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Protective effect of DNA-mediated immunization with a combination of SAG1 and IL-2 gene adjuvant against infection of Toxoplasma gondii in mice
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作者 陈观今 陈海峰 +1 位作者 郭虹 郑焕钦 《Chinese Medical Journal》 SCIE CAS CSCD 2002年第10期8-12,142,共6页
To characterize the immune response induced by SAG1 encoding plasmid combined with IL 2 gene adjuvant in mice and to assess the protective effect of this vaccination against toxoplasmosis Methods Mice were co inje... To characterize the immune response induced by SAG1 encoding plasmid combined with IL 2 gene adjuvant in mice and to assess the protective effect of this vaccination against toxoplasmosis Methods Mice were co injected intramuscularly with plasmid encoding Toxoplasma gondii SAG1 plus murine IL 2 expression vector at a dose of 100 μg Booster immunizations were employed 2 more times at 3 week interval As controls, mice were inoculated with PBS or empty plasmid pcDNA3 Humoral and cellular responses were assayed using ELISA for the determination of Ab, Ab isotype and IFN γ, as well as IL 4 To detect the integration and dissemination of DNA in the injected mice, PCR and in situ hybridization were performed All mice were then infected with highly virulent RH tachyzoites of Toxoplasma gondii intraperitoneally Results Significant increases in specific IgG levels were observed in mice after immunization three times with SAG1 expression plasmid With respect to the IgG isotype, co inoculation of IL 2 expression plasmid enhanced the level of IgG2a and the production of IFN γ Challenging mice by vaccinating with combined plasmids with RH tachyzoites resulted in prolonged survival Conclusion Humoral and cytokine responses elicited by SAG1 DNA immunization can be modulated by co inoculation with IL 2 expression plasmid The use of DNA vaccine in combination with an appropriate cytokine gene to prevent T gondii infection warrants further investigation 展开更多
关键词 DNA immunization · Toxoplasma gondi · surface antigen 1 (SAG1) · gene adjuvant · IL 2
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Enhancing DNA vaccine potency against hantavirus by co-administration of interleukin-12 expression vector as a genetic adjuvant 被引量:4
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作者 ZHENGLan-yan MOULing +2 位作者 LINSong LURun-ming LUOEn-jie 《Chinese Medical Journal》 SCIE CAS CSCD 2005年第4期313-319,共7页
Background The heavy incidence and mortality of hemorrhagic fever with renal syndrome, as well as no specific drugs in curing the disease, clearly indicate the need for development Of the more effective hantavirus vac... Background The heavy incidence and mortality of hemorrhagic fever with renal syndrome, as well as no specific drugs in curing the disease, clearly indicate the need for development Of the more effective hantavirus vaccine. Refining the DNA vaccination strategy to elicit more clinically efficacious immune responses is now under intensive investigation. In the present study, we examined the effects of using an interleukin-12 expression plasmid as a genetic