Comparison of methods for determination of immunogenicity of recombinant human papillomavirus vaccine

目的评价重组人乳头瘤病毒(HuMAn PAPIllOMAVIruS,HPV)疫苗免疫原性2种检测方法的相关性。方法应用间接酶联免疫吸附(ElISA)法和假病毒中和(PSEudOVIrIOn-bASEd nEuTrAlIzATIOn ASSAy,PbnS)法对26份阴性血清样本、34份自然感染HPV16或HPV18的血清样本及30份接种HPV16/18型双价疫苗后7个月的血清样本进行抗体滴度检测,并采用PEArMAn进行相关性分析。结果 3组样本的抗HPV16型和HPV18型抗体滴度的2次检测结果比值显示,试验室内重复性较好;2种方法检测血清样本抗HPV16型抗体滴度的总体相关系数为0.826,抗HPV18型为0.921;自然感染HPV的血清样本抗HPV16型抗体滴度的相关系数为0.519,抗HPV18型为0.613;接种HPV16/18型双价疫苗后的血清样本抗HPV16型抗体滴度的相关系数为0.671,抗HPV18型为0.879。结论间接ElISA法和PbnS法在检测血清样本中抗HPV16/18型抗体滴度时均具有较好的重复性和相关性,其中以接种疫苗后的血清样本中抗体滴度相关性最佳,为HPV疫苗免疫原性的检测及剂量探索试验提供了参考依据。Objective To assess the correlation between two methods for determination of immunogenicity of recombinant human papillomavirus(HPV) vaccine.Methods Enzyme-linked immunosorbent assay(ELISA) and pseudovirion-based neutralization assay(PBNS) were used to measure the anti-HPV 16 or anti-HPV 18 antibody levels in 26 negative serum samples,34 serum samples naturally infected with HPV16 or HPV18 and 30 serum samples 7 months after immunization with HPV16 / 18 bivalent vaccine,between which the correlation was analyzed by Pearman method.Results The ratio of results of two tests for HPV16 and HPV18 antibody titers in three groups of samples showed high intra-reproducibility.The total correlation coefficient of test results of HPV16 antibody titers by two methods was 0.826,while that of HPV18 antibody was 0.921.The correlation coefficient for HPV 16 antibody titer in naturally infected samples was 0.519,while that for HPV18 antibody titer was 0.613.However,the correlation coefficient for HPV 16 antibody titer in serum samples after immunization with HPV16 / 18 bivalent vaccine was 0.671,while that for HPV 18 antibody titer was 0.879.Conclusion Both ELISA and PBNS showed high reproducibility and correlation for determination of HPV16 / 18 antibody titers in serum samples,especially for those in samples after vaccination.It provided a reference for determination of immunogenicity and optimization of dosage of HPV vaccine.国家高技术研究发展计划(2012AA02A402); Themajorpro-jectofInternationalScienceandTechnologyCollaborativeProgram(2010DFB30100

pJAK2 polypeptide,an antagonist of suppressors of cytokine signaling-1,can enhance the antitumor effect of dendritic cells

