猴疟原虫感染后免疫功能的变化

<正> 猴疟原虫感染后其免疫功能处于全面低下和紊乱状态,本文对猴接种疟原虫后的红细胞免疫、血小板免疫、淋巴细胞免疫反应进行了较全面的动态观察,现将资料报道如下。 研究对象和方法 一、研究对象:体重3kg左右恒河猴6只,每只血传接种食蟹猴疟原虫B株14万～54万个,在感染前及感染后9天、22天计数原虫,计算方法是制成白细胞厚片(红细胞已破坏处理),显微镜下计     

血传接种食蟹猴疟原虫B株后获好的感染蚊的血餐时间探讨

５只恒河猴血传接种食蟹猴疟原虫Ｂ株后，在原虫血症显著期（原虫密度≥２‰）供１１０批大劣按蚊血餐（１批／日），共获最佳感染蚊５２批。较好感染蚊１０批、不佳感染蚊２７批，阴性２１批。在各猴上获得好的（最佳和较好）感染蚊的批数和时间与其原虫血症消长类型（可分３型）有关。Ａ型（初峰为最高峰的３峰形）：初峰顶前２～４ｄ为蚊媒最佳或较好感染时间，次２峰顶及其前后１－２ｄ为最佳感染时间。Ｂ型（初峰非最高峰的３峰形）：初峰顶及其前后１－２ｄ为最佳感染时间，次１和欢２峰期各天基本上均为最佳感染时间。Ｃ型（双峰形）：初峰顶及其前１－３ｄ、次峰顶及其前１－３ｄ和峰顶后１ｄ为最佳或较好感染时间。     

甲型肝炎灭活疫苗(SH株)接种食蟹猴的免疫原性研究

目的 研究甲型肝炎灭活疫苗(SH株)食蟹猴的免疫原性.方法 甲型肝炎病毒SH株接种人胚肺二倍体细胞(MRC-5),培养、收获的病毒液,经纯化、灭活后铝佐剂吸附制成甲型肝炎灭活疫苗.采用食蟹猴进行疫苗的免疫原性研究.结果 接种疫苗后的食蟹猴,体内产生抗HAV抗体和中和抗体,抗核抗体结果为阴性.结论该疫苗具有良好的免疫原性.     

抗恶性疟原虫单克隆抗体的分析

在两次细胞融合中,经2—4次克隆化后已建成28株抗恶性疟原虫(FCC—1/HN)的杂交瘤。其中针对裂殖子和分裂体抗原的McAb有9株,仅对裂殖体特异的有4株,对裂殖体和滋养体期原虫产生荧光反应的有15株。 应用间接荧光抗体技术,检查这28株McAb对恶性疟原虫(安徽株),间日疟原虫、食蟹猴疟原虫,诺氏疟原虫、伯氏疟原虫、约氏疟原虫和鸡疟原虫的交叉反应。结果,仅有93D4和94B5属株特异性McAb,其余各株均对恶性疟原虫(安徽株)有明显荧光反应。有5     

Ad5-HIVgag免疫的食蟹猴中腺病毒中和抗体与Gag特异性细胞免疫反应的研究

目的 观察不同剂量的重组腺病毒疫苗Ad5-HIVgag反复肌内注射免疫食蟹猴后,食蟹猴体内产生的腺病毒中和抗体水平及Gag特异性细胞免疫反应水平,初步探讨腺病毒中和抗体的产生对Gag特异性细胞免疫反应的影响.方法 将24只食蟹猴随机分为4组：对照组、低剂量组、中剂量组、高剂量组.每3周免疫1次,连续免疫5次.免疫前及免疫后不同时间点用中和实验检测动物体内腺病毒中和抗体水平,用Elispot方法检测Gag特异性细胞免疫反应水平.结果 初次免疫后3周3个实验组动物都检测到了较高水平的腺病毒中和抗体,在初次免疫后8周达到高峰,恢复期结束时略有下降.初次免疫后5周三个实验组动物都检测到了Gag特异性细胞免疫反应,除初次免疫后12周反应水平有所下降,其他时间点细胞反应呈逐渐增高趋势,但各剂量组间未见明显的量效关系.结论 在一定的剂量范围内反复多次应用Ad5-HIVgag免疫食蟹猴后,可产生针对腺病毒的中和抗体,随着免疫次数的增多Gag特异性细胞免疫反应有增高趋势,Ad5疫苗免疫后中和抗体的产生对Ad5疫苗反复应用诱导的Gag特异性细胞免疫反应抑制作用并不明显.     