adjuvant to enhance the immune responses induced by a DNA vaccine based on the S gene encoding nucleocapsid protein against hantavirus. Methods BALB/c mice were immunized three times by intramuscular inoculations of DNA vaccine encoding of hantanvirus nucleoeapsid protein alone or in combination with a plasmid expressing murine interleukin-12 (pcIL-12). Booster immunizations were employed 2 times at 2-week interval. To evaluate the Immoral and cellular immune responses, antigen-specific lymphocyte proliferation and antibody production were assayed by MTT method and ELISA respectively. The level of interleukin-4 and interferon-gamma in the splenic lymphocytic cultured supernatant were detected with ELISA kit at day 5, 10, 17, 35 and 42 after primary immunization. Results Antigen-specific IgG antibodies was increased markedly at day 17 in the experiment groups and reached a plateau after day 35. As pcIL-12 co-injected, a significant inhibition of antigen-specific IgG levels was displayed over the period and the antibody mean titre was decreased to only about 1: 50 at day 42 after primary immunization, significantly lower than the group immunized with pcDNA3.1 + S alone, in which the mean titre was about 1:70. Interferon-gamma was increased remarkably by the co-injection of pcIL-12 compared with the injection of pcDNA3.1 + S alone. However, the production of interleukin-4 was inhibited by pcIL-12 coinjection. Furthermore, pcIL-12 co-injection efficiently enhanced antigen-specific lymphocyte proliferation. Conclusion Humoral and cytokine responses elicited by pcDNA3.1 + S inoculation can be modulated by coinoculation with pcIL-12 and efficiently induced Th1-dominant immune responses. 展开更多
关键词 DNA vaccines HANTAVIRUS gene adjuvant INTERLEUKIN-12 nucleocapsid proteins
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猪白细胞介素4的基因克隆及其在囊尾蚴DNA疫苗中的应用 被引量:10
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作者 吴丹 郭瀛军 +1 位作者 王庆敏 孙树汉 《第二军医大学学报》 CAS CSCD 北大核心 2002年第11期1192-1194,共3页
目的:克隆猪白细胞介素4(IL-4)cDNA,观察其在抗囊尾蚴免疫中的疫苗佐剂作用。方法:通过RT-PCR法获得带有5'AUG侧翼翻译优化突变序列的猪IL-4 cDNA,测序后重组入真核表达载体pcDNA,在体外瞬时表达的基础上与囊尾蚴保护性抗原DNA疫苗... 目的:克隆猪白细胞介素4(IL-4)cDNA,观察其在抗囊尾蚴免疫中的疫苗佐剂作用。方法:通过RT-PCR法获得带有5'AUG侧翼翻译优化突变序列的猪IL-4 cDNA,测序后重组入真核表达载体pcDNA,在体外瞬时表达的基础上与囊尾蚴保护性抗原DNA疫苗联合免疫仔猪。结果:获得的猪IL-4 cDNA序列与文献报道的完全一致,在体外瞬时表达中获得了有生物学活性的猪IL-4。其与囊尾蚴保护性抗原DNA疫苗联合应用可有效提高个体体液免疫应答水平。结论:构建了一高效表达猪IL-4的真核表达质粒,初步实验证实了其在抗囊尾蚴DNA疫苗中的佐剂效应。 展开更多
关键词 猪白细胞介素4 基因克隆 囊尾蚴 DNA疫苗 佐剂作用
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LTB-hpaA融合基因原核表达产物的鉴定及其免疫保护作用 被引量:2
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作者 谈潘莉 严杰 +1 位作者 李立伟 毛亚飞 《浙江大学学报(理学版)》 CAS CSCD 北大核心 2006年第1期95-100,共6页
分别从幽门螺杆菌(Helicobacter pylori,Hp)临床菌株Y06和大肠杆菌44851株DNA中扩增1tB和hpaA基因,构建1tB-hpaA融合基因厦其原核表达系统,鉴定其表达产物的免疫原性、佐荆活性及其对Hp SS1株感染小鼠的保护作用.采用PCR分别从上... 分别从幽门螺杆菌(Helicobacter pylori,Hp)临床菌株Y06和大肠杆菌44851株DNA中扩增1tB和hpaA基因,构建1tB-hpaA融合基因厦其原核表达系统,鉴定其表达产物的免疫原性、佐荆活性及其对Hp SS1株感染小鼠的保护作用.采用PCR分别从上述菌株DNA中扩增hpaA基因和1tB基因片段,构建1tB-hpaA融合基因,T—A克隆后测定核苷酸序列.采用质粒pOE32和宿主菌E.coliM15构建1tB-hpaA融合基因表达系统,并用不同浓度的IPTG诱导表达.采用western blot和GM1—ELISA鉴定表达产物的抗原性、免疫反应性和佐荆活性.建立HP SS1株感染BALB/e小鼠模型,检测rLTB-HpaA的免疫保护效果.采用ELISA检测125例胃、十二指肠粘膜活检标本尿素酶阳性患者血清中HpaA抗体和41株Hp临床菌株HpaA表达情况.与Gen Bank登录的相关序列比较,所构建的ltB-hpaA融合基因核苷酸序列同源性分别为94.25%~99.71%和95.38%~99.19%.所构建的表达系统pQE32-1tB-hpaA—M15的目的重组蛋白(rLTB-HpaA)表达量高达细菌总蛋白的1/3左右.rLTB-HpaA能与Hp全菌抗体发生结合反应,免疫家兔能产生特异性抗体.GM3-ELISA结果显示rLTB-HpaA能与牛GM1结合.rHpaA免瘦小鼠的保护率仅为66.7%,rLTB-HpaA免疫小鼠的保护率可增至83.3%.81.6%患者血清(102/125)HpaA抗体检测结果阳性,所有菌株均含有HpaA.本文成功地构建了ltB-hpaA融合基因高效原核表达系统,所表达的rLTB-HpaA有良好的免疫原性和佐剂活性,并对Hp感染小鼠有较强的免疫保护作用. 展开更多
