mRNA疫苗的开发及临床研究进展

随着mRNA稳定性和安全高效的递送系统的研究日渐成熟,近年来,mRNA疫苗在肿瘤个体化疫苗中取得了较大进展,因其生产工艺简单、在细胞内表达抗原、安全性优于DNA疫苗等特点,是一种很有前途的新型疫苗。为了解全球mRNA疫苗的开发与研究现状,在此重点对mRNA疫苗的分子设计、递送系统、临床研究现状进行了分析和综述,为后续mRNA疫苗的开发和研究提供参考依据。     

汉坦病毒疫苗研制进展

本文从脑组织灭活疫苗、培养细胞单双价灭活疫苗、基因重组疫苗、DNA疫苗、减毒活疫苗、佐剂、灭活方法、免疫策略等方面概述了汉坦病毒疫苗研制近况。     

恶性疟原虫抗原表位的亚单位疫苗与DNA疫苗的联合免疫

构建了含有恶性疟原虫抗原基因 ( AWTE)的真核表达质粒 p CMV- AWTE,以及能在大肠杆菌中得到分泌性表达的原核表达质粒 p MC0 5 ,表达的蛋白 AWTE保持了疟原虫抗原的抗原性。将 p CMV- AWTE以及 AWTE两者混合各 1 0μg鼻腔免疫小鼠 ,一次后诱导机体产生了较高水平的体液免疫及细胞免疫     

核酸疫苗的研究及应用进展

核酸疫苗(DNA疫苗)是20世纪90年代发展起来的一种新型疫苗。核酸疫苗不仅可引起体液免疫反应,而且能诱导高水平的细胞免疫应答,尤其是细胞毒T淋巴细胞(CTL)反应,被认为在病毒、细菌、寄生虫等病原体感染的防治中具有更大的优势。核酸疫苗既可作为病毒、细菌或寄生虫的预防疫苗,也可作为非感染性疾病如肿瘤病的治疗用疫苗。     

重组乙型肝炎疫苗(CHO细胞)鉴别试验的方法研究

目的：建立重组乙型肝炎疫苗(CHO细胞)鉴别试验的方法。方法：取重组乙型肝炎疫苗(CHO细胞)以1：10比例加入lmol/L盐酸调节pH值,采用ELISA双抗体夹心法检测,同时设以氢氧化铝、未处理的重组乙型肝炎疫苗、狂犬疫苗、出血热疫苗,以及试剂盒的阴、阳性作为对照。结果：狂犬疫苗、出血热疫苗及氢氧化铝对照为阴性,直接检测重组乙型肝炎疫苗(CHO细胞)呈弱阳性,加入盐酸调节pH值后原倍样品呈弱阳性,10倍稀释后呈强阳性。采用该方法检测5批疫苗均呈强阳性。组内平行试验,组间重复试验。结论：将重组乙型肝炎疫苗(CHO细胞)调节pH值后10倍稀释,用ELISA双抗体夹心法检测呈阳性,而且稳定可靠,该方法可用于该疫苗的鉴别试验。     

Laboratory assays of different types of field trial typhoid vaccines and relationship to efficacy in man

Pittman, Margaret (National Institutes of Health, Bethesda, Md.), and Howard J. Bohner. Laboratory assays of different types of field trial typhoid vaccines and relationship to efficacy in man. J. Bacteriol. 91:1713-1723. 1966.-Antibody responses of rabbits to H, O, and Vi antigens did not differentiate vaccine K (acetone-killed and dried) from vaccine L (heat-phenolized and dried) relative to human efficacy. A mouse protection assay in which intraperitoneal vaccination and challenge suspended in mucin were used showed vaccine K to be 3.69 times more potent than vaccine L; with subcutaneous vaccination, vaccine K was only 0.78 as potent. With the challenge suspended in saline, the effect of the route of vaccination was accentuated. An old U.S. reference vaccine, heat-phenolized, induced the same types of response as vaccine L. In the assay with intraperitoneal vaccination and mucin-suspended challenge, the potency of vaccine K relative to vaccine L and the potencies of Polish vaccines, P, N, and T, relative to vaccine K were directly correlated with the efficacies of the vaccines for man, as reported for the recent World Health Organization cooperative field trials in British Guiana, Yugoslavia, and Poland. This assay gave a high potency for alcohol-treated vaccine V relative to vaccine L, but the values did not reflect relative efficacy in the USSR field trial; the subcutaneous vaccination assay more closely reflected their human efficacy. An analysis suggested that vaccine K had a mouse protective factor not present in vaccine L and that vaccine V may have had a third factor. The influence of a few variable factors on the assay with intraperitoneal vaccination and mucin-suspended challenge was studied briefly.     

