Effects of epitopes combination and adjuvants on immune responses to anti-Alzheimer disease DNA vaccines in mice

Alzheimer disease （AD） is a neurodegenerative disorder characterized by neuropathological hallmarks including deposits of the β-amyloid peptide （AssP）. Studies have shown that immunization with Aβ42 peptide reduces both the spatial memory impairments and Alzheimer disease-like neuropathologic changes in Alzheimer disease transgenic mice, but can cause side effect of a cell-mediated autoimmune meningoencephalitis. Recently, some studies showed that DNA vaccination could be used to generate an antibody response to Aβ without the adverse cell-mediated immune effect. In the current study, we generate four DNA vaccine plasmids （pV-GE1, pV-GE2, pV-GE3, and pV-GE4） against Alzheimer disease by separately fusing Aβ epitope sequences （coding for EFGH, DAEFGH, EFGH＋EFGH, and EFGH＋DAEFGH） with IgG heavy chain coding region of mouse. Meanwhile, the full-length gene Aβ encoding plasmid （pV-Aβ）, empty vector （pVAX） and synthetic AssP were also included as control. The sera of BALB/c mice immunized via intramuscular with plasmids and peptide were tested by indirect     

Regulating Effects of Novel CpG Chitosan-nanoparticles on Immune Responses of Mice to Porcine Paratyphoid Vaccines

INTRODUCTION CpG oligodeoxynucleotide is a newly emerging powerful adjuvant inducing a broad array of immune responses to a wide variety of antigens. Previous studies showed that CpG motifs activate B cells and dendritic cells (DC), trigger immune cascade…     

HPV16 E6 DNA疫苗诱导小鼠抗宫颈癌主动免疫

目的：探讨HPV16 E6 DNA疫苗能否在体内诱导小鼠抗宫颈癌主动免疫应答。 方法：将HPV16 E6基因转染615小鼠宫颈癌U14,建立高表达E6的U14细胞株即E6+U14;制备HPV16 E6 DNA疫苗;随后分组进行体内预防和治疗E6+U14移植同系小鼠试验,设立对照组;观察各组小鼠肿瘤大小,计算抑瘤率、生存时间;计算淋巴和肺转移率;体外分别检测经HPV16 E6 DNA疫苗免疫后的小鼠T 淋巴细胞对E6+U14和U14的杀伤作用。结果：HPV16 E6 DNA疫苗预防和治疗组小鼠各时期瘤体大小均显著低于对照组（均P<0.001 ）,两组抑瘤率均大于70%;平均生存时间均大于对照组（均P<0.001）;预防组淋巴转移率低于对照组（P<0.05）;HPV16 E6 DNA疫苗免疫小鼠T淋巴细胞在不同效靶比,对E6+U14的杀伤效率均明显高于对U14者（均P<0.001）。结论：HPV16 E6 DNA疫苗能诱导机体产生针对HPV16 E6阳性的宫颈癌细胞主动免疫应答及特异性细胞毒淋巴细胞,提示该疫苗可能对临床治疗宫颈癌有效。     

Epitope DNA vaccines against tuberculosis: spacers and ubiquitin modulates cellular immune responses elicited by epitope DNA vaccine

Cell-mediated immune responses are crucial in the protection against tuberculosis. In this study, we constructed epitope DNA vaccines (p3-M-38) encoding cytotoxic T lymphocyte (CTL) epitopes of MPT64 and 38 kDa proteins of Mycobacterium tuberculosis. In order to observe the influence of spacer sequence (Ala-Ala-Tyr) or ubiquitin (UbGR) on the efficacy of the two CTL epitopes, we also constructed DNA vaccines, p3-M-S(spacer)-38, p3-Ub (UbGR)-M-S-38 and p3-Ub-M-38. The immune responses elicited by the four DNA vaccines were tested in C57BL/6 (H-2b) mice. The cytotoxicity of T cells was detected by LDH-release method and by enzyme-linked immunospot assay for epitope-specific cells secreting interferon-gamma. The results showed that DNA immunization with p3-M-38 vaccine could induce epitope-specific CD8 CTL response and that the spacer sequence (AAY) only enhanced M epitope presentation. The protein-targeting sequence (UbGR) enhanced the immunogenicity of the two epitopes. The finding that defined spacer sequences at C-terminus and protein-targeting degradation modulated the immune response of epitope string DNA vaccines will be of importance for the further development of multi-epitope DNA vaccines against tuberculosis.     

