马来丝虫感染沙鼠治疗前后血清抗体水平的动态研究

应用马来丝虫和牛丝虫(Setaria cervi)成虫冰冻切片抗原作间接荧光抗体试验(IFAT)及免疫酶染色试验(IEST),均能显示马来丝虫感染沙鼠治疗前后血清IgG及IgM水平的消长情况。两者高峰分别在感染后12～14wk及2～6wk。IgG水平与感染时间长短有密切关系,但与感染度无显著相关。感染沙鼠治后6个月抗体下降。认为用同种和异种抗原作IFAT及IEST,对感染沙鼠均具有诊断和考核疗效的价值。     

班氏丝虫病人治疗前后血清抗体水平及免疫学诊断方法的研究

应用马来丝虫成虫冰冻切片抗原免疫酶染色技术(IEST),检测班氏丝虫微丝蚴阳性病人107例及治疗后一年微丝蚴阴转者35例和治后三年者112例血清特异IgG、IgE和IgM抗体水平,结果表明:治前病人血清IgG抗体滴度≥1:40者阳性率为96.3％,阳性GMRT为116.6;IgE抗体滴度≥1:20者阳性率为86.9％,阳性GMRT为37.1;IgM抗体滴度≥1:20者阳性率为88.8％,阳性GMRT为31.0,该三种抗体从阳性滴度     

日本血吸虫感染小鼠组织内的抗原,抗体定位

应用间接荧光抗体试验(IFAT)检测小鼠感染血吸虫后肝、肠内抗原,结果感染后4wk未检出抗原,6wk抗原水平显著升高,10—12wk达到高峰、其几何均数倒数分别高达512和483。组织内虫卵肉芽肿的病变,亦最为显著,而肝内虫卵肉芽肿的直径显著大于肠内的。用直接荧光抗体试验(DFAT)及IFAT测定小鼠感染血吸虫后肝、肠组织内抗体,结果在感染6wk后,三种抗体(IgG、IgM和IgA)均达低水平,10—12wk均显著上升,而肝、肠组织内IgG抗体则非常显著地高于IgM和IgA的。提示血吸虫虫卵肉芽肿的免疫应答,主要是IgG抗体。     

日本血吸虫感染后IgG和IgM抗体反应动态观察

本文用5种日本血吸虫抗原动态观察了实验感染小鼠血清特异性IgG和IgM抗体反应,并检测了慢性血吸虫病患者血清IgG和IgM抗体。结果显示,感染小鼠的IgG反应对SEA、JEU和JWU3种抗原的反应最强,感染后第6wk急剧上升,第7wk达到高峰,并持续在较高水平至观察结束的第16wk;小鼠的IgM抗体反应以对SEA和JEU两种虫卵抗原的反应最强,第4wk后显著上升,第7—8wk达到高峰。以后逐渐下降。检测125例慢性血吸虫病患者血清,IgG抗体阳性率以SEA、JEU、JWU为最高(88.8％—95.2％),IgM抗体阳性率以SEA最高(65.6％)。上述结果可为免疫诊断、血清流行病学调查及考核防治效果提供科学参考资料。     

抗独特型抗体替代马来丝虫感染期幼虫表面抗原

应用D1E5单克隆抗体(AB1,为小鼠IgM McAb,能识别马来丝虫感染期幼虫表面抗原决定簇)发现有一期特异性表面抗原仅存在于马来丝虫第二期后期和第三期幼虫。感染动物后2～3天内该抗原即从虫体表面丧失。它可能与丝虫传播有关和/或可以作为免疫攻击的靶抗原,然而分离和鉴定这种抗原的工作均未成功,因此制备含有D1E5独特型“内影像”的兔抗独特型抗体来替代该抗原。将AB1免疫兔产生抗独特型抗体(AB2),继用经典竞争试验检测AB2抗独特型特异性,结果表明AB2能特异性抑制AB1与马来丝虫幼虫表面结合,而不能抑制犬恶     

间日疟传播阻断疫苗Pvs25和Pvs28重组蛋白免疫活性的实验观察

目的观察间日疟传播阻断疫苗候选抗原Pvs25和Pvs28重组蛋白免疫小鼠后抗体产生情况,从而探讨此两种候选抗原的免疫活性。方法用酵母菌表达的Pvs25和Pvs28重组蛋白皮下免疫DBA/2小鼠;采用ELISA方法,分别对各组小鼠免疫前后不同时间点血清中特异性IgG及其亚类IgG1、IgG2a的水平进行动态检测。结果初次免疫后第28dIgG及其亚类IgG1、IgG2a均出现有意义的升高(P<0.01),至第112dIgG及其亚类IgG1一直持续于较高的水平(P<0.01);而IgG2a水平则于初次免疫后第42d开始下降,但至112d仍高于免疫前及对照组(P<0.05)。结论Pvs25和Pvs28重组蛋白免疫DBA/2小鼠后可产生较高水平的特异性IgG抗体,并以IgG1亚类为主。     

