树Qu实验感染后血清中抗基孔肯雅病毒IgM和IgG抗体检查

成年树Ｑｕ经人工感染基孔肯雅病毒后，能产生２－６天的病毒血症。感染后第６天能产生特异性ＩｇＭ抗体，第１４－２１天为高峰，以后逐渐下降，感染后第１２天，ＩｇＧ抗体开始出现，３０－６０天为高峰，并持续不降，表明树Ｑｕ对基孔肯雅病毒敏感。     

白纹伊蚊经实验感染后涎腺中基孔肯雅病毒和乙型脑炎病毒抗原检测

用基孔肯雅和乙型脑炎病毒经口感染白纹伊蚊，采用免疫荧光试验对雌蚊涎腺作病毒抗原检查，结果于感染后第８￣１４天基孔肯雅的阳性为５０％￣９１．６７％，乙型脑炎为７０％￣８０％。试验认为，白纹伊蚊在感染后第６￣８天即可传播基孔肯雅和乙型脑炎病毒。     

严重急性呼吸综合征患者外周血病毒载量和抗体水平动态监测

目的 动态监测严重急性呼吸综合征(SARS)患者外周血SARS病毒载量和抗体含量变化。方法 选择3例因同一传染源感染的SARS患者为研究对象，应用实时荧光定量逆转录，聚合酶链反应(RT—PCR)检测患者外周血PBMCs中病毒载量，用酶联免疫检测技术检测血清SARS病毒IgG、IgM含量。结果 3例患者分别在出现症状后第3～11日、第1～13日、第1～9日检测到SARS冠状病毒核酸，第5～7日病毒载量达到高峰。IgM抗体在患者起病后第7～9日即可被检测到，IgG抗体在患者起病后第7～14日即可被检测到，高峰在第10～20日。结论 SARS病毒载量存在自限性消长过程。SARS抗体IgG、IgM均较早出现，IgG维持时间比IgM长。     

肾综合征出血热病毒感染野兔的实验研究

捕自湖北的5只草兔(Lepus capensis)和2只华南兔即短耳兔(Lepus sinensis),经肌肉、眼睑双途径人工感染HFRS(肾综合征出血热)病毒后,其血中在感染后第8天即可查到HFRS IgG抗体,并于16～20天达到高峰;血浆和血白细胞中HFRS抗原分别在感染后第3～7天和6～11天检出;尿和粪便中特异性抗原分别在第7～20天和8～28天检出,但口腔洗漱液中则在感染40天内未检出阳性。用抗原阳性的粪、尿标本接种乳小白鼠,于第一代就发生感染。感染野兔的脾、肾、大肠和小肠可检出特异性抗原,尤以脾和小肠多见。     

中国首例输入性基孔肯雅热临床分析

目的 了解2008年我国首例输入性基孔肯雅热病例的流行病学、临床、病原学特点及预后转归.方法 对1例成年男性患者流行病学及临床资料进行回顾性分析,并采用ELISA和胶体合法检测患者血清基孔肯雅病毒IgM(CHIKV-IgM)抗体,实时荧光PCR法检测基孔肯雅病毒核酸(CHIKV RNA).结果 该患者临床表现为发热起病,有头痛、全身肌肉及关节酸痛,全身充血性斑丘疹,WBC及PLT减少.发病第9天CHIKV-IgM阳性;第3、5,7、9天CHIKV RNA阳性;RT-PCR扩增产物为575个核苷酸碱基片段,经序列测序结果与基孔肯雅病毒核酸序列同源性高达99%.患者经对症支持等治疗痊愈出院.结论 该患者为我国首例输入性基孔肯雅热确诊病例,本病临床表现与登革热相似.     

