点组织与血清中蛋白质电泳谱的变化。分别提取东方田鼠感染日本血吸虫(0、7、12、25 d) 和小鼠感染日本血吸虫(0、7、12、45 d)后的肝脏、脾脏、肺脏蛋白及血清进行聚丙烯酰胺凝胶电泳,对结果比较分析并进行组织病理形态学观察。结果显示,小鼠感染日本血吸虫第7和12天后肝脏150 ku附近蛋白质条带较东方田鼠表达量明显增高;东方田鼠在感染日本血吸虫后40~50 ku附近蛋白质条带较正常东方田鼠表达明显上调;感染第7天后肺脏40 ku附近蛋白质条带较其他时间点变化明显;小鼠感染日本血吸虫第7、12和45天后血清中IgG类蛋白质表达量较东方田鼠显著增高。东方田鼠和小鼠在日本血吸虫感染后不同时点间的组织及血清蛋白组分存在差异,东方田鼠肝脏和血清中150 ku附近蛋白条带较其他动物特异,蛋白质表达丰度高,该蛋白可能与东方田鼠抗日本血吸虫相关。     

东方田鼠感染日本血吸虫前后血常规和血清生化指标的动态变化

东方田鼠具有天然抗日本血吸虫病特征,为了探索东方田鼠抗日本血吸虫机制,本研究分析了东方田鼠和小鼠感染日本血吸虫尾蚴前后血液细胞以及生化指标的动态变化。东方田鼠和BALB/c鼠定量感染日本血吸虫尾蚴,于感染后第0、1、3、5、8、12、16、21、28、35、42 d分别采集抗凝血和血清,进行血常规和血清生化分析。血常规检测结果表明,东方田鼠感染后第3~16 d,WBC数量显著升高(Neu数量显著升高),而小鼠血常规检测结果正常;血清生化结果显示,东方田鼠感染日本血吸虫后ALB、HDL-C显著升高,小鼠各指标变化不明显。本研究结果显示东方田鼠天然免疫细胞Neu和血清ALB可能与其抗日本血吸虫特性相关,对补体杀伤日本血吸虫机制研究具有重要的指导意义。     

东方田鼠组织与血清中抗日本血吸虫相关蛋白的分离与筛选

本试验以兔、牛、人及感染日本血吸虫后小鼠血清蛋白为对照,分别提取东方田鼠和小鼠的心脏、肝脏、脾脏、肺脏、肾脏、肌肉、皮肤、骨髓和脑组织蛋白,进行聚丙烯酰胺凝胶电泳,综合比较和筛选东方田鼠组织及血清中潜在的抗日本血吸虫相关蛋白组分。结果表明,随着感染日龄的增加,小鼠在感染日本血吸虫后12和45 d,其血清中85 ku附近蛋白质条带较东方田鼠表达量明显上调,且有继续升高的趋势;东方田鼠肝脏及血清中150 ku组分蛋白电泳条带与其他动物相同组织中相应蛋白质在表达量存在显著差异,蛋白质表达丰度高,该蛋白质组分可能与东方田鼠抗日本血吸虫作用相关。     

东方田鼠抗日本血吸虫病现象的观察

本试验分别应用１０００条、２００条和４０条日本血吸虫尾蚴人工攻击感染东方田鼠,金黄地鼠和Ｂａｌｂ／ｃ小鼠,在攻击后第１、２、３、４、７、９、１２、１５、１６、１８、１９、２１、２２、２４、４２天时分别将其剖杀,集虫、观察。结果观察到血吸虫在东方田鼠体内不能发育成熟,在攻击后１９天时虫体已全部死亡。而黄金地鼠和Ｂａｌｂ／ｃ小鼠的虫体回收率分别为６５％和６０．６％。在虫体移行时,东方田鼠肺组织和肝组织     

