人巨细胞病毒IgG抗体(ELISA)检测试剂盒稳定性观察

目的观察人巨细胞病毒(human cytomegalovirus,HCMV)IgG抗体(ELISA)检测试剂盒的稳定性。方法将HCMV IgG抗体(ELISA)检测试剂盒分别以不同时间放置于2~8℃和37℃,检测试剂盒的外观、阳性参考品符合率、阴性参考品符合率、重复性和检测限,并对其进行稳定性(实时稳定性、加速破坏稳定性、开瓶稳定性、运输稳定性)的观察及初步应用。结果 3批实验用试剂盒实时稳定性和3批试生产试剂盒加速破坏稳定性、开瓶稳定性、运输稳定性试验中,外观均合格,阴性参考品符合率及阳性参考品符合率均为100%,最低检出限参考品均能检出。3批实验用试剂盒实时稳定性检测重复性良好,CV值均≤15.00%。3批试生产试剂盒在37℃条件下放置6 d后,CV值分别为7.94%、7.16%、4.66%;开瓶后分别置于2~8℃冰箱0w、1w、2w、3w、4w,CV值均≤1 5.00%;运输至上海,CV值分别为2.59%、3.12%、2.37%;运输至海南,CV值分别为4.89%、2.65%、2.33%。样品反复冻融5次后,试剂盒检测稳定性良好。结论 HCMV IgG抗体(ELISA)检测试剂盒具有良好的稳定性,在2~8℃条件下至少可保存15个月。     

人巨细胞病毒疫苗研究进展

人巨细胞病毒(Human cytomegalovirus,HCMV)是一类在自然界普遍存在的具有严格种属特异性的病毒,属β疱疹病毒亚科.Zsofia Gyulai等[1]对HCMV感染作了流行病学调查,发现体内抗病毒体液免疫反应主要针对gB;而细胞免疫反应主要针对pp65、pp150.根据HCMV感染的特点和机制,以上述几种结构糖蛋白设计候选基因工程疫苗,已得到了广泛的研究,包括基因工程亚单位疫苗、质粒载体DNA疫苗、病毒载体DNA疫苗、合成蛋白(肽)疫苗,另外还有Towne减毒活疫苗.在国外,有的疫苗已完成Ⅱ期临床试验研究[2].本文现就HCMV疫苗的种类及优缺点、相关佐剂、展望进行综述.     

人巨细胞病毒疫苗研究进展

人巨细胞病毒(Human cytomegalovirus, HC-MV)是一类在自然界普遍存在的具有严格种属特异性的病毒,属β疱疹病毒亚科。Zsofia Gyulai等[1]对HCMV感染作了流行病学调查,发现体内抗病毒体液免疫反应主要针对gB;而细胞免疫反应主要针对pp65、pp150。根据HCMV感染的特点和机制,以上     

人巨细胞病毒疫苗研究进展

人巨细胞病毒（HCMV）基因在优生优育方面的重要作用,一直受到人们的广泛重视。迄今,随着免疫缺陷病人的增多,HCMV感染对人体健康的危害也日趋严重,发展有效,安全的疫苗即成为防治HCMV疾病的重要手段。本文就国外学者对HCMV疫苗的研究状况做了简要介绍。     

人巨细胞病毒IgG酶联免疫检测试剂盒的研制

为研制人巨细胞病毒 (HCMV)IgG酶联检测试剂盒 ,将HCMV接种人二倍体细胞 ,收获的病毒经纯化后用作包被抗原 ,辣根过氧化物酶 (HRP)标记羊抗人IgG为检测抗体 ,采用间接ELISA制备酶联免疫检测试剂盒并进行检定。该试剂盒操作简便、特异性强 ,稳定性、线性及精密性符合体外诊断试剂的要求。     

人巨细胞病毒的多聚酶链反应研究进展

人巨细胞病毒是疱疹病毒科的一个成员，在全世界呈广泛分布，现在发现该病毒不只在新生儿，免疫抑制及免疫缺陷患者中导致广泛的临床症状甚至死亡，而且与人类三大疾病－－心血管疾病，恶性肿瘤及糖尿病有关，因此在当前人类疾病防治中ＣＭＶ感染的研究成为一个热门课题。本文就病毒感染的多聚酶链反应诊断方法的研究进展作一介绍。     

