疫苗,DNA

摘要： DNA疫苗技术近年来迅速发展,但当被实际应用于人体试验时免疫原性不足仍是其最大的弱点.国内外科研工作者尝试了多种策略来解决这一问题,包括重组质粒载体的构建及编码抗原的基因优化,电穿孔疫苗免疫、接种方案优化、与能增强转染基因表达和疫苗诱导免疫的分子佐剂共免疫等.熟悉DNA疫苗的作用机制能更好地利用宿主天然免疫应答以增强疫苗的免疫原性.该综述总结了DNA疫苗免疫效应增强技术的新进展,以及这些进展对DNA疫苗未来发展方向可能产生的影响.%In recent years,DNA vaccine technologies have developed rapidly.Unfortunately,it shows inadequate immunogenicity when applying to human subjects.Many different strategies have been tested to solve this problem by scholars at home and abroad,including synthetic plasmid construction and their encoded antigen gene optimization,electroporation-mediated vaccine delivery,optimization of vaccination program as well as co-delivery with molecular adjuvants which collectively enhance both transgene expression and the elicitation of vaccine-induced immunity.Better understanding of the mechanisms of DNA vaccines has also enabled better use of the intrinsic host response to improve vaccine immunogenicity.The recent advances in DNA vaccine immunity enhancing technologies and their possible impact on future directions of DNA vaccine development are summarized in this article.

摘要： 目的 构建pIRES2-MLAA34-HSP70重组质粒,并检测其免疫效果.方法 采用逆转录PCR(RT-PCR)的方法提取急性单核细胞白血病相关抗原基因MLAA-34和热休克蛋白(HSP) 70基因,设计特异性重叠引物,以重叠延伸PCR(SOE-PCR)技术扩增MLAA34-HSP70融合基因,构建核酸疫苗pIRES2-MLAA34-HSP70.将核酸疫苗免疫BALB/c小鼠,检测小鼠脾淋巴细胞对U937细胞的杀伤作用及小鼠脾细胞悬液中白细胞介素(IL)-2、IL-4和γ干扰素(IFN-γ)水平.结果 扩增出MLAA34-HSP70融合基因2 956 bp,成功构建了核酸疫苗pIRES2-MLAA34-HSP70,经鉴定与预期结果一致;脾淋巴细胞杀伤活性结果显示,核酸疫苗pIRES2-MLAA34-HSP70对U937细胞的杀伤效率明显高于其他实验组及对照组(P＜0.01);核酸疫苗pIRES2-MLAA34-HSP70组细胞因子IL-2、IL-4和IFN-γ水平亦明显高于其他实验组及对照组(P＜0.01).结论 成功构建了pIRES2-MLAA34-HSP70核酸疫苗,该核酸疫苗能诱发强烈的体液免疫,增强机体对肿瘤细胞的免疫应答,对MLAA34阳性细胞具有特异性杀伤作用.%Objective To construct the DNA vaccine pIRES2-MLAA34-HSP70,and to detect its immune effect.Methods The acute monocytic leukemia associated antigen gene MLAA-34 and heat-shock protein (HSP)70 gene were extracted by using RT-PCR.The specific overlapping primer was designed,and the fusion gene MLAA34-HSP70 was amplified by using SOE-PCR technique.Then the DNA vaccine pIRES2-MLAA34-HSP70 was constructed,and BALB/c mice were immunized with this DNA vaccine.The splenic lymphocyte killing activity was detected by using MTT,levels of IL-2,IL-4 and IFN-γ were also detected by using ELISA.Results The MLAA34-HSP70 gene (2 956 bp) and the DNA vaccine pIRES2-MLAA34-HSP70 was amplified and constructed successfully.The killing efficiency of DNA vaccine pIRES2-MLAA34-HSP70 in U937 cells was significantly higher than that in other experimental groups and control group (P＜0.01),and levels of IL-4,IL-2 and IFN-γin DNA vaccine pIRES2-MLAA34-HSP70 group were significantly higher than those in the other experimental groups and control group (P＜0.01).Conclnsion The DNA vaccine pIRES2-MLAA34-HSP70 is constructed successfully.It is shown that the DNA vaccine induces strong humoral immunity,which could enhance immune responses to tumor cells and specificlly kill MLAA34 positive cells.

