日本血吸虫Sj31核酸疫苗联合IFN-γ重组质粒诱导小鼠保护性免疫的研究

目的　探讨干扰素 γ重组质粒对日本血吸虫组织蛋白酶B核酸疫苗在小鼠抗血吸虫作用的影响。　方法　将小鼠干扰素 γ基因PCR扩增片段克隆入真核表达载体pCDNA3 1以构建重组真核表达质粒pCDNA3 1 IFN γ ,并与日本血吸虫组织蛋白酶B真核表达质粒VR10 12 Sj3 1一同免疫小鼠。小鼠分为 4组 ,其中实验组每鼠同时肌注VR10 12 Sj3 1及pCDNA3 1 IFN γ各 10 0 μg ,3个对照组分别为VR10 12 Sj3 1肌注 10 0 μg ,pCDNA3 1 IFN γ肌注 10 0 μg和载体VR10 12及pCHAN3 1肌注各 10 0 μg。共免疫 3次 ,每次间隔 2周。于末次免疫后两周免疫组化检测表达质粒在小鼠肌细胞的表达 ,于末次免疫后 3周经小鼠皮肤攻击感染 40± 1条日本血吸虫尾蚴。 45d后杀小鼠计算减虫率。　结果　VR10 12 Sj3 1及pCDNA3 1 IFN γ均在小鼠肌细胞表达 ,日本血吸虫Sj3 1核酸疫苗联合IFN γ重组质粒免疫可诱导小鼠产生 2 7 3 7%的减虫率 ,与日本血吸虫Sj3 1核酸疫苗单独免疫组比较减虫率显著 (P <0 0 5 )。　结论　IFN γ表达质粒能增强日本血吸虫组织蛋白酸B核酸疫苗的抗日本血吸虫的作用     

不同载体的日本血吸虫DNA疫苗诱导小鼠免疫效果的观察

目的　观察不同真核表达载体的日本血吸虫 DNA疫苗的免疫效果 ,筛选合适的血吸虫 DNA疫苗。方法　将小鼠分成 5组 ,于第 0、 3、 5周将日本血吸虫真核表达载体 p BK- Sj2 3 ,p BK- Sj2 6以及 p CD- Sj2 3 ,p CD- Sj2 6,分别接种小鼠股四头肌 ,第 9周每组小鼠以 40± 2条血吸虫尾蚴攻击感染 ,攻击感染 6周后 ,剖杀小鼠 ,计算减虫率和减卵率 ,比较含血吸虫同一抗原分子的不同载体诱导小鼠的抗攻击感染能力。结果　发现疫苗 p CD- Sj2 3和 p CD- Sj2 6诱导小鼠的减虫率和减卵率分别为 3 0 .5 0 %和 3 3 .0 2 % ;疫苗 p BK- Sj2 3和 p BK- Sj2 6诱导小鼠的减虫率和减卵率分别为 18.2 4%和 2 1.70 % ,含血吸虫同一抗原分子的不同载体诱导小鼠的抗攻击感染能力具有显著性差异。结论　由真核表达载体p CD构建的血吸虫 DNA疫苗比由真核表达载体 p BK构建的血吸虫 DNA疫苗具有更好的抗血吸虫感染的能力     

日本血吸虫多价DNA疫苗pBK—Sj26（Sj32）—Sj23免疫效果的观察

为了观察血吸虫病多价DNA疫苗的保护力,将小鼠分成5组：空白对照组、空质粒对照组、单价抗原DNA疫苗pBK-CMV-Sj23组、多价抗原DNA疫苗pBK-CMV-Sj26-Sj23和pBK-CMV-Sj32-Sj32组。大量提取各组质粒DNA后,各组于0、3、5周在BALB／c小鼠股四头肌注射相应质粒DNA,9周用血吸虫尾蚴攻击感染,15周剖杀小鼠计算减虫率及减卵率。结果显示：与对照组比较,实验组小鼠减虫率及减卵率及极显著性差异（P＜0.01）;与单价pBK-CMV-Sj23组比较,多价DNA疫苗组的减虫率及减卵率有显著性差异,提示血吸虫多价DNA疫苗诱导小鼠对血吸虫的保护力优于单价DNA疫苗。     