高流行区恶性疟患者的抗恶性疟原虫抗体与免疫状态的关系

用戊二醛固定、空气干燥的单层感染红细胞作抗原,以RESA-IFA试验检测恶性疟患者的RESA抗体可反映患者对恶性疟原虫感染的临床免疫状况。患者的RESA抗体滴度与年龄及在疫区居住时间呈正相关(r_s分别为0.531和0.313,P均<0.05);与原虫密度呈负相关(r_s=0.353,P<0.05)。但全虫抗原及环状体抗原检出的抗体滴度与上述指标似无关。     

菲立磁胞内标记食蟹猴骨髓基质细胞

目的 探索利用菲立磁和转染试剂体外磁性标记食蟹猴骨髓基质细胞（BMSCs）方法的可行性。方法 无菌条件下取食蟹猴骨髓,梯度密度离心法分离获取BMSCs,使用菲立磁-多聚赖氨酸复合物（FE-PLL）标记BMSCs,普鲁士蓝染色、电镜和台盼蓝排除实验等方法鉴定FE-PLL标记食蟹猴BMSCs的效率和细胞的活力,倒置相差显微镜和免疫细胞化学方法检测BMSCs的增殖和分化能力。结果 菲立磁可以高效率地标记BMSCs,标记效率在99％左右。光镜和电镜下BMSCs胞质内分别可见细小的蓝色铁颗粒和许多包裹铁颗粒的囊泡。FE-PLL标记对BMSCs的活力、增殖和分化等能力没有明显的影响。结论 菲立磁可以用来体外标记食蟹猴BMSCs。     

艾塞那肽微球在食蟹猴体内的免疫原性及药代动力学比较研究

目的:考察艾塞那肽微球单次皮下注射给予食蟹猴的免疫原性及药代动力学特征,并与市售药Byduren比较。方法:选用16只食蟹猴,雌雄各半,分性别随机分成受试物组和市售品组(n=8),分别单次皮下注射给予受试物/市售品0.44mg·kg-1。于给药前、给药后2、4、6、8、10w采集全血,分离血浆,采用ELISA法分析血浆中抗艾塞那肽抗体产生情况;于给药前(0h)、给药后0.5、1、3、6、12h,1、2、3、5、7、10、14、17、21、24、28、31、35、38、42、45、49、52、56、63、70d采集全血,分离血浆,采用ELISA法分析血浆中艾塞那肽浓度,WinnonLin计算各药代参数。结果:给药后2w受试物组1只动物产生抗药抗体,给药后4w受试物、市售品分别有4只、3只动物抗药抗体产生,至给药后70d,两组各有7只动物产生抗药抗体,且抗体的滴度影响血浆艾塞那肽浓度,滴度大于1:125的动物血浆药物浓度及系统暴露量明显升高。受试物、市售品皮下注射给予食蟹猴后,血药浓度随时间的变化趋势基本一致,给药后的前6h有一个初始释放,此后药物低浓度释放,药物的主要释放在给药后约10d开始,28d左右达峰,至给药后70d两组动物艾塞那肽仍持续释放。结论:单次皮下注射给予食蟹猴后,艾塞那肽微球在两组动物体内具有相似的免疫原性和相似的药代特征。     

恶性疟原虫保护性抗原复合基因——痘苗病毒重组活疫苗株免疫猴血清对恶性疟原虫的体外抑制试验

用重组痘苗病毒、非重组对照痘苗病毒免疫猴血清及正常对照猴血清进行了恶性疟原虫的体外培养抑制试验。结果培养渡中加人重组痘苗病毒免疫猴血清后，疟原虫感染率逐渐下降，第4天后原虫感染率维持在较低水平(0．1％以下)，第12天后血片中没有原虫检出；加入非重组对照痘苗病毒免疫猴血清的培养瓶中疟原虫感染率的变化与正常对照猴血清无明显的差异，说明重组痘苗病毒免疫猴血清对体外培养的恶性疟原虫具有一定抑制作用。     