摘要： 目的：研究LIGHT作为免疫佐剂在EB病毒核抗原1（EBNA1）DNA疫苗诱导免疫反应中的作用。方法以Balb/c小鼠肝脏cDNA为模板，PCR扩增构建真核表达载体pcDNA3.1-LIGHT；转染细胞，验证LIGHT蛋白的表达；采用电脉冲方法，以LIGHT质粒为佐剂与EBV DNA疫苗联合免疫小鼠，测定血清中EBNA1特异性抗体；分离和收集小鼠脾脏淋巴细胞，经EBNA1抗原刺激后，测定细胞因子干扰素-γ（IFN-γ）的分泌水平。结果 PCR扩增出LIGHT目的片段，大小为720 bp；蛋白质免疫印迹法（Western blotting）鉴定结果：在蛋白Marker相对分子质量26×103上方有单一清晰目的条带，表明有LIGHT蛋白表达；免疫小鼠血清测定实验结果：LIGHT联合DNA疫苗实验组的抗体滴度是单独DNA疫苗免疫组的3倍；脾淋巴细胞因子测定结果表明LIGHT联合免疫组能明显增强IFN-γ的分泌。结论 LIGHT作为免疫佐剂，可依赖其介导的信号通路对抗体的产生和相关细胞因子的分泌具有显著促进作用。%Objective To study the effect of LIGHT as an adjuvant in the immune responses induced by EB viral nuclear antigen1(EBNA1) DNA vaccine. Methods The liver cDNA of Balbc/c mouse served as the template. The eukaryotic expression vector pcDDNA3.1-LIGHT was constructed by PCR amplification;the cell transfection was conducted,the LIGHT protein expres-sion was verified;by adopting the electric pulse method,the mouse was immunized with the LIGHT plasmid as the adjuvant com-bined with EBV DNA vaccine. The specific antibody of serum EBNA1 was detected. The mouse spleen lymphocytes were separat-ed and collected. After stimulation by EBNA1 antigen ,cytokine IFN-γsecretion level was detected. Results The LIGHT target fragment was amplified by PCR,which was about 720 bp,the Western blot identification results showed that a single clear target band was above 26 ×103,demonstrating that LIGHT protein was expressed;the serum detection results in the immune mouse showed that the antibody titer in the LIGHT combined DNA vaccine experimental group was 3 times of that in the single DNA vac-cine immune group;the spleen lymphocyte factor detection results showed that the LIGHT combined immune group could signifi-cantly increase the IFN-γsecretion. Conclusion LIGHT as the adjuvant has significant promoting effect in the generation of an-tibody and secretion of related cytokines by depending on its mediated signal pathway.

摘要： 目的 比较甲型流感病毒分支状多肽疫苗最佳免疫途径.方法 通过化学方法合成(M2e) 4-tuftsin,与Al(OH)3佐剂混合,肌注途径免疫小鼠;与JY佐剂混合,滴鼻免疫小鼠,观察两种途径的免疫效果.结果 肌内注射免疫途径诱导产生抗体的能力高于滴鼻免疫,(M2e) 4-tuftsin滴鼻免疫诱导细胞免疫的能力最强,(M2e)4-tuftsin通过肌肉注射和滴鼻免疫途径均对小鼠有一定的保护效果,但肌注免疫效果最好.结论 分支状M2e多肽肌内注射比滴鼻免疫保护效果好.%Objective To compare the immune effects induced with universal influenza A vaccine based on branched M2e polypeptides by two immunization routes in mice.Methods The M2e monomer and (M2e)4-tuftsin were synthesized using chemical method.One group of mice were immunized intramuscularly (i.m.) with 10 μg of (M2e)4-tuftsin plus aluminum adjuvant.And the other group of mice were intranasally immunized with the same amount of polypeptides plus JY adjuvant.Then the immune effects of different immune pathways were observed.Results Intramuscular route induced higher level of M2 specific antibody than intranasal route.Intramuscular injection of (M2e)4-tuftsin was the most effective in stimulating T cell responses.Both intramuscular and intranasal vaccinations (M2e) 4-tuftsin had protective effect,but intramuscular vaccination could provide the most efficient protective immunity.Conclusion The data indicated that the intramuscular vaccination could be the better way for immunization than intranasal vaccination.