日本血吸虫多价DNA疫苗pBK-Sj26(Sj32)-Sj23免疫效果的观察

为了观察血吸虫病多价DNA疫苗的保护力,将小鼠分成5组空白对照组、空质粒对照组、单价抗原DNA疫苗pBK-CMV-Sj23组、多价抗原DNA疫苗pBK-CMV-Sj26-Sj23和pBK-CMV-Sj32-Sj23组.大量提取各组质粒DNA后,各组于0、3、5周在BALB/c小鼠股四头肌注射相应质粒DNA,9周用血吸虫尾蚴攻击感染,15周剖杀小鼠计算减虫率及减卵率.结果显示与对照组比较,实验组小鼠减虫率及减卵率有极显著性差异(P<0.01);与单价pBK-CMV-Sj23组比较,多价DNA疫苗组的减虫率及减卵率有显著性差异.提示血吸虫多价DNA疫苗诱导小鼠对血吸虫的保护力优于单价DNA疫苗.     

核酸疫苗Sj31BIN联合IL-12免疫抗日本血吸虫攻击感染的实验研究

目的 :在证明日本血吸虫重组蛋白酶B核酸疫苗Sj31BIN具有抗生殖免疫作用的基础上 ,联合使用IL 12 ,观察IL 12是否具有佐剂效果。方法 :分别用Sj31BIN +IL 12和IL 12免疫Balb/C小鼠。攻击感染后 6周计数成虫负荷和组织内虫卵数及肝脏表面虫卵结节数。结果 :Sj31BIN +IL 12免疫小鼠可降低成虫发育率。肝组织减卵率为5 9 74% ;肠组织减卵率为 5 9 6 0 % ;肝脏表面虫卵结节减少率为 71 30 % ;单独使用IL 12有一定的免疫保护作用。结论 :Sj31BIN +IL 12能诱导小鼠产生较强的抗生殖免疫作用。     

日本血吸虫组织蛋白酶B DNA疫苗不同免疫途径免疫保护效果研究

目的:观察本室所构建日本血吸虫大陆株31kDa组织蛋白B DNA疫苗在BALB/c小鼠体内的不同免疫途径的免疫保护效果.方法:用纯化的空白质粒载体VR1012、重组质粒VR1012-Sj31免疫BALB/c鼠,将36只6～8周龄BALB/c鼠随机分为3组,每组12只,对照组(A组)肌肉注射空白质粒载体VR1012,实验组(B组)皮下注射重组质粒VR1012-Sj31,C组肌肉注射重组质粒VR1012-Sj31.质粒剂量均为100μg,每隔2w免疫1次,共免疫3次,第3次免疫后第21d,每只鼠经腹部感染10条尾蚴,42d后剖杀计数各小鼠成虫数和每克肝卵数,观察诱导产生的减虫和减卵效果,并用ELISA分析小鼠血清中的抗体.结果:ELISA分析表明,第3次免疫后小鼠出现特异性IgG抗体.与空白质粒对照组比较,2个实验组的减虫率分别为15.0%(P＞0.05)和25.0%(P＜0.05),减卵率分别为38.22%和54.90%(P＜0.001).与皮下免疫组相比,VR1012/Sj31肌肉免疫组的减卵率分别为26.99%(P＜0.001).结论:DNA疫苗VR1012-Sj31能诱导小鼠产生一定水平的抗日本血吸虫感染保护作用,肌肉注射的保护效果大于皮下注射.     