人巨细胞病毒与肿瘤研究进展

人巨细胞病毒（HCMV）是目前已知最大的β疱疹病毒。HCMV感染具有持续性和潜在性,感染率在全世界范围内都很高,并随着年龄的增长而升高,女性感染率高于男性,其主要的传播途径有垂直传播和性传播等。近年来的研究显示在人类胶质瘤、结直肠癌、乳腺癌和前列腺癌及小细胞型肝癌等多种类型的肿瘤组织中都存在HCMV感染和病毒基因表达,并与肿瘤的恶性程度有相关性,这提示HCMV可能在人类某些类型肿瘤的形成和发展过程中扮演重要角色,有可能成为一种新的人类肿瘤相关病毒。HCMV基因产物可通过多种细胞信号通路抑制细胞凋亡、促进细胞增殖、侵袭、转移和血管生成,并形成独特的免疫逃避机制对抗机体免疫反应。深入研究HCMV与肿瘤的病因学关系及其作用机制,可为肿瘤的临床防治提供新思路。     

巨细胞病毒感染的治疗研究进展

人巨细胞病毒(HCMV)为DNA双螺旋病毒,属疱疹病毒科β亚科,是疱疹病毒科中基因组最大的病毒。HCMV通常呈隐性感染,正常人感染HCMV并无临床症状,一旦人体的免疫机能发生变化(如肿瘤、AIDS、妊娠、新生儿和器官移植),HCMV即从潜伏态转化为原发性或继发性的增殖感染,引起严重的临床症状甚至死亡。因此,HCMV的预防和治疗越来越受到关注,就CMV感染的预防及治疗方法进行了综述。     

孕妇尿液人巨细胞病毒DNA筛查及血液HCMV抗体测定的对照研究

目的对于孕妇尿液进行HCMV—DNA筛查,减少和有效地避免胎儿及新生儿的HCMV感染。HCMV．DNA筛查同时进行HCMV抗体检测,明确诊断HCMV感染的灵敏、准确方法。方法应用荧光定量PCR（FQ．PCR）方法进行尿液HCMV-DNA测定。血液HCMV IgM、IgG抗体检测应用酶联免疫（ELISA）方法。结果筛查6568例孕妇尿HCMV．DNA,阳性273例,阳性率为4．2％。孕妇在12—20周者,阳性率为10．3％;孕妇在21～30周者,阳性率为33．3％;孕妇在31～39周者,阳性率为56．4％。在273例尿液HCMV—DNA阳性者中,56例同时进行血液HCMV IgM、IgG抗体检测,Igi抗体阳性者2例,IgG抗体阳性者22例。结论在孕妇HCMV感染的筛查与诊断中FQ．PCR是灵敏准确的方法。孕妇中HCMV—DNA筛查是保证母婴健康,提高人口素质的保障。     

人巨细胞病毒抗原血症

人巨细胞病毒（ＨＣＭＶ）抗原血症试验是近年来报道的一种早期,快速诊断活动性ＨＣＭＶ感染的新方法。本文就其方法学,敏感性与特异性及实用价值作一综述。     

更昔洛韦联合大剂量丙种球蛋白治疗婴儿巨细胞病毒性肝炎的疗效观察

目的:研究更昔洛韦联合大剂量丙种球蛋白治疗婴儿巨细胞病毒性(CMV)肝炎的临床疗效,为婴儿CMV肝炎的抗菌治疗提供理论依据。方法:选取我院自2009年7月至2016年1月收治的103例CMV肝炎婴儿,根据随机数字表法分为治疗组58例,对照组45例。两组患儿均常规给予退黄、保肝药物治疗,对照组在此基础上给予更昔洛韦治疗,治疗组给予大剂量丙种球蛋白联合更昔洛韦治疗。对比观察两组患儿治疗后黄疸消退情况、肝功能变化、血CMV-IgM和尿CMV-DNA转阴情况、肝和脾B超变化、治疗效果及不良反应情况。结果:治疗组患儿黄疸消退时间、血CMV-IgM和尿CMV-DNA转阴时间均短于对照组(P0.05);治疗组肝功能指标包括总胆红素(TBIL)、直接胆红素(DBIL)、丙氨酸氨基转移酶(ALT)、γ-谷氨酰转肽酶(GGT)水平均较对照组和治疗前降低(P0.05);治疗后两组肝、脾明显减小,且治疗组减小更显著,差异有统计学意义(P0.05);治疗组中血CMV-IgM转阴率、尿CMV-DNA转阴率、治疗总有效率均高于对照组(P0.05);治疗组不良反应发生率低于对照组(P0.05)。结论:更昔洛韦联合大剂量丙种球蛋白治疗婴儿CMV肝炎临床效果良好,可以有效缓解黄疸症状,恢复肝脏功能,肝、脾回缩明显,不良反应少,值得推广。     