摘要： Objective To develop a therapeutic minicircle DNA plasmid of HBV and investigate their immune activity in vitro and in vivo. Methods Two genes were amplified, which separately encoded HBV middle envelope protein (preS2.S) as a vaccine and human interleukin (IL)-2/interferon (IFN)γfusion protein as an adjuvant by PCR from plasmids pcNDA3.1/preS2.S and pcDNA3.1/hIL-2-IFNγ. Then two genes were cloned into eukaryotic vector ZY781 and the parent plasmid preS2.S/ZY781and hIL-2-IFN γ/ZY781 were obtained respectively. The new plasmids were analyzed by restriction endonuclease and DNA sequencing. The parent plasmids were induced to produce minicircle pMCS2.S and pMCIIF by arabinose. The minicircle plasmids were transfected into COS-7 cells in vitro with LipofectamineTM 2000. At 24, 48 and 72 h after transfection, the supernatants were quantified by ELISA. The two minicircle plasmids were injected into BALB/c mice by electroporation in situ together. When the mice were killed 4 weeks later, specific humoral and cellular immune responses were examined. Results The gene segments and inserting direction of pMCS2.S and pMCIIF were correct by genetic analysis. At 48 h after transfection, the concentration of HBsAg, IL-2 and IFNγwere 60.3±7.8 ng/ml, 17.7±1.9 ng/ml and 10.3 ±0.9 ng/ml, respectively, which were the peak values. Protective antibodies of HBsAb were produced, and co-injection of minicircle pMCS2.S together with adjuvant pMCIIF resulted in more evident immune responses. There was strongly specific humoral and cell immunity by only 5μg plasmid per mice. Conclusions The minicircle pMCS2.S and pMCIIF are constructed successfully, and in-duce strongly specific immune activity in vitro and in vivo.%目的 构建HBV的治疗性微环质粒并进行体内外实验研究. 方法 采用聚合酶链式反应扩增HBV包膜蛋白preS2.S抗原基因和hIL-2-IFNγ融合基因,将2个基因分别克隆于ZY781载体,得到亲本质粒preS2.S/ZY781和hIL-2-IFNγ/ZY781,测序正确后,用阿拉伯糖诱导生成微环质粒pMCS2.S和pMCIIF. 转染微环质粒于COS-7细胞株,用酶联免疫吸附法定量法检测24、48、72 h时目的蛋白的表达情况.双质粒联合在体电脉冲注射BALB/c小鼠,4周后处死小鼠,检测特异性的体液和细胞免疫应答. 结果 微环质粒pMCS2.S和pMCIIF转染COS-7细胞后,目的蛋白HBsAg、白细胞介素-2、干扰素 γ在不同时间均有表达,在48 h达峰值,分别为(60.3±7.8)、(17.7±1.9)、(10.3±0.9) ng/ml. 微环质粒pMCS2.S能诱导小鼠产生HBsAb的抗体保护,佐剂pMCIIF可显著增强其免疫效果,在低剂量5μg/只就能诱导较强特异性细胞和体液免疫. 结论 成功构建了HBV微环质粒pMCS2.S及其佐剂微环质粒pMCIIF,其有较好的体内外活性.

摘要： 西方马脑炎是由西方马脑炎病毒感染引起的,经蚊虫传播的急性人兽共患传染病,可引起严重的脑损伤.此外,西方马脑炎病毒也被视为潜在的生物战剂(生物恐怖剂).目前西方马脑炎尚无获批疫苗和有效药物,研发有效疫苗用于相关人群的免疫接种很有必要.传统灭活病毒疫苗具有良好的免疫保护效能,但为了避免其制备过程中病毒培养对昂贵生物安全3级实验室的依赖以及操作者潜在感染的风险,研究者采用新的生物学技术研发西方马脑炎亚单位疫苗、DNA疫苗、嵌合体疫苗和重组活载体疫苗等新型疫苗.本文对疫苗的研究进展进行综述.