日本血吸虫复合DNA疫苗的组织表达及免疫保护效果研究

目的 :观察本课题组所构建日本血吸虫大陆株复合DNA疫苗VR10 12 SjGST和VR10 12 SjGST Sj32在小鼠肌肉组织中的表达 ,并进行免疫保护效果测定。方法 :用纯化质粒免疫昆明鼠 :4 8只小鼠分为 4组 ,两个对照组的小鼠分别于股四头肌注射生理盐水 10 0 μl或空质粒VR10 12 10 0 μg,两个实验组的小鼠则同法分别注射VR10 12 SjGST和VR10 12 SjGST Sj32各 10 0 μg。末次免疫后 ,每组解剖两只小鼠 ,以间接免疫荧光法观察SjGST、Sj32在肌肉组织中的表达。其余每只鼠经腹部感染 10条尾蚴 ,4 5d后剖杀计数各小鼠成虫数和肝卵数。结果 :小鼠肌肉组织表达出抗原特异性蛋白质抗原分子。与生理盐水组比较 ,两个实验组的减虫率分别为 33.9%和及 2 7.14 %(均为P <0 .0 5 ) ,减卵率分别为 6 1.86 %和 6 8.87% (均为P <0 .0 0 1)。与VR10 12 SjGST组相比 ,VR10 12 SjGST Sj32组的减卵率为 18.5 1% (P <0 .0 5 )。结论 :DNA疫苗VR10 12 SjGST和VR10 12 SjGST Sj32能在组织中正常表达 ,能诱导小鼠产生一定水平的抗日本血吸虫感染保护作用 ,复合疫苗的保护效果要大于单价疫苗     

日本血吸虫DNA多价疫苗SjGST-FABP/pcDNA3的构建及其保护性免疫研究

目的构建日本血吸虫DNA多价疫苗SjGST-FABP/pcDNA3,用以免疫小鼠,观察其在小鼠抗血吸虫感染中的免疫保护作用。方法根据质粒pGEX-4T-1中SjGST-ORF和SjFABP基因序列,利用基因重组、PCR等技术将SjGST和SjFABP编码基因拼接在一起,得到融合基因SjGST-FABP,将融合基因SjGST-FABP定向克隆到pcDNA3多克隆位点上,转化大肠杆菌,经质粒扩增和DNA序列测定后,进行小鼠动物免疫和日本血吸虫尾蚴攻击感染及免疫保护性评价。结果成功构建了日本血吸虫DNA多价疫苗SjGST-FABP/pcDNA3。免疫小鼠获得42.39%的减虫率和56.09%肝减卵率（P〈0.05）。结论日本血吸虫DNA多价疫苗SjGST-FABP/pcDNA3可诱导部分抗血吸虫尾蚴攻击感染的免疫保护效果,具有疫苗研究与开发价值。     

日本血吸虫Calpain的DNA疫苗诱导小鼠免疫保护性研究

目的　研究日本血吸虫中国大陆株Calpain的DNA疫苗对Balb/c小鼠的免疫保护性作用。方法　大量制备pVAC质粒DNA和pVAC -CalpainDNA疫苗 ,Balb/c小鼠 5 0只 ,随机均分为对照组和实验组 ,对照组每只小鼠的股四头肌注射接种 10 μgpVAC质粒DNA ,实验组以同样方式注射 10 μgpVAC -CalpainDNA疫苗 ,第 3周加强免疫一次。第 4周每只小鼠经腹部皮肤感染 ( 2 5± 1)条尾蚴 ,感染 6周后剖杀 ,从门静脉灌流收集计成虫数、碎脾计T淋巴细胞总数、从各鼠肝脏的同一部位切下小块肝组织 ,置 5 %KOH溶液消化后 ,计算每克肝组织虫卵数 ,比较减虫率、减雌率和减卵率。于免疫前、第一次免疫后 2周和感染后 2周分别从尾静脉采血 ,用ELISA测定抗体水平。结果　与对照组相比 ,实验组获得 36.84%的减虫率、36.36%的减雌率和 43.31%的减卵率 ,感染后实验组小鼠IgG抗体水平升高幅度大于对照组 ,对照组T淋巴细胞数显著多于实验组。结论　CalpainDNA疫苗可诱导小鼠产生较好的免疫保护作用 ,具有较大的发展潜力     

日本血吸虫DNA疫苗辅以FQ2诱导小鼠保护性免疫的实验研究

目的探讨日本血吸虫Sj26GSTDNA疫苗辅以佐剂FQ2共同作用对小鼠的免疫保护作用。方法采用FQ212"g/只分别与Sj26GSTDNA疫苗混合经股四头肌接种BALB/c小鼠,共接种3次,同时设Sj26GSTDNA疫苗组与生理盐水对照组,接种后4周以日本血吸虫尾蚴攻击感染,感染后6周剖杀小鼠,计数减虫率和减卵率。结果FQ2 疫苗组的减虫率与减卵率分别为34.24%与41.12%,而疫苗组的减虫率与减卵率各为28.76%与29.03%,结果显示FQ2 疫苗组的减虫率与减卵率均高于疫苗组,减卵率差异显著(P<0.05)。结论FQ2对Sj26GSTDNA疫苗有明显的增效作用,提高了DNA26Kda疫苗对小鼠的保护作用。     