轮状病毒灭活疫苗和减毒活疫苗序贯免疫的体液免疫应答效果

目的:评价采用轮状病毒灭活疫苗进行初始免疫,减毒活疫苗进行加强免疫的序贯免疫方案的体液免疫应答效果。方法:将实验小鼠随机分为4组(口服疫苗组、序贯疫苗组、口服对照组及序贯对照组),按相应方案免疫后,ELISA检测血清轮状病毒特异性IgG和IgA、肠道轮状病毒特异性IgA;微量中和实验检测血清病毒特异性中和抗体;同时采用ELISA分析口服活疫苗后病毒排出情况。结果:与对照组相比,序贯疫苗组小鼠产生的轮状病毒特异性血清IgG、IgA、中和抗体及肠道IgA水平显著升高。与口服疫苗组相比,序贯疫苗组的免疫方案诱发的轮状病毒特异性血清IgG、IgA、中和抗体水平显著升高,肠道IgA水平两组间没有显著差异。同时,与口服疫苗组相比,序贯疫苗组中轮状病毒灭活疫苗进行的初始免疫未影响第一次口服活疫苗后病毒的排出量和排出时间,但序贯疫苗组第二次口服活疫苗后病毒的排出量迅速减少,排毒时间快速缩短,与口服疫苗组第三次服苗后病毒的排出量和排出时间相似。结论: 轮状病毒灭活疫苗和减毒活疫苗序贯免疫可有效诱发小鼠全身和黏膜局部的体液免疫应答,该方案将有可能成为轮状病毒疫苗临床应用的候选方案。     

表位疫苗及相关技术研究进展

表位疫苗是用抗原表位制备的疫苗,是近年来新兴的一种疫苗研制技术,也是今后最具开发前景的疫苗技术之一,在肿瘤、病毒等疾病的防治中有着自身独特的优势。本文详细阐述了T表位和B表位的筛选和鉴定方法、表位疫苗的载体研究及表位疫苗在肿瘤、病毒和微生物感染中的应用等,对表位疫苗的最新研究进展进行了综述。     

The characteristics of the reactogenicity and immunological activity of a new cholera bivalent chemical vaccine based on the results of controlled trials]

The reactogenic properties and immunological potency of modified cholera chemical vaccine (choleragen-toxoid + O-antigens Inaba and Ogawa) were tested in 278 volunteers aged 18 years and over in comparison with those of a commercial batch of monovalent cholera vaccine (choleragen-toxoid + O-antigen Inaba). The cholera vaccine, enriched with O-antigen Ogawa, was found to be safe; vaccination with this vaccine was not accompanied by the development of systemic and local reactions whose frequency and intensity met the requirements for the reactogenic properties of commercial cholera vaccine. The immunological potency of the bivalent vaccine with respect to strain Inaba was not inferior to that of the commercial vaccine; at the same time in persons immunized with the new preparation the titers of vibriocidal antibodies to strain of serovar Inaba were five-fold higher. The conclusion on the expediency of using cholera chemical vaccine enriched with O-antigen Ogawa was made.     

核酸疫苗制备工艺研究进展

核酸疫苗是指被用作疫苗的带有编码外源蛋白抗原基因的真核表达质粒,它能较安全地表达目的蛋白,是具有较大发展潜力的一类生物制品。研究核酸疫苗的制备工艺,对于规模化生产成本低、免疫效果好的疫苗具有重要的现实意义。核酸疫苗规模化生产工艺的流程主要包括核酸疫苗的构建、工程菌的发酵、菌体裂解、分离纯化及产品检验等关键环节,而每个环节对核酸疫苗的质量都有较大的影响。     

Evaluation of the reactogenicity and immunogenic potency of combined vaccine "Bubo-Kok" in the immunization of children against diphtheria, tetanus, pertussis and viral hepatitis B

Combined vaccine "Bubo-Kok" is characterized by safety and high immunological activity. The number of postvaccinal reactions in children aged 1 and 2 years, immunized with vaccine "Bubo-Kok", was not statistically different from those in groups of children immunized with adsorbed DPT vaccine, as well with such vaccine in combination with vaccine against hepatitis B. After the completion of the primary course of immunization 100% of children had protective antibody titers against diphtheria, tetanus and hepatitis B. Antibody titers against pertussis, equal to or exceeding protective titers, were found in more than 70% of immunized children. The immunogenic potency of vaccine "Bubo-Kok" with respect to all its components was not inferior to that of adsorbed DPT vaccine and vaccine against hepatitis B, when introduced simultaneously in different areas of the body. Vaccine "Bubo-Kok" successfully passed state trials and was recommended for registration.     