Structural Characterization of Anti-Inflammatory Immunoglobulin G Fc Proteins

Immunoglobulin G (IgG) is a central mediator of host defense due to its ability to recognize and eliminate pathogens. The recognition and effector responses are encoded on distinct regions of IgGs. The diversity of the antigen recognition Fab domains accounts for IgG's ability to bind with high specificity to essentially any antigen. Recent studies have indicated that the Fc effector domain also displays considerable heterogeneity, accounting for its complex effector functions of inflammation, modulation, and immune suppression. Therapeutic anti-tumor antibodies, for example, require the pro-inflammatory properties of the IgG Fc to eliminate tumor cells, while the anti-inflammatory activity of intravenous IgG requires specific Fc glycans for activity. In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc C_H2 domain include terminal α2,6-linked sialic acids. We used chemoenzymatic glycoengineering to prepare fully disialylated IgG Fc and solved its crystal structure. Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the C_H2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc.     

The Raw Material Equivalents of International Trade

This article aims at estimating the raw material equivalents (RMEs)—the upstream used material flows required along the production chain—of imports and exports for some Latin American countries: Brazil, Chile, Colombia, Ecuador, and Mexico. Furthermore, the United States is included in the analysis as a reference for a high‐income economy. The RME concept and the empirical evidence are articulated by use of an input?output methodology. Results are set out for the year 2003 for each of the countries and in time series for the years 1977, 1986, 1996, and 2003 in the case of Chile. The findings show not only the physical dimensions behind direct material traded but also how the previous exporter (importer) position of a country (based on standard material flow analysis indicators) deteriorates, alleviates, or changes. Implications for material consumption indicators, such as direct material consumption (DMC) and raw material consumption (RMC), are also drawn. The results suggest basing the discussion of material flows on a broader set of indicators to obtain a more comprehensive picture of the implications of international trade and its impacts on the environment.     

干热处理法对人免疫球蛋白某些特性的影响

人免疫球蛋白制品以蔗糖为热稳定剂经６０℃８０小时和７２℃８０小时干热处理后理化性质的改变结果表明,６０℃８０小时干热处理后ＩｇＧ各成份相对含量,重新溶解后外观等结果与干热处理前基本相同,经选择不同种类糖作热稳定剂比较,以７－８％蔗糖浓度稳定效果较好     

免疫球蛋白基因的转录调控

免疫球蛋白(Ig)的表达是B细胞特异性和发育阶段特异性的事件.Oct2、NF-kB和HLH蛋白与Ig基因细胞特异转录有关.Oct2含有POU结构域,属POU转录调控因子家族;NF-kB有rel-like结构域,属rel-like转录调控因子家族;HLH蛋白特征性结构为螺旋-环-螺旋(helix-loop-helix,HLH)结构域,已报道的HLH蛋白都与转录调节有关.     

Exploring variation in binding of Protein A and Protein G to immunoglobulin type G by isothermal titration calorimetry

Bacterial Protein A (PrtA) and Protein G (PrtG) are widely used for affinity purification of antibodies. An understanding of how PrtA and PrtG bind to different isotypes of immunoglobulin type G (IgG) and to their corresponding Fc fragments is essential for the development of PrtA and PrtG mimetic ligands and for the establishment of generic processes for the purification of various antibodies. In this paper, the interactions between the two IgG-binding proteins and IgG of two different subclasses, IgG1 and IgG4, as well as their analogous Fc fragments have been studied by isothermal titration calorimetry. The results indicate that both protein ligands bind IgG and Fc fragments strongly with Ka values in the range of 10(7) -10(8) M(-1) and for both ligands, the interaction with both IgG isotypes is enthalpically driven though entropically unfavorable. Moreover, variation in the standard entropic and standard enthalpic contribution to binding between the two isotypes as well as between IgG and Fc fragment implies that the specific interaction with PrtA varies according to IgG isotype. In contrast to PrtA, PrtG bound to F(ab')(2) fragment with a Ka value of 5.1 × 10(5) M(-1) ; thus underscoring the usefulness of PrtA as a preferred ligand for generic antibody purification processes.     