细胞因子对HCV基因疫苗诱生免疫反应的增强作用

目的 探讨细胞因子对丙型肝炎病毒（ＨＣＶ）核心（Ｃ）基因疫苗诱生免疫应答强度的增强作用。方法 将包含ＨＣＶ－Ｃ蛋白的基因片段插入真核表达载体ｐｃＤＮＡ３中,构建重组质料ｐｃＤＮＡＨＣＶ－Ｃ,将此重组质粒单独或与包含鼠ＩＬ－２或ＩＬ－１２基因的表达质粒共免疫ＢＡＬＢ／ｃ小鼠,ＥＬＩＳＡ法检测ＨＣＶ Ｃ特异性抗体产生情况,以ｐｃＤＮＡＨＣＶ－Ｃ转梁小鼠ＳＰ２／０细胞,表达ＨＣＶ Ｃ抗原为靶细胞,进行     

Humoral immune response to HCV core DNA vaccine in mice

DNAvaccine,alsocallednucleicacidvaccine,isarecombinanteukatyoticexpressionplasmidencodingcertainantigenofpathogen.DNAvaccinecanexpressantigenandinducecellularandhumoralimmuneresponsesaftervaccinationbyintramuscularinjectionorplasmid--coatedmicroparticlebombardment.Itshightemperaturestability,lowexpenseofmassproductionandtheabilitytoinducestrongimmuneresponsesevenfortheuseintherapyhavemadeDNAvaccineanexcitingwayfornewvaccinedevelopment['J.InordertofurnishevidenceforHCVDNAvaccinesuitablefo…     

小鼠对HBV DNA疫苗的免疫应答及细胞因子的免疫佐剂效应

目的 :观察编码小鼠 IL- 2及 IL- 12的真核表达载体 ,对 HBV DNA疫苗诱导 Balb/ c小鼠 (H- 2 d )免疫应答的调节作用及其对稳定表达 HBs Ag的小鼠肥大细胞瘤 P815细胞 (P 815 - HBV- S)成瘤性的影响。　方法 :肌内注射HBV DNA疫苗及细胞因子真核表达载体 ;背部皮下接种 P 815 - HBV - S细胞 ,观察成瘤情况 ;EL ISA法测定血清抗HBs;4h5 1 Cr释放法检测小鼠脾细胞 CTL 活性。　结果 :免疫 8周后 ,以 p CR3.1- S、p CR3.1- S+IL- 2及 p CR3.1-S+IL- 12免疫的小鼠血清 ,45 0 nm A值分别为 0 .87± 0 .1、1.98± 0 .17及 1.6 7± 0 .15 ,均较 p CR3.1组显著提高(P<0 .0 5 )。CTL 细胞杀伤活性分别为 (5 0 .5± 6 .4) %、(6 1.9± 7.1) %及 (73.3± 8.8) % ,后两组与 p CR3.1- S组比较差异均显著 (P<0 .0 5 )。脾细胞悬液经抗 CD4+ 单克隆抗体处理后 ,CTL细胞杀伤活性分别为 (48.3± 5 .9) %、(5 6 .2± 7.5 ) %和 (75 .6± 9.1) % ,抗 CD8+ 单克隆抗体处理后分别为 (10 .6± 1.4) %、(13.6± 1.9) %和 (16 .9±2 .3) %。对照组成瘤率为 10 0 % ,p CR3.1- S组小鼠成瘤率为 12 .5 % ,p CR3.1- S+IL- 12真核表达载体组成瘤率为0 ,对照组平均存活期和 6周后存活率分别为 2 8.4天和 0天。后两组分别 >38.2天及 45     

