多价精子抗原表位肽的设计与免疫效应研究

目的 开发可供两性使用的多价合成嵌合肽疫苗。方法 选择小鼠精子／睾丸特异性抗原SP17、Cyritestin中特异性氨基酸序列与牛核糖核酸酶非限制性T细胞表位作为骨架，按一定连接方式设计、合成新抗原35肽，并与弗氏佐剂混悬后免疫雌性BALB／C小鼠。结果 在430A固相自动肽合成仪上成功地合成35肽，并经HPLC纯化、质谱鉴定证实其纯度达95％以上；免疫后小鼠血清特异性IgG抗体滴度最高达1：6000，阴道粘膜冲洗液中特异性IgA抗体滴度最高达1：300；小鼠脾脏淋巴细胞增殖率达50％左右，淋巴细胞分泌的INF－γ和IL－4分别为60、108μg/ml；实验组BALB／c雌鼠妊娠率平均降至60％，每胎产仔数平均降低58％－71％。结论 含有两种特异性精子抗原中特异性氨基酸序列的新抗原可激发小鼠产生较理想的特异性体液免疫和粘膜免疫，并使小鼠受孕率下降，为研制适宜于两性的多价抗生育疫苗提供了实验基础。     

卡介苗对哮喘豚鼠的影响

目的 研究卡介苗对哮喘豚鼠的防治作用。方法 24只豚鼠随机分为阳性对照组、地塞米松组、卡介苗组及阴性对照组,每组6只,除阴性对照组用生理盐水外,其余三组每只腹腔内注射鸡卵蛋白100mg,2周后用1％鸡卵蛋白激发致敏,每次30min,每日1次,持续3d,地塞米松组每次激发前肌注地塞米松2mg/kg,卡介苗组每次激发前肌往卡介苗30mg/kg,阳性对照组每次激发前30min肌注生理盐水2ml,最后用乌拉坦麻醉,检验动脉血气分析参数,嗜酸粒细胞(EOS)计数等参数,所有参数用t检验或卡方检验分析。结果 阳性对照组与其它三组比较,在豚鼠的发病程度、动脉血气分析、外周血EOS计数、支气管肺泡灌洗液中EOS计数、支气管病理切片每20个高倍视野EOS计数均有明显差异(P<0.05或P<0.01)。结论 卡介苗能够抑制动物模型中的哮喘反应。     

西安市麻疹疫苗免疫接种情况的调查

目的　为了掌握西安市婴儿麻疹疫苗 (以下简称MV)基础免疫接种情况 ,以进一步提高免疫成功率。方法　在西安市城区和郊区随机抽查 5 7名 0岁组健康婴儿 ,采用酶联免疫吸附法 (ELISA)测定血清麻疹IgG抗体水平。结果　免疫前MV阳性率为 2 8 0 7% ,GMT 1∶4;免疫后MV阳性率为 98 2 5 % ,GMT 1∶1666;免疫前后阳性率、抗体平均滴度经统计分析 ,差异有非常显著性 (P <0 .0 1) ;对MV免疫后不同性别、不同地区的抗体阳性率 ,平均滴度进行统计分析 ,差异无显著性 (P >0 0 5 )。结论　婴儿在出生后 8个月 ,母体胎传抗体逐渐衰减 ,致MV阳性率极低 ,只需按现行的免疫程序及时有效地接种MV ,就能获得保护抗体 ,达到控制麻疹发病的目的。同时也说明MV的免疫成功与否不受性别和地区的影响。     

乙型肝炎病毒多表位治疗性抗原基因的合成、表达及抗原性研究

目的研究乙型肝炎病毒多表位治疗性疫苗基因的合成、表达及其产物的抗原性。方法设计、并合成乙型肝炎病毒多表位抗原基因BPT,克隆入融合表达载体pWR450-1,构建重组质粒PWR/BPT。在大肠杆菌中表达乙型肝炎多表位抗原蛋白并纯化该蛋白,并用Western-blotting方法初步检测该抗原蛋白的抗原性。结果成功构建了融合表达载体PWR/BPT,在大肠杆菌中表达出乙型肝炎多表位抗原蛋白,Western-blotting 检测显示该蛋白具有良好抗原性。结论HBV多表位治疗性抗原基因的设计是成功的,其在大肠杆菌中表达的重组融合蛋白具有良好的抗原性,可能是较理想的HBV治疗性疫苗候选物。     

卡介苗配用腺苷接种提高巨噬细胞抗结核杆菌效能的研究

目的　通过腺苷 (ADO)改善卡介苗 (BCG)初次免疫接种效果 ,提高巨噬细胞抗结核杆菌毒力株效能。方法　将BALB/C小鼠随机分BCG组、ADO组和对照组。对BCG、ADO两组分别一次性皮内注射卡介苗 0 1ml。对ADO组腹腔注射ADO 30mg/(kg·d) ,对BCG组、对照组腹腔注射生理盐水 0 1ml/d ,共 5d。 6周后 ,三组均经血管接种人型结核分枝杆菌标准毒力株 1× 10 6CFU ,BCG组、ADO组于结核杆菌感染后第 10、2 0天和 30天分批取材 ,对照组于第 30天取材。将肺、脾用多聚甲醛固定 ,石蜡包埋 ,HE染色与抗酸染色 ,进行原位细胞凋亡检测。结果　对照组结核杆菌感染后第30天 ,肺组织大部分实变 ,以中性粒细胞浸润为主 ,结核结节多 ,肺实变与肺泡隔重度增宽处弥漫分布大量菌体 ;BCG组于感染后第 10～ 30天肺泡隔增宽 ,逐渐加重至肺大部分实变 ,间质内以淋巴细胞浸润为主 ,结核结节较少 ,肺内大量菌体 ;而ADO组肺实变和肺泡隔重度增宽的面积较小 ,间质内以单核细胞浸润为主 ,结核结节很少 ,肺内菌体明显减少。结论　ADO促进BCG诱导的免疫记忆性反应中能引起单核 巨噬细胞的数量增多和杀菌能力的增强     

