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1.
Human immunodeficiency virus infection of human-PBL-SCID mice   总被引:32,自引:0,他引:32  
Severe combined immunodeficient (SCID) mice reconstituted with human peripheral blood leukocytes (hu-PBL-SCID mice) have inducible human immune function and may be useful as a small animal model for acquired immunodeficiency syndrome (AIDS) research. Hu-PBL-SCID mice infected with human immunodeficiency virus-1 (HIV-1) contained virus that was recoverable by culture from the peritoneal cavity, spleen, peripheral blood, and lymph nodes for up to 16 weeks after infection; viral sequences were also detected by in situ hybridization and by amplification with the polymerase chain reaction (PCR). Mice could be infected with multiple strains of HIV-1, including LAV-1/Bru, IIIB, MN, SF2, and SF13. HIV-1 infection affected the concentration of human immunoglobulin and the number of CD4+ T cells in the mice. These results support the use of the hu-PBL-SCID mouse for studies of the pathogenesis and treatment of AIDS.  相似文献   

2.
目的 观察外周血 4 种细胞因子(IL-2、IL-4、IL-15及IL-21)在HIV感染者/AIDS患者(HIV/AIDS)和正常人表达水平的区别及其与HIV RNA病毒载量和CD3+、CD4+细胞数量的相关性,分析其与HIV感染的关系和临床意义。方法 40例未经抗病毒治疗的HIV/AIDS 和31名正常人,采用ELISA法检测外周血4种细胞因子(IL-2、IL-4、IL-15及IL-21),使用流式细胞计数法检测外周血CD3+、CD4+ T 淋巴细胞数量,实时聚合酶链反应测定血清中HIV RNA水平。结果 未经抗病毒治疗的HIV/AIDS外周血IL-2、IL-15及IL-21水平低于正常人,而IL-4水平高于正常人(P<0.05)。CD3+、CD4+细胞数量和各细胞因子水平显示出一定的相关关系,IL-2和IL-15水平均与HIV-1 RNA载量呈负相关。结论 提示外周血4种细胞因子都可能参与了HIV感染的致病机制,并与HIV RNA病毒载量及CD3+、CD4+细胞数量相关。  相似文献   

3.
4.
A murine model in which neurotropic retroviral infection can be studied over short periods of time was developed. Microinjection of Cas-Br-E virus into midgestation mouse embryos caused paralysis and death within 25 days after birth, in contrast to virus-infected neonates which develop disease only after 4 months. To evaluate whether antiviral drugs could cross the placental barrier and influence the course of the disease, the drug 3'-azido-3'-deoxythymidine (AZT) was administered to infected embryos through the drinking water of pregnant females. AZT treatment markedly retarded the onset and course of virus-induced central nervous system disease, permitting animals to survive beyond 4 months of age. These results are evidence for effective antiviral treatment during gestation and in the perinatal period and are of potential significance for the management of maternal transmission of the acquired immune deficiency syndrome (AIDS) virus.  相似文献   

5.
Infection of the SCID-hu mouse by HIV-1   总被引:44,自引:0,他引:44  
  相似文献   

6.
The study of human hematopoietic cells and the human immune system is hampered by the lack of a suitable experimental model. Experimental data are presented showing that human fetal liver hematopoietic cells, human fetal thymus, and human fetal lymph node support the differentiation of mature human T cells and B cells after engraftment into mice with genetically determined severe combined immunodeficiency. The resultant SCID-hu mice are found to have a transient wave of human CD4+ and CD8+ T cells and human IgG (immunoglobulin G) in the peripheral circulation. The functional status of the human immune system within this mouse model is not yet known.  相似文献   

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8.
Confirmed infection with HTLV-II (human T cell leukemia virus type II) has been described only in rare cases. The major limitation to serological diagnosis of HTLV-II has been the difficulty of distinguishing HTLV-II from HTLV-I (human T cell leukemia virus type I) infection, because of substantial cross-reactivity between the viruses. A sensitive modification of the polymerase chain reaction method was used to provide unambiguous molecular evidence that a significant proportion of intravenous drug abusers are infected with HTLV, and the majority of these individuals are infected with HTLV-II rather than HTLV-I. Of 23 individuals confirmed by polymerase chain reaction analysis to be infected with HTLV, 21 were identified to be infected with HTLV-II, and 2 were infected with HTLV-I. Molecular identification of an HTLV-II--infected population provides an opportunity to investigate the pathogenicity of HTLV-II in humans.  相似文献   

9.
Chronic hepatitis B virus (HBV) infection is a major cause of liver disease. Only interferon-alpha and the nucleosidic inhibitors of the viral polymerase, 3TC and adefovir, are approved for therapy. However, these therapies are limited by the side effects of interferon and the substantial resistance of the virus to nucleosidic inhibitors. Potent new antiviral compounds suitable for monotherapy or combination therapy are highly desired. We describe non-nucleosidic inhibitors of HBV nucleocapsid maturation that possess in vitro and in vivo antiviral activity. These inhibitors have potential for future therapeutic regimens to combat chronic HBV infection.  相似文献   

