自体淋巴细胞免疫重建对白血病瘤苗的加强作用

目的观察在免疫重建过程中应用白血病瘤苗对DBA/2小鼠的一般状况、抵御L1210细胞攻击以及小鼠保护性免疫功能的影响。方法用同系小鼠脾脏淋巴细胞对照射后的免疫缺陷鼠进行免疫重建,同时用灭活的L1210细胞皮下注射免疫DBA/2小鼠,7d后用L1210细胞皮下攻击小鼠,观察小鼠的一般状况、成瘤率、瘤块生长情况。通过ELISA法测定外周血IFN-γ含量,以评估细胞免疫作用的强弱;通过病理切片了解瘤块中心坏死区及周围浸润细胞情况。结果与对照组相比,免疫重建+瘤苗可以有效提高单纯应用瘤苗所产生的保护性免疫功能,IFN-γ含量明显升高;用L1210细胞攻击,成瘤小鼠瘤块生长缓慢,瘤周单个核细胞浸润明显,瘤块内部肿瘤细胞坏死明显,生存期大大延长。结论在免疫重建的同时应用瘤苗,可以大大加强瘤苗的保护性免疫作用。     

Construction and characterization of hGM-CSF-expressing K-562 cell line

Objective The whole process of vaccine preparation is time-consuming and technically challenging. Here the hGM-CSF-engineered K-562 cell line was constructed to simplify tumor vaccine preparation process. Methods The eukaryocyte expressing plasmid pcDNA3. 1/GM-CSF was first constructed and its accuracy was verified through sequencing. The pcDNA3. 1/GM-CSF was transfected into COS-7 cells to verify GM-CSF expression and cytokine activity using TF-1 cell line. Then the plasmid was transfected into K-562 cell line using liposome method, and was selected under G-418 and sub-cloned by limiting dilution. GM-CSF product from the monoclone GM-CSF-K-562 cell lines was quantified using ELISA method. Results We successfully constructed the hGM-CSF eukaryocyte expressing plasmid and hGM-CSF expressing K-562 cell line. Conclusion The construction of K-562/GM-CSF line will simplify the preparation of tumor vaccine, thus facilitating the application of tumor vaccination therapy in clinical application.     

改进的自适应分组个体重构免疫算法

针对免疫算法收敛速度慢,有可能陷入局部寻优情况,提出了一种改进的自适应分组个体重构免疫算法.在个体重构的实现上采用了分组进行,同时对重构算法进行了合理的改进,既保证了收敛速度,同时也保证了全局寻优的过程.仿真实验也表明了这一改进算法在收敛速度和寻优能力方面较原算法有较大的改善.     

新型疫苗治疗急性白血病及多发性骨髓瘤的临床研究

目的　探讨新型疫苗主动治疗急性白血病及多发性骨髓瘤的疗效及毒副作用。方法　应用基因工程生产的细胞因子和急性白血病及多发性骨髓瘤肿瘤细胞制备的疫苗 ,主动免疫治疗急性白血病 7例、多发性骨髓瘤37例。结果　 37例急性白血病患者中化疗未缓解或化疗不能耐受者 2 9例 ,经疫苗治疗达到完全缓解者 5例占17.3% ,部分缓解者 6例占 2 0 .7% ,微效者 4例占 13.8%。总有效率 5 1.8%。化疗缓解组 8例疫苗治疗后维持缓解达 8～ 13个月复发。多发性骨髓瘤 7例 ,其中 1例完全缓解 ,5例部分缓解。治疗中未发现明显副作用。结论　应用新型疫苗治疗化疗无效或化疗不能耐受的急性白血病及多发性骨髓瘤具有肯定疗效。且经济简便 ,无明显毒副作用     

结核分枝杆菌Ag85A基因DNA疫苗的构建和免疫原性研究

目的　构建编码结核分枝杆菌蛋白Ag85A基因为基础的DNA疫苗。 方法　采用聚合酶链反应从结核杆菌H37Rv株基因组中 ,扩增出分泌蛋白Ag85A成熟蛋白的编码基因 ,用限制性内切酶消化后 ,插入克隆载体 pGEM TEasy中。经酶切鉴定与序列测定证实后 ,以亚克隆法构建于真核表达载体 pcDNA3.1的相应酶切位点。重组质粒肌注免疫小鼠 ,4周后用ELISA法检测抗体滴度。结果　结核分枝杆菌H37Rv株Ag85A成熟蛋白的编码基因经序列测定证实无突变 ;经BamHⅠ和EcoRⅠ双酶切鉴定证实克隆基因正确插入载体 pcDNA3.1。ELISA法检测几何平均滴度为 1∶10 0 0。结论　以Ag85A成熟蛋白的编码基因为基础的DNA疫苗的成功构建和表达 ,为进一步研究其在结核病与肿瘤防治方面的作用奠定了基础     

