细粒棘球绦虫原头节在NIH株小鼠体内发育的组织学及宿主细胞反应观察

本文的观察结果表明:接种后1周,原头节的皮层内已有实质细胞长入,虫周有较大空隙,宿主细胞反应轻微。死亡原头节则已被大量炎细胞紧密包围;2～4周,大部分原头节的实质组织消失而形成细粒棘球蚴生发层,宿主细胞反应基本消失。而死虫周围则有夏科-雷登氏结晶体出现;2个月后,生发层外均已出现角质层,死亡崩解虫周的细胞反应开始减轻;4～6个月,细粒棘球蚴囊不断增大,并在生发层内陆续出现不育囊结构,虫周纤维组织较少,而解体的原头节内均已有大量胶原及网状纤维长入。对生发层的组织发生学以及适宜中间宿主对原头节的细胞反应特点进行了简要讨论。     

细粒棘球绦虫原头节在NIH小鼠体内发育的组织化学研究

本文报道了细粒棘球绦虫原头节在NIH小鼠体内发育过程的组织化学变化,结果如下: 一、糖原:感染后1～2周,原头节体内糖原含量明显减少或消失;4周～6个月,生发层内出现糖原并陆续增加。 二、核酸及蛋白质:感染后1～4周,原头节体内的核酸及蛋白质含量均有逐渐减少的趋势;2～3个月时,生发细胞内的上述生化物质含量仍然较少;感染后4～6个月,凡在芽状增殖的生发层内,其聚集生发细胞中均含有相当丰富的核酸及蛋白质成分。 三、AKP、ATP及ACP酶感染后1～2周,原头节体内AKP及ATP酶的活力逐渐减弱;但在4周～6个月,生发层内的酶活力有所增强;原头节皮层内的ACP活力则在感染后1～2周才开始显现;但4周后活力减弱,感染后2～6个月,生发层内ACP的活力又逐渐增强。 对细粒棘球绦虫不同时期幼虫的上述生化物质动态变化的生理意义,进行了扼要的讨论。     

棘球蚴感染中的免疫逃避相关分子

棘球蚴可以依赖有效的免疫逃避机制在宿主体内长期生存,并造成慢性感染。包囊囊壁、原头节和囊液等所含抗原物质多而复杂,在逃避宿主的免疫应答中起重要作用。棘球蚴逃避宿主免疫反应机制分为主动免疫逃避与免疫调节两种:棘球蚴通过抑制补体的激活,耗竭特异性抗体,影响T细胞活性,下调B细胞功能等有效地避开宿主的免疫反应。棘球蚴的EgTeg蛋白和囊液中的抗原B等能明显抑制中性粒细胞的趋化作用;刺激宿主产生TH2型细胞因子,激发非保护性的免疫应答;干扰单核细胞分化和调节树突状细胞的表型,逃逸宿主免疫监视。     

大鼠感染泡球蚴后外周血及脾脏中滤泡辅助T淋巴细胞的变化

目的分析泡球蚴感染宿主后外周血和脾脏中滤泡辅助T淋巴细胞(Tfh)水平与包虫病进展的关系。方法 20只SD大鼠随机分为正常对照组和模型组,每组10只,模型组开腹直视下在右肝接种约2000个原头节,对照组不做任何处理,3个月后麻醉处死SD大鼠,取外周血和脾脏细胞,以CD4~+CXCR5~+PD1~+为Tfh细胞的标记,使用流式细胞术检测外周血和脾脏中Tfh细胞的水平。使用t检验比较两组之间Tfh细胞的差异。结果大鼠感染泡球蚴3个月后,肝脏可见明显病灶,HE染色显示病灶中可见原头节。泡球蚴感染模型组外周血中CD4~+CXCR5~+PD1~+Tfh细胞在CD4~+细胞中的平均占比为25.63%±3.47%,正常对照组为11.12%±2.94%,2组比较差异有统计学意义(t=10.230,P 0.001),模型组CD4~+CXCR5~+PD1~+Tfh细胞在所有细胞中所占的比例较正常对照组下降显著(0.08%±0.02%vs 0.18%±0.05%,t=5.520,P 0.001);在所有细胞中,模型组脾脏中CD4~+CXCR5~+PD1~+Tfh细胞所占的比例(3.00%±0.42%)显著低于正常对照组(5.30%±1.40%)(t=4.769,P 0.001)。结论外周血中Tfh细胞与包虫病进展密切相关,有望成为反应泡球蚴感染的指标。     