摘要： 目的 研究不同剂量结核病DNA疫苗电转染后的免疫原性.方法 40只BALB/c小鼠通过随机数字表法分为8组,每组5只.其中4组分别用100 μl生理盐水和10 μg、50 μg、100 μg结核分枝杆菌Ag85A DNA肌内注射免疫小鼠;另4组用100 μl生理盐水和10 μg、50 μg、100 μgAg85ADNA肌内注射加电转染免疫小鼠.每2周免疫1次,共3次.最后1次免疫后2周杀死小鼠.用ELISA方法检测小鼠脾细胞培养上清液中γ干扰素(IFN-γ)和白细胞介素4(interleukin 4,IL-4)水平;用流式细胞术检测小鼠外周血单个核细胞(PBMC)分泌IFN-γ的辅助性T细胞(helper T cell,Th)1细胞百分比、分泌IL-4的Th2细胞百分比,以及Th1与Th2的细胞比值.用SAS 9.2软件处理数据,实验数据以“-x±s”表示,对有关数据进行两因素析因设计的方差分析,两两比较采用t检验,P＜0.05为差异有统计学意义.结果 免疫结束后2周,小鼠脾淋巴细胞分泌IFN-γ水平,50 μg[(646.05±342.53) pg/ml]和100 μgAg85ADNA肌内注射组[(738.61±372.68) pg/ml]显著高于生理盐水组[(1.73±3.88)pg/ml](f值分别为4.065、4.647,P值均＜0.05)和10 μg Ag85A DNA组[(87.83±120.82)pg/ml](t值分别为3.513、4.094,P值均＜0.05);10 μg Ag85A DNA电转染组[(357.06±105.18) pg/ml]显著高于生理盐水组(t=2.247,P＜0.05),高于100 μg Ag85ADNA电转染组[(86.08±135.73) pg/ml],但差异无统计学意义(t=1.706,P＞0.05);50 μg Ag85ADNA电转染组[(648.60±439.41)pg/ml]显著高于生理盐水组(t=4.081,P＜0.05)和100 μg Ag85A DNA电转染组(t=3.539,P＜0.05).与直接肌内注射组IFN-γ水平[(87.83±120.82) pg/ml]比较,10 μg Ag85A DNA电转染组增高3倍[(357.06 pg/ml)/(87.83 pg/ml)]; 50 μg Ag85A DNA肌内注射组[(646.05±342.53) pg/ml]与电转染组[(648.60±439.41)pg/ml]比较,差异无统计学意义(t=-0.016,P＞0.05);100 μg Ag85A DNA电转染组[(86.08±135.73)pg/ml]较肌内注射组[(738.61±372.68) pg/ml]降低88.35％(t=4.105,P＜0.05).小鼠PBMC分泌IFN-γ的Th1细胞百分比,不同剂量Ag85A DNA肌内注射组[10 μg Ag85A DNA:(1.39±0.84)％;50 μg Ag85A DNA:(1.55±0.33)％;100 μg Ag85A DNA:(2.13±0.47)％]和DNA电转染组[10 μg Ag85A DNA:(1.42±0.47)％; 50 μg Ag85A DNA:(1.88±0.51)％; 100 μg Ag85ADNA:(1.43±0.68)％]均高于生理盐水组[(0.65±0.31)％](t值分别为2.002、2.431、4.015、2.084、3.332和2.105,P值均＜0.05),但不同剂量Ag85A DNA电转染组与肌内注射组之间差异均无统计学意义(t值分别为0.081、0.901和-1.91,P值均＞0.05).小鼠PBMC分泌IL-4的Th2细胞百分比,不同剂量Ag85A DNA肌内注射组[10 μg Ag85A DNA:(1.42±1.18)％; 50 μg Ag85A DNA:(1.14±0.78)％; 100 μg Ag85A DNA:(1.24±0.76)％]和DNA电转染组[10 μg Ag85A DNA:(1.19±1.09)％; 50 μg Ag85A DNA:(2.06±0.96)％;100 μgAg85A DNA:(1.47±0.65)％]均显著低于生理盐水电转染组[(4.14±2.55)％](t值分别为-3.392、-3.738、-3.616、-3.676、-2.599和-3.325,P值均＜0.05),但不同剂量DNA肌内注射组与DNA电转染组之间差异无统计学意义(t值分别为0.284、-1.139和-0.292,P值均＞0.05).结论 DNA电转染免疫可以增强低剂量DNA疫苗的免疫应答,使用少量的DNA疫苗就可以产生较好的免疫效果.

