共查询到20条相似文献,搜索用时 78 毫秒
1.
《中国实验动物学报》2006,(3)
两种SARS候选疫苗攻毒效果评价SARS疾病流行已经过去3年多了,但是对SARS防治的研究还在继续。加拿大的科学家在病毒遗传学杂志上报道了他们新开发的两种抗SARS病毒疫苗,并利用129S6/SvEv小鼠模型比较了它们的有效性。这两种疫苗,一个为灭活疫苗(用β-丙内酯,beta-propiolactone灭活),另一个为两种腺病毒载体重组的疫苗,分别表达SARS-CoV的N蛋白和S蛋白(命名为Ad S/N)。评价方式包括血清中和抗体滴度、细胞免疫反应和对SARS-CoV在肺部复制的抑制作用。灭活疫苗经皮下注射129S6/SvEv小鼠,而Ad S/N疫苗分别采取鼻腔吸入和肌肉… 相似文献
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为探讨SARS-CoV的M蛋白的免疫学特性以及M蛋白作为SARS-CoV病毒疫苗组分的可行性和必要性.分别用pET-15b和pET-22b在大肠杆菌中表达SARS-CoV的M蛋白,亲和层析纯化后作为抗原应用.同时,将M蛋白的编码基因克隆进分泌型真核表达载体pSecTagB中得到重组质粒pSecM作为DNA疫苗,免疫BALB/c小白鼠、制备SARS-CoV M蛋白的抗血清.并用纯化后的M蛋白建立的SARS-CoV M抗体ELISA检测技术研究所构建的M-DNA疫苗的免疫效果.结果表明两种重组M蛋白在大肠杆菌中均以可溶性形式得到高效表达,经与华大产的用灭活SARS全病毒制备的SARS-CoV抗体ELISA检测试剂盒比较实验,证明该原核表达的重组M蛋白能与SARS确诊病人血清以及M-DNA免疫鼠血清发生特异性抗原抗体反应.这两种重组M蛋白有可能作为抗原组分用于临床SARS-CoV检测中;所构建的SARS-CoV的M基因核酸疫苗能在小鼠体内产生特异性抗体,提示M蛋白在SARS-CoV疫苗尤其是组分疫苗的研制中应加以考虑,为DNA疫苗的开发提供了依据. 相似文献
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为探讨SARS-CoV的M蛋白的免疫学特性以及M蛋白作为SARS-CoV病毒疫苗组分的可行性和必要性.分别用pET-15b和pET-22b在大肠杆菌中表达SARS-CoV的M蛋白,亲和层析纯化后作为抗原应用.同时,将M蛋白的编码基因克隆进分泌型真核表达载体pSecTagB中得到重组质粒pSecM作为DNA疫苗,免疫BALB/c小白鼠、制备SARS-CoV M蛋白的抗血清.并用纯化后的M蛋白建立的SARS-CoV M抗体ELISA检测技术研究所构建的M-DNA疫苗的免疫效果.结果表明:两种重组M蛋白在大肠杆菌中均以可溶性形式得到高效表达,经与华大产的用灭活SARS全病毒制备的SARS-CoV抗体ELISA检测试剂盒比较实验,证明该原核表达的重组M蛋白能与SARS确诊病人血清以及M-DNA免疫鼠血清发生特异性抗原抗体反应.这两种重组M蛋白有可能作为抗原组分用于临床SARS-CoV检测中;所构建的SARS-CoV的M基因核酸疫苗能在小鼠体内产生特异性抗体,提示M蛋白在SARS-CoV疫苗尤其是组分疫苗的研制中应加以考虑,为DNA疫苗的开发提供了依据. 相似文献
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新型冠状病毒作为严重急性呼吸道综合征暴发流行的病原体(SARS-CoV),目前仍无抗SARS-CoV的特效药物,因此疫苗的研究为预防SARS再次流行具有重大意义.目前研制的SARS-CoV疫苗有SARS-CoV灭活疫苗、SARS-CoV DNA疫苗、SARS-CoV腺病毒载体疫苗、重组SARS-CoV刺突蛋白-减毒痘苗病毒等.体外及体内动物攻毒保护实验结果显示,所研制的SARS疫苗均可诱生体液和细胞免疫应答,并对免疫动物具有保护性作用.本文将近期有关研究结果作了综述. 相似文献
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针对SARS冠状病毒重要蛋白的siRNA设计(英) 总被引:4,自引:0,他引:4
RNA干涉(RNA interference, RNAi)是一种特异性地导致转录后基因沉默的现象,在哺乳动物细胞中小分子干扰RNA双链体(small interfering RNA duplexes, siRNA duplexes)可以有效地诱导RNAi现象,为一些疾病的治疗开辟了新的途径.针对SARS冠状病毒(SARS coronavirus, SARS-CoV)中编码5个主要蛋白质的基因,用生物信息学的方法设计了348条候选siRNA靶标.在理论上,相应的siRNA双链体能特异地抑制SARS-CoV靶基因的表达,同时不会影响人体细胞基因的正常表达,这为进一步siRNA类药物的实验研究提供了理论基础. 相似文献
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SARS-CoV是引起严重急性呼吸道综合症(SARS)的病原体.更多地了解SARS-CoV的基因组、蛋白结构以及它与其它冠状病毒的关系,将有助于SARS疾病的防治. 相似文献
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严重急性呼吸综合征-冠状病毒(SARS-CoV)作为肆虐我国及世界的SARS病原已被确认.为数众多的分子生物学、发病机制、实验室诊断及药物和疫苗研究正在蓬勃兴起,国内外的研究热潮已逐步走向深入.本期刊出了数篇文章,介绍了有关SARS的不同领域. 相似文献
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SARS-CoV感染动物模型的建立可加深对SARS病原学的了解和发病机制的研究,加速实验室诊断技术的建立、抗病毒药物的筛选和疫苗的开发,同时也有助于给该病一个更精确的定义。可以说SARS动物模型的建立,不但是SARS研究的瓶颈问题,其应用更是贯穿SARS研究的整个过程。到目前为止,已经报道有4种非人灵长类动物(恒河猴、食蟹猴、绒猴、非洲绿猴)和6种啮齿类动物(大鼠、小鼠、豚鼠、田鼠、仓鼠、转基因鼠),以及雪貂、家猫等可以作为SARS动物模型用于实验研究,并已经开始利用动物模型进行疫苗和药物的安全性和有效性评价。本文就已报道的各类SARS动物模型进行综述,并根据动物模型和SARS患者的比对,提出动物模型建立的技术要点。 相似文献
11.
Mina Nakauchi Hiroaki Kariwa Yasuhiro Kon Kentaro Yoshii Akihiko Maeda Ikuo Takashima 《Microbiology and immunology》2008,52(12):625-630