感染日本血吸虫前后东方田鼠血清补体的动态变化和杀虫作用

东方田鼠是迄今为止发现的唯一一种具有天然抗日本血吸虫病特性的哺乳动物,但具体抗病机制尚不明确。本研究应用组织培养法比较了东方田鼠和BALB/c鼠感染日本血吸虫尾蚴后1、3、8、12、15 d的皮肤、肺脏和肝脏中童虫回收情况,结果表明东方田鼠对日本血吸虫的杀伤主要发生在感染后1～3 d和8～12 d。应用RT-PCR和ELISA检测东方田鼠和BALB/c鼠的补体相关分子C3、C9、CD59和DAF的转录水平,结果显示BALB/c小鼠各指标变化不明显,东方田鼠感染后8 d各因子转录水平显著升高。体外杀虫试验结果显示,东方田鼠正常血清和灭活补体血清孵育的日本血吸虫童虫死亡率分别为85.50%±7.15%和60.99%±8.61%,差异具有显著统计学意义(P 0.001)。结果表明东方田鼠补体对日本血吸虫童虫具有显著杀伤效果,东方田鼠早期对日本血吸虫的杀伤作用可能和补体相关。     

东方田鼠肝脏裂解物筛选日本血吸虫成虫噬菌体展示cDNA文库

东方田鼠对血吸虫具有天然抗性。为寻找东方田鼠抗血吸虫抗性相关靶基因，探索其分子机理，我们将日本血吸虫童虫与东方田鼠不同组织／器官裂解物共培养，比较它们对血吸虫童虫的杀伤效果，之后，以杀伤效果较好的肝脏裂解物为探针，以亲和淘洗方法筛选日本血吸虫成虫（44d）噬菌体展示cDNA文库。经三轮筛选后，随机挑取92个噬菌体克隆进行序列测定及生物信息学分析，获得了19个有效EST序列，其中15条EST序列与已知基因或表达序列标签同源，4个EST序列与已知基因或表达序列标签均无同源性，为新的表达序列标签。对19个有效EST编码蛋白的功能预测结果显示，其中6个为卵壳蛋白（Eggshell protein）相关基因，10个EST及其编码蛋白的功能与血吸虫基因表达调控及蛋白质加工修饰相关。研究结果提示，东方田鼠直接或间接地影响血吸虫基因表达，进而影响血吸虫的发育，可能是东方田鼠具有抗血吸虫特性的一个重要分子机理。     

东方田鼠净化的研究初报

东方田鼠隶属仓鼠科(Cricetidae)、田鼠亚科(Microti-nae)、田鼠属(Microtus)。早在上世纪50年代人们就注意到东方田鼠对日本血吸虫具有天然抗感染性〔1〕,近年来随着许多学者对实验室饲养和繁殖的东方田鼠从其遗传学、免疫学等多方位有了较为系统的观察研究〔2~5〕,以期推动东方田鼠抗病基因的重大突破。但要充分利用东方田鼠这一种质资源,则必须对东方田鼠进行实验动物化〔6~8〕,通过将其净化,为抗日本血吸虫机制研究提供安全、理想的动物模型。而东方田鼠的净化工作尚无先例,因此净化方案的确立要综合多方面的因素考虑,既科学又有可操…     

东方田鼠IgG和IgG3对日本血吸虫抗原反应特性的研究

本文对东方田鼠(Microtus fortis)总IgG和IgG3对日本血吸虫抗原的反应特性进行了研究.纯化东方田鼠IgG,用间接ELISA法检测不同浓度东方田鼠总IgG和IgG3对日本血吸虫童虫可溶性抗原(soluble schistosomula antigen,SSA)、成虫可溶性抗原(soluble adult worm antigen,SWA)、虫卵可溶性抗原(soluble egg antigen,SEA)、童虫不可溶性抗原(insoluble schistosomula antigen,ISSA)、雄虫不可溶性抗原(insoluble male worm antigen,ISMA)、雌虫不可溶性抗原(insoluble female worm antigen,ISFA),以及基因工程抗原rSj14、rSj23-HLD、rSJ28GST、rSjPP的反应情况.结果显示,HRP标记免抗东方田鼠IgG抗体和羊抗小鼠IgG3抗体作为二抗,A值变化规律一致.该结果显示东方田鼠总IgG和IgG3与日本血吸虫抗原可能存在非特异性反应.     