青蒿琥酯对小鼠免疫功能的影响

本文观察蒿琥酯对正常小鼠免疫器官重量、血清溶菌酶水平、红细胞免疫粘附功能及ＩＬ－２产生的影响，结果表明青蒿琥酯能减轻胸腺重量，降低血清溶菌酶水平，增加红细胞Ｃ３ｂ受体花环率及抑制ＣｏｎＡ诱导的脾细胞ＩＬ－２产生。青蒿琥酯抑制ＩＬ－２产生可能是其免疫抑制的重要机制。     

The potential role of immunological adjuvants in influenza vaccines

Aqueous influenza vaccines seldom induce long-persistent protection against epidemic risk, and often fail to immunize up to one-third of the recipients, especially when there has been no previous immunological experience of the serotype of virus contained in the vaccine. Attempts to improve the situation by increasing the size or number of doses are limited by expense, toxicity and the time available in face of an oncoming epidemic. The future use of isolated viral antigens as vaccines may be limited by similar disadvantages.     

MUC1基因疫苗和GM-CSF对小鼠的细胞免疫和体液免疫应答

目的:观察MUC1基因疫苗和GM-CSF共免疫对小鼠特异性细胞免疫和抗体水平的影响.方法:采用股四头肌肌肉注射,将构建的MUC1基因疫苗pcDNA3.1-MUC1免疫雌性Balb/c小鼠,每3 wk 1次,共3次.MUC1基因加GM-CSF组于每次基因免疫后1,3,5 d,皮下注射GM-CSF 100 μL.最后1次基因免疫后第3周接种表达MUC1的EMT6小鼠乳腺癌细胞.2 wk后观察、记录肿瘤的生长情况.流式细胞仪检测外周血CD4 和CD8 淋巴细胞亚群的百分比和比值.ELISA方法检测小鼠血清MUC1特异性抗体的水平.4 h 51Cr释放法检测小鼠脾特异性CTL的杀伤活性.结果:淋巴细胞亚群:与pcDNA3.1对照组相比,MUC1基因疫苗预防组的CD8 T淋巴细胞升高,差异有统计学意义(P《0.01),CD4 T淋巴细胞升高,差别无统计学意义;加GM-CSF佐剂预防组与单独MUC1疫苗预防组相比,CD4 T淋巴细胞升高,差异有统计学意义(P《0.01),CD8 T淋巴细胞升高差别无统计学意义.小鼠血清MUC1抗体水平:与对照组相比,MUC1基因疫苗预防组抗体水平升高,差异有统计学意义(P《0.05);加佐剂组与预防组相比,抗体水平升高但差别无统计学意义.在不同效靶比时, MUC1基因疫苗加GM-CSF组与单独MUC1基因免疫组相比较,特异性CTL对EMT6靶细胞的杀伤率差异显著(P《0.05).结论:MUC1基因疫苗可诱导小鼠产生特异性CD8 T淋巴细胞及体液免疫应答, GM-CSF对小鼠CD4 T淋巴细胞具有一定的调节作用.     

重组蛋白CTB表位对HCV多表位DNA疫苗的小鼠中免疫应答的影响

将一12aa的CTB表位连接于丙型肝炎病毒（Hepatitis C virus,HCV)复合多表位抗原基因PCX的5'端,再克隆于真核表达载体pcDNA3中,获得重组质粒pcDNA3/CTB-PCX,利用表达IL-12的pWRG/mIL-12作为佐剂,肌注免疫小鼠后6W,用重组蛋白GZ-PCX加强免疫1次,第6W时可检测到较高水平的抗GZ-PCXIgG,最高滴度达1：10^3,与对照pcDNA3/PCX相比无显升高,提示CTB表位并无明显的增强PCX基因特异性体液免疫应答的作用。重组蛋白GZ-PCX可有效提高抗体水平,促进CD^8 T细胞增殖。免疫小鼠可诱发针对GZ-PCX融合蛋白的迟发性超敏反应（DTH）,免疫后小鼠体重正常,肝脾未见明显肿大。具有良好安全性。     