10.
目的:建立可表达血清淀粉样P成分(SAP)转基因小鼠模型,以研究SAP基因的功能。方法:采用基因重组技术将SAP基因插入到pCMV-Tag5A真核表达载体上。经BciVI限制性内切酶酶切后,琼脂糖凝胶电泳回收2.7 kb目的片断,受精卵原核显微注射法将其注入雄原核,制备转基因小鼠。采用PCR法在基因水平筛选SAP转基因小鼠阳性小鼠;免疫沉淀法结合免疫印迹法在蛋白水平检测SAP基因在PCR阳性转基因小鼠中的表达情况。结果:成功构建了pCMV-Tag5A/mSAP真核表达载体。经显微注射法将2.7 kb线性目的基因片断注射入受精卵雄原核,移植入代孕母鼠,所生127只小鼠,其中PCR鉴定获得10只转基因阳性小鼠。免疫沉淀法和免疫印记法随机检测83#♂founder、7#♀founder与C57 BL/6J小鼠回交F1代PCR阳性转基因小鼠血清中外源SAP基因蛋白水平的表达,发现均有表达。结论:该研究以C57BL/6J小鼠为研究对象,成功地产生能稳定遗传SAP基因的过表达转基因小鼠模型,它将有利于研究SAP基因在炎症、淀粉样沉淀、自身免疫系统中的作用。  相似文献   

11.
连翘酯苷对IFN-α和Mx1表达的影响   总被引:3,自引:0,他引:3  
 【目的】探讨连翘酯苷作用于感染PCV2的小鼠模型后,对IFN-α和Mx1表达的影响,以评价连翘酯苷的抗病毒作用。【方法】复制PCV2感染小鼠模型。以(2、4、10 mg?mL-1)连翘酯苷分为高、中、低 3个剂量组,并设空白对照组和病毒对照组。给药12、24、36 h后,分别提取小鼠肺组织总RNA和总蛋白,应用实时荧光定量PCR检测IFN-α、Mx1 mRNA含量,采用Western blotting检测Mx1蛋白的表达水平。【结果】空白对照组小鼠肺组织中Mx1和IFN-α无表达,病毒对照组少量表达。随着连翘酯苷浓度的升高,IFN-α和Mx1 mRNA水平逐步升高,两者的表达量呈现平行相关;高剂量组中IFN-α和Mx1表达量最高(P<0.001);随着药物作用时间的延长表达量降低,12 h时达到最高(P<0.001),其后逐渐下降。【结论】昆明小鼠适于PCV2感染模型的建立。中药连翘酯苷能显著上调IFN-α和Mx1的表达,并在给予高剂量药物后12 h发挥显著的抗病毒作用。Mx1蛋白与病毒的感染密切相关,可用于病毒感染的早期诊断,及具有较好的应用前景。  相似文献   

12.
The article reports the efficacy assessment for the Allokin-alpha (Allokin) preparation by modeling the experimental viral infection in vivo in poultry (day-old chickens). The experimental infection included the following steps: vaccination using the vaccine HVT strain, the intramuscular injection of Allokin, and the preparation of feather follicle extracts. The specific antigen was detected in the feather follicle epithelium using the method of gel diffusion precipitation reaction (DPR). A decrease in the number of positive DPR test results for HVT antigen was observed in the case of Allokin-alpha treatment. It has been demonstrated that the optimal time for the Allokin-alpha administration was on the tenth day after the vaccination with HVT as soon as the number of positive DPR results was 33%, while the number of positive results was 50% when the tested preparation was administered 5 and 15 days after the vaccination (samples were collected on day 28 after the vaccination). Since Allokin-alpha has been successfully used to treat human papillomavirus for 15 years, it may also work well as an antiviral agent in the experimental viral infection simulation by the inoculation of vaccine HVT strain; the preparation is not species specific.  相似文献   

13.
应用PCR方法调查实验大小鼠螺杆菌感染情况   总被引:2,自引:0,他引:2  
通过聚合酶链反应(PCR)对上海地区实验大鼠、小鼠的螺杆菌携带情况进行调查,并用5种啮齿动物螺杆菌特异性16SrRNA基因引物对螺杆菌属阳性样品进一步鉴定,为我国实验动物微生物等级及监测标准的制定提供参考依据。结果表明:实验大鼠阳性率为70.3%(71/101),其中清洁级、SPF级分别为69.6%(48/69)和71.9%(23/32);实验小鼠阳性率为35.8%(126/352),其中清洁级、SPF级阳性率分别为51.5%(52/101)和29.5%(74/251)。5种啮齿动物螺杆菌特异性16SrRNA基因引物进一步分析,结果显示在携带螺杆菌的107只小鼠及68只大鼠中,主要携带的是H.rodentium、H.hepaticus和H.bilis 3种。上海地区实验大、小鼠皆存在不同程度的螺杆菌感染,PCR法可用于实验大小鼠螺杆菌感染状况的初步调查和流行病学监测。  相似文献   