食蟹猴疟原虫B株子孢子感染后的虫血症及按蚊血餐感染情况

用食蟹猴疟原虫Ｂ株子孢子感染的６只恒河猴，初发期虫血症自然消长曲线２只猴为双峰形、４只猴为多峰形。虫血症显著期（原虫密度≥２‰）最短者为１３ｄ，最长者为３７ｄ。在虫血症显著期供１１３批（１批／日）蚊（大劣按蚊或斯氏按蚊）血餐，获得感染好的蚊虫（卵囊均数＝５－３２７．７，胃感染率＝６５－１００％）５８批、平均为９．７批。感染好的蚊批分布：双峰形的初峰期为１４．２９％（２／１４），次峰期为８５．７１％（１２／１４）；多峰形的初峰、次１和次２峰期分别为２２．７３％（１０／４４）、４５．５％（２０／４４）和３１．８２％（１４／４４）。结果提示对蚊感染性较强的血餐时间主要分布在次１和次２峰期。     

恒河猴实验感染庚型肝炎病毒的实验研究

目的研究庚型肝炎病毒（ＨＧＶ）在恒河猴中的实验感染状态。方法用一名ＨＧＶＲＮＡ阳性、ＨＢＶ、ＨＣＶ均阴性的健康献血员血浆实验感染２只恒河猴，并取第一代猴感染后６周的血再感染１只第二代恒河猴，然后用以第二代猴感染６周后血继续感染２只第三代恒河猴。分别用逆转录聚合酶链反应（ＲＴ－ｎＰＣＲ）检测受感染猴血清中的ＨＧＶＲＮＡ，并每周抽血测定血清中丙氨酸转氨酶（ＡＬＴ）。结果感染１周后猴血清ＨＧＶＲＮＡ阳转，最长持续阳性２８周以上。不同感染个体血清ＡＬＴ水平有明显差异，其中１号猴有短期轻度升高，５号猴血清ＡＬＴ较长时间在１００Ｕ／Ｌ以上。肝活检发现，感染后１６周猴肝组织出现明显的病毒性肝炎样病理改变。进一步对该献血员血浆和感染后猴血清中的ＨＧＶ５’端部分非编码区基因ＰＣＲ产物进行测序，结果显示感染用献血员血浆和猴血清中ＨＧＶ序列与国外株ＨＧＵ４４４０２的同源性分别为９８３３％和９５８３％；与ＨＧＵ３６３８０株的同源性分别为９２５０％和８９１７％；感染猴血清中ＨＧＶ序列与献血员ＨＧＶ序列同源性为９５８３％。结论恒河猴对ＨＧＶ敏感，可以做为实验模型动物     

Response to primary infection with Herpesvirus saimiri in immunosuppressed juvenile and newborn squirrel monkeys.

Immunosuppression of juvenile squirrel monkeys with combined azathioprine, prednisolone, and antilymphocyte globulin resulted in decreased antibody responses to viral antigens after primary infection with Herpesvirus saimiri (HVS). The virus was repeatedly isolated from the oropharynx of immunosuppressed monkeys but not from untreated infected controls. Thus immune factors are important in inhibiting shedding of HVS from the oropharynx. HVS could be isolated from the peripheral blood lymphocytes of infected control monkeys but not from the lymphocytes of immunosuppressed monkeys. Immunosuppressed monkeys also had decreased percentages of lymphocytes capable of forming rosettes with sheep erythrocytes. These results indicate that the immunosuppressive agents had inhibitory effects on lymphocytes (presumably thymus derived) capable of being latently infected with HVS. Antibody responses in newborn monkeys infected with HVS were delayed compared with juvenile monkeys. Treatment of newborn monkeys with antilymphocyte globulin had no suppressive effect on antibody responses to HVS.     

Experimental infection of green monkeys with adenoassociated virus

Primary infection and reinfection with adeno-associated virus type 4 (AAV-4) was reproduced in green monkeys experimentally infected with AAV-4 in mixture with adenovirus. Wide dissemination of the satellite virus in animals was observed. AAV-4 and its antigen were detectable 5 to 23 days after inoculation. In monkeys infected with a mixture of AAV-4 and adenovirus or with one of them the infection was accompanied by a marked fever persisting from the 5th to the 20th day after inoculation. The infected monkeys showed an intensive rise of homologous antibody titer most marked on the 10th-15th day after inoculation with AAV-4. AAV-4 and its antigen were detected in smears from conjunctival and tonsillar mucosa, rectal specimens in the time course of the infectious process, as well as from the trachea, lungs, liver, spleen, intestines and kidneys of the sacrificed monkeys. Besides, AAV-4 antigen was found in cells of the tonsils and blood leukocytes of the sacrificed monkeys. No virus or its antigen were found in the brain and heart tissues. Virions of adeno-associated virus were found by electron microscopic examinations of kidney cells of one of 3 monkeys infected with AAV-4.     