SARS‐CoV has four major structural proteins: the N, S, M, and E proteins. To investigate the mechanism of SARS‐CoV assembly, we cloned the genes encoding these four proteins into the eukaryotic expression vector pCAGGS and transfected them into 293T cells. When all four expression vectors were co‐transfected VLP formed, as confirmed using electron microscopy. Using a rabbit polyclonal antibody specific to the N protein, N‐protein‐containing particles similar in size to the VLP were also observed by immunoelectron microscopy, indicating that the VLP contained the N protein. Co‐immunoprecipitation analyses demonstrated an interaction between the N and M proteins, suggesting that N protein binds directly to M protein to be incorporated into VLP. 相似文献
12.
Tseng CT Sbrana E Iwata-Yoshikawa N Newman PC Garron T Atmar RL Peters CJ Couch RB 《PloS one》2012,7(4):e35421
Background
Severe acute respiratory syndrome (SARS) emerged in China in 2002 and spread to other countries before brought under control. Because of a concern for reemergence or a deliberate release of the SARS coronavirus, vaccine development was initiated. Evaluations of an inactivated whole virus vaccine in ferrets and nonhuman primates and a virus-like-particle vaccine in mice induced protection against infection but challenged animals exhibited an immunopathologic-type lung disease.Design
Four candidate vaccines for humans with or without alum adjuvant were evaluated in a mouse model of SARS, a VLP vaccine, the vaccine given to ferrets and NHP, another whole virus vaccine and an rDNA-produced S protein. Balb/c or C57BL/6 mice were vaccinated IM on day 0 and 28 and sacrificed for serum antibody measurements or challenged with live virus on day 56. On day 58, challenged mice were sacrificed and lungs obtained for virus and histopathology.Results
All vaccines induced serum neutralizing antibody with increasing dosages and/or alum significantly increasing responses. Significant reductions of SARS-CoV two days after challenge was seen for all vaccines and prior live SARS-CoV. All mice exhibited histopathologic changes in lungs two days after challenge including all animals vaccinated (Balb/C and C57BL/6) or given live virus, influenza vaccine, or PBS suggesting infection occurred in all. Histopathology seen in animals given one of the SARS-CoV vaccines was uniformly a Th2-type immunopathology with prominent eosinophil infiltration, confirmed with special eosinophil stains. The pathologic changes seen in all control groups lacked the eosinophil prominence.Conclusions
These SARS-CoV vaccines all induced antibody and protection against infection with SARS-CoV. However, challenge of mice given any of the vaccines led to occurrence of Th2-type immunopathology suggesting hypersensitivity to SARS-CoV components was induced. Caution in proceeding to application of a SARS-CoV vaccine in humans is indicated. 相似文献13.