Detection of disease-specific augmentation of abnormal immunoglobulin G in sera of patients with rheumatoid arthritis

Galactose-free immunoglobulin G (IgG), which is known to be higher in the sera of patients with rheumatoid arthritis, was prepared from IgG of healthy volunteers using enzymes. Its reactivity to lectins was analyzed. The galactose-free IgG showed no reactivity to Ricinus communis agglutinin 120 but displayed greater reactivity to concanavalin A and Lens culinaris lectin than did intact human IgG. Then, IgG in serum samples was bound to protein A immobilized on a nitrocellulose membrane, and its reactivity to biotinylated concanavalin A was measured with streptavidin-conjugated horseradish peroxidase. When the reactivity to concanavalin A of IgG in sera from healthy individuals and patients with rheumatoid arthritis (RA), osteoarthritis, systemic lupus erythematosus, or hepatic disease was compared, higher levels were shown in patients with RA, notably in 60% of the seronegative patients and 80% of the early phase patients. Therefore, it was suggested that augmentation of the abnormal IgG in sera was highly specific to patients with RA and that this novel serum test could be very useful for an accurate diagnosis of this disease.     

Conformational Destabilization of Immunoglobulin G Increases the Low pH Binding Affinity with the Neonatal Fc Receptor

Crystallographic evidence suggests that the pH-dependent affinity of IgG molecules for the neonatal Fc receptor (FcRn) receptor primarily arises from salt bridges involving IgG histidine residues, resulting in moderate affinity at mildly acidic conditions. However, this view does not explain the diversity in affinity found in IgG variants, such as the YTE mutant (M252Y,S254T,T256E), which increases affinity to FcRn by up to 10×. Here we compare hydrogen exchange measurements at pH 7.0 and pH 5.5 with and without FcRn bound with surface plasmon resonance estimates of dissociation constants and FcRn affinity chromatography. The combination of experimental results demonstrates that differences between an IgG and its cognate YTE mutant vary with their pH-sensitive dynamics prior to binding FcRn. The conformational dynamics of these two molecules are nearly indistinguishable upon binding FcRn. We present evidence that pH-induced destabilization in the CH2/3 domain interface of IgG increases binding affinity by breaking intramolecular H-bonds and increases side-chain adaptability in sites that form intermolecular contacts with FcRn. Our results provide new insights into the mechanism of pH-dependent affinity in IgG-FcRn interactions and exemplify the important and often ignored role of intrinsic conformational dynamics in a protein ligand, to dictate affinity for biologically important receptors.     

A comparative study of the effect of modification of the surface of human platelets on the receptors for aggregated immunoglobulins and for ristocetin-von Willebrand factor

The receptors for aggregated immunoglobulin G (IgG) (an Fc receptor) and for ristocetin-von Willebrand factor on human platelets were studied by means of various modifications of the platelet surface. The expression of these receptors was measured by the agglutination of platelets to ristocetin in the presence of von Willebrand factor, which is part of the factor VIII complex, and by the binding of aggregated IgG coupled to ³H-labelled diazobenzen. Treatment of platelets with chymotrypsin, trypsin, papain and pronase which removed protein and glycoprotein from the platelet under conditions where the release reaction was inhibited caused loss of the expression of the receptor for ristocetin-von Willebrand factor and an enhancement of that for aggregated IgG. Induction of membrane changes with ADP and of the release reaction with the ionophore A23187 abolished agglutination to ristocetin-von Willebrand factor but did not alter the receptor for aggregated IgG. Possible contributions of unspecific membrane changes, produced by protease treatment of platelets, to the modification of receptor expression were eliminated by the use of formaldehyde-treated platelets. Trypsin, papain and pronase destroyed the ability of these platelets to agglutinate to ristocetin-von Willebrand factor but produced no change in the binding of aggregated IgG. Therefore, the receptor for ristocetin-von Willebrand factor is truly sensitive to proteolysis while the Fc receptor is not, but is partially masked by protease-sensitive material.