葎草花粉变应原的研究进展与展望

变态反应性疾病是世界范围内重大卫生问题之一,近50%的变态反应性疾病患者对草类花粉过敏。葎草在我国分布广泛,是引起花粉症的主要致敏物之一。葎草花粉症患者呈逐年上升趋势,已受到人们广泛关注。本文从葎草花粉的致敏状况、主要致敏蛋白组分、葎草花粉症诊断和治疗方法的建立以及主要变应原重组等方面进行综述。     

INTRAMUSCULAR VERSUS INTRADERMAL HEPATITIS B REVACCINATION IN HEALTHY NON-RESPONDER CHILDREN： A 5-YEAR PROSPECTIVE RANDOMIZED STUDY

HepatitisBvaccine,introducedovertwenty yearsago,hasshownexcellentrecordsofsafety, immunogenicityandprotectiveefficacy.Recentre viewsindicatedthatwithhelpofimmunologic memoryaprimaryvaccineseriescanprotectre spondersfromhepatitisBvirus(HBV)clinically sig…     

NINE-YEAR EFFICACY OF HEPATITIS B VACCINATION WITHLOW-DOSE IN THE THIRD INJECTION

Nowthescheduleofvaccinationagainsthep-atitis BisusuallythreedosesoflOUg(or2oUg)',,.Itisusedasgoldstandard,asitislargelyappliedandrecommendedbyvaccinemanufactqrers.Inor-dertousethevaccinemorerationallyandtoobtainsafeimmunizationwithreducedcosts,manystud-iescompareddifferentscheduleswithvariantdosages.Buttheconclusionsdidn'tmatchwitheachother-Andfornolong-termfollowingup,theresultscan'tbeputintopractice.Inadouble-blindandrandomizedtrial,start-edinl986,weprovidedvaccineofvariantdosagesinthethir…     

葎草花粉主要变应原核酸疫苗pcDNA3.1-LC2的构建及其免疫原性鉴定

目的 构建葎草花粉主要变应原核酸疫苗pcDNA-LC2并鉴定其免疫原性.方法 将pTripIEx2-LC2质粒用EcoRⅠ和HindⅢ双酶切得到LC2葎草花粉变应原基因片段,将其插入pcDNA3.1(-)真核表达载体,在鉴定重组质粒构建成功后,大量制备去内毒素核酸疫苗pcDNA3.1-LC2,应用pcDNA3.1-LC2核酸疫苗免疫小鼠,取免疫血清行双向免疫扩散实验,分析其免疫原性.结果 通过测序确认构建pcDNA3.1-LC2中的672bp的编码框碱基与原序列一致,没有突变及缺失.应用pcDNA3.1-LC2免疫BALB/c小鼠,经双向免疫扩散试验验证pcDNA3.1-LC2在小鼠体内可以表达并产生特异性抗体.结论 成功的构建了葎草花粉主要变应原核酸疫苗pcDNA3.1-LC2,pcDNA3.1- LC2可在小鼠体内表达,并能产生葎草花粉变应原特异性抗体,具有良好的免疫原性.     

HPV-6/11 L1/E6、HPV-6/11 L1/E7嵌合DNA疫苗质粒的构建

目的　构建HPV 6 / 11L1/E6、HPV 6 / 11L1/E7预防、治疗性DNA疫苗质粒。方法　运用PCR扩增HPV 6 /11L1、E6、E7序列 ,PCR产物连接至pGEMT Easy质粒 ,测序鉴定正确后 ,分别连接至真核表达载体pVAX ,再用酶切、连接而构建成HPV 6 / 11L1 E6 /E7 pVAX质粒。运用电穿孔法将所获得质粒转染COS 7细胞 ,免疫组化检测蛋白表达情况。结果　所构建质粒的插入目的片段测序正确 ;免疫组化检测在胞浆胞核可见棕黄色点状阳性产物沉积。结论　所构建的HPV 6 / 11L1 E6 /E7 pVAX融合蛋白表达质粒可在体外表达L1 E6 /L1 E7蛋白 ,为今后进行DNA疫苗的动物实验及临床实验研究做好准备     

SIX-YEAR EFFICACY OF HEPATITIS B REVACCINATION:A DOUBLE-BLIND,PLACEBO-CONTROLLED AND RANDOMIZED FIELD TRIAL

Alargenumberofstudiesshowedthatanti--HBsinducedbyhepatitisBvaccinedeclinedrapidlysoonafterthebasiccourseofimmunization,andfellbelowdetectablelevelorprotectivelimit(10IU/LorS/NM10)in20%~50%ofthevaccineesbetween3--5years',.,'.Forthisreason,manyresearcherssuggestedaboosterdoseduringthisperiod.Seeingthatthemajorityoftheboosterdoserecipientshadananamnesticresponsewithapromptincreaseofanti--HBs,theefficacyofrevaccinationwasconsideredtobeperfect".'>.However,thepersistenceofanti-HBsandtheprotecti…     