Immune responses against Schistosoma japonicum after vaccinating mice with a multivalent DNA vaccine encoding integrated membrane protein Sj23 and cytokine interleukin-12

Background The vaccination of mice with DNA encoding single candidate antigens has failed to induce significant protection against Schistosoma japonicum ( S. japonicum) challenge infections. In this study, we evaluated the feasibility of using a multivalent DNA vaccine which co-expressed S.japonicum integral membrane protein Sj23 and murine cytokine IL-12 to induce protective immune responses.Methods The plasmid pVIVO2-IL12-Sj23, a eukaryotic expression vector expressing Sj23 and murine IL-12 simultaneously, was constructed, identified, and tested for expression in vitro. Its ability to protect against S. japonicum challenge infections was analyed according to worm reduction rate and egg reduction rate after vaccination of BALB/c mice. The serum levels of specific IgG antibody were determined by enzyme-linked-immuno sorbent assay (ELISA) and Western blot analysis. Using cultured spleen cells, IFN-γ and IL-4 post-stimulation were quantified by ELISA. The phenotypes of splenocyte populations were analyzed by flow cytometry (FCM).Results The plasmid DNA pVIVO2-IL12-Sj23 was proven to express well in vitro by transient transfection of HEK-293 cells. Immunization resulted in a worm reduction rate of 45. 53% and egg reduction rate of 58.35%. ELISA and Western blot analysis indicated that immunized mice generated specific IgG against Sj23. Spleen cells showed significant increases in IFN-γ but decreases in IL-4.No significant differences in CD4^ and CD8^ subgroup ratios were observed after the challenges.Conclusions The multivalent DNA vaccine pVIVO2-1L12-Sj23 is sufficient to elicit moderate but highly significant levels of protective immunity against challenge infections. Cytokine IL-12, as a gene adjuvant, was able to enhance the Thl responses and, hence, the protective immunity.     

Construction and Expression of DNA Vaccine pIRES-Sj97-Sj14-Sj26 and Its Immunogenicity in Mice

To find a new preventive strategy for the infection of Schistosoma japonica, plasmid pIRES-Sj97-Sj 14-Sj26 that contains fatty binding protein （Sj 14）, GST （Sj26） and paramyocin （Sj97） that are expressed on the membrane, was constructed. RT-PCR was used to detect the expression of Sj 14 mRNA, Sj26 mRNA and Sj97 mRNA in the Hela cells, the indirect immunofluorescent test was employed for the detection of the expression of trans-membrane Sj26 after the plasmid was transfected into Hela cells. Fifty BALB/c mice were randomly divided into 5 groups and plRES-Sj97-SjI4-Sj26 plasmid DNA, plRES-Sj 14-Sj26 plasmid DNA, plRES-Sj26 plasmid DNA, plRES blank vector and normal saline were respectively injected into the quadriceps muscles of thigh Eight weeks after the immunization the mice were killed and significantly higher level of IgG was detected in the plRES-Sj97-Sj 14-Sj26 group as compared with the plRES blank vector, normal saline and plRES-Sj26 groups （P〈 0.01） and the plRES-Sj 14-Sj26（P〈0.05）. Single splenocyte suspension was prepared to detected the level of IFN-T by ELISA and the lymphocyte stimulating index （SI） by MTT. SI was significantly higher of in the plRES-Sj97-Sj 14-Sj26 group than in other groups （P〈 0.01）, while the IFN-T level was significantly higher the plRES-Sj97-Sj 14-Sj26 group than in plRES blank vector and normal saline groups （P〈0.01）, but no significant differences were found when compared with plRES-Sj 14-Sj26 and plRES-Sj26 groups. Flow cytometery showed that the percentages of CD4＋ and CD8＋ T cells were much higher in the plRES-Sj97-Sj 14-Sj26 group （P〈 0.01, P〈0.05）. It was concluded that plRES-Sj97-Sj 14-Sj26 vaccine may induce stronger immune response in BALB/c mice.     