马来丝虫感染沙鼠血清中特异性抗体水平的动态研究

应用马来丝虫和牛丝虫成虫冰冻切片抗原作IFAT和IEST检测马来丝虫感染长爪沙鼠血清中抗体IgG和IgM水平的动态变化,结果分别在感染后12～14周及2～6周达高峰。感染沙鼠IgG水平与感染时期长短密切相关,但与感染度无关。认为IFAT和IEST,特别是后者,可望成为丝虫病诊断中较为理想的方法;牛丝虫抗原可望能代替马来丝虫抗原用于丝虫病血清诊断中。     

细粒棘球绦虫Eg95基因疫苗和重组抗原诱导小鼠免疫应答的比较研究

目的观察比较细粒棘球绦虫Eg95重组抗原和基因疫苗诱导小鼠的免疫应答状况。方法实验组和对照组小鼠分别注射Eg95重组抗原(rEg95)、费氏佐剂(FCA)、pcDNA3-Eg95基因疫苗、pcDNA3质粒和生理盐水,收集各组血清用酶联免疫吸附(ELISA法)检测抗体IgG和IgG2a亚类水平;采集脾细胞用四甲基偶氮唑盐试验(MTT法)检测免疫小鼠的脾淋巴细胞增殖反应。结果rEg95免疫组小鼠在第二次免疫后开始检测到抗Eg95抗原的IgG,并随着免疫次数的增多,血清抗体效价升高,在第1次免疫后第10周时,免疫抗体滴度可达到1∶25,600。Eg95基因疫苗免疫的小鼠产生抗体滴度随免疫次数的增加而升高,最高可达1∶3,200,但是低于Eg95重组蛋白免疫小鼠产生的抗体滴度水平。pcDNA3-Eg95免疫组产生IgG2a亚类抗体水平明显高于对照组和rEg95组。在第四次免疫后,进行淋巴细胞转化试验,MTT法检测证实rEg95和pcD-NA3-Eg95免疫的小鼠,其脾细胞均可在体外被特异性刺激增生。结论细粒棘球绦虫Eg95重组抗原和基因疫苗均可诱发小鼠产生特异性免疫应答。     

晚期丝虫病患者血清特异性抗体及抗体亚型的分析

目的评价晚期丝虫病患者血清抗丝虫抗体及抗体亚型的免疫学特性.方法用马来丝虫成虫和微丝蚴抗原,以ELISA法检测120份血清标本抗丝虫特异性抗体和抗体亚型,FPT方法进行丝虫皮内试验.结果病原学检查120份受试者均为非微丝蚴血症.80例晚期丝虫病患者FPT试验阳性率为95%(76/80),晚期丝虫病患者血清抗微丝蚴和成虫特异性抗体阳性率分别为82.5%(66/80)和80%(64/80).流行区对照抗体阳性率为10%(2/20),非流行区正常人则均为阴性.晚期丝虫病患者血清抗体亚型以IgG2为主,阳性率92.5%,其它亚型分别为IgG11.25%、IgG322.5%、IgG46.25%.结论晚期丝虫病患者血清中存在抗丝虫特异性抗体,抗体亚型主要是IgG2,它也可能是晚期丝虫病患者特异的抗体亚型.     

诊断人体班氏丝虫感染的抗体测定法

本文作者用指状腹腔丝虫成虫抗原对几组丝虫感染者和非感染者进行了班氏丝虫微丝蚴表面抗原和可溶性抗原与血清抗体(IgG和IgE)反应水平的比较研究。     

马来丝虫感染沙鼠治疗前后血清抗体水手的动态研究

Indirect immunofluorescent antibody test (IFAT) and immunoenzymatic staining technique (IEST), using frozen section of Brugia malayi and Setaria cervi adult worms, were applied to detect levels of IgG and IgM antibodies in jirds infected with Brugia malayi before and after treatment. Both methods could show the dynamic of antibodies during the course of infection. The peak of IgG and IgM antibodies were at the 12-14th week and 2-6th week after infection respectively. A high correlation was observed between the levels of IgG antibody and the period of infection, whereas the antibody titer had no relation with the density of infection. During 6 months after treatment, the levels of antibodies detected by IFAT and IEST using two antigens decreased to low titre. It is considered that IFAT and IEST using heterologous and homologous antigens could be equally used for the serodiagnosis and the evaluation of cure in filariasis.     