棘球蚴病与囊尾蚴病免疫诊断中交叉反应的动物实验观察

在包虫病和囊虫病免疫诊断中普遍存在交叉反应现象。由于病人来源差异,对较全面了解其机理带来了一定限制。为此我们分别用绵羊细粒棘球蚴囊液(EgCF)、原头节(EgPS)和猪囊尾蚴囊液(CcCF)、头节及囊壁(ccSCW)免疫小白鼠,一个月后用 EgCF 和 CcCF。致敏的血球测其抗体水平及交叉反应情况,现将结果报告如下。     

细粒棘球蚴在NIH小鼠体内发育的组织学及组织化学的进一步观察

本文继续观察感染后6～24个月的细粒棘球蚴在NIH小鼠体内发育的组织学及组织化学变化。结果表明,在鼠体内出现育囊的时间为感染后7～8个月,囊液内见到游离原头节及子囊的时间分别为8及10个月,并发现细粒棘蚴体内的糖原、DNA、RNA、碱性蛋白质的含量,AKP、ACP及ATP酶的活力,均以生发层的芽状突起部分及原头节内的较丰富和较强。     

旋毛虫感染病兔抗体动态的研究

以旋毛虫成囊期幼虫的盐水浸出液为抗原，用ＥＬＩＳＡ方法检测人工感染旋毛虫病兔ＩｇＧ抗体的反应动态。结果表明：１，特异性抗体水平与感染度呈正相关且重感染组的特异性抗体比轻感染组早３ｄ检出。２．抗体水平随感染时间的延长而增高，从感染后７７ｄ起明显下降但可持续４个月之久。     

旋毛虫感染病兔抗体动态的研究

以旋毛虫成囊期幼虫的盐水浸出液为抗原，用ＥＬＩＳＡ方法检测人工感染旋毛虫病兔ＩｇＧ抗体的反应动态。结果表明：１．特异性抗体水平与感染度呈正相关且重感染组的特异性抗体比轻感染组早３ｄ检出。２．抗体水平随感染时间的延长而增高，从感染后７７ｄ起明显下降但可持续４个月之久。     

水牛体内日本血吸虫成虫存活与抗体水平变化关系的观察

在非血吸虫病疫区选取22头水牛,实验感染血吸虫尾蚴后不作任何处理,观察减虫率至4年半达97.4％,至5年为100％;抗体水平感染后3～6个月达高峰,1年后开始下降,7～8年才全部转阴.每g肝脏虫卵数感染后2个月为496个,至感染后8年仅为0.2个.直肠粘膜虫卵数感染后2个月4,300个,感染后7年半未发现虫卵.感染后6～7年肝脏和直肠粘膜检查全部为变黑死卵.实验结果表明COPT环沉率在5％以上,IHA滴度在1:10以上提示水牛体内有活虫的可能,抗体水平下降和治疗关系不明显,主要与宿主体内虫卵数的减少有关.     

人体细粒棘球蚴接种小鼠腹腔的早期发育的观察

人体肺或肝内棘球蚴的生发囊或原头节接种杂交小鼠腹腔,最初可见游离的原头节周围有大量中性细胞及少数嗜酸粒细胞,原头节结构模糊,生发囊外有明显的细胞反应带。存活原头节的结构多集聚于一点,并残留少数顶突钩。这种原头节可向不同方向形成有角质层外被的突起,突起可脱离原头节,形成有角质层的新个体,但未见内侧有生发层。根据实验所见,作者认为:     

Cytokine response and outcome of infection depends on the infective dose of parasites in experimental infection by Echinococcus granulosus

We here analysed whether the cytokine responses in early and late experimental infection with Echinococcus granulosus depend on the dose of parasites to which the host is exposed. To this purpose Balb/c mice were inoculated intraperitoneally (i.p.) with either 500 or 2000 protoscoleces. Splenocytes of mice were obtained at days 3, 7, 14 and 21 and also on week 37 post-infection and cultured in vitro with protoscolex antigens. Type-1 and type-2 cytokines were analysed in supernatants by ELISA. Results showed that the inoculation of 500 protoscoleces induced an early type-0 and a late type-2 cytokine response, whereas the inoculation of 2000 protoscoleces induced an early type-2 and a late type-0 cytokine response. Parasite growth was lower in the group inoculated with the low infective dose. These results indicate that the cytokine response during the infection by the helminth E. granulosus depends on the dose of parasites to which the host has been exposed.     