麻疹、腮腺炎、风疹三联减毒活疫苗的安全性和免疫原性研究

为了评价国产麻疹、腮腺炎、风疹三联减毒活疫苗(MMR)的安全性和免疫原性,按整体随机抽样原则,以进口同类疫苗和国产各单价疫苗作为对照,开展现场临床观察;比较不同疫苗组免疫后的反应率、抗体阳转率、保护率及几何平均滴度(GMT)。试验组与对照组接种后,除了试验疫苗的中、强发热反应率高于进口对照疫苗的发热反应率(8.60%与2.00%)外,未见其它有显著差异的不良反应。试验组麻疹免后抗体阳转率高于进口对照疫苗(99.5%与94.6%),麻疹抗体GMT也高于单价麻疹对照疫苗的GMT;试验疫苗与进口MMR疫苗的风疹抗体阳转率、腮腺炎抗体阳转率相比,均无显著性差异。实验研究结果显示,试验MMR疫苗与进口MMR疫苗具有相似的临床反应及良好的免疫原性。     

Current Status of Measles and Oral Poliovirus Vaccines

Live attenuated measles vaccine, accompanied by a dose of gamma globulin to reduce systemic reactions, has given a high degree of protection, probably long lasting. Further attenuated vaccine gives promise of achieving the same result without the use of gamma globulin. Inactivated vaccine has not been shown to give durable immunity, but a schedule of killed vaccine followed by live vaccine has provided protection with minimal reactions. Inactivated vaccine can probably be combined with other antigens.Sabin oral poliovirus vaccines of all three types have been highly effective in preventing paralytic illness and reducing the spread of virulent strains. Because of the rare occurrence, chiefly in adults, of paralytic cases considered to be probably vaccine-associated, though no proof was possible, it has been recommended in Canada that initial immunization with Salk vaccine be continued and that all infants and children should subsequently receive trivalent Sabin vaccine.     

Antibody Response of Guinea Pigs to Trivalent Parainfluenza Virus Vaccine Prepared from Embryonated Eggs

A trivalent parainfluenza virus vaccine has been tested in guinea pigs. The parainfluenza 2 virus vaccine component was superior in the magnitude of antibody titers, and in the ability to convert animals serologically after two doses of an undiluted or a 10-fold diluted vaccine. The parainfluenza 1 virus vaccine gave a higher percentage of conversion than parainfluenza 3 virus vaccine after administration of two doses of either undiluted or 10-fold diluted vaccine.     

Immunological activity of a Proteus vaccine administered to volunteers simultaneously with a pyoimmunogen and adsorbed staphylococcal anatoxin

The comparative study of the immunological activity of Proteus vaccine prepared from soluble antigenic complexes was made after the immunization of volunteers with this vaccine used in the form of a single preparation and in combination with pyoimmunogen (Pseudomonas aeruginosa vaccine) and/or adsorbed staphylococcal toxoid. The injection of the vaccine in the form of a single preparation and in different combinations increased the ingestion of Proteus cells by neutrophils. The injection of Proteus vaccine simultaneously with pyoimmunogen and staphylococcal toxoid ensured the intensive phagocytosis of staphylococci. All combinations with Proteus vaccine, used in this investigation, stimulated the intensive formation of antibodies to Proteus vaccine strain and Re-glycolipid. Proteus vaccine introduced in combination with adsorbed staphylococcal toxoid essentially stimulated the synthesis of anti-alpha-staphylolysin.     

轮状病毒四价灭活疫苗的制备及在小鼠的免疫原性研究

制备轮状病毒四价灭活疫苗,观察其在小鼠体内的抗体应答情况。实验采用轮状病毒原液经凝胶过滤层析纯化,灭活后配制四价疫苗,肌肉注射免疫小鼠,ELISA法测定小鼠血清IgA、IgG。将单价G1、G2、G3、G4型灭活病毒原液及四价轮状病毒灭活疫苗免疫小鼠后,均可刺激产生针对RV的高水平的特异性IgG抗体,但IgA应答较弱。表明四价轮状病毒灭活疫苗在小鼠中具备很好的免疫原性。     

Effect of adjuvants on responses to skin immunization by microneedles coated with influenza subunit vaccine

Recent studies have demonstrated the effectiveness of vaccine delivery to the skin by vaccine-coated microneedles; however there is little information on the effects of adjuvants using this approach for vaccination. Here we investigate the use of TLR ligands as adjuvants with skin-based delivery of influenza subunit vaccine. BALB/c mice received 1 μg of monovalent H1N1 subunit vaccine alone or with 1 μg of imiquimod or poly(I:C) individually or in combination via coated microneedle patches inserted into the skin. Poly(I:C) adjuvanted subunit influenza vaccine induced similar antigen-specific immune responses compared to vaccine alone when delivered to the skin by microneedles. However, imiquimod-adjuvanted vaccine elicited higher levels of serum IgG2a antibodies and increased hemagglutination inhibition titers compared to vaccine alone, suggesting enhanced induction of functional antibodies. In addition, imiquimod-adjuvanted vaccine induced a robust IFN-γ cellular response. These responses correlated with improved protection compared to influenza subunit vaccine alone, as well as reduced viral replication and production of pro-inflammatory cytokines in the lungs. The finding that microneedle delivery of imiquimod with influenza subunit vaccine induces improved immune responses compared to vaccine alone supports the use of TLR7 ligands as adjuvants for skin-based influenza vaccines.     