摘要： Objective To identify the transcription of eukaryotic coexpression vector pMAGEA3-IRES-SEA by electroporation method in muscle of mice. Methods The eukaryotic coexpression vector was constructed with staphylococcal endotoxin A(SEA) and human melanoma-associated antigen gene A3 (MAGE-A3) originating from laryngocarcinoma by genetic recombinant method in advance. Unilateral hindlimb skeletal muscles of BALB/C mice were immuned with eukaryotic expression plasmid pMAGEA3-IRES-SEA by electrotransfection method,one time every two week, each immuned of 50μg, a total of three times. Two weeks after the final immunization, their gene transcriptions were identified at the muscle organization of the injection site by fluorescence quantitative PCR (qRT-PCR) assay. Results The SEA and MAGE-A3 genes had been transcripted in experimental mice skeletal muscle. Conclusions The constructed pMAGEA3-IRES-SEA eukaryotic expression plasmid can be effectively transcripted in mice by electroporation method. This work provided a theoretical basis for the expression of mouse in vivo transcription protein and the application of the plasmid to clear laryngocarcinoma by inducing the cellular immunity and humoral immunity in organism.%目的：检测真核质粒pMAGEA3-IRES-SEA经过电穿孔转染在小鼠中的转录。方法将葡萄球菌肠毒素A（staphylococcal endotoxin A，SEA）和喉癌来源的黑色素瘤抗原A3（melanoma-associated antigen A3， MAGE-A3），构建成基因共表达的真核质粒 pMAGEA3-IRES-SEA。将pMAGEA3-IRES-SEA用电转染的方法免疫BALB/C小鼠单侧股四头肌，每2周1次，每次注射50μg，共免疫3次。末次免疫2周后，取注射部位组织，用荧光定量PCR（qRT-PCR）法检测目的基因在注射部位肌肉中的转录情况。结果在实验小鼠注射部位的骨骼肌内检测到 SEA、MAGE-A3基因转录mRNA。结论所构建的pMAGEA3-IRES-SEA真核表达质粒，可通过电转染的方式，在小鼠体内能有效地转录。这为研究该质粒通过小鼠的体内转录蛋白的表达，诱导机体细胞免疫、体液免疫来清除喉癌，奠定了理论基础。

摘要： 目的 研制鼠伤寒沙门菌细菌影,负载防龋DNA疫苗,探寻增强防龋DNA疫苗黏膜免疫效能的方法.方法将pREP4质粒和噬菌体PhiX基因E表达质粒同时转入鼠伤寒沙门菌减毒株J357中,加入异丙基硫代半乳糖苷(isopropyl β-D-1-thiogalactopyranoside,IPTG)诱导,收集洗涤,负载防龋DNA疫苗,分4组经鼻免疫小鼠,分别为:细菌影+pGJGLU/VAX组,细菌影负载5μg防龋DNA疫苗pGJGLU/VAX;细菌影+pVAX1组,细菌影负载5μg空载体pVAX1;pGJGLU/VAX组,布比卡因包裹5 μg pGJGLU/VAX; pVAX1组,布比卡因包裹5μg pVAX1.酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法检测唾液抗体产生结果.结果 ELISA结果显示经鼻黏膜免疫鼠伤寒沙门菌J357细菌影负载的防龋DNA疫苗pGJGLU/VAX(细菌影+pGJGLU/VAX组)后,诱导了显著的唾液特异性抗葡聚糖结合区的IgA抗体,(x)±sx为(0.367 ±0.086) A/μg,与细菌影+pVAX1组[(0.122±0.077)A/μg]、pGJGLU/VAX组[(0.068±0.068)A/μg]和pVAX1组[(0.089±0.089) A/μg]相比,差异均有统计学意义(P值分别为0.028、0.012和0.030).结论 成功制备了鼠伤寒沙门菌细菌影,负载防龋DNA疫苗后经鼻黏膜途径免疫小鼠能有效提高免疫效能.%Objective To develop an anti-caries DNA vaccine-loaded Salmonella typhimurium(St) ghost and enhance the efficacy of immune responses induced by anti-caries DNA vaccine via mucosal route.Methods Both pREP4 and PhiX gene E expression plasmids were transformed into StJ357 and then induced with isopropyl β-D-1-thiogalactopyranoside (IPTG).The bacterial ghosts (BG) were collected after wash and loaded with anti-caries DNA vaccine pGJGLU/VAX.Mice were divided into four groups and immunized through the nasal route with pGJGLU/VAX-loaded BG(Group Ghost + pGJGLU/VAX),pVAX1-loaded BG (Group Ghost + pVAX1),pGJGLU/VAX-Bupivacaine complex (Group pGJGLU/VAX) and pVAX1-Bupivacaine complex (Group pVAX1),respectively.Enzyme-linked immunosorbent assay (ELISA) was used to evaluate the immune responses.Results ELISA results showed that group Ghost + pGJGLU/ VAX had significantly higher level of specific anti-GLU SIgA antibody [(0.367 ± 0.086) A/μg] compared with group Ghost + pVAX1 [(0.122 ± 0.077) A/μg],Group pGJGLU/VAX [(0.068 ± 0.068) A/μg] or Group pVAX1 [(0.089 ±0.089) A/μg] (P =0.028,0.012 or 0.030,respectively).Conclusions St ghost was developed successfully,which enhanced the efficacy of immune responses induced by anti-caries DNA vaccine pGJGLU/VAX via the nasal route.