LONG-TERM EFFICACY AND IMMUNOLOGICAL MEMORY OF PLASMA-DERIVED HEPATITIS B VACCINE 11 YEARS AFTER INITIAL INOCULATION

The length of protection of hepatitis B vaccineand the effective time for vaccine booster doseshave not been defined.Most of studies suggestedthatthe vaccine was effective for8to1 0 years[1～ 5] .However,in most cases,the study design was notrigorous. Many were cross- sectional studies per-formed before and aftervaccination,orstudies thatcompared the vaccinees with the general popula-tion.A few of studies as by Hadler[1] et al .werebased on randomized,double- blind and placebo-controlled tria…     

THREE－YEAR FOLLOW－UP AFTER A SINGLE BOOSTER DOSE OF VACCINE IN THE NONRESPONDERS AND HYPORESPONDERS TO HEPATITIS B VACCINE

ＴＨＲＥＥ－ＹＥＡＲＦＯＬＬＯＷ－ＵＰＡＦＴＥＲＡＳＩＮＧＬＥＢＯＯＳＴＥＲＤＯＳＥＯＦＶＡＣＣＩＮＥＩＮＴＨＥＮＯＮＲＥＳＰＯＮＤＥＲＳＡＮＤＨＹＰＯＲＥＳＰＯＮＤＥＲＳＴＯＨＥＰＡＴＩＴＩＳＢＶＡＣＣＩＮＥ（王学良）（徐慧文）（隋秀芬）（门伯媛）...     

乙型肝炎疫苗无弱应答者复种效果观察

目的　观察乙肝疫苗无弱应答者复种后近、远期效果。方法　严格筛选无弱应答者 ,用3针加倍剂量全程接种 ,随机分组观察皮内和肌肉接种效果。于接种首针后 1、2、8个月随访采血 ,检测乙肝三项指标 ,做近期观察。结果　皮内接种效果不如肌肉接种 ;接种 3针似不能取得比 1、2针更好的效果 ;8个月观察抗体应答率虽高达 90 % ,但第 8个月时已见抗体水平下降 ,与一般人群接种应答规律显著不同。结论　从近期效果看 ,复种是否能取得较好的远期效果 ,尚待观察研究。     

THE CONSTRUCTION AND PRELIMINARY APPRAISEMENT OF HSV-2 gD GENE DNA VACCINE

Ｈｅｒｐｅｓｓｉｍｐｌｅｘｖｉｒｕｓｔｙｐｅ 2 (ＨＳＶ 2 ) ,ｏｎｅｋｉｎｄｏｆｄｏｕｂｌｅｓｔｒａｎｄｓＤＮＡｖｉｒｕｓ ,ｃａｎｃａｕｓｅｓｅｒｉｏｕｓｗｏｒｌｄｗｉｌｄｉｎｆｅｃｔｉｏｎｓ.Ｈｅｒｐｅｓｓｉｍｐｌｅｘｖｉｒｕｓｇｅｎｏｍ ｉｃＤＮＡｃｏｄｅｓｆｏｒａｐｐｒｏｘｉｍａｔｅｌｙ 80 ｐｒｏｔｅｉｎｓ .Ｍｏｓｔｏｒａｌｌａｒｅｅｘｐｒｅｓｓｅｄｄｕｒｉｎｇｌｙｔｉｃｉｎｆｅｃｔｉｏｎ ,ｍａｎｙａｒｅｆｏｕｎｄｉｎｉｎｔａｃｔｖｉｒｉｏｎｓａｎｄａｔｌｅａｓｔ 11ｈａｖｅｂｅｅｎｉｄｅｎｔ…     

PRELIMINARY STUDY OF A NOVEL HUMAN PAPILLOMAVIRUS TYPE 16L1／E6-E7 CHIMERIC RECOMBINANT DNA VACCINE

Objective Preparations of HPV16 L1/E6 and L1/E7 prophylactic and therapeutic DNA vaccines. Methods The nucleotides within HPV16 E6 and E7 genes, which are responsible for viral transforming activity, were mutated by mage primer site-directed mutagenesis method. The correctly mutated E6 and E7 fragments were separately cloned into an eukaryotic expression vector pVAX1, together with HPV16 L1 gene, generating chimeric recombinants plasmids 1MpVAX1-L1E6, 2MpVAX1-L1E6, 1MpVAX1-L1E7, 2MpVAX1-L1E7 and 3MpVAX1-L1E7. CHO cells were transiently transfected with the individual DNA vaccines by calcium phosphate method. Target protein expressions in the extracts of the transfected cell lines were measured by ELISA and immunohistochemistry, with HPV16 L1 and E6 specific monoclonal antibodies. Results ELISA assays showed the P/N ratios in the cell extracts transfected with L1E6 and L1E7 plasmids were more than 2.1. Immunohistochemistry revealed brownish precipitant signal in cytoplasm and nuclei of the transfected cells. Conclusion Successful constructions of prophylactic and therapeutic DNA vaccine plasmids lay solid foundation for future animal experiment and clinical trial.     