低压缺氧和亚低温复合因素对小鼠免疫功能的影响

目的 :研究模拟高空低压缺氧和亚低温复合环境 ,对小鼠血浆神经肽含量和免疫功能的影响 .方法 :采用放免法检测高空缺氧和亚低温复合因素作用下 ,小鼠血浆 β 内啡肽 (β EP)和亮氨酸脑啡肽 (L EK)的含量 ;采用MTT法检测ConA或LPS诱导的脾细胞增殖反应 ,胸腺细胞IL 1和脾细胞IL 2的活性 .结果 :7km高空缺氧室温组 (2 2± 2 )℃血浆β EP含量 (ng·L-1)显著高于地面室温组 (395± 6 6vs 2 71±37,P <0 .0 1) ,7km 5℃组血浆 β EP含量显著低于 7km室温组 (2 83± 5 9vs 2 71± 37,P <0 .0 1) .地面 5℃组血浆L EK含量显著高于相应的室温组 (5 75± 88vs 4 0 5± 6 5 ,P <0 .0 1) .7km 10℃组ConA或LPS诱导的脾细胞增殖 (MBq)反应显著低于 7km室温组 (6 2 .5± 6 .8vs 2 .1± 0 .2 ,5 8.8± 8.1vs 30 .6± 3.9,P <0 .0 1) ;7km 5℃组ConA或LPS诱导的脾细胞增殖反应显著高于地面 5℃组 (2 3.3± 3.8vs 4 4 .8±4 .4 ,31.9± 0 .5vs 6 6 .0± 2 .7,P <0 .0 1) .7km 10℃组IL 1活性显著高于 7km室温组 (2 .88± 0 .4 9vs1.5 1± 0 .31,P <0 .0 1) ,7km 5℃组IL 1活性显著低于 7km室温组 (1.2 8± 0 .32vs 1.5 1± 0 .31,P <0 .0 1) .7km 10℃和 5℃组小鼠IL 2活性低于 7km室温组 ,但无显著差异 .结论 :缺氧 1     

亚硒酸钠对荷瘤小鼠免疫系统的影响

研究亚硒酸钠对荷瘤机体的免疫调节作用。方法：实验分别测定了亚硒酸钠＋肿瘤组，肿瘤对照组和正常对照组动物的脾淋巴细胞转化率，ＡＮＡＥ＾＋淋巴细胞百分率和免疫器官指数，并在光镜下对脾脏和胸腺的形态学变化作了观察。     

CEA串连表位-HSP70融合基因疫苗诱导小鼠的表位特异性免疫应答

目的：观察癌胚抗原(CEA)串联表位-HSP70融合基因疫苗诱导的免疫应答，为开发新型肿瘤特异性基因疫苗奠定基础。方法：在已经构建含有变异热休克蛋白(HSP)序列的基础上，插入重组CEA串联表位的编码片段获得CEA串联表位-HSP融合基因疫苗。3次肌肉注射免疫Balb/c小鼠，设立注射生理盐水的阴性对照组、注射氢氧化铝佐剂混悬CEA串联表位的阳性对照组及注射pCITriCEA_625-667-mtHSP70的实验组。FCM分析脾脏T细胞亚群；体外培养脾细胞，ELISA检测培养上清中干扰素γ(IFNγ)的相对含量；同时测定小鼠血清CEA特异性抗体的滴度。结果：阴性对照组脾细胞中CD3⁺和CD4⁺T细胞分别为55.1%±6.1%和30.2%±4.1%；实验组脾细胞中CD3⁺和CD4⁺T细胞分别为78.7%±9.2%和48.9%±4.7%，两组比较差异有显著性(P<0.01)。其体外特异性抗原肽诱导的IFNγ分泌处于本底水平，可视为生理性参数，体外非特异性和特异性的IFNγ分泌分别增加3和6倍(P<0.01)。血清中CEA表位特异性抗体滴度阴性对照组为0，阳性对照组＜1∶500,而融合基因疫苗免疫组达到1∶4 000。结论：CEA串联表位-HSP融合基因疫苗在体内诱导以激发辅助性T细胞为特征的免疫应答。     