目的:探讨细胞因子信号转导抑制因子-1(SuPPrESSOrS Of CyTOkInE SIgnAlIng1,SOCS1)拮抗物PJAk2多肽(氨基酸序列号为1001-1013)参与树突状细胞(dEndrITIC CEllS,dCS)的体外诱导培养后对dCS抗肿瘤作用的影响。方法:采集健康人外周血,离心获得单个核细胞,用重组人粒细胞巨噬细胞集落刺激因子(rECOMbInAnT HuMAn grAnulOCyTE-MACrOPHAgE COlOny STIMulATIng fACTOr,rHgM-CSf)及重组人白细胞介素-4(rECOMbInAnT HuMAn InTErlEukIn-4,rHIl-4)诱导dCS,第5天分为4组:单纯dCS培养(对照)组、抗原负载(lySATE-dCS)组、多肽修饰(PJAk2-dCS)组和抗原与多肽共培养(lySATE+PJAk2-dCS)组,第6天各组加入肿瘤坏死因子-α(TuMOr nECrOSIS fACTOr-AlPHA,Tnf-α)促成熟。倒置显微镜下观察dCS形态;fCM法检测dCS表型;乳酸脱氢酶(lACTATE dEHydrOgEnASE,ldH)细胞毒实验检测各组细胞毒性T淋巴细胞(CyTOTOXIC T lyMPHOCyTE,CTl)对胃癌细胞bgC-823的靶向杀伤作用;ElISA法检测白细胞介素-12(InTErlEukIn-12,Il-12)和γ干扰素(InTErfErOn-γ,Ifn-γ)的水平。结果:与未加入诱导剂组相比,各组均成功诱导出成熟dCS,均高表达Cd80、Cd83、Cd86和人类白细胞dr抗原(HuMAn lEukOCyTE AnTIgEn dr,HlA-dr),但以lySATE+PJAk2-dCS组的表达水平最高。在10:1~30:1的效靶比范围内,CTl杀伤作用与效靶比呈正相关。当效靶比为30:1时,对照组的CTl杀伤率达(19.77±2.34)%,低于其他3组(P<0.01),而lySATE+PJAk2-dCS组较lySATE-dCS组及PJAk2-dCS组都高(P<0.05)。lySATE+PJAk2-dCS组培养上清液中Il-12及Ifn-γ的分泌水平明显高于对照组(P<0.01)。结论:SOCS1拮抗物PJAk2多肽(1001-1013)可增强dCS对胃癌细胞的抗原递呈及特异性抗肿瘤作用。Objective:To investigate the effect of pJAK2 polypeptide,an antagonist of SOCS1(suppressors of cytokine signaling 1),on antitumor effect of in vitro cultivation-induced DCs(dendritic cells).Methods:Peripheral blood was collected from the healthy volunteers,and the PBMCs(peripheral blood mononuclear cells)were isolated.DCs were induced by rhGM-CSF(recombinant human granulocyte-macrophage colony-stimulating factor)and rhIL-4(recombinant human interleukin-4).On the fifth day,DCs were divided into four groups:control group,Lysate-DCs group,pJAK2-DCs group,and Lysate + pJAK2 DCs group.On the sixth day,TNF-α(tumor necrosis factor-alpha)was added into each group.The morphological features of DCs were observed under an inverted microscope;the phenotypes were detected by FCM(flow cytometry);the killing effect of CTLs(cytotoxic T lymphocytes)on gastric cancer BGC-823 cells was evaluated by LDH(lactate dehydrogenase)cytotoxicity test;the concentrations of IL-12(interleukin-12)and IFN-γ(interferon-γ)were detected by ELISA(enzyme-linked immuno sorbent assay).Results:Mature DCs presented typically morphological and phenotypic features;the DCs in Lysate + pJAK2-DCs group had the highest expression levels of CD80,CD83,CD86 and HLA-DR(human leukocyte antigen DR).When the ratio of effectors to target cells ranged from 10:1 to 30:1,the killing activity of CTLs had a positive correlation with the ratio.When the ratio of effectors to target cells was 30:1,the killing activity of CTLs in the control group was(19.77±2.34)%,which was lowest as compared with the other groups(P < 0.01),meanwhile the killing activity of CTLs in Lysate + pJAK2-DCs group was higher than those in Lysate-DCs and pJAK2-DCs groups(P < 0.05).The levels of IL-12 and IFN-γ secretion in Lysate + pJAK2-DCs group were apparently higher than those in the control group(P < 0.01).Conclusion:An antagonist of SOCS1,pJAK2 polypeptide,can enhance the ability of antigen presentation and specific antitumor effect of DCs on gastric cancer cells.南京军区医学科技创新课题资助项目(编号:10MA068); 福建省自然科学基金面上项目(编号:2010D013); 厦门市科技计划创新项目(编号:3502z20104014

Effect of Helicobacter pylori on the function of peripheral blood monocyte-derived dendritic cells in gastric cancer patients