ASSOCIATION OF GENETIC BACKGROUND WITH NON-ANDHYPORESPONSIVENESS TO HEPATITIS B VACCINE

ItwasestimatedthatafterthebasicimmunizationseriesofhepatitisBvaccine,about5%--10%ofthevaccinesfailedtodevelopdetectableanti--HBsorhadaminimalantibodyresponse(anti--HBsS/Nratiolessthan10),andbecameso-callednonrespondersandhyporesponders.Forthesepeoplethevaccinationwasa~.nailynoteffectiveandprotective.Accordingtothepublisheddata.thepoorresponsivenesswasrelatedtovariousfactorsandgeneticswasoneofthevariablesconcernedtl.2).However,theassociationofgeneticbackgroundwiththepoorresponsivenessstillla…     

经直肠豚鼠志贺菌痢疾模型的改良

目的改良经直肠途径建立豚鼠志贺菌痢疾动物模型的方法。方法 20只受试豚鼠根据感染前24 h禁食或不禁食处理分为2组,8只非禁食组参照文献经直肠灌注致病性福氏志贺菌sf301菌株,12只禁食组经直肠分别灌注PBS(5只)和sf301菌株(7只),观察受试豚鼠的感染症状,通过组织病理学方法证实其发病与否。结果非禁食组豚鼠仅有1只(1/8)在感染后24 h内表现出明显的体重减轻、发热等典型的菌痢症状。禁食组经直肠灌注sf301菌株的豚鼠(6/7)在感染后24 h内有显著的发热症状,体重减轻则发生在感染后的24~48 h内;灌注PBS的豚鼠(0/5)无异样症状。组织病理学结果显示,发病豚鼠结肠末端的黏膜以及黏膜下层遭到明显的破坏,肠壁增厚,出现水肿、糜烂、大量中性粒细胞浸润等。结论成功建立了改良的经直肠豚鼠志贺菌痢疾动物模型。     

白血病瘤苗对正常小鼠免疫作用的观察

目的　观察自制的一种白血病瘤苗对C57BL/ 6小鼠一般状况 、抵御FBL 3细胞的攻击及小鼠免疫功能的影响。方法　用自制的白血病疫苗 ,内含灭活的FBL 3细胞 +不完全弗氏佐剂 (incompletefoundadjuvant,IFA) +细胞因子(rIL 2、rIL 6、rGM CSF) ,皮下注射免疫C57BL/ 6小鼠 ,用FBL 3细胞皮下注射攻击小鼠 ,观察小鼠一般状况、成瘤率、瘤块生长情况 ;用病理学方法了解瘤块及疫苗注射部位病理变化 ;用MTT比色法检测小鼠巨噬细胞 (macrophage ,Mφ)及细胞毒性T淋巴细胞 (cytotoxicTlymphocyte ,CTL)杀伤活性。 结果　与对照组相比 ,发现自制成的白血病瘤苗能有效的提高小鼠的细胞免疫功能 ,Mφ及CTL杀伤活性明显提高 ,用FBL 3细胞攻击 ,小鼠成瘤率低 ,成瘤小鼠瘤块生长缓慢 ,瘤周单个核细胞浸润明显 ,瘤块内瘤细胞坏死明显。结论　所用白血病疫苗制备简单 ,疗效肯定 ,副作用小 ,有必要进一步研究     

THE CONSTRUCTION OF LEUKARYOTIC EXPRESSION PLASMID PCDNA3/HPV16E6 AND EXPERIMENTAL OBSERVATION OF NAKED DNA INJECTED INTO ANIMAL

Cervicalcancerisacommonmalignancyofmarriedwomen.HighriskHPV(HPV16,18)infectioniscloselyassociatedwithcervicalcancer(l).HPV16DNAcanbedetectedinapproximately50%~90?rvicalcarcinomastZ).HPV16E6andE7geneshasbeenprovedtobetransforminggenes.TheirgeneproductscanbindtoandinactivateP53andPRbrespectively,playingacausalroleinthedevelopmentofcervicalcarcinomat27.AlotofinvestigationindicatethatHPV16E6/E7wereconsistentlyandhighlyexpressedincervicalcarcinomas,andthatE6/E7proteinscouldinducebothhu…