Effect of prior vaccination with a seasonal trivalent influenza vaccine on the antibody response to the influenza pandemic H1N1 2009 vaccine: a randomized controlled trial

Vaccination with the non-adjuvanted split-virion A/California/7/2009 influenza vaccine (pandemic H1N1 2009 vaccine) began in October 2009 in Japan. The present study was designed to assess the effect of prior vaccination with a seasonal trivalent influenza vaccine on the antibody response to the pandemic H1N1 2009 vaccine in healthy adult volunteers. One hundred and seventeen participants aged 22 to 62 were randomly assigned to two study groups. In Group 1 (the priming group), participants were first vaccinated with the seasonal trivalent influenza vaccine followed by two separate one-dose vaccinations of the pandemic H1N1 2009 vaccine, whereas in Group 2 (the non-priming group), the participants were first vaccinated with one dose of the pandemic H1N1 2009 vaccine, followed by simultaneous vaccination of the seasonal trivalent vaccine and the second dose of the pandemic H1N1 2009 vaccine. The participants in Group 2 had a seroprotection rate (SPR) of 79.7% and a seroconversion rate (SCR) of 79.7% in the hemagglutination-inhibition test after the first dose of the pandemic H1N1 2009 vaccine, indicating that the pandemic H1N1 2009 vaccine is sufficiently immunogenic. On the other hand, the participants of Group 1 had a significantly weaker antibody response, with a SPR of 60.8% and a SCR of 58.5%. These results indicate that prior vaccination with the seasonal trivalent influenza vaccine inhibits the antibody response to the pandemic H1N1 2009 vaccine. Therefore, the pandemic H1N1 2009 vaccine should be administered prior to vaccination with the seasonal trivalent influenza vaccine.     

Investigations on the efficacy of monovalent Pseudomonas aeruginosa vaccine for oral administration in Pseudomonas aeruginosa experimental infection

Therapeutic efficacy of Pseudomonas aeruginosa vaccine for oral use (10(10) killed germs/ml), prepared from strain 4922, belonging to serotype XV, by Meitert-Meitert scheme, on 4 experimental models in mice (pneumonia, infected burn, septicaemia and urinary tract infection) was studied in comparison with monovalent Ps. aeruginosa vaccine serotype XV (10(9) killed germs/ml) for subcutaneous use and also with associated administration of the two vaccine variants. Mice immunization by using vaccine for oral use was performed by 0.5 ml vaccine per day, for 10 days and vaccine for subcutaneous use was administrated in a volume of 0.5 ml x 2, at 3 days interval. Mice immunization by using the two vaccine types, in association was concomitantly performed and in the same quantity as for separate immunization. In experimental pneumonia, Ps. aeruginosa vaccine for oral use protected mice in 35% of cases, those with infected burns were protected in 33.3% of cases, those with septicemia--in 96.6% of cases and those with urinary tract infection in 50% of cases. As compared to Ps. aeruginosa vaccine for subcutaneous use, the results obtained by vaccine for oral use are less favourable but associated administration of both vaccine variants led to superior results. Thus, in experimental pneumonia, it was obtained a surviving rate of 65% for animals immunized with both vaccine types, in comparison with 50% for animals immunized with vaccine for subcutaneous use only, and in Ps. aeruginosa infected burn, it was obtained a recovering rate of 79.1% for the animals immunized by using both vaccines, in comparison with 70.8% surviving for animals immunized with vaccine for subcutaneous use. In experimental septicaemia and urinary tract infection, combined use of both vaccine variants determined animals surviving and recovering in percents similar to those obtained by separate administration of vaccine for subcutaneous use (in septicemia--100% protection; in urinary tract infection--75% protection).     

丙型肝炎病毒（HCV）实验性疫苗的研究进展

丙型肝炎病毒是引起输血相关肝炎及慢性肝炎、肝硬化、肝癌的主要病原,目前尚无有效的治疗与预防手段。本文将综述HCV感染所引起的机体免疫应答及近年来实验性疫苗（主要是DNA疫苗、病毒载体疫苗及联合疫苗）的研究进展。