2型登革病毒临床分离株感染Balb/c小鼠后Th2类主要细胞因子的动态变化

目的观察3种细胞因子在登革病毒(DEN)_2两次感染过程中的整个消涨动态,以探讨DEN_2临床分离株感染Balb/c小鼠后其血浆中白介素(IL)-2、IL-5及IL-6产生及相关关系。方法用不同量DEN_2临床分离株经皮下多点注射,建立Balb/c小鼠感染模型,并用双抗体夹心酶联免疫吸附实验(ELISA)检测感染后不同时间各组动物血浆中IL-2、IL-5及IL-6浓度,观察其产生动态。结果各组实验动物初次感染B株后,各组IL-2、IL-5和IL-6水平较正常组均无明显增加;再次感染后均有显著增高。IL-2水平于再次感染后第4天(初次感染后第20天)达高峰,各组均值为(101522．44±10465．375)pg/ml,其后逐渐下降;再次感染后,Balb/c小鼠体内IL-5水平第1、2组于第1天(初次感染后第16天)达高峰,较正常组差异有统计学意义(X~2=35．65,P＜0．05);再次感染后各组小鼠IL-6水平于第1～2天达高峰,其中第3组峰值最高,且维持高水平时间较长,并与正常饲养对照比较差异有统计学意义(X~2=32．86,P＜0．05)。结论DEN_2临床分离株再次感染可使Balb/c小鼠产生IL-2、IL-5及IL-6增加,在再次感染阶段Th2应答发挥重要作用。     

汉坦病毒气溶胶感染实验动物的特征和抗体反应

观察汉坦病毒气溶胶传播感染和受试动物抗体反应的特征。方法我们用感染的黑线姬鼠排泄物形成的自然病毒气溶胶和人工发生的病毒气溶胶进行实验。结果离乳小鼠和乳小鼠在自然病毒气溶胶中暴露10天，血清中荧光抗体效价分别为1：80和1:20，到第25天时可达1：16；黑线姬鼠在暴露15天时，血清中荧光抗体效价为1：20，暴露35天仅为1：40。在人工病毒气溶胶感染乳小鼠的实验中，感染后的第12天，血清中ELISA抗体效价为1：10，第21天达到1：160。结论结果表明这三种实验动物对汉坦病毒气溶胶感染均敏感，而且，汉坦病毒气溶胶浓度高低对感染没有明显地影响。随着鼠龄的增大，对病毒感染的抵抗力也增大。这为研究汉坦病毒的气溶胶免疫提供了科学依据。     

抗体捕获ELISA检测IgM抗体在新疆出血热病人早期诊断中的应用

采用抗人μ链抗体包被,捕获病人血清标本中的IgM抗体,再加已知的特异性病毒抗原与IgM抗体结合,用抗新疆出血热病毒单克隆抗体酶结合物与病毒抗原结合病人血清中IgM抗体的方法,测定急性期患者特异性IgM抗体,探讨其早期诊断的价值。对8例发病后不同时期的XHF患者进行检测的结果,发病第5天即可检出IgM抗体且滴度已达较高水平,于第14天达高峰后开始下降,但阳性可持续70天以上。而IgG抗体对照检测显示虽然在1/3急性期病人发病第6天即可检出,但滴度很低,第14日后迅速升高,至第70天仍然保持很高滴度。实验结果表明用本法检测IgM抗体进行早期诊断的意义可与病毒分离及特异性病毒抗原的检测结果相比。为XHF的早期诊断提供了一种特异性和敏感性高,并且稳定可靠的诊断工具。     

猴疟感染过程特异性IgM抗体的变化

以食蟹猴疟原虫猕猴动物模型,用间接荧光抗体试验法(IFAT)观察疟疾感染过程中特异性IgM抗体的变化,共观察无复燃或复发的短原虫血症期(15天内,5只猴)、长原虫血症期(26～37天,2只猴)及出现复燃(5只猴)3种类型的疟疾感染。疟疾IgM抗体初次出现于子孢子感染后13～16天或阳性血感染后3～9天,抗体滴度高峰出现于子孢子感染后17～25天或阳性血感染后12～26天。尔后抗体逐渐降低,抗体消失时间在原虫血症短者为感染后2个月,在原虫血症长者约3个月,复燃出现在感染终止后约1个月。结果表明,疟疾特异性IgM抗体的出现与存留,是新近疟疾感染的一个佐证。     