Pathogenesis of Rift Valley fever in rhesus monkeys: role of interferon response

Summary Rhesus monkeys inoculated intravenously with Rift Valley fever (RVF) virus presented clinical disease syndromes similar to human cases of RVF. All 17 infected monkeys had high-titered viremias but disease ranged from clinically inapparent to death. Three (18%) RVF virus-infected monkeys developed signs of hemorrhagic fever characterized by epistaxis, petechial to purpuric cutaneous lesions, anorexia, and vomiting prior to death. The 14 remaining monkeys survived RVF viral infection but, 7 showed clinical signs of illness characterized by diminished food intake, cutaneous petechiae, and occasional vomiting. The other 7 monkeys showed no evidence of clinical disease. All monkeys had detectable serum interferon 24–30 h after infection, but 4 of 7 monkeys that did not develop clinical illness had serum interferon titers within 12h after infection. In lethally infected macaques, indices of hepatic function and blood coagulation were abnormal within 2 days, implicating early pathogenetic events as critical determinants of survival. Serum transferase values were elevated in proportion to severity of clinical disease and outcome of infection. Both myocardial damage and laboratory evidence consistent with disseminated intravascular coagulation were present in fatal infections. All surviving monkeys developed neutralizing antibodies to RVF virus 4–7 days after infection, and this coincided with termination of viremia. Two fatally infected monkeys were viremic until death on days 6 and 8, and the third cleared viremia on day 5 and developed antibody on day 6 but died on day 15. There was a significant correlation between a delayed interferon response and mortality, suggesting that the early appearance of interferon was influential in limiting the severity of disease.     

Replication of Herpesvirus saimiri in cultured lymphocytes of infected owl monkeys (Aotus trivirgatus). An electron microscopic and immunofluorescent study.

The lymphocytes of five owl monkeys infected with Herpesvirus saimiri (HVS) and three control monkeys were studied. Lymphocytes were separated on Ficoll-Hypaque gradients, incubated in suspension cultures, and prepared for electron microscopic and immunofluorescent study at 24, 48, and 72 hours after the beginning of culture (AC). Buffy coats of whole blood immediately after bleeding were also prepared for study. At the time of the study, four of the five infected monkeys had died of malignant lymphoma and lymphocytic leukemia. HVS virions were demonstrated by electron microscopy and HVS antigens by immunofluorescence in 1 to 20 per cent of the lmyphocytes from infected monkeys in two of five cultures at 24 hours AC, four of five at 48 hours AC, and all five at 72 hours AC. There was quite good agreement between the electron microscopic and the immunofluorescent data. None of the control monkey cultures and none of the buffy coat preparations contained HVS virions or antigens. By electron microscopy the great majority of the virus particles were nucleocapsids within the nuclei of lymphocytes. Very seldom were enveloped virions observed. There was some evidence to suggest that the higher the percentage of lymphocytes containing HVS the poorer the prognosis for the monkey. HVS was isolated from all five infected monkeys by cocultivation of lymphocytes with Vero cells. Control lymphocyte cocultivations were negative.     

Shigella flexneri infection: a histopathologic study of colonic biopsies in monkeys infected with virulent and attenuated bacterial strains

