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目的探讨马抗SARSCoV中和性抗体是否对SARSCoV感染有防治作用。方法用SARSCoV(BJ01株)免疫马匹,从免疫马血清中制备IgGs及其F(ab’)2片段。通过细胞病变法(CPE)、MTT法和空斑形成实验(PFU)等方法,在体外培养的VeroE6细胞中检测F(ab’)2片段对SARSCoV感染的预防性和治疗性作用。再在Balb/c小鼠体内模型中,用CPE法和定量RTPCR方法检测该抗体的保护性效应。结果CPE、MTT和PFU三种方法证实来自三匹马的血清F(ab’)2的中和滴度均达到1∶1600以上。体内实验证实,50μg的F(ab’)2片段可以完全中和1×104TCID50SARSCoV。结论马抗SARSCoV抗体在体内外均能有效地中和SARSCoV和预防其感染宿主细胞,为马抗体将来在大的灵长类动物或人体内进行实验提供实验依据。 相似文献
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Vaccine design for severe acute respiratory syndrome coronavirus 总被引:2,自引:0,他引:2
Severe acute respiratory syndrome (SARS) is an emerging infectious disease caused by a new coronavirus (SARS-CoV). Recent studies suggest that SARS-CoV is zoonotic and may have a broad host range besides humans. Although the global outbreak of SARS has been contained, there are serious concerns over its re-emergence and bioterrorism potential. As a part of preparedness, development of a safe and effective vaccine is one of the highest priorities in fighting SARS. A number of candidate vaccines, using a variety of approaches, are under development. The first vaccine tested in clinical trial is made from the inactivated form of SARS-CoV. Several live attenuated, genetically engineered or vector vaccines encoding the SARS-CoV spike (S) protein have been in pre-clinical studies. These vaccine candidates are effective in terms of eliciting protective immunity in the vaccinated animals. However, caution should be taken with the safety of whole virus or full-length S protein-based immunogens in humans because they may induce harmful immune or inflammatory responses. We propose to use the receptor-binding domain (RBD) of SARS-CoV S protein (residues 318--510) for developing a safe and effective subunit SARS vaccine, as it is not only a functional domain that mediates virus-receptor binding but also a major neutralization determinant of SARSCoV. It has been demonstrated that the RBD of SARS-CoV S protein contains multiple conformational epitopes capable of inducing highly potent neutralizing antibody responses and protective immunity. 相似文献
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Marie Lin Hsiang-Kuang Tseng Jean A Trejaut Hui-Lin Lee Jun-Hun Loo Chen-Chung Chu Pei-Jan Chen Ying-Wen Su Ken Hong Lim Zen-Uong Tsai Ruey-Yi Lin Ruey-Shiung Lin Chun-Hsiung Huang 《BMC medical genetics》2003,4(1):1-7
Background
The human leukocyte antigen (HLA) system is widely used as a strategy in the search for the etiology of infectious diseases and autoimmune disorders. During the Taiwan epidemic of severe acute respiratory syndrome (SARS), many health care workers were infected. In an effort to establish a screening program for high risk personal, the distribution of HLA class I and II alleles in case and control groups was examined for the presence of an association to a genetic susceptibly or resistance to SARS coronavirus infection.Methods
HLA-class I and II allele typing by PCR-SSOP was performed on 37 cases of probable SARS, 28 fever patients excluded later as probable SARS, and 101 non-infected health care workers who were exposed or possibly exposed to SARS coronavirus. An additional control set of 190 normal healthy unrelated Taiwanese was also used in the analysis.Results
Woolf and Haldane Odds ratio (OR) and corrected P-value (Pc) obtained from two tails Fisher exact test were used to show susceptibility of HLA class I or class II alleles with coronavirus infection. At first, when analyzing infected SARS patients and high risk health care workers groups, HLA-B*4601 (OR = 2.08, P = 0.04, Pc = n.s.) and HLA-B*5401 (OR = 5.44, P = 0.02, Pc = n.s.) appeared as the most probable elements that may be favoring SARS coronavirus infection. After selecting only a "severe cases" patient group from the infected "probable SARS" patient group and comparing them with the high risk health care workers group, the severity of SARS was shown to be significantly associated with HLA-B*4601 (P = 0.0008 or Pc = 0.0279).Conclusions
Densely populated regions with genetically related southern Asian populations appear to be more affected by the spreading of SARS infection. Up until recently, no probable SARS patients were reported among Taiwan indigenous peoples who are genetically distinct from the Taiwanese general population, have no HLA-B* 4601 and have high frequency of HLA-B* 1301. While increase of HLA-B* 4601 allele frequency was observed in the "Probable SARS infected" patient group, a further significant increase of the allele was seen in the "Severe cases" patient group. These results appeared to indicate association of HLA-B* 4601 with the severity of SARS infection in Asian populations. Independent studies are needed to test these results. 相似文献6.