小鼠实验感染不同种旋毛虫后血清IgG抗体水平的变化

目的观察小鼠感染不同种旋毛虫后血清IgG抗体水平的变化。方法将50只小鼠随机分成5组（每组10只），分别感染旋毛虫（T1）、乡土旋毛虫（T2）、布氏旋毛虫（T3）、伪旋毛虫（T4）及纳氏旋毛虫（T7），每只感染300条幼虫，感染后1～6周每周尾部静脉采血，6周后每2周尾部静脉采血，至感染后20周，用旋毛虫肌幼虫ES抗原ELISA检测血清中抗旋毛虫IgG抗体水平。结果小鼠感染旋毛虫、乡土旋毛虫、布氏旋毛虫及纳氏旋毛虫后3～5周，血清IgG抗体水平快速升高，至感染后第8周达高峰，此后旋毛虫和乡土旋毛虫感染小鼠的血清IgG抗体水平缓慢下降，布氏旋毛虫及纳氏旋毛虫感染小鼠的血清IgG抗体水平迅速下降；小鼠感染伪旋毛虫后3～5周血清IgG抗体水平快速升高，至第16周达高峰，之后缓慢下降。5种旋毛虫感染小鼠的血清IgG抗体水平差异具有显著性（P〈0．05）。结论5种旋毛虫感染小鼠的血清IgG抗体水平和动态变化不同；旋毛虫肌幼虫ES抗原可用于其他4种旋毛虫（乡土旋毛虫、布氏旋毛虫、伪旋毛虫、纳氏旋毛虫）感染的血清学诊断及流行病学调查。     

Induction of indoleamine 2,3-dioxygenase in mouse lung during virus infection.

Indoleamine 2,3-dioxygenase [indoleamine: oxygen 2,3-oxidoreductase (decyclizing)] activity in the supernatant fraction (30,000 X g, 30 min) of mouse lung homogenate increased approximately 120-fold after infection with PR8 influenza virus. Both specific and total enzyme activities started to increase linearly from the 5th day after infection, reached the highest level around the 11th day, and then gradually decreased to normal values in about 3 weeks. Other enzymes in the lung, such as certain lysosomal enzymes and monoamine oxidase, did not change significantly throughout the experiments. The time course of the increase in the enzyme activity was quite different from that of virus replication in the lung (a peak by the 3rd day and persistence until the 9th day) or that of serum antibody content (started to rise on the 9th day). Rather, it appeared to be closely related to the infiltrations of mononuclear and lymphocytic cells. When mice were exposed to a higher dose of virus and did not recuperate, the time course of the increase of the enzyme activity was essentially identical to that seen with a low concentration of virus. A maximum stimulation of the enzyme activity in the lung occurred on the 9th day after infection; the increase was approximately 100-fold. However, serum antibody content was slight and virus titer in the lung remained high.     

Recently introduced Aedes albopictus in Corsica is competent to Chikungunya virus and in a lesser extent to dengue virus

Aedes albopictus has been established in Europe for some decades rendering temperate countries vulnerable to tropical diseases. The Italian chikungunya (CHIK) outbreak in the summer of 2007 demonstrated that indigenous transmission of CHIK was possible in Europe. To estimate the risk of a CHIK outbreak in Corsica, we assessed the vector competence of A. albopictus established in the island since 2006 towards a CHIK variant (E1-A226V). A dengue serotype 2 virus was also tested. Experimental infections showed that A. albopictus was highly competent to CHIK virus (disseminated infection rates ranging from 75% to 100%) and to a lesser extent, to dengue virus (12.5–68.8%). Moreover, A. albopictus ensured a high level of viral replication and was able to transmit the virus as early as 2 days after ingestion of infected blood with around 1 000 viral RNA available in salivary glands. The risk for a local transmission of CHIK is thus likely in Corsica, if other parameters determining the vector capacity of A. albopictus are suitable.     

不同途径感染弓形虫小鼠在脑内形成包囊的研究

目的用3种不同途径感染Fukaya株弓形虫速殖子,观察弓形虫感染小鼠慢性期病变特点及虫体在脑内的成囊过程,以期寻求一个稳定、易建的慢性感染动物模型,为弓形虫病的病理诊断提供依据,并对弓形虫的致病机理加深理解。方法弓形虫Fukaya株速殖子(5×104个)分别以腹腔、皮下和口服途径感染小鼠,给适量药物使之形成慢性感染,并与不给药组做对照,于感染后第3d、6d1、4d2、1d、28d、42d和90d,取脑进行间接免疫酶染色,统计脑内虫体数量及形成包囊情况。结果不给药组第3d、6d与给药组第6d,3种感染途径的腹腔含虫数比较:均为腹腔感染>皮下感染>经口感染。比较小鼠脑内虫体分布:腹腔感染组,第28d的脑内虫体达到高峰,此时脑内包囊最多。皮下感染组,第21d的脑内虫体最多,脑内包囊也多。经口感染组,第21d脑内虫体最多,但脑内偶见包囊。经口感染Fukaya株速殖子似不易成囊。经腹腔、皮下和口服感染虫体的小鼠存活率在第42d分别为100%、66%、80%。结论弓形虫感染慢性期小鼠脑多被累及。腹腔与皮下感染弓形虫Fukaya株速殖子比口服易成囊,经腹腔感染方式建立弓形虫慢性感染动物模型较稳定。     