目的:研究严重急性呼吸系统综合征冠状病毒(SARS-CoV)N蛋白对甘油三酯和总胆固醇含量的影响。方法:检测分析144例SARS患者甘油三脂和总胆固醇含量在发病后的变化;将小鼠随机分组,分别注射生理盐水和SARS-CoV N蛋白,连续给药9 d后检测小鼠血清中甘油三酯和总胆固醇含量的变化。结果:SARS患者的甘油三酯和总胆固醇含量随发病时间有升高变化(P0.05),在发病40 d前后甘油三酯和总胆固醇含量超标的病例数占本时间段内的病例数的比例显著高于其他时间段,而且超标的含量也明显升高。SARS-CoV N蛋白使小鼠体重明显高于对照组(P0.05),甘油三酯含量在39和51 d明显高于对照组(P0.05),总胆固醇含量在21和39 d也明显高于对照组(P0.05)。结论:SARS-CoV N蛋白可以升高小鼠的甘油三酯和总胆固醇的含量,其升高最为明显的时间段与SARS患者的甘油三酯和总胆固醇含量升高的时间段基本一致。由此推测,SARS-CoV N蛋白可能是促使SARS患者发病后甘油三酯和总胆固醇含量升高的重要原因。 相似文献
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重症急性呼吸综合征(SARS)是由SARS冠状病毒(SARS-CoV)引起的一种急性传染病,在其序列被测出后几个月内人们就找到了SARS-CoV的受体血管紧张素转换酶2(ACE2)。因病毒受体与病毒入侵细胞密切相关,因而有必要深入研究ACE2与SARS-CoV之间的关系。本文总结了ACE2在各组织器官的分布及功能,分析了ACE2基因的变异与病毒进入及SARS疾病严重程度之间的关系、ACE2基因的表达水平与病毒进入及SARS疾病严重程度之间的关系。这些研究将为理解SARS-CoV与ACE2之间的相互作用及设计针对ACE2的抗SARS药物提供重要的理论依据。 相似文献
15.
Sirika Pillay Ann Meyers Anna‐Lise Williamson Edward P. Rybicki 《Biotechnology progress》2009,25(4):1153-1160
A baculovirus‐insect cell expression system potentially provides the means to produce prophylactic HIV‐1 virus‐like particle (VLP) vaccines inexpensively and in large quantities. However, the system must be optimized to maximize yields and increase process efficiency. In this study, we optimized the production of two novel, chimeric HIV‐1 VLP vaccine candidates (GagRT and GagTN) in insect cells. This was done by monitoring the effects of four specific factors on VLP expression: these were insect cell line, cell density, multiplicity of infection (MOI), and infection time. The use of western blots, Gag p24 ELISA, and four‐factorial ANOVA allowed the determination of the most favorable conditions for chimeric VLP production, as well as which factors affected VLP expression most significantly. Both VLP vaccine candidates favored similar optimal conditions, demonstrating higher yields of VLPs when produced in the Trichoplusia ni Pro? insect cell line, at a cell density of 1 × 106 cells/mL, and an infection time of 96 h post infection. It was found that cell density and infection time were major influencing factors, but that MOI did not affect VLP expression significantly. This work provides a potentially valuable guideline for HIV‐1 protein vaccine optimization, as well as for general optimization of a baculovirus‐based expression system to produce complex recombinant proteins. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 相似文献
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The recent swine H1N1 influenza outbreak demonstrated that egg-based vaccine manufacturing has an Achille's heel: its inability to provide a large number of doses quickly. Using a novel manufacturing platform based on transient expression of influenza surface glycoproteins in Nicotiana benthamiana, we have recently demonstrated that a candidate Virus-Like Particle (VLP) vaccine can be generated within 3 weeks of release of sequence information. Herein we report that alum-adjuvanted plant-made VLPs containing the hemagglutinin (HA) protein of H5N1 influenza (A/Indonesia/5/05) can induce cross-reactive antibodies in ferrets. Even low doses of this vaccine prevented pathology and reduced viral loads following heterotypic lethal challenge. We further report on safety and immunogenicity from a Phase I