封闭繁殖的成年东方田鼠的生物学特性Ⅰ.心率、血压的检测和正常值的估计

东方田鼠 ( Microtus fortis)是进行神经行为比较研究的理想材料。更为重要的是 ,该鼠长期栖息在日本血吸虫疫区 ,与其他啮齿动物不同 ,是目前所知的唯一对日本血吸虫具有天然抗感染性的啮齿动物 [1]。若将东方田鼠按实验动物的要求进行培育 ,实现实验动物化 ,将为生物医学提供一种极有价值的实验动物新品种。本研究对封闭饲养的成年东方田鼠的心率、血压进行了测定 ,以提供其基本的生理学数据。1　材料与方法1 .1　动物　自中国科学院长沙农业现代化研究所引进。封闭繁殖 4～ 5代的成年东方田鼠 2 0对 ,饲养在 40 cm× 2 8cm× 1 5cm的塑…     

东方田鼠肺脏裂解物筛选日本血吸虫成虫噬菌体展示cDNA文库

目的寻找与东方田鼠抗性相关的血吸虫靶基因。方法以东方田鼠肺脏裂解物为探针,亲和淘洗筛选日本血吸虫成虫（44d）噬菌体展示cDNA文库。三轮筛选后．随机挑取76个阳性噬菌体克隆进行序列测定及生物信息学分析。结果共获得了13个有效EST序列,其中8条EST序列与已知基因或表达序列标签同源,5个EST序列与已知基因或表达序列标签均无同源性,为新的表达序列标签。功能预测结果显示．这13个有效EST编码蛋白主要与血吸虫基因表达调控及蛋白质加工修饰相关。结论东方田鼠直接或间接地影响血吸虫基因表达,进而影响血吸虫的发育,可能是东方田鼠具有抗血吸虫特性的一个重要分子机理。     

Use of an enzyme-linked immunosorbent assay to measure antibody levels in turkey breeder hens, eggs, and progeny following natural infection or immunization with a commercial Bordetella avium bacterin

Twelve large white turkey hens were immunized with a commercially available Bordetella avium bacterin. Hens and eggs were tested using an enzyme-linked immunosorbent assay (ELISA) to determine the response to the bacterin. Three hundred poults were then obtained from two commercial flocks, the hens of one flock having been immunized with the same bacterin used on the group of 12 turkeys. Titers of the poults were monitored for 7 weeks, and poults were challenged by exposure to infected poults at 1, 7, 14, and 21 days post-hatch. Hens produced an antibody response following immunization, with a parallel antibody response being detected in eggs. Maternal antibodies were present in poults from immunized hens. Poult titers declined to near the level of poults from unimmunized hens by 14 days of age. Poults from immunized hens challenged at 1 and 7 days were resistant to development of clinical disease and gross lesions, whereas all poults from unimmunized hens exhibited clinical signs and gross lesions. After 14 days, the resistance of both groups to development of clinical disease, became near equal, neither group being affected as severely as the unimmunized hens challenged at days 1 and 7. Six commercial turkey breeding flocks and their progeny that had not been vaccinated for B. avium and had no history of B. avium infection were evaluated with the B. avium ELISA. There were variations between the flocks, with poult titers reflecting those found in the hens.     

Characterization of a novel vaccine candidate and serine proteinase inhibitor from Schistosoma japonicum (Sj serpin)