柯萨奇病毒性心肌炎小鼠的自然病程观察

目的以组织病理学观察CVB3感染鼠的自然病程及此过程中IL-2、IL-10的变化。方法对昆明种小鼠采用CVB3腹腔注射染毒后不同时间观察心、肝、脾、肾的病理改变及检测血清中IL-2和IL-10。结果心肌炎小鼠的各脏器出现不同程度的病理改变,IL-2I、L-10随病程的发展呈现出一定的变化:即接种CVB3后第4天IL-2水平明显低于正常均值,随着病程的发展IL-2水平呈升高趋势,第9-14天IL-2水平明显高于正常值,第10天IL-2水平达高峰,第18天IL-2水平接近正常;随着病程的发展IL-10水平逐渐下降,第9-14天IL-10水平明显低于正常均值,此时心肌病变程度加重。结论病毒性心肌炎是一类全身感染性疾病,心外器官病变不容忽视,IL-2和IL-10在此过程中起不同的作用。     

Antibody responses in Japanese volunteers after immunization with yellow fever vaccine

To monitor the development of specific and cross-reactive antibody response in twenty Japanese volunteers after vaccination with live yellow fever vaccine. Serum samples were collected on various days after vaccination and examined for hemagglutination inhibition (HI) antibodies against yellow fever virus (YFV), Japanese encephalitis virus (JEV) and dengue virus (DV), neutralizing antibodies against YFV and JEV, and IgM antibodies against YFV. None of the volunteers had been previously immunized with this vaccine. Fifteen of 20 had pre-vaccinated with JEV 7 to 40 years before. Ten of the 20 had neutralizing antibodies against JEV before immunization. None of the 20 had detectable antibodies against YFV or DV before vaccination. On day 10th after the vaccination, neutralizing antibodies to YFV were detected in 6 of 19 volunteers and IgM antibodies against YFV were detected in 7 of 19. On day 14th, HI, neutralizing, and IgM antibodies against YFV were detected in all the tested sera. Neutralizing antibodies against JEV were developed in 2 volunteers and HI antibodies against JEV were increased in 3 of 6 volunteers respectively. On day 29th, cross-reactive HI antibodies for JEV and DV were detected in all the tested sera. The results indicate that YF vaccine induces YFV-specific antibodies in all the tested volunteers and that it also induces HI antibodies cross-reactive for JEV and DV. The YF vaccine has a strong immunogenicity because it is a live vaccine, and induces antibody against YFV predominantly. The international certificate of yellow fever vaccination becomes valid 10 days after vaccination. On day 14th after vaccination, we detected neutralizing antibodies against YFV from all tested volunteers, however, only 6 of 19 volunteers had detectable neutralizing antibody on the 10th day after vaccination. Therefore, the vaccine may not be perfectly effective on day 10th after the vaccination.     

Variation among goegraphic strains of Aedes albopictus in susceptibility to infection with chikungunya virus.

Two types of variation were observed when the susceptibility of 16 different geographic strains of Aedes albopictus to oral infection with chikungunya (CHIK) virus was studied. One was differential susceptibility to infection with the virus. The other was variation in the quantity of virus present in infected mosquitoes after a standard incubation period. Mean virus titers of infected mosquitoes of different geographic strains varied almost 1,000-fold. Attempts to develop increasingly resistant or susceptible mosquito lines through genetic selection were unsuccessful. Infection rates did not change significantly despite 3 to 6 generations of selective inbreeding. In contrast, crosses between strains of high and low CHIK susceptibility yielded hybrid mosquitoes with infection rates and mean virus titers intermediate between those of the parent colonies. These data suggest that at least one factor controlling the susceptibility of A. albopictus to CHIK infection is genetic. Two lines of A. albopictus with a marked difference in susceptibility to oral infection with dengue viruses were equally susceptible to oral infection with chikungunya virus.     