Construction and expression of DNA vaccine pIRES-Sj97-Sj14-Sj26 and its immunogenicity in mice

Summary To find a new preventive strategy for the infection of Schistosoma japonica, plasmid pIRES-Sj97-Sj14-Sj26 that contains fatty binding protein (Sj14), GST (Sj26) and paramyocin (Sj97) that are expressed on the membrane, was constructed. RT-PCR was used to detect the expression of Sj14 mRNA, Sj26 mRNA and Sj97 mRNA in the Hela cells, the indirect immunofluorescent test was employed for the detection of the expression of trans-membrane Sj26 after the plasmid was transfected into Hela cells. Fifty BALB/c mice were randomly divided into 5 groups and pIRES-Sj97-Sj14-Sj26 plasmid DNA, pIRES-Sj14-Sj26 plasmid DNA, pIRES-Sj26 plasmid DNA, pIRES blank vector and normal saline were respectively injected into the quadriceps muscles of thigh. Eight weeks after the immunization the mice were killed and significantly higher level of IgG was detected in the pIRES-Sj97-Sj14-Sj26 group as compared with the pIRES blank vector, normal saline and pIRES-Sj26 groups (P<0.01) and the pIRES-Sj14-Sj26(P<0.05). Single splenocyte suspension was prepared to detected the level of IFN-γ by ELISA and the lymphocyte stimulating index (SI) by MTT. SI was significantly higher of in the pIRES-Sj97-Sj14-Sj26 group than in other groups (P<0.01), while the IFN-γ level was significantly higher the pIRES-Sj97-Sj14-Sj26 group than in pIRES blank vector and normal saline groups (P<0.01), but no significant differences were found when compared with pIRES-Sj14-Sj26 and pIRES-Sj26 groups. Flow cytometery showed that the percent-ages of CD4+ and CD8+ T cells were much higher in the pIRES-Sj97-Sj14-Sj26 group (P< 0.01, P<0.05). It was concluded that pIRES-Sj97-Sj14-Sj26 vaccine may induce stronger immune response in BALB/c mice. This project was supported by grants from National Natural Sciences Foundation of China (No. 30471603).     

Construction and Expression of Bivalent Membrane-anchored DNA Vaccine Encoding Sj14FABP and Sj26GST Genes

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjc14FABP and Sjc26GST genes and identify their expression in vitro, Sj 14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human plaeental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj 14. The expression of Sl14 and Sl26 genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and pIRES-Sj26-Sj 14 were successfully constructed and the expression of modified Sj 14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.     

Construction and Expression of Bivalent Membrane-anchored DNA Vaccine Encoding Sj14FABP and Sj26GST Gene

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjc14FABP and Sjc26GST genes and identify their expression in vitro, Sj 14 and Sj26 genes were ob- tained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sjl4 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 （IL-2） upstream Sj 14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyi-terminal of human placental alkaline phosphatase （PLAP） downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid plRES, resulting in another new recombinant plasmid plRES-Sj26-Sj14. The expression of Sj 14 and Sj26 genes was detected by RT-PCR and indirect immunofluorescent assays （IFA） when the plasmid plRES-Sj26-Sj 14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and plRES-Sj26-Sj 14 were successfully constructed and the expression of modified Sj 14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj 14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.     

rSj26-Sj32-Immunogold-Dipstick试剂对慢性日本血吸虫病的诊断价值研究

目的研究rSj26-Sj32-Immunogold-Dipstick试剂对慢性日本血吸虫病的诊断价值。方法用rSj26-Sj32融合蛋白和日本血吸虫成虫抗原(SjAWA)Immunogold-Dipstick法检测慢性日本血吸虫病患者血清,同时以华支睾吸虫病、卫氏并殖吸虫病、泡型棘球蚴病、囊性棘球蚴病、乙型肝炎、肺结核患者和健康人血清作为对照,比较两种抗原检测抗体效果的差异。结果该法检测慢性日本血吸虫病的敏感性和特异性分别为92.50%和97.67%,诊断该病的阳性预告值、阴性预告值及诊断效率分别为97.37%、93.33%和95.18%,并且与其他疾病患者血清均无交叉反应。结论 rSj26-Sj32-Immunogold-Dipstick试剂可用于慢性日本血吸虫病的免疫诊断。     

