吸附无细胞百白破-重组乙型肝炎(CHO细胞)联合疫苗的中试工艺

目的探讨吸附无细胞百白破-重组乙型肝炎(CHO细胞)联合疫苗(DTaP-HBV)的中试工艺。方法通过不同的配比条件,探索DTaP-HBV联合疫苗的最佳中试工艺。以此工艺制备3批DTaP-HBV,进行各项指标的检定及稳定性试验。结果配方以无细胞百日咳疫苗原液18μgPN/ml,精制白喉类毒素25Lf/ml,精制破伤风类毒素7Lf/ml,重组乙肝疫苗原液20μg/ml为宜;DTaP-HBV的稀释液选用0.85%NaCl溶液吸附效果较好,各抗原间无干扰作用;不同配合方式对四联疫苗的效力影响不大。3批DTaP-HBV的各项检定指标均符合《中国药典》三部(2005版)要求,且稳定性良好。结论已筛选出DTaP-HBV联合疫苗的最佳中试工艺,以此工艺制备的疫苗安全、稳定、有效。     

A+C群脑膜炎球菌多糖疫苗大规模接种后的速发型接种反应观察

目的观察A+C群脑膜炎球菌多糖疫苗大规模接种后的速发接种反应。方法以A+C群脑膜炎球菌多糖疫苗为观察组,伤寒Vi多糖疫苗为对照组,按组群随机分层配对的原则,将观察现场分为108个组群,观察组和对照组各分配54个组群进行接种。建立接种反应监测系统,按统一表格和方法对两组的速发接种反应进行监测。结果两组共接种34 543人,其中A+C群脑膜炎球菌多糖疫苗接种18 167人,伤寒Vi多糖疫苗接种16 376人。A+C群脑膜炎球菌多糖疫苗速发接种反应率为0.44‰;伤寒Vi多糖疫苗速发接种反应率为0.79‰,二者差异无统计学意义;速发接种反应均出现在接种后15 min内,最早的出现在接种后5 min,速发接种反应中有2例为异常反应,但未出现严重反应。结论A+C群脑膜炎球菌多糖疫苗速发接种反应发生率低,预后良好。     

免疫增强性重建流感病毒体在疫苗研究中的应用

免疫增强性重建流感病毒体(IRIV)是重建的流感病毒囊膜,其保持了由病毒囊膜糖蛋白血凝素介导的天然病毒的细胞结合及膜融合特性,但缺乏病毒遗传物质。越来越多的证据表明,IRIV可作为佐剂、流感疫苗及抗原呈递载体,在疫苗的研究中发挥重要作用。本文就IRIV作为佐剂、流感疫苗和抗原呈递载体的应用进展作一综述。     

分子生物学技术在黄病毒属病毒疫苗研制中的应用及该类疫苗的研究进展

黄病毒科黄病毒属包括70多种病毒,其中约40多种与人类疾病相关,大部分为虫媒病毒。其中对人类致病的重要病原主要有登革病毒、乙型脑炎病毒、黄热病毒、蜱传脑炎病毒等。分子生物学技术的发展为黄病毒属病毒疫苗的设计带来了新的策略。本文对当前分子生物学技术如感染性克隆技术在黄病毒属病毒疫苗研制中的应用以及嵌合疫苗、蛋白亚单位疫苗和DNA疫苗的研究进展作一综述。     

冻干甲型肝炎减毒活疫苗的接种反应及免疫原性观察

目的观察冻干甲型肝炎减毒活疫苗的接种反应和免疫原性。方法采用随机、开放式试验设计,选择出生后未接种过甲肝疫苗的18月龄以上的儿童和18～35周岁的成人,接种冻干甲肝减毒活疫苗1针。接种后30min观察即时反应,24、48和72h进行局部和全身反应观察,免后30d采血,分离血清,进行免疫原性检测。结果562名观察对象接种疫苗后均未发生局部反应,全身反应仅表现为轻度发热,有35人次(6.23%)发生发热反应。471名完成免疫原性检测的观察对象免后抗-HAV阳性率为99.8%,保护性抗体阳性率为96.4%,抗-HAVGMC水平为527.4mIU/ml,较免前增长30.3倍。儿童组和成人组免后保护性抗体阳性率分别为97.0%和95.9%,抗-HAVGMC水平分别为84.1和2119.1mIU/ml。2组免前免后抗体阳性率和GMC水平差异均有统计学意义。277名易感者免后抗体阳性率为99.6%,保护性抗体阳性率为93.9%,GMC水平为92.6mIU/ml,儿童组免后抗体阳性率明显高于成人组,免后抗-HAVGMC水平则明显低于成人组,2组之间免后抗体阳性率和GMC水平差异均有统计学意义。结论国产冻干甲型肝炎减毒活疫苗接种后副反应轻微,免疫原性良好,适于推广应用。     