绿脓杆菌MSHA菌毛株菌苗与白血病免疫应答反应

绿脓杆菌(MSHA)菌苗是一种具有广谱抗感染效果的免疫调节剂。我们观察了20例白血病患者多次疫苗主动免疫的免疫应答反应,14例病人血中广谱抗体效价升高,临床上有很好的治疗反应,6例抗体效价无变化,而临床症状加重,最后均死于败血症。上述结果提示白血病病人对疫苗的免疫应答反应,在白血病缓解中起着重要作用,对临床早期准确判断白血病患者的预后很有意义。     

变形链球菌表面蛋白和葡糖基转移酶基因疫苗对唾液变形链球菌和牙菌斑的影响

目的　了解变形链球菌表面蛋白和葡糖基转移酶基因疫苗单独及联合免疫对定菌鼠唾液变链菌和牙面菌斑的影响。方法　 2 8d龄 Wistar大鼠 3 6只 ,随机分为 pc DNA3 - pac组、pc DNA3 - gtf B组、pc DNA3 - pac联合pc DNA 3 - gtf B组、变形链球菌灭活全菌组、pc DNA3空载体组和 PBS液组 ,进行三次双侧颌下腺腺周注射免疫 ,建立定菌鼠模型 ,作诱龋实验 3个月。唾液变链菌计数和菌斑计分。结果　唾液变链菌菌落计数和牙面菌斑计分在 pc D-NA3与 PBS组最高 ,其次为单基因疫苗免疫组 ,联合基因疫苗和灭活全菌细胞免疫组最低 ,各组间有显著性差异 ( P<0 .0 5 )。结论　 pc DNA3 - gtf B和 pc DNA3 - pac具有明显的免疫抑菌作用 ,联合基因疫苗免疫优于单基因疫苗     

An adenovirus type 5 (AdV5) vector encoding an envelope domain III-based tetravalent antigen elicits immune responses against all four dengue viruses in the presence of prior AdV5 immunity

Dengue is a mosquito-borne viral disease caused by four antigenically distinct serotypes of dengue viruses (DENVs). This disease, which is prevalent in over a hundred tropical and sub-tropical countries of the world, represents a significant global public health problem. A tetravalent dengue vaccine capable of protecting against all four DENV serotypes has been elusive so far. Current efforts are focused on producing a tetravalent vaccine by mixing four monovalent vaccine components. In this work, we have utilized a discrete carboxy-terminal region of the major DENV envelope (E) protein, known as domain III (EDIII), which mediates virus entry into target cells and contains multiple serotype-specific neutralizing epitopes, to create a chimeric tetravalent antigen. This antigen derived by in-frame fusion of the EDIII-encoding sequences of the four DENV serotypes was expressed using a replication-defective recombinant human adenovirus type 5 (rAdV5) vaccine vector. This rAdV5 vector induced cell-mediated immune responses and virus-neutralizing antibodies specific to each of the four DENVs in mice. Interestingly, anti-AdV5 antibodies did not suppress the induction of DENV-specific neutralizing antibodies. We observed that anti-AdV5 antibodies in the sera of immunized mice could promote uptake of a rAdV5-derived reporter vector into U937 cells, suggesting that pre-existing immunity to AdV5 may in fact facilitate the uptake of rAdV5 vectored vaccines into antigen presenting cells. This work presents an alternative approach to developing a single component tetravalent vaccine that bypasses the complexities inherent in the currently adopted four-in-one physical mixture approach.     

Novel IgE peptide-based vaccine prevents the increase of IgE and down-regulates elevated IgE in rodents

BACKGROUND: Immunotherapy with anti-IgE antibodies for treatment of allergy is promising but a short half-life and extremely high cost limit its application. OBJECTIVE: We sought to develop IgE vaccines that induce longer-lasting auto-antibodies to neutralize self-IgE as an alternative therapy. METHODS: The vaccine was made by conjugating three synthetic peptides corresponding to human IgE receptor-binding sites to a carrier, hepatitis B surface antigen. To test the immunogenicity of the vaccine, rats were immunized with the vaccine or hepatitis B surface antigen as control. Serum IgG titres to human IgE and the IgE of other species were measured. The inhibition by rat antisera of the binding of human IgE to its receptor was assessed by ELISA, flow cytometry analysis, and passive cutaneous anaphylaxis (PCA), and its ability to recognize receptor-bound IgE was examined. The in vivo effect of the vaccine was evaluated in trichosanthin-sensitized mice and rats. In the preventative study, vaccination started before sensitization commenced, while in the treatment study, vaccination started after sensitization. Sensitized mice and rats receiving injections of the carrier served as controls. Trichosanthin-specific IgE was measured using PCA. RESULTS: Sera from vaccine-immunized rats contained high titre antibodies that reacted with soluble and plate-bound but not with receptor-bound human IgE; they also reacted with mouse, rat, and dog IgE. Furthermore, the sera inhibited the binding of human IgE to its receptor in a dose-dependent manner. In preventative and treatment studies, serum trichosanthin-specific IgE levels were significantly reduced in vaccinated groups compared with controls. CONCLUSION: Antibodies against self-IgE can be induced by IgE peptide-based vaccines, which are effective in preventing the increase of IgE and in down-regulating IgE in sensitized animals.