Serine proteinase inhibitors (serpins) represent an important superfamily of endogenous inhibitors that regulate proteolytic events active in a variety of physiological functions. Immunological screening of a Schistosoma japonicum adult worm cDNA expression library with sera of Microtus fortis, a naturally resistant vertebrate host, has identified one clone that encoded for a sequence homologous to those of the serpin superfamily. The full-length sequence encoding S. japonicum serpin (Sj serpin) was amplified from adult worm cDNA by using 5'-RACE-PCR and subsequently cloned into the prokaryotic expression vector pET28c. The full-length Sj serpin fusion-protein with his-tag was expressed in E. coli, purified by affinity chromatography and used to immunize New Zealand white rabbits. Sj serpin is located on the tegument in S. japonicum adult worms. C57BL/6 mice immunized with Sj serpin induced the production of high levels of specific IgE and IgG1 subclass antibodies as well as a marked IL-4 response. Lymphocyte surface marker analysis revealed proliferation of CD19 expressing B cells, indicating a predominant Th2-type response to Sj serpin. Immunized mice developed moderate protection against infection of S. japonicum as demonstrated by a 36 and 39% reduction in the recovery of adult worms and eggs, respectively. These data suggested a role for Sj serpin as a vaccine candidate or as a novel target for anti-schistosome drugs.     

Ovine coccidiosis: heavy infection in young lambs increases resistance without causing disease

Seven groups of twin lambs, kept with their dams on pasture, were given single oral inoculations of 10,000 oocysts of Eimeria ovinoidalis and 10,000 E crandallis at one, two, four, seven, 14, 21 or 28 days after birth, respectively. All were then challenged with 100,000 of each species at 42 days of age. An eighth group was challenged without having received the earlier 'immunising' inoculum, and a ninth group was not inoculated at all. Bodyweights, faecal consistency, oocyst output, and serum coccidial antibody levels were monitored up to 12 weeks of age. No clinical response was detected to inoculation up to four days of age. Loosening of faeces and a slight setback in weight-gain occurred in lambs inoculated seven, 14 and 21 days after birth; inoculation 28 days after birth caused severe diarrhoea and weight loss. Challenge at 42 days caused severe coccidiosis with 50 per cent mortality in the 'unimmunised' group. In those that had received 'immunising' inoculations, the challenge at 42 days caused some diarrhoea and some weight loss, but much less than in the 'unimmunised' lambs. The later the 'immunisation', the less severe was the disease attributable to the challenge at 42 days. Serum antibody levels correlated fairly closely with resistance to the disease. It was concluded that very young lambs were resistant to the pathogenic effects of some coccidia, but were able to respond to them immunologically.     

大肠杆菌油佐剂灭活苗免疫雏鸡抗体消长规律的研究

本试验用鸡源致病性大肠埃希氏菌Ｏ２血清型制成的油佐剂灭活苗对１５日龄雏鸡进行免疫接种，以测定其抗体消长规律；对免疫后不同抗体滴度的雏鸡进行了攻毒试验．结果表明，免疫１周后可监出抗体，２～３周抗体达到高峰，抗体可持续１０周左右．抵抗同型大肠杆菌攻霉的最低抗体滴度为１：１６～１：３．免疫保护期可达２个目以上．     

Duration of immunity induced in chickens by an attenuated live Salmonella enteritidis vaccine and an inactivated Salmonella enteritidis/typhimurium vaccine

The aim of this study was to examine the duration of immunity of different vaccination schemes using the S. enteritidis live vaccine Gallivac Se and the S. enteritidis-S. typhimurium inactivated vaccine Gallimune Se+St. Three groups of Lohman Brown chickens were used. Group one was vaccinated three times orally with Gallivac Se at weeks one, seven and 13 of age. Group two was vaccinated twice orally with Gallivac Se in weeks one and seven and once i.m. with Gallimune Se+St in week 14 of age. A third group was not vaccinated and served as the control group. Eight randomly selected chickens from each of the three groups were challenged with a nalidixic acid resistant S. enteritidis PT4 strain in weeks 24, 51 and 71 of age and the same number of animals were challenged with a S. typhimurium DT 104 strain in weeks 26, 54 and 73 (75) of age.The chickens were euthanised seven days post challenge and the number of challenge strain organisms (log10 cfu) in the liver and on caecal mucosa was determined.The quantitative investigation of the challenge strain in the liver and caecal mucosa revealed a statistically significant (p < 0.05) lower challenge strain burden in the vaccinated groups compared with the non-vaccinated control group up to week 71 (73) of age. The protective effects were demonstrated for both challenge strains.