Macaca fascicularis monkeys were orally infected with live virulent Shigella flexneri wild-type strains of either serotype Y (S. flexneri SFL1), 2a (S. flexneri M4243) or 1b (S. flexneri SFL27). Clinical signs of shigellosis varied from mild watery diarrhea (SFL1) to dysentery (M4243, SFL27), with a fatal outcome in two monkeys (SFL27). Colonoscopy showed slight pathologic changes in monkeys infected with SFL1, and pronounced changes in monkeys infected with SFL27. In colonic biopsies the most severe acute inflammation, with surface epithelial erosions and ulcerations, was seen after infection with SFL27, followed by SFL1, and M4243. The live S. flexneri serotype Y vaccine strain SFL114, derived from SFL1 and attenuated because of an inactivated aroD gene and hence auxotrophic for p-aminobenzoic acid, caused no diarrheal illness in 14 monkeys. In colonic biopsies, SFL114 only elicited a slight acute inflammatory reaction. Vaccinated monkeys were protected against clinical disease when challenged with any one of the three virulent S. flexneri wild-type strains. Histopathologically, the acute inflammation was of less intensity than that seen in non-vaccinated monkeys. A good correlation between clinical signs, endoscopic findings and the degree of acute inflammation was demonstrated for monkeys vaccinated with SFL114 and challenged with either SFL1 or SFL27.     

中国株庚型肝炎病毒(HGV)感染猕猴的实验研究

应用０．５ｍｌ含中国株庚型肝炎病毒（ＨＧＶ）的血清接种５只猕猴，５只猕猴于接种后１周血清ＨＧＶＲＮＡ均阳转；ＨＧＶＲＮＡ滴度在猴体内可高达１∶１０５，较接种血清中ＨＧＶＲＮＡ滴度（１∶１０２）高出许多，提示ＨＧＶ在猕猴体内复制。其中４只猕猴血清抗－ＨＧＶ阳转；４只出现ＡＬＴ异常。后用其中１只猕猴感染后４５天的血清给另２只猕猴接种，该２只猴也出现血清ＨＧＶＲＮＡ和抗－ＨＧＶ阳转及ＡＬＴ异常。本研究表明，中国猕猴有可能作为ＨＧＶ感染的动物模型。     

Serotype-specific and cross-reactive neutralizing antibody responses in cynomolgus monkeys after infection with multiple dengue virus serotypes

Neutralizing antibody responses were examined in monkeys after dengue virus infections. In monkeys that had been infected once or twice with DENV-2, neutralizing antibody was cross-reactive with all four serotypes after secondary or tertiary infection with DENV-3. In monkeys that had been inoculated with DENV-1 and DENV-2 in the primary and secondary infections, neutralizing antibody titers did not increase after tertiary infection with DENV-3. These results indicate that antibody responses after secondary and tertiary infections with different serotypes are cross-reactive with all four serotypes, consistent with what has been observed in humans, and suggest that monkeys are useful for determining neutralizing antibody responses.     

Simian varicella virus infects ganglia before rash in experimentally infected monkeys

Monkeys experimentally infected with simian varicella virus (SVV) develop rash 10-14 days later. However, the route and the time of ganglionic infection are unknown. Using PCR, we analyzed DNA extracted from tissues of 13 monkeys 5 to 60 days after either intratracheal or intravenous inoculation with SVV. SVV DNA was detected in ganglia from four of five monkeys sacrificed 6 to 7 days after intratracheal inoculation. Further, analysis of ganglia from monkeys sacrificed at 10 days revealed that intravenous inoculation produced a higher proportion of SVV DNA-positive ganglia (63%) than that after intratracheal inoculation (13%), pointing to the role of hematogenous spread in ganglionic infection. Like other organs, monkey ganglia become infected with SVV before the appearance of rash.     

Response to experimental infection with mumps virus in rhesus monkeys

Experiments were conducted in which rhesus monkeys were infected with mumps virus by inoculation into the parotid gland. Virus was isolated from buccal swab specimens from animals up to 6 days after inoculation. Immunofluorescence staining for mumps antigen was positive, and infectious virus was recovered from tissue biopsies of inoculated glands. Histological examination of biopsied glands revealed lymphocytic infiltration of the tissue. Virus was not isolated from urine, buffy coat, or biopsies of thyroid and spleen. In other experiments, animals were infected by inoculation into the thyroid gland. Virus was not isolated from buccal swabs, urine specimens, or thyroid tissues from these monkeys. All inoculated animals responded immunologically to mumps virus regardless of route of infection. Previously infected animals did not shed virus when challenged with mumps virus. A histological response to challenge in glandular tissue was observed which suggested a hypersensitivity to mumps antigen. The response was qualitatively identical to that in acutely infected tissue but more intense. None of the animals infected developed antibodies to thyroid tissue regardless of route of injection of virus or site of challenge of immune monkeys.