分析HP阳性和HP阴性的胃癌患者外周血单核细胞来源树突状细胞(MOdCS)功能的差异性及其临床意义。方法:用尿素14C呼气试验对解放军第一七四医院2011年1月至2012年10月收治的84例胃癌患者进行HP感染状态鉴定,分别采集HP阳性和阴性胃癌患者外周血,分离外周血单个核细胞,采用经典方法(rHgM-CSf、rHIl-4联合rHTnf-α)诱导产生dCS,采用流式细胞仪检测dCS表型,采用ldH释放法检测dCS致敏T细胞对胃癌细胞的毒性杀伤作用,采用ElISA方法检测细胞因子Il-12、Ifn-γ的分泌水平。结果:两组MOdCS成熟过程形态变化无差异,HP阳性组MOdCS表面标记分子Cd1A、Cd80、Cd83、Cd86和HlA-dr平均表达百分率均高于HP阴性组,其中Cd80、Cd83、Cd86的表达水平差异有统计学意义,Cd1A、HlA-dr差异无统计学意义。HP阳性组dCS致敏T淋巴细胞对胃癌细胞杀伤率和Il-12、Ifn-γ的分泌水平均高于HP阴性组(P<0.05)。结论:HP感染状态不影响胃癌患者外周血MOdCS成熟过程形态变化,HP感染的胃癌患者MOdCS成熟和活化水平更高。Objective:This study aimed to compare and analyze the functional differences between peripheral blood monocyte-derived dendritic cells(DCs) of Helicobacter pylori-positive and H.pylori-negative patients with gastric cancer.Methods:H.pylori infection was detected in 84 patients with gastric cancer in our hospital from January 2011 to October 2012 by the14C-urea breath test.DCs were generated from monocytes isolated by an adherent method from the two groups of patients and cultured in the presence of rhIL-4,rhGM-CSF,and rhTNF-α.Furthermore,the expression of surface marker molecules was determined by fluorescence-activated cell sorting analysis.The cytotoxicity of DCs pulsed T cells against gastric carcinoma cell was assessed by the lactate dehydrogenase-releasing assay.The secretion of IL-12 and IFN-γ in the supernatant was determined by enzyme-linked immunosorbent assay.Results:No difference was observed in the morphological change of the maturation process.The mean expression of CD1a,CD80,CD83,CD86,and HLA-DR molecules in DCs of H.pylori-infected patients was higher than that in DCs of H.pylori-negative group,and the differences were statistically significant except for CD1a and HLA-DR.The cytotoxicity activities,IL-12 release,and IFN-γ release in the H.pylori-positive group were significantly higher than those in the H.pylori-negative group(P<0.05).Conclusion:H.pylori infection has no effect on the morphological change of the maturation process of monocyte-derived DCs.These data clearly demonstrate that monocyte-derived DCs of H.pylori-infected patients with gastric cancer can induce stronger maturation and activation than those of H.pylori-negative patients.南京军区医学科技创新课题(编号:10MA068); 福建省自然科学基金资助项目(编号:2010D013); 厦门市科技计划创新项目(编号:3502z20104014)资助~

HPV1618假病毒中和抗体检测方法的验证

通讯作者：李长贵 E-mail：[email protected] 作者简介：赵慧（1980-），女，助理研究员 主要从事病毒疫苗质量控制工作目的 应用假病毒中和法建立检测血清HPV16/18中和抗体滴度检测方法并进行验证。方法 分别采用不同批次假病毒以及不同代次细胞对不同滴度的HPV16/18阳性血清进行多次平行检测，考察这些因素对检验结果的影响；同时通过对抗HPV16/18双价阳性血清、抗HPV16单价阳性血清和抗HPV18单价阳性血清的检测进一步评估中和抗体检测法的准确性、特异性及重复性。结果 不同批次假病毒和不同代次细胞对检验结果的影响均在四倍范围内，此外该检测法的准确性、特异性、重复性均在可接受标准范围之内。结论建立的假病毒法可满足中和抗体效价检测的要求，可用于评价疫苗的免疫效果。国家高技术研究发展计划（2012AA02A402）；国际科技合作项目 (2010DFB30100

Women's education in Meiji Japan and the development of Christian girls' schools

published_or_final_versionHistoryDoctoralDoctor of Philosoph

天然腐植酸肥料是目前最理想的有机肥料

&lt;正&gt; 我国是世界上的农业大国,农业发展历史悠久。民以食为天,农业是基础,至今仍然十分重要。 国家把农业放在十分重要的地位。诸如增加对农业发展的经济投入,加强农业科学研究和技术成果推广工作,对农产品价格进行的调整,对农村的改革,实行持续稳定,

有机多元活性腐殖酸液肥肥效试验

采用相同浓度的有机多元活性腐殖酸叶面肥，对苹果进行１、２、３次喷施，以喷水为对照，观测其对苹果叶片、果实膨大、着色和产量等的效应。结果表明，有机多元活性腐殖酸叶面肥能增加叶面积和重量，防止干旱落果，果实着色率增加３％～１６％，果实径级&ge;７０ｍｍ以上的增加１６．６％，与对照相比喷肥各处理产量分别增加２４．５％、４５．８％和１２３．５％

微机电光开关

对微机械光开关的特点和应用作了简要的论述，并对近年国外几种典型微机械光开关的结构、原理和研制方法作了简要的介绍和评述。这类新型的开关器件的主要优点是串话小，插入损耗小、消光比高，对波长和偏振不敏感，且体积小，造价低等，在光纤通信和光纤测试系统中具有广阔的应用前景

伤员转运机器人自平衡机构的动力学分析

为了提高复杂灾变环境中伤员的救援效率和成功率，提出一种具有3R1T的4-UPS/PS并联机构作为转运机器人的自平衡系统，利用螺旋理论对自由度进行分析，并利用修正的G-K公式检验。采用封闭矢量法求解各驱动杆杆长与动平台位姿间的关系。推导了自平衡补偿机构的动能表达式、势能表达式，利用拉格朗日法建立了自平衡补偿器的动力学模型，通过Adams仿真软件对自平衡过程进行仿真分析运动性以及驱动力。实验结果表明，机构运动性能优越，驱动性能良好，具有很强的可控性和稳定性，适用于救援转运机器人的自平衡系统