摘要： 核酸疫苗是将编码某种抗原的外源基因(DNA或RNA)直接导入动物体细胞内,通过宿主细胞表达系统合成抗原,诱导宿主产生对该抗原的免疫应答,以达到防治疾病目的的一类新型疫苗.研究表明,核酸疫苗不仅具有良好的免疫原性和安全性,而且由于核酸更易于修饰和改造,使核酸疫苗较以往蛋白疫苗具有更大的灵活性.在实际生产中,核酸疫苗的制备和纯化工艺较为简单,生产周期短,生产成本低,且易于制成多价疫苗,因此极有可能成为下一个成功应用于人类的新型疫苗.此文对核酸疫苗的研究进展做一综述.%Nucleic acid vaccine is a new type of vaccine,which can deliver foreign genes (DNA or RNA) encoding antigens of interest directly into animal cells,produce antigens through host cell expression system,and further induce immune response in the host.It is reported that nucleic acid vaccine is better than protein vaccine because it is immunogenic and safe,and easy to be modified and manufactured.In practical production,nucleic acid vaccine has many advantages,such as easy preparation and purification,short production cycle,low production cost,and easy to form multivalent vaccines.Therefore,it is hopeful that nucleic acid vaccine will become a successful vaccine for human use.This review summarizes research progress of nucleic acid vaccine.

摘要： 一种HIV多表位DNA疫苗和HIV复合多表位DNA疫苗，属于生物工程领域，特别是涉及一种HIV复合多表位DNA疫苗。本疫苗基因全长634bp，优选了HIV基因组中九个高度保守的免疫优势表位，一个HIV-1分离株共有抗体表位，另外加入了一个MHC非限制性的辅助性T淋巴细胞表位，各表位间用三个丙氨酸连接，同时在上述融合基因的中引入ER信号肽引导序列。以HIV-1CNB gag基因为模板，利用PCR方法获得全长693bp的HIV衣壳蛋白P24后插入权利要求1所述基因中，获得了以p24核心蛋白为载体分子的全新HIV复合多表位DNA疫苗。

摘要： 本发明涉及以成纤维激活蛋白α为靶点肿瘤DNA疫苗和病毒载体疫苗领域。具体地说，本发明涉及重组DNA载体CpVR‑FAP、重组腺病毒Ad‑FAP和重组痘病毒MVA‑FAP疫苗，以及DNA疫苗和病毒载体疫苗以及病毒载体疫苗之间免疫方式的优化组合或联合环磷酰胺在制备抗肿瘤疫苗中的用途。

摘要： 本发明涉及肿瘤DNA疫苗和病毒载体疫苗领域。具体地说，本发明涉及重组DNA载体VR?S8和VR?MS、重组腺病毒Ad?S8和Ad?MS及重组痘病毒MVA?MS疫苗，以及DNA疫苗和病毒载体疫苗以及病毒载体疫苗之间免疫方式的优化组合在制备抗肿瘤疫苗中的用途。