乙型脑炎减毒活疫苗诱导小鼠的细胞免疫应答

目的探讨乙型脑炎(简称乙脑)减毒活疫苗在小鼠中诱导细胞免疫应答的能力,评价疫苗的免疫原性。方法以乙脑减毒活疫苗免疫BALB/c小鼠,制备小鼠脾淋巴细胞,在体外以重组蛋白NS3-1和NS3-2进行刺激,采用酶联斑点法(ELISPOT)检测分泌细胞因子的效应T细胞的频数。结果乙脑减毒活疫苗免疫小鼠的脾淋巴细胞在体外经重组蛋白NS3-1和NS3-2刺激后,可诱导高频数的分泌IFNγ的CD4+和CD8+T淋巴细胞及低频数的分泌IL-4的淋巴细胞;而乙脑灭活疫苗仅诱导低频数的分泌IFNγ和IL-4的T淋巴细胞。结论乙脑减毒活疫苗可在小鼠中有效地诱导NS3-1和NS3-2抗原特异性T淋巴细胞应答,且应答水平显著高于乙脑灭活疫苗。     

一种新的求解配电网重构问题的免疫遗传算法

针对遗传算法求解配电网重构问题存在的不足,建立了以网损最小为目标函数的配电网重构数学模型,提出了一种新的免疫遗传算法。该算法的关键在于疫苗库的构建和免疫算子的设计。疫苗库可自动建立和更新,免疫算子由接种疫苗和免疫检测组成。另外,采用了基于基本环路的编码方法、高频变异和大选择压的锦标赛选择算子。IEEE 33和IEEE 69系统的仿真测试结果表明：该算法符合配电网重构问题的特点,能有效抑制进化中的退化和波动现象,在确保解的质量的同时,具有很快的收敛速度;与传统遗传算法和相关文献中的同类算法相比,该算法在效率和性能方面具有优越性。     

乳酸菌用作口服疫苗表达载体的应用研究进展

乳酸菌是人和大多数动物肠道内的常见细菌,被公认为绿色安全级微生物,可以在机体内粘附、定植和复制,益生作用和免疫原性使其成为最有前景的活菌载体。利用基因工程手段插入外源基因制备重组乳酸菌口服疫苗,能够持续刺激机体分泌特异性抗原蛋白,多个基因的插入甚至能达到一免治多病的目的。口服免疫效果优于注射免疫,治疗成本也大大降低。该文论述了乳酸菌活载体疫苗的优势,对乳酸菌表达载体在动物疫病防控及代谢产物调控上的应用等方面的研究进行综述,并对其应用前景进行分析和展望。     

The Versatile Manipulations of Self-Assembled Proteins in Vaccine Design

Protein assemblies provide unique structural features which make them useful as carrier molecules in biomedical and chemical science. Protein assemblies can accommodate a variety of organic, inorganic and biological molecules such as small proteins and peptides and have been used in development of subunit vaccines via display parts of viral pathogens or antigens. Such subunit vaccines are much safer than traditional vaccines based on inactivated pathogens which are more likely to produce side-effects. Therefore, to tackle a pandemic and rapidly produce safer and more effective subunit vaccines based on protein assemblies, it is necessary to understand the basic structural features which drive protein self-assembly and functionalization of portions of pathogens. This review highlights recent developments and future perspectives in production of non-viral protein assemblies with essential structural features of subunit vaccines.     

Babesia Bovis Ligand-Receptor Interaction: AMA-1 Contains Small Regions Governing Bovine Erythrocyte Binding

Apical membrane antigen 1 is a microneme protein which plays an indispensable role during Apicomplexa parasite invasion. The detailed mechanism of AMA-1 molecular interaction with its receptor on bovine erythrocytes has not been completely defined in Babesia bovis. This study was focused on identifying the minimum B. bovis AMA-1-derived regions governing specific and high-affinity binding to its target cells. Different approaches were used for detecting ama-1 locus genetic variability and natural selection signatures. The binding properties of twelve highly conserved 20-residue-long peptides were evaluated using a sensitive and specific binding assay based on radio-iodination. B. bovis AMA-1 ectodomain structure was modelled and refined using molecular modelling software. NetMHCIIpan software was used for calculating B- and T-cell epitopes. The B. bovis ama-1 gene had regions under functional constraint, having the highest negative selective pressure intensity in the Domain I encoding region. Interestingly, B. bovis AMA-1-DI (¹⁰⁰YMQKFDIPRNHGSGIYVDLG¹¹⁹ and ¹²⁰GYESVGSKSYRMPVGKCPVV¹³⁹) and DII (³⁰²CPMHPVRDAIFGKWSGGSCV³²¹)-derived peptides had high specificity interaction with erythrocytes and bound to a chymotrypsin and neuraminidase-treatment sensitive receptor. DI-derived peptides appear to be exposed on the protein’s surface and contain predicted B- and T-cell epitopes. These findings provide data (for the first-time) concerning B. bovis AMA-1 functional subunits which are important for establishing receptor-ligand interactions which could be used in synthetic vaccine development.