A型口蹄疫病毒VP1免疫活性肽基因串联片段的化学合成及克隆与表达

A型口蹄疫病毒（FMDV）VP1蛋白的141～160和200～213位氨基酸序列是决定FMDV免疫原性的抗原决定簇片断．人工合成一个免疫活性肽基因的串连片段（20AA－14AA-20AA）、全部选用大肠杆菌偏爱密码子．在5’端带有EcoR1位点和一个起始密码子ATG，在3’端带有BamH1位点和一个终止密码子TGA．利用这两个酶切位点把该基因连接到pWR590质粒上．并筛选高表达的菌株．结果表明该融合蛋白在大肠杆菌JM101中能高表达．经斑点ELISA实验证明．大肠杆菌细胞中表达的融合蛋白有A型FMDV抗原活性．     

简化免疫程序对肉仔鸡生产性能及免疫效果的影响

对肉仔鸡利用简化免疫程序免疫后的生产性能及免疫效果进行了观察。结果表明：对肉仔鸡采用ＮＤ－ＩＢＤ活、死苗一次性免疫接种，具有ＮＤ及ＩＢＤ抗体产生早、高而持久的特点，足以保护肉仔鸡整个饲养周期。减少了因免疫接种引起的应激反应，从而使其生产性能得到显著提高。     

疫苗研制及应用的研究现状与展望

主要从灭活疫苗、减毒活疫苗、重组亚单位疫苗以及核酸疫苗四个方面综述了近年来疫苗研制及应用的现状、存在的问题,并对理想的新一代疫苗作了展望。     

HIV-1C亚型DNA疫苗诱导小鼠免疫反应的研究

 构建含中国流行株HIV-1 C亚型核心蛋白gag基因的重组质粒pVAX-gag,并在体外进行了表达与鉴定.同时构建了含此gag基因的原核表达质粒pGEX-gag,表达纯化并鉴定重组蛋白Gag.以质粒pVAX-gag免疫Balb/C小鼠后,用ELISpot和流式细胞仪检测其细胞免疫反应.再以纯化后的重组蛋白Gag作为包被抗原,用ELISA检测其体液免疫反应.结果显示重组质粒pVAX-gag免疫小鼠后可有效地诱导机体产生细胞免疫和体液免疫反应,且免疫剂量和免疫效果存在一定的正相关性.重组原核表达质粒pGEX-gag的表达产物能与抗p24单克隆抗体发生特异性反应,可用于抗HIV抗体检测.     

Repeated epitope in the recombinant epitope-peptide could enhance ELDKWA-epitope-specific antibody response

Based on the hypothesis suggested by us that epitope-vaccine may be a new strategy against HIV mutation, we have studied several neutralizing epitopes on HIV envelope proteins. However we do not know whether a repeated epitope in a recombinant epitope-peptide can enhance epitope-specific antibody response or not. ELDKWA-epitope (aa669-674) on the C-domain of HIV-1 gp41 is a neutralizing epitope defined by the monoclonal antibody (mAb) 2F5 with broad neutralizing activity. In this study, we designed and prepared a series of the recombinant epitope-peptides bearing 1, 4 and 8 copies of ELDKWA-epitope respectively. In the comparison of the antisera induced by the three recombinant antigens, an obviously increased titre of ELDKWA-epitope-specific antibody was observed in the case of four and eight repeated epitopes. In flow cytometry analysis, the epitope-specific antibodies in both antisera showed stronger activity to bind the transfected CHO-WT cells that stably express HIV-1 envelope glycoprotein on the cell surfaces. These experimental results indicated that repeated epitope in the recombinant epitope-peptide could enhance ELDKWA-epitope-specific antibody response, which could contribute to designing an effective recombinant epitope-vaccine.     

Preparation, characterizationand preliminary in vivo studies of inactivated SARS-CoV vaccine

A large quantity of SARS-CoV virus was proliferated in Veto cells, inactivated with β-ptopiolactone, then purified by Sepharose 4FF column chromatography to prepare inactivated vaccine. The vaccine was identified by Western blot, mass spectrographic analysis, ELISA and electton microscopy. The vaccine with or without aluminum hydtoxide adjuvant was inoculated into female BALB/c mice at different dosages. The result showed that the antibodies to SARS-CoV were induced in the mice. The antibody levels induced by the vaccine with aluminum hydroxide were higher than those without aluminum hydroxide.     

卵透明带DNA避孕疫苗pVAX1—pZP3α的构建及其在体外的培养

目的：构建pVAX1-pZP3α，探讨其在体外的表达，研制卵透明带DNA避孕疫苗，方法：利用基础工程的方法构建卵透明带DNA避孕疫苗pVAX1-pZP3α,用脂质体法转染Hela细胞并采用RT-PCR和Westem blot检测卵透明带抗原pZP3α的体外表达，结果：构建了卵透明带DNA避孕疫苗pVAX1-pZP3α，并在体外表达系统检测到卵透明带抗原pZP3α mRNA和pZP3α的表达。结论：正确构建了卵透明带DNA避孕疫苗pVAX1-pZP3α。     

豚鼠气单胞菌灭活疫苗的安全性和免疫防治

对鲤鱼竖鳞病疫苗的安全性、免疫保护力和免疫效价进行了系统性的研究,并进行了口服免疫养殖应用实验。结果表明,该疫苗口服无毒性,注射途径的免疫保护率为８８．９％,免疫效价为２５００;口服途径的免疫保护率为８０％,免疫效价为２０００;而对照组的免疫效价为１５。养殖应用实验证明,口服疫苗可使鲤鱼竖鳞病的自然发病率降低５０％。     

病例对照研究评价疫苗效力的样本含量

应用预设和调节可信区间宽度的方法，提出了病例对照研究评价疫苗效力VE所需样本含量的计算公式．由公式计算的样本含量能使研究者掌握由样本所获得的VE点估计值接近总体真实值的程度，从而能较为准确地判断研究结果的实际意义。     

复合稳定剂增加脊髓灰质炎糖丸疫苗的稳定性

该研究改进了现行脊髓灰质炎糖丸疫苗的稳定性，由１ｍｏｌ·Ｌ－１ＭｇＣｌ２稳定性改为１ｍｏｌ·Ｌ－１ＭｇＣｌ２＋０．５ｍｏｌ·Ｌ－１磷酸盐缓冲液复合稳定剂（１ｍｏｌ·Ｌ－１ＭｇＣｌ２＋０．５ｍｏｌ·Ｌ－１ＰＢ）．采用复合稳定剂后，大大增加了疫苗的稳定性．２２℃保存时间比１ｍｏｌ·Ｌ－１ＭｇＣｌ２稳定剂延长了一周，即保存时间从１２ｄ延长至１８～２１ｄ．疫苗于４℃保存了６个月，滴度降低甚微．在３７℃保存３ｄ，疫苗的滴度降仍符合ＷＨＯ规程的要求