旋毛虫病治疗前后血清抗体的观察

应用旋毛虫幼虫冰冻切片抗原免疫酶染色试验（ＩＥＳＴ）观察旋毛虫病治疗前和阿苯达唑治疗后血清抗体水平的变化。结果表明，感染后（治疗前）第２ｗｋ开始出现阳性反应，第４ｗｋ后阳性率均达到９０％以上；治疗后抗体阳性率第１ｗｋ为１００％，第３ｗｋ开始下降，第５ｗｋ后抗体滴度维持于阳性低水平，１２ｗｋ后未见阳性反应。此现象可作为一种治疗有效的客观指标，对旋毛虫病近期及远期疗效考核均有一定价值。     

EB病毒潜伏膜蛋白2的B细胞表位免疫原性研究

目的 分析EB病毒(EBV)潜伏膜蛋白2(LMP2)B淋巴细胞表位的免疫原性.方法 利用生物信息学技术筛选出EBV LMP2可能的优势B淋巴细胞表位LMP2199-209、LMP2318-322和LMP2381-391,将其相应基因片段分别克隆至pET32a(+)载体并转化至大肠埃希菌BL21(DE3)菌株诱导表达,表达产物经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白免疫印迹分析鉴定,通过Ni-NTA离子螯合亲和层析柱纯化.分别以纯化的表位蛋白作为免疫原,皮下多点注射BALB/c小鼠,设pET32a(+)蛋白和PBS为对照.分别以3个表位蛋白作包被抗原,ELISA法检测第0、3、6周小鼠血清中相应表位特异性抗体IgG,并于免疫后第6周检测各免疫组小鼠血清抗体识别天然EBV抗原的能力.结果 在原核表达体系中分别获得了表位LMP2199-209、LMP2318-322和LMP2381 391融合表达产物.纯化的表位蛋白免疫小鼠,血清中分别能检测到相应的表位特异性抗体IgG,其抗体水平随着免疫次数的增加而呈现逐渐升高的趋势,表位LMP2318-322免疫组第3、6周小鼠血清抗体显著高于pET32a(+)蛋白对照组(F=493.85、773.99,均P＜0.05),LMP2381 391免疫组第3、6周抗体水平亦显著高于pET32a(+)蛋白对照组(F=926.33、309.14,均P＜0.05),而表位LMP2199 209免疫组至第6周特异性抗体高于pET32a(+)蛋白对照组(F=87.27,P＜0.05).3个表位蛋白免疫小鼠血清抗体IgG均能识别EBV天然抗原,以表位LMP2199-209和LMP2381-391免疫组抗EBV-IgG生成水平尤为显著.结论 筛选的EBV LMP2可能的优势B淋巴细胞表位LMP2199-209、LMP2318-322和LMP2381-391通过原核表达体系中制备的表位蛋白,具有较好的免疫原性,可进一步用于EBV感染及其相关肿瘤表位疫苗的研究.     

旋毛虫肌肉期幼虫分泌排泄物中46-58kD抗原诱发小鼠保护性免疫的研究

为探讨旋毛虫肌肉期幼虫分泌排泄物（ＴｓＬ１ＥＳ）中４６５８ｋＤ抗原的保护性免疫作用，本文用该纯化抗原组分加福氏完全佐剂（ＦＣＡ）腹腔注射免疫昆明小鼠３次，继以旋毛虫感染性幼虫３００条攻击感染，感染后第７天计数小鼠肠道成虫数、第３０天肌肉幼虫数和血清抗体ＩｇＧ滴度。结果：４６５８ｋＤ抗原免疫鼠的肠道成虫减虫率、肌肉幼虫减虫率和血清抗体ＩｇＧ的几何平均倒数滴度分别为４１８％、４６１％和２８４１２；与ＴｓＬ１ＥＳ诱导的保护性免疫力（４８３％、５０２％和３４５８６）水平接近；显著高于佐剂对照组（４８％、８３％和７４８６）。表明：ＴｓＬ１ＥＳ中４６５８ｋＤ抗原组分可产生明显的保护性免疫作用     