初次及再次感染肺炎衣原体Icr小鼠局部的免疫反应

目的:肺炎衣原体(Cpn)是一种常见的人类呼吸道致病菌,感染多为持续或慢性感染,机体感染后可引起以细胞免疫(CMI)为主的免疫反应.文中使用Icr小鼠建立初次及再次感染Cpn的动物模型,研究小鼠肺部及脾脏在初次及再次感染Cpn后的炎症机制及CMI反应. 方法:初次感染(PT)组鼻内接种Cpn 105IFU/只.再次感染(ST)组4周后重复接种同样剂量的Cpn.对照组接种PBS 0.05ml/只.在预设时间点处死小鼠后取肺及脾,分离出肺单核细胞及脾的淋巴细胞.用Cpn刺激,体外培养3d.MTT法测定细胞增殖指数;ELISA法测定IL-4、IL-10、IFN-γ. 结果:体外CMI显示用Cpn刺激后在PT组小鼠肺中出现一个较弱的增殖反应,并无IFN-γ的分泌.在ST组的肺及脾中则均引起以IFN-γ为主的细胞因子的分泌(P<0.01).IL-4在2组小鼠肺中的测定基本为0.IL-10在PT组肺中无表达,在ST组中稍有升高,平均为0.4ng/ml.在ST组脾中IL-4及IL-10均在35d后有升高趋势,与PT组比较均有显著性差异(P<0.05). 结论:Th1型CMI引起的IFN-γ产生和淋巴反应增加是再次感染的特征.     

乙肝病毒表面抗原基因疫苗诱导小鼠细胞及体液免疫应答的研究

为观察乙肝病毒 Ｈ Ｂｓ Ａｇ 基因疫苗免疫小鼠后的细胞及体液免疫应答,将该疫苗定向克隆于质粒ｐｃ Ｄ Ｎ Ａ３ 巨细胞病毒启动子下游,构建能在真核细胞内表达的重组基因疫苗ｐｃ Ｄ Ｎ Ａ Ｈ Ｂｓ Ａｇ 。将此疫苗通过多点肌肉注射免疫 Ｂａｌｂ／ Ｃ 小鼠后第六周,实验组小鼠均出现抗－ Ｈ Ｂｓ Ｉｇ Ｇ 阳性,其抗体水平明显高于对照组,同时 Ｔｈ１ 免疫应答相关的细胞因子 Ｉ Ｌ２ 及γ干扰素水平也明显高于对照组,表明本文构建的基因疫苗ｐｃ Ｄ Ｎ Ａ３ Ｈ Ｂｓ Ａｇ 能诱导 Ｂａｌｂ／ Ｃ 小鼠产生良好的细胞及体液免疫应答。     

HIV-1 DNA vaccine with adjuvant cytokines induces specific immune responses against HIV-1 infection in mice

There is mounting evidence that the induction ofstrong mucosal and cell-mediated immuneresponses is key element to consider in constructing efficacious HIV-1 vaccine. Therapeutic vaccines that induce high levels of CTL specific to HIV are currently being developed worldwide. To induce strong and stable cell-mediated immune responses against HIV infection, immunoregulatory molecules including cytokines and chemokines have been used in the immunization protocols, and some of them were found …