核酸疫苗Sj31BIN联合IL-12免疫抗日本血吸虫攻击感染的实验研究

目的：在证明日本血吸虫重组蛋白酶B核酸疫苗Sj31BIN具有抗生殖免疫作用的基础上,联合使用IL-12,观察IL-12是否具有佐剂效果。方法：分别用Sj31BIN IL-12和IL-12免疫Balb／C小鼠。攻击感染后6周计数成虫负荷和组织内虫卵数及肝脏表面虫卵结节数。结果：Sj31BIN IL-12免疫小鼠可降低成虫发育率。肝组织减卵率为59．74％;肠组织减卵率为59．60％;肝脏表面虫卵结节减少率为71．30％;单独使用IL-12有一定的免疫保护作用。结果：Sj31BIN IL-12能诱导小鼠产生较强的抗生殖免疫作用。     

日本血吸虫（中国大陆株）Sj16基因的体外扩增及克隆

为了构建日本血吸虫（中国大陆株)Sj16基因原核表达重组质粒pGEX-4T-1-Sj16。根据曼氏血吸虫Sm16基因已知序列设计合成一对引物,用PCR技术从日本血吸虫（中国大陆株）成虫cDNA库中扩增Sj16基因;将Sj16基因定向克隆入pGEX-4T-1;转化感受态BL21/DE3菌;用酶切,PCR扩增鉴定筛选得到的重组阳性克隆。结果表明,从日本血吸虫（中国大陆株）成虫cDNA库中获取Sj16基因,重组质粒中含有Sj16基因。结果提示,成功构建日本血吸虫（中国大陆株）Sj16基因原核表达重组质粒pGEX-4T-1-Sj16,为进一步研究Sj16基因功能打下基础。     

日本血吸虫成虫脲溶42 kD蛋白的分离及其初步诊断应用

目的 :分离日本血吸虫成虫脲溶抗原中有免疫学活性的抗原分子 ,探讨其诊断血吸虫病的效果。方法 :利用SDS PAGE及免疫印迹法分析日本血吸虫成虫脲溶抗原 ,采用电泳层析法分离有免疫活性的 4 2kD蛋白 ,ELISA分析 4 2kD蛋白诊断血吸虫病的灵敏度和特异性。结果 :电泳层析获得了具有免疫学活性的 4 2kD蛋白 ;分离的 4 2kD蛋白诊断血吸虫病的灵敏度和特异性分别为 88.0 9%和 97.0 6 %。结论 :日本血吸虫成虫脲溶 4 2kD蛋白是一种日本血吸虫的血清学抗原 ,可用于血吸虫病诊断     

初探日本血吸虫SWA及SEA对Ⅰ型糖尿病小鼠血糖的抑制作用

目的：探讨日本血吸虫SWA及SEA对实验性Ⅰ型糖尿病小鼠血糖的抑制作用。方法：制备日本血吸虫可溶性成虫抗原（Soluble worm antigen ,SWA）与可溶性虫卵抗原（Soluble egg antigen ,SEA）并构造实验性Ⅰ型糖尿病小鼠模型。把24只糖尿病实验小鼠随机平分为A、B、C组（每组8只）。A组实验小鼠从腹部皮下多点免疫注射日本血吸虫SWA ;B组实验小鼠从腹部皮下多点免疫注射日本血吸虫SEA ;C组实验小鼠用PBS进行腹部皮下多点免疫,3组小鼠每周免疫1次,连续免疫4周后,采小鼠尾静脉血,检测实验小鼠的血糖变化。结果：B组实验小鼠的血糖在4周左右为（9．85＆#177;1．17）mmol/L ,有明显降低,相比C组小鼠血糖（12．14＆#177;1．25）mmol/L具有显著性差异（P＜0．01）,但A组实验小鼠血糖在4周左右为（12．11＆#177;0．67）mmol/L与C组小鼠血糖无显著差异（P＞0．05）。结论：日本血吸虫SEA 对实验性Ⅰ型糖尿病小鼠的血糖具有一定的抑制作用,其机制可能是日本血吸虫SEA刺激了机体的免疫应答,导致T h1反应向T h2反应偏移。