免疫酶染色试验诊断日本血吸虫病价值的研究

应用感染30条血吸虫尾蚴后7wk的鼠肝组织内虫卵冰冻切片抗原作免疫酶染色试验,采取单盲法检测606例经粪检确诊的日本血吸虫病人和310名健康人,阳性率和假阳性率分别为89.9％和4.2％;检测239例已治疗过的日本血吸虫病人,阳性率为24.7％,转阴率为75.3％;检测10例肺吸虫病人及60例华支睾吸虫病人,交叉反应率为0～6.7％。对不同感染期及不同感染度的血吸虫病兔血清中特异性抗体的动态检测,能显示抗体的消长,免疫酶染色的反应性和反应强度与感染期和感染度密切相关。本文进一步证明该法具有较高的诊断效果,并有疗效考核的价值。     

Inhibition of arteriosclerosis by T-cell depletion in normocholesterolemic rabbits immunized with heat shock protein 65.

Previous studies in our laboratory have shown that arteriosclerotic changes can be induced in normocholesterolemic rabbits by immunization with mycobacterial heat shock protein (hsp) 65. To further investigate the immunologic mechanisms underlying such vascular lesions, 39 male New Zealand White rabbits were treated by triple immunization with fortified Freund's complete adjuvant containing 5 mg/mL Mycobacterium tuberculosis as a source of hsp65 and simultaneous immunosuppressive therapy twice per week with either anti-CD3 monoclonal antibody (1 mg/kg) and prednisolone (1 mg/kg) or prednisolone (1 mg/kg) alone. Sixteen weeks after the first immunization the animals were killed, and as expected, severe arteriosclerotic lesions in the intima of the aortic arch were found in 9 of 10 immunized rabbits. However, only 1 of 10 rabbits immunized and immunosuppressed with the combined anti-CD3 monoclonal antibody and prednisolone treatment showed a single moderate lesion in the aorta, whereas 5 of 9 rabbits immunized and immunosuppressed by prednisolone treatment alone showed lesions, albeit mild. In conclusion, the early inflammatory stages of arteriosclerotic lesions induced by immunization with hsp65 can be inhibited by immunosuppressive therapy with anti-CD3 monoclonal antibody.     

In Vitro Stimulation of Sensitized Lymphocytes by Herpes Simplex Virus and Vaccinia Virus

Splenic lymphocytes from rabbits immunized with herpes simplex virus (HSV) were incubated in vitro with ultraviolet light-inactivated HSV, and the degree of lymphocyte transformation was determined by measurement of the incorporation of [(3)H]thymidine into acid-insoluble material. Lymphocytes from immunized rabbits were stimulated as much as 30-fold, whereas lymphocytes from control rabbits were unaffected. Lymphocyte sensitization occurred within 3 days after immunization; sensitized lymphocytes could still be detected 120 days after immunization. The antigenicity of the ultraviolet light-inactivated crude virus pool was found to reside primarily in the virion. Infectious virus, in comparison with inactivated virus, produced less lymphocyte stimulation. Studies on the interaction of the humoral and cellular immune responses showed that incubation of anti-HSV antibody with HSV antigens did not reduce the capacity of the viral antigens to stimulate sensitized lymphocytes. Other experiments revealed that lymphocytes from both the spleen and peripheral blood of animals immunized with vaccinia virus could be stimulated by vaccinia, but not by HSV. Conversely, lymphocytes from animals immunized with HSV could not be stimulated by vaccinia. The transformation of sensitized lymphocytes by viral antigens appears to be a simple, highly specific, and quantitative in vitro technique for the study of the cellular immune response to viral infections.     

Maternal immunization with a herpes simplex virus type 2 replication-defective virus reduces visceral dissemination but not lethal encephalitis in newborn mice after oral challenge

Herpes simplex virus (HSV) causes devastating infections in newborns. Maternal immunization is one potential strategy to reduce neonatal HSV disease. Female mice were immunized with an HSV type 2 (HSV-2) replication-defective mutant (HSV-2 dl5-29, which is defective for the genes UL5 and UL29) and then mated. Protection was evaluated in newborn mice after a virulent HSV-2 oral challenge. Heightened neonatal susceptibility was observed to a thymidine kinase-negative HSV-2 strain (HSV-2 186DeltaKpn) that is highly attenuated in adult mice. Maternal immunization with HSV-2 dl5-29 and HSV-2 186DeltaKpn reduced visceral spread of infectious challenge virus in pups after challenge at either 1 day or 1 week of age but did not prevent replication at the site of entry, spread to the central nervous system, or lethal encephalitis. No protection was seen in pups born to mock-immunized mothers or to mothers immunized with a UV-inactivated wild-type HSV-2 strain. Levels of protection correlated with levels of passively transferred maternal HSV-2-specific IgG antibody.