Synat Keam Dewi Megawati Shailesh Kumar Patel Ruchi Tiwari Kuldeep Dhama Harapan Harapan 《Reviews in medical virology》2020,30(5)
The outbreak of coronavirus disease 2019 (COVID‐19) and pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), has become a major concern globally. As of 14 April 2020, more than 1.9 million COVID‐19 cases have been reported in 185 countries. Some patients with COVID‐19 develop severe clinical manifestations, while others show mild symptoms, suggesting that dysregulation of the host immune response contributes to disease progression and severity. In this review, we have summarized and discussed recent immunological studies focusing on the response of the host immune system and the immunopathology of SARS‐CoV‐2 infection as well as immunotherapeutic strategies for COVID‐19. Immune evasion by SARS‐CoV‐2, functional exhaustion of lymphocytes, and cytokine storm have been discussed as part of immunopathology mechanisms in SARS‐CoV‐2 infection. Some potential immunotherapeutic strategies to control the progression of COVID‐19, such as passive antibody therapy and use of interferon αβ and IL‐6 receptor (IL‐6R) inhibitor, have also been discussed. This may help us to understand the immune status of patients with COVID‐19, particularly those with severe clinical presentation, and form a basis for further immunotherapeutic investigations. 相似文献
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Recombinant protein-based assays for detection of antibodies to severe acute respiratory syndrome coronavirus spike and nucleocapsid proteins 下载免费PDF全文
Haynes LM Miao C Harcourt JL Montgomery JM Le MQ Dryga SA Kamrud KI Rivers B Babcock GJ Oliver JB Comer JA Reynolds M Uyeki TM Bausch D Ksiazek T Thomas W Alterson H Smith J Ambrosino DM Anderson LJ 《Clinical and Vaccine Immunology : CVI》2007,14(3):331-333
Recombinant severe acute respiratory syndrome (SARS) nucleocapsid and spike protein-based immunoglobulin G immunoassays were developed and evaluated. Our assays demonstrated high sensitivity and specificity to the SARS coronavirus in sera collected from patients as late as 2 years postonset of symptoms. These assays will be useful not only for routine SARS coronavirus diagnostics but also for epidemiological and antibody kinetic studies. 相似文献
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Ksiazek TG Erdman D Goldsmith CS Zaki SR Peret T Emery S Tong S Urbani C Comer JA Lim W Rollin PE Dowell SF Ling AE Humphrey CD Shieh WJ Guarner J Paddock CD Rota P Fields B DeRisi J Yang JY Cox N Hughes JM LeDuc JW Bellini WJ Anderson LJ;SARS Working Group 《The New England journal of medicine》2003,348(20):1953-1966
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Evaluation of real-time reverse transcriptase PCR and real-time loop-mediated amplification assays for severe acute respiratory syndrome coronavirus detection 总被引:6,自引:0,他引:6 下载免费PDF全文
Poon LL Wong BW Chan KH Ng SS Yuen KY Guan Y Peiris JS 《Journal of clinical microbiology》2005,43(7):3457-3459
We compared the performance of a recently established real-time loop-mediated amplification (LAMP) assay with the one from a highly sensitive quantitative PCR assay. None of these assays produced false-positive results in this study. For samples isolated from patients within the first 3 days of disease onset, the detection rate of the quantitative PCR assay was higher (14 of 15 were positive) than the LAMP assay (9 of 15 were positive). By contrast, the detection rates of these assays toward specimens sampled from patients with more than 3 days of illness were similar (32 of 44 for PCR and 33 of 44 for LAMP were positive). The simpler operation of LAMP might be a possible solution for on-site diagnosis. 相似文献
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Zhao G Ni B Jiang H Luo D Pacal M Zhou L Zhang L Xing L Zhang L Jia Z Lin Z Wang L Li J Liang Y Shi X Zhao T Zhou L Wu Y Wang X 《Viral immunology》2007,20(1):197-205