clinical study of the plant-made H5 VLP vaccine in healthy adults 18-60 years of age who received 2 doses 21 days apart of 5, 10 or 20 μg of alum-adjuvanted H5 VLP vaccine or placebo (alum). The vaccine was well tolerated at all doses. Adverse events (AE) were mild-to-moderate and self-limited. Pain at the injection site was the most frequent AE, reported in 70% of vaccinated subjects versus 50% of the placebo recipients. No allergic reactions were reported and the plant-made vaccine did not significantly increase the level of naturally occurring serum antibodies to plant-specific sugar moieties. The immunogenicity of the H5 VLP vaccine was evaluated by Hemagglutination-Inhibition (HI), Single Radial Hemolysis (SRH) and MicroNeutralisation (MN). Results from these three assays were highly correlated and showed similar trends across doses. There was a clear dose-response in all measures of immunogenicity and almost 96% of those in the higher dose groups (2 × 10 or 20 μg) mounted detectable MN responses. Evidence of striking cross-protection in ferrets combined with a good safety profile and promising immunogenicity in humans suggest that plant-based VLP vaccines should be further evaluated for use in pre-pandemic or pandemic situations. TRIAL REGISTRATION: ClinicalTrials.gov NCT00984945. 相似文献
17.
Yeu-Chun Kim Fu-Shi Quan Richard W. Compans Sang-Moo Kang Mark R. Prausnitz 《AAPS PharmSciTech》2010,11(3):1193-1201
Mortality due to seasonal and pandemic influenza could be reduced by increasing the speed of influenza vaccine production
and distribution. We propose that vaccination can be expedited by (1) immunizing with influenza virus-like particle (VLP)
vaccines, which are simpler and faster to manufacture than conventional egg-based inactivated virus vaccines, and (2) administering
vaccines using microneedle patches, which should simplify vaccine distribution due to their small package size and possible
self-administration. In this study, we coated microneedle patches with influenza VLP vaccine, which was released into skin
by dissolution within minutes. Optimizing the coating formulation required balancing factors affecting the coating dose and
vaccine antigen stability. Vaccine stability, as measured by an in vitro hemagglutination assay, was increased by formulation with increased concentration of trehalose or other stabilizing carbohydrate
compounds and decreased concentration of carboxymethylcellulose (CMC) or other viscosity-enhancing compounds. Coating dose
was increased by formulation with increased VLP concentration, increased CMC concentration, and decreased trehalose concentration,
as well as increased number of dip coating cycles. Finally, vaccination of mice using microneedles stabilized by trehalose
generated strong antibody responses and provided full protection against high-dose lethal challenge infection. In summary,
this study provides detailed analysis to guide formulation of microneedle patches coated with influenza VLP vaccine and demonstrates
effective vaccination in vivo using this system. 相似文献
18.
Shih‐Yeh Lin Chia‐Tsui Yeh Wan‐Hua Li Cheng‐Ping Yu Wen‐Chin Lin Jyh‐Yuan Yang Hsueh‐Ling Wu Yu‐Chen Hu 《Biotechnology and bioengineering》2015,112(10):2005-2015
19.