Immune response against Chlamydia trachomatis inoculation in mice--changes in interferon activity, antibody titers and weight of the spleen as parameters of infection

We examined the immune response against Chlamydia trachomatis (serovar L2) inoculation in mice by measuring the serum interferon (IFN) level, 2'-5'A synthetase (2-5A(S] activity, antibody titers (IgM, IgG) and the spleen weight as parameters of infection. The interferon activity was detected 6 hrs (400 U/ml) and the activity peaked 12 hrs (450 U/ml) after inoculation, and then gradually decreased thereafter (24 hrs: 12 U/ml, 36 hrs: undetectable). It was found that Chlamydia trachomatis induces IFN as well as bacteria. To monitor the behavior of IFN action after serum IFN was cleared, 2-5A(S) activity in the spleen cell extract was measured. It was found that the activity reached its peak 1 to 2 days after inoculation and then decreased as well as in infectivity of Chlamydia trachomatis. The activity however was almost not detected in sera of mice after inoculation of heat-inactivated Chlamydial organism (56 degrees C, 10 min). This may indicate that intact Chlamydial organisms were required for induction of IFN. IFN induced in mice was stable in pH 2.0 treatment and IFN induced by Newcastle disease virus inhibited growth of Chlamydia trachomatis in L929 cell cultures in a dose-dependent manner. The weight of the spleen gradually increased and reached its peak (2- to 3-fold of the control) in 3 to 5 days after inoculation, and then gradually decreased to the control level. IgM and IgG antibodies to Chlamydia trachomatis were detected by immunofluorescence method and enzyme-linked immunosorbent assay, respectively. The antibody IgM was detected as early as 2 days and reached its peak 3 to 4 days after inoculation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Systemic and intestinal immune responses to HIV-2287 infection in Macaca nemestrina

Nonhuman primate models of human AIDS have been used successfully to evaluate candidate vaccines and infection intervention therapies. Successes of pathogenicity studies in primate models have been limited because of the varied infection outcomes and characteristic low number of study animals. The acutely pathogenic HIV-2(287)--Macaca nemestrina model has shown promise both in antiviral drug evaluation and in pathogenicity studies. Here we describe virus replication, spread, and host responses during the first 28 days of HIV-2(287) infection. Focusing on 18 macaques from a larger 27-macaque study, we report changing virus loads, CD4(+) cell depletions, and antibody responses both systemically and in the mucosa of the small intestine. After intravenous inoculation, blood and intestinal tissue were collected from pairs of macaques at 12 hr and 1, 2, 4, 6, 10, 14, 21, and 28 days postinfection. Specimens were examined for evidence of infection by quantitative cultures, in situ hybridization, lymphocyte subset monitoring, and antibody production. The data were presented serially as though all samples were collected from a single macaque. The highest blood virus loads were detected between days 10 and 14 and subsequently decreased through day 28. This coincided with a significant increase in ileum mucosa virus loads on day 10, which became undetectable by day 28. The lowest levels of CD4(+) cells were observed on days 21 and 28 in blood and ileum mucosa. CD4(+):CD8(+) cell ratios in blood and ileum dropped dramatically after day 10 to lowest levels by day 28. Intestinal virus loads were inversely correlated with CD4(+) cell and virus-specific antibody levels in the ileum after day 6. These results underscore the suitability of this model for pathogenicity studies as well as the importance of the intestinal lymphoid tissues as an initial site of virus replication and cell destruction during the acute, asymptomatic stage of AIDS development.     

神经生长因子对脑缺血再灌注大鼠突触素表达的影响

为研究大鼠脑缺血再灌注后突触素p38表达的变化规律及其与神经生长因子的关系,取大鼠72只,随机分为假手术组,人工脑脊液组和神经生长因子治疗组,采用线栓法建立大脑中动脉缺血再灌注模型,应用免疫组织化学方法观察P38的表达,结果发现,脑缺血再灌注3天后P38表达增高,第7天达高峰,之后逐渐降低,第21天降至对照组水平,应用外源性神经生长因子后,P38表达较对照组无明显升高（P＞0．05）,提示中枢神经系统损伤后,神经元具有再生和修复的可塑性,外源性神经生长因子对P38的调节有待于进一步研究。