Induction and maintenance of immune effector cells in the gastric tissue of mice orally immunized to Helicobacter pylori requires salivary glands

BACKGROUND & AIMS: Helicobactor pylori mostly colonizes the gastric mucus that contains salivary antibodies. We studied the role of saliva in the induction and maintenance of gastric immunity conferred by oral vaccination against H. pylori. METHODS: C57BL/6 mice underwent a sialoadenectomy before and after intragastric immunization using whole-cell sonicates of H. pylori and cholera toxin as an adjuvant. At 1 and 6 months after oral inoculation, we assessed the density of the H. pylori colonizing the stomach, specific antibodies in gastric secretion and sera, and the constituents of cellular infiltrates in the tissue. RESULTS: A sialoadenectomy before, but not after, immunization abrogated protection by the vaccination at 1 month after inoculation. Protected mice had more neutrophils, plasma cells, and lymphocytes, but fewer eosinophils, in the gastric tissue than nonprotected mice. Protected mice had a greater increase of immunoglobulin (Ig) G1 specific to H. pylori than IgG2a in sera. At 6 months after inoculation, oral immunization was less effective in mice who had a sialoadenectomy than in control immunized mice. The antibody titers in both gastric secretion and in sera did not correlate with the density of bacteria colonizing the stomach. CONCLUSIONS: It is suggested that, in intragastric immunization against H. pylori, saliva is necessary for both the induction and maintenance of optimal immunity in the stomach. Effective immunity was associated with an increased number of neutrophils and lymphocytes in gastric tissue.     

Induction of protective immunity for Bordetella pertussis by nasal inoculation of pertussis vaccine in mice

Although nasal vaccination has emerged as an interesting alternative to intramuscular or oral vaccination, knowledge is scarce about the immune responses after such immunization. In the present study, we inoculated purified Pertussis Toxin (PT) and Filamentous Haemagglutinin (FHA) with or without adjuvant (kayexalate), or Diphtheria acellular Pertussis Tetanus (DaPT) combined vaccine to mice intranasally three times every four weeks to investigate the references of the immunoresponses between nasal and intramuscular vaccination. The levels of pertussis specific serum IgG antibodies (Abs) and secretory IgA Abs in the nasal wash were measured by ELISA, and cytotoxic T cell activities were examined by proliferative response, and compared with the result from intramuscular inoculation. We also studied the efficacy of adjuvant in the nasal vaccination. The intramuscular inoculation of pertussis vaccine induced serum IgG antibodies and cellular immunity against PT and FHA, but did not induce local IgA antibodies. On the other hand, the nasal inoculation induced both serum and local antibody responses. Moreover, it also induced significant cellular immunity to pertussis antigen. In nasal vaccination, the inoculation with adjuvant was superior to inoculation without adjuvant for the induction of both humoral and cellular immunity.     

Antibody response of Echinococcus granulosus infected mice: Protoscolex specific response during infection is associated with decreasing specific IgG1/IgG3 ratio as well as decreasing avidity

The antibody response was followed during 68 weeks in 17 Balb/c mice intraperitoneally (i.p.) infected with Echinococcus granulosus protoscoleces (PSC) and in three mice i.p. immunized with dead PSC. Titres of antibodies recognizing peptidic and glucidic PSC epitopes, as well as their isotypic and avidity profiles were followed by ELISA. In addition, antigen recognition patterns were analysed by immunoblot. The response against carbohydrate epitopes was dominant in infected and immunized mice but stronger in the first group. Infected mice showed similar profiles of specific IgG and IgM with maximum titres from week 38 to 53. Although IgG1 and IgG3 were the predominant antibody subclasses, the ratio of IgG1/IgG3 antibody titres as well as antibody avidity decreased during the experiment, encompassing a decrease in recognition of peptidic epitopes. Immunized mice did not show significant levels of specific IgM and, after week 15, showed IgG titres lower than the infected mice. IgG1 was the predominant IgG subclass during all the experiment with background levels of IgG3. The mean Ab avidity was high and showed no significant changes during immunization. Different patterns of response were thus produced by dead and developing live parasites. Although high avidity IgG1 antibodies were early found in both cases, lower avidity IgG3 antibodies were increasingly produced afterwards only in infected animals. The isotype switch and avidity decrease observed only during infection are consistent with a possible parasitic mechanism to evade host immunity .     

抗疟治疗对宿主保护性免疫影响的实验研究

目的探讨药物治疗对宿主保护性免疫建立的影响。方法用不同剂量氯喹或青蒿琥酯治疗约氏疟原虫(非致死型)感染的BALB/c小鼠。待自愈组清除疟原虫后30d,用约氏疟原虫(非致死型和致死型)以及伯氏疟原虫再次感染。以吉姆萨薄血膜染色法观察小鼠虫体血症水平,采用ELISA法检测脾细胞培养上清中IFN-γ水平和血清中特异性抗体水平。结果自愈组和各治疗组小鼠于初次感染早期均产生高水平IFN-γ,随之抗约氏疟原虫(非致死型)特异性IgG抗体水平显著升高,且无明显差异。对约氏疟原虫(非致死型和致死型)再次攻击小鼠均呈完全抵抗,少数出现低水平一过性虫体血症。而异种疟原虫攻击时小鼠全部感染并死亡。结论氯喹或青蒿琥酯不同剂量治疗不影响小鼠保护性免疫的建立和免疫记忆的维持,特异性IgG抗体是宿主抵御再感染主要的免疫效应分子。     