关键词 幽门螺杆菌/hpaA基因 大肠杆菌/ltB基因 融合基因 克隆/表达 免疫原性/佐荆活性 动物感染模型 免疫保护
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大肠杆菌不耐热肠毒素B亚单位基因改建及其表达产物粘膜免疫佐剂活性的研究(英文) 被引量:1
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作者 孙菊云 谈潘莉 +1 位作者 胡野 毛亚飞 《中国人兽共患病学报》 CAS CSCD 北大核心 2006年第5期385-390,共6页
目的改建大肠杆菌不耐热肠毒素B亚单位(LTB)基因序列以提高重组LTB(rLTB)原核表达量,确定重组改建LTB(rrLTB)粘膜免疫佐剂活性。方法按大肠杆菌偏爱密码子合成全长LTB基因的核苷酸序列。构建pET32a-rLTB原核重组表达质粒及pET32a-rLTB-E... 目的改建大肠杆菌不耐热肠毒素B亚单位(LTB)基因序列以提高重组LTB(rLTB)原核表达量,确定重组改建LTB(rrLTB)粘膜免疫佐剂活性。方法按大肠杆菌偏爱密码子合成全长LTB基因的核苷酸序列。构建pET32a-rLTB原核重组表达质粒及pET32a-rLTB-E.coliBL21DE3表达系统,提取重组质粒并测定其插入的rLTB基因序列。采用SDS-PAGE和BioRad凝胶成像分析系统鉴定rrLTB表达产量,并与未改建的重组LTB(roLTB)比较。采用GM1-ELISA确定rrLTB和roLTB结合牛GM1的能力。分别以幽门螺杆菌重组尿素酶B亚单位(rUreB),检测rrLTB和roLTB对rUreB对幽门螺杆菌SS1株感染BALB/c小鼠保护作用及产生特异性S-IgA的增强效应。结果pET32a-rLTB-E.coliBL21DE3经1 mmol/LIPTG诱导后,rrLTB表达量约占细菌总蛋白的35.4%,为roLTB(2.8%)的12.6倍。GM1-ELISA结果证实rrLTB和roLTB均能与牛GM1结合。单一rUreB对感染小鼠的免疫保护率为66.7%,但与rrLTB或roLTB合用时,保护率可至91.6%,并可提高感染小鼠特异性S-IgA产生量(P<0.01)。结论改建的LTB基因能明显提高表达量,所表达的rrLTB仍保持较强的粘膜免疫佐剂活性。 展开更多
关键词 大肠杆菌 LTB基因/改建 重组蛋白/表达 粘膜免疫 佐剂活性
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Th1 cytokine-based immunotherapy for cancer 被引量:4
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作者 Hong-Mei Xu 《Hepatobiliary & Pancreatic Diseases International》 SCIE CAS 2014年第5期482-494,共13页
Cytokine-based immunotherapy is executed by harnessing cytokines to activate the immune system to suppress tumors. Th1-type cytokines including IL-1, IL-2, IL-12 and granulocyte-macrophage colony-stimulating factor ar... Cytokine-based immunotherapy is executed by harnessing cytokines to activate the immune system to suppress tumors. Th1-type cytokines including IL-1, IL-2, IL-12 and granulocyte-macrophage colony-stimulating factor are potent stimulators of Th1 differentiation and Th1-based antitumor response. Many preclinical studies demonstrated the antitumor effects of Th1 cytokines but their clinical efficacy is limited.Multiple factors influence the efficacy of immunotherapy for tumors. For instance immunosuppressive cells in the tumor microenvironment can produce inhibitory cytokines which suppress antitumor immune response. Most studies on cytokine immunotherapy focused on how to boost Th1 response; many studies combined cytokine-based therapy with other treatments to reverse immunosuppression in tumor microenvironment.In addition, cytokines have pleiotropic functions and some cytokines show paradoxical activities under different settings.Better understanding the physiological and pathological functions of cytokines helps clinicians to design Th1-based cancer therapy in clinical practice. 展开更多
关键词 cytokine gene therapy adjuvant therapy
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