Equine anti-severe acute respiratory syndrome-associated coronavirus F(ab')(2) has been verified to protect mice from infection with severe acute respiratory syndrome-associated coronavirus (SARS-CoV). However, before potential clinical application, the antibody needs to be tested in as many animal models as possible to ensure its safety and efficiency. In this study, after verification by various methods that the golden Syrian hamster constitutes a model susceptible to SARS-CoV infection, we confirmed that the antibody could protect animals completely from SARS-CoV infection in the preventive setting. More importantly, the antibody could reduce viral titers or copies by approximately 10(3)- to 10(4)-fold in animal lung after virus exposure, compared with negative control. These data provide further evidence to warrant clinical studies of this antibody in the treatment and prevention of SARS. 相似文献
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Detection of severe acute respiratory syndrome coronavirus in stool specimens by commercially available real-time reverse transcriptase PCR assays 下载免费PDF全文
Louie L Simor AE Chong S Luinstra K Petrich A Mahony J Smieja M Johnson G Gharabaghi F Tellier R Willey BM Poutanen S Mazzulli T Broukhanski G Jamieson F Louie M Richardson S;Ontario Laboratory Working Group for the Rapid Diagnosis of Emerging Pathogens 《Journal of clinical microbiology》2006,44(11):4193-4196
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Tao Pu Chen Ding Yadong Li Xiaojuan Liu Haiwei Li Jinmei Duan Heng Zhang Yanwei Bi Wei Cun 《Journal of medical virology》2020,92(9):1609-1614
Since the outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in humans in late 2019, it has rapidly spread worldwide. To identify the biological characteristics of SARS-CoV-2 in a normal laboratory environment (biosafety level 2 [BSL-2]), a lentiviral-based nucleocapsid was used to carry the spike protein of SARS-CoV-2 onto the surface of pseudoviral particles as a surrogate model to evaluate the infective characterization of SARS-CoV-2. This study indicated that SARS-CoV-2 has extensive tissue tropism for humans and may infect monkeys and tree shrews but not rodents. More importantly, the use of pseudoviral particles in this study allows rapid assessment of neutralizing antibodies in serum in a BSL-2 laboratory. This study will provide a quick and easy tool for evaluating neutralizing antibodies in the serum of recovering patients and assessing the potency of candidate vaccines. 相似文献
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SARS肺的病理鉴别诊断 总被引:2,自引:4,他引:2
目的 分析严重急性呼吸综合征(severe acute respiratory syndrome,SARS)肺部病变的临床病理形态学特征及与其他肺部炎性疾病的鉴别诊断。方法 对3例行尸体解剖的SARS病例的肺组织进行组织形态学、免疫组织化学和电镜下超微结构的观察,对其临床表现、病理形态学特征及主要鉴别诊断进行探讨,并与病理资料完整的16例病毒性肺炎、13例间质性肺炎的肺部病变进行鉴别诊断。结果 死亡的SARS病人肺部病变主要表现为弥漫性的肺部损伤,但是各处病变轻重不一。肺泡腔内可见细颗粒样或泡状水肿液,其中可见脱落的肺泡上皮细胞。肺泡间隔表面可见透明膜覆盖,有的区域肺泡腔内出现机化性改变,肺泡腔和肺问质内可见大量的巨噬细胞浸润。同时,肺间质内的小动脉壁出现明显的损伤性改变。结论 SARS的肺部病理形态学特征可与其他肺部炎症性疾病鉴别。 相似文献
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Nitric oxide (NO) has been shown to suppress Japanese encephalitis virus (JEV) RNA synthesis, viral protein accumulation,
and virus release from infected cells. In this article, the potential viral structural proteins as the activators of NO product
were studied at the molecular level. First, the genomic region encoding the JEV structural proteins was cloned into a prokaryotic
expression vector pET for high-level expression. After purification, these JEV recombinant proteins were added to macrophages
to examine the productions of NO and pro-inflammatory mediators. In this study, the recombinant core protein, but not envelope
(E), could trigger NO and pro-inflammatory mediators (TNF-α, IL-6, and IL-12) productions on macrophages. And their effects
were about 85–95% relative to LPS-stimulated macrophages in a dose-dependent manner. Meanwhile, the rCore-2D could up regulate
promoters of IL-8 and TNF-α via EGFP expression in reporter plasmid (IL-8p–EGFP and TNF-αp–EGFP)-transfected cells by flow
cytometric analysis. These results suggest that JEV core protein could regulate pro-inflammatory mediators and NO production,
and may play a crucial role in the innate immunity for the host to restrict the initial stage of JEV infection. 相似文献