复合鼠疫疫苗F1+ rV的免疫学研究

目的通过由提取的鼠疫F1抗原和重组鼠疫V抗原免疫小鼠,评价鼠疫亚单位疫苗的免疫效果,为鼠疫疫苗研制奠定基础。方法将60只小鼠随机分为3组实验组,免疫后不同时间点进行血清ELISA检测、ELISPOT、MTT 细胞增殖以及流式细胞分析。结果ELISA检测结果显示F1+rV+Al(OH)3组有较强的体液免疫应答;ELISPOT、MTT 细胞增殖结果显示F1+rV+Al(OH)3组在不同时间点经F1、rV抗原刺激后分泌IFN γ的脾脏淋巴细胞数均高于其余各组。结论鼠疫双组分疫苗能引起小鼠明显的体液免疫和细胞免疫,Al(OH)3佐剂能够显著提高其应答水平。     

EB病毒融合基因Z2A重组BCG的免疫学特性研究

目的对EB病毒融合基因Z2A构建的重组BCG进行鉴定及免疫学研究。方法采用Westernblot方法检测重组BCG中Z2A基因的表达;用重组BCG免疫小鼠,用ELIsA方法检测小鼠血清特异性抗体水平,采用乳酸脱氢酶法检测小鼠的细胞免疫水平,评价重组BCG的免疫效果。结果重组BCG表达的蛋白能被血清BZLFl抗体（或LMP2A抗体）识别;ELISA检测表明,重组BCG能刺激机体产生抗体,当重组BCG细菌数为5×10-8个／只时,抗体滴度最高为17600（186．5±6．7Pg／ml）;BCG对照组及PBS对照组对GT39细胞的杀伤率分别为（32．5±2．7）％和（22．8±2．3）％,重组BCG组为（62．7±6．2）％,差异有统计学意义（P〈0．05）。结论EB病毒融合基因Z2A重组BCG免疫效果良好,能刺激小鼠产生体液免疫和细胞免疫反应。     

小鼠感染泡球蚴后细胞因子水平的变化

目的　观察人工接种泡球蚴(EM)昆明小鼠体内6种细胞因子水平的动态变化,研究泡球蚴病的免疫学机制。　方法　实验组和对照组小鼠分别腹腔接种泡球蚴及生理盐水,持续观察260d,收集各组血清用ELISA法检测血清中白细胞介素2(IL2)、γ干扰素 (IFNγ)、肿瘤坏死因子α(TNFα)、IL4、IL5、IL10水平。　结果　实验组6种细胞因子水平始终高于正常对照组,其中辅助性T细胞1(Th1)类细胞因子IL2水平在感染后 80d达到峰值,感染140d后迅速降低;TNFα水平在感染后40d较对照组明显上升,感染100d左右达到峰值,140d后迅速下降;IFNγ在感染80d后达到峰值,140d后缓慢下降;而在80d以前,辅助性T细胞2(Th2)类细胞因子IL4、IL5、IL10维持在较低水平;100d后,这3类细胞因子明显上升,其中IL4、IL10水平在100d达到峰值,IL5水平在140d达到峰值,以后维持在较高水平。　结论　Th2细胞介导的体液免疫与Th1细胞介导的细胞免疫共同参与了宿主抗棘球蚴免疫。感染早期以Th1细胞介导的细胞免疫应答为主,感染中晚期转化为以Th2细胞介导的体液免疫为主。     

广州管圆线虫感染小鼠血清细胞因子及IgG亚类的动态观察

目的观察小鼠感染广州管圆线虫后机体免疫的动态变化。方法分别采集感染前、感染后第1、3、7、18d的小鼠血清,用ELISA方法检测血清中细胞因子IL-2、IL-4以及特异性免疫球蛋白G（IgG）亚类水平。结果广州管圆线虫感染的小鼠血清中IL-2的水平较感染前呈逐渐下降趋势,IL-4的水平与感染前小鼠相比先下降后呈升高趋势。抗体IgG1的水平与感染前小鼠相比明显升高,IgG2a的水平与感染前小鼠相比未见明显变化。结论小鼠Th1型免疫应答较弱,Th2型免疫应答增强。表明小鼠感染广州管圆线虫后机体细胞免疫较弱,体液免疫较强。