Tanja Smith Martha M. O'Kennedy Daniel B.R. Wandrag Modupeore Adeyemi Celia Abolnik 《Plant biotechnology journal》2020,18(2):502-512
The efficacy, safety, speed, scalability and cost‐effectiveness of producing hemagglutinin‐based virus‐like particle (VLP) vaccines in plants are well‐established for human influenza, but untested for the massive poultry influenza vaccine market that remains dominated by traditional egg‐grown oil‐emulsion whole inactivated virus vaccines. For optimal efficacy, a vaccine should be closely antigenically matched to the field strain, requiring that influenza A vaccines be updated regularly. In this study, an H6 subtype VLP transiently expressed in Nicotiana benthamiana was formulated into a vaccine and evaluated for efficacy in chickens against challenge with a heterologous H6N2 virus. A single dose of the plant‐produced H6 VLP vaccine elicited an immune response comparable to two doses of a commercial inactivated H6N2 vaccine, with mean hemagglutination inhibition titres of 9.3 log2 and 8.8 log2, respectively. Compared to the non‐vaccinated control, the H6 VLP vaccine significantly reduced the proportion of shedders and the magnitude of viral shedding by >100‐fold in the oropharynx and >6‐fold in the cloaca, and shortened oropharyngeal viral shedding by at least a week. Despite its potency, the cost of the antigenic mismatch between the inactivated H6N2 vaccine and challenge strain was evident not only in this vaccine's failure to reduce viral shedding compared to the non‐vaccinated group, but its apparent exacerbation of oropharyngeal viral shedding until 21 days post‐challenge. We estimate that a kilogram of plant leaf material can produce H6 VLP vaccines sufficient for between 5000 and 30 000 chickens, depending on the effective dose and whether one or two immunizations are administered. 相似文献
20.
Protective immunity and antibody-secreting cell responses elicited by combined oral attenuated Wa human rotavirus and intranasal Wa 2/6-VLPs with mutant Escherichia coli heat-labile toxin in gnotobiotic pigs 下载免费PDF全文
Yuan L Iosef C Azevedo MS Kim Y Qian Y Geyer A Nguyen TV Chang KO Saif LJ 《Journal of virology》2001,75(19):9229-9238
Two combined rotavirus vaccination regimens were evaluated in a gnotobiotic pig model of rotavirus infection and disease and were compared to previously tested rotavirus vaccination regimens. The first (AttHRV/VLP2x) involved oral inoculation with one dose of attenuated (Att) Wa human rotavirus (HRV), followed by two intranasal (i.n.) doses of a rotavirus-like particle (2/6-VLPs) vaccine derived from Wa (VP6) and bovine RF (VP2) rotavirus strains. The 2/6-VLPs were coadministered with a mutant Escherichia coli heat-labile toxin, LT-R192G (mLT) adjuvant. For the second regimen (VLP2x/AttHRV), two i.n. doses of 2/6-VLPs+mLT were given, followed by one oral dose of attenuated Wa HRV. To compare the protective efficacy and immune responses induced by the combined vaccine regimens with individual rotavirus vaccine regimens, we included in the experiments the following vaccine groups: one oral dose of attenuated Wa HRV (AttHRV1x and Mock2x/AttHRV, respectively), three oral doses of attenuated Wa HRV (AttHRV3x), three i.n. doses of 2/6-VLPs plus mLT (VLP3x), three i.n. doses of purified double-layered inactivated Wa HRV plus mLT (InactHRV3x), mLT alone, and mock-inoculated pigs. The isotype, magnitude, and tissue distribution of antibody-secreting cells (ASCs) in the intestinal and systemic lymphoid tissues were evaluated using an enzyme-linked immunospot assay. The AttHRV/VLP2x regimen stimulated the highest mean numbers of intestinal immunoglobulin A (IgA) ASCs prechallenge among all vaccine groups. This regimen induced partial protection against virus shedding (58%) and diarrhea (44%) upon challenge of pigs with virulent Wa HRV. The reverse VLP2x/AttHRV regimen was less efficacious than the AttHRV/VLP2x regimen in inducing IgA ASC responses and protection against diarrhea (25% protection rate) but was more efficacious than VLP3x or InactHRV3x (no protection). In conclusion, the AttHRV/VLP2x vaccination regimen stimulated the strongest B-cell responses in the intestinal mucosal immune system at challenge and conferred a moderately high protection rate against rotavirus disease, indicating that priming of the mucosal inductive site at the portal of natural infection with a replicating vaccine, followed by boosting with a nonreplicating vaccine at a second mucosal inductive site, may be a highly effective approach to stimulate the mucosal immune system and induce protective immunity against various mucosal pathogens. 相似文献