摘要： 本发明涉及肿瘤DNA疫苗和病毒载体疫苗领域。具体地说，本发明涉及重组DNA载体VR-S8和VR-MS、重组腺病毒Ad-S8和Ad-MS及重组痘病毒MVA-MS疫苗，以及DNA疫苗和病毒载体疫苗以及病毒载体疫苗之间免疫方式的优化组合在制备抗肿瘤疫苗中的用途。

摘要： 本发明属于疫苗领域，具体涉及一种疫苗递送系统、疫苗递送系统的应用和制备的DNA疫苗。本发明首先提供了一种多肽，所述多肽与多聚物形成递送系统能够与表达病毒抗原的DNA结合，并在生物体内递送，递送效率高、安全性好，能实现病毒抗原的DNA的高效表达，全面均衡的诱导体液及粘膜免疫应答，对DNA疫苗的研发和应用具有重要的意义；其次，以新型冠状病毒SARS‑CoV‑2的Spike蛋白为抗原，制备了新型冠状病毒SARS‑CoV‑2的DNA疫苗，实现了新型冠状病毒SARS‑CoV‑2DNA疫苗的体内递送，同时能够诱导血清中IgG抗体和肺部sIgA抗体的产生，在肺部形成效应记忆T细胞，用于DNA疫苗的制备。

摘要： 一种HIV多表位DNA疫苗和HIV复合多表位DNA疫苗，属于生物工程领域，特别是涉及一种HIV复合多表位DNA疫苗。本疫苗基因全长634bp，优选了HIV基因组中九个高度保守的免疫优势表位，一个HIV－1分离株共有抗体表位，另外加入了一个MHC非限制性的辅助性T淋巴细胞表位，各表位间用三个丙氨酸连接，同时在上述融合基因的中引入ER信号肽引导序列。以HIV－1CNB gag基因为模板，利用PCR方法获得全长693bp的HIV衣壳蛋白P24后插入权利要求1所述基因中，获得了以p24核心蛋白为载体分子的全新HIV复合多表位DNA疫苗。

摘要： 本发明公开了一种非洲猪瘟p30、p54、p72和B602L的DNA疫苗和亚单位疫苗及其制备方法与应用。所述的DNA疫苗包括编码非洲猪瘟病毒p30、p54、p72和B602L蛋白的四个重组pCAGGS真核表达质粒；亚单位疫苗包括利用重组杆状病毒表达系统的三个蛋白：单独表达的非洲猪瘟病毒p30蛋白、非洲猪瘟病毒p54蛋白，联合表达的非洲猪瘟病毒p72蛋白和非洲猪瘟病毒B602L蛋白，以及药学上可以接受的佐剂。针对亚单位疫苗及核酸疫苗免疫后仅能提供部分保护或者无保护力的问题，为提高亚单位疫苗和核酸疫苗的免疫效果，通过猪源及昆虫细胞的密码子优化，且采取“鸡尾酒”式混合免疫，用以提高疫苗的免疫效果。

摘要： 本发明涉及以成纤维激活蛋白α为靶点肿瘤DNA疫苗和病毒载体疫苗领域。具体地说，本发明涉及重组DNA载体CpVR‑FAP、重组腺病毒Ad‑FAP和重组痘病毒MVA‑FAP疫苗，以及DNA疫苗和病毒载体疫苗以及病毒载体疫苗之间免疫方式的优化组合或联合环磷酰胺在制备抗肿瘤疫苗中的用途。

摘要： 本发明涉及肿瘤DNA疫苗和病毒载体疫苗领域。具体地说，本发明涉及重组DNA载体VR-S8和VR-MS、重组腺病毒Ad-S8和Ad-MS及重组痘病毒MVA-MS疫苗，以及DNA疫苗和病毒载体疫苗以及病毒载体疫苗之间免疫方式的优化组合在制备抗肿瘤疫苗中的用途。

摘要： 本发明涉及肿瘤DNA疫苗和病毒载体疫苗领域。具体地说，本发明涉及重组DNA载体VR-S8和VR-MS、重组腺病毒Ad-S8和Ad-MS及重组痘病毒MVA-MS疫苗，以及DNA疫苗和病毒载体疫苗以及病毒载体疫苗之间免疫方式的优化组合在制备抗肿瘤疫苗中的用途。