14.
根据狂犬病病毒核蛋白基因保守区序列设计1对引物,建立了用于狂犬病病毒特异性核酸检测的RT-PCR技术。该技术可从狂犬病病毒CVS株、8202株和SRV9株的含毒细胞培养物及鼠脑组织中.扩增出443bp的核酸片段,检出核酸的敏感性约为3pg。对30份不同种动物脑组织的检测结果与小鼠脑内接种试验(MIT)的测定结果完全吻合,但前者可在3h内直接对组织匀浆进行诊断,具有快速、简便和敏感的优点。  相似文献   

15.
用微卫星引物对近交系小鼠进行遗传监测   总被引:6,自引:0,他引:6       下载免费PDF全文
目的 :近交系小鼠广泛地应用于医学和生物学研究领域 ,无品系污染是其基本要求 ,采用 1 0对微卫星引物对 BALB/ C-nu-nu、DBA/ 2、SCID、T73 9、TA2 、6 1 5六种近交系小鼠进行遗传监测 ,试图寻找相关品系遗传监测的基因位点和应用于实际监测。方法 :在确定实验材料符合要求的情况下 ,采用聚合酶链式反应扩增短串联重复序列 ,选择多态性位点作为相关品系的遗传监测标记。结果 :除 1个位点表现为单态性外 ,其余9个微卫星位点表现出多态性 ,其中 D2 Nds3、D3Mit1 5、D3Mit1 7、D3Mit1 8等 4个位点多态性显著 ,这 4个位点适宜用于小鼠遗传监测。上述结果说明微卫星 DNA多态性标记适用于近交系小鼠遗传监测 ,有助于遗传监测从表现型过渡到 DNA水平。  相似文献   

16.
为了建立一套适宜于白僵菌(Beauveria bassiana)退化研究的RAPD-PCR反应体系及反应程序,通过采用L16(45)正交试验及退火温度和循环次数的单因素优化对反应体系中的各因素进行优化组合。结果表明:20μL PCR反应体系及反应程序中各因素优化组合为,10×Buffer 2μL,MgCl2(25 mmol/L)2.4μL,4种dNTP(各2.5mmol/L)0.8μL,随机引物(10μmol/L)1.4μL,TaqDNA聚合酶(5 U/μL)0.4μL,模板DNA(10 mg/L)1μL。反应条件为,94℃预变性2 min,94℃变性30 s,38℃退火40 s,72℃延伸1 min,循环次数40次,72℃延伸5 min。  相似文献   

17.
通过制备抗致病性大肠杆菌特异性IgY及富含特异性IgY的功能性卵黄粉,进行小鼠的攻毒保护试验,探讨功能性卵黄粉对由致病性大肠杆菌引起的疾病的防治作用.结果显示:(1)致病性大肠杆菌与特异性IgY混合后,菌体出现凝集,且病原菌的运动力明显受抑制;(2)浓度为1.3×109CFU·mL-1的致病性大肠杆菌K88、K99和9...  相似文献   

18.
陈佳  吕芬  吴崇超  彭舟  魏博  杨辉  陈伟  熊盛 《安徽农业科学》2013,41(8):3323-3326
[目的]研究蓝藻抗病毒蛋白N(CVN)衍生物(LCVN)对永生化上皮角质形成细胞(HaCaT)的毒性及LCVN凝胶的制备和体外活性检测。[方法]用MTT法测定LCVN对HaCaT细胞株的半数毒性浓度(CC50)以及LCVN对流感病毒毒株A/HK/8/68(H3N2)的半数抑制浓度(IC50);用流式细胞检测法检测LCVN对HaCaT细胞周期与细胞凋亡的影响;用酶联免疫吸附法(ELISA)检测LCVN凝胶与HIV包膜蛋白GP120的相互作用。[结果]培养24和48 h后,LCVN对HaCaT的CC50分别为(9.25±1.21)与(1.94±0.12)μmol/L,将LCVN做成凝胶后,与HIV外膜蛋白GP120的结合在一定浓度范围内具有剂量关系,LCVN凝胶对A/HK/8/68(H3N2)毒株的IC50为(98.20±0.03)nmol/L。[结论]LCVN对HaCaT细胞的毒性明显小于CVN,当做成凝胶以后LCVN对流感病毒A/HK/8/68(H3N2)具明显抑制作用,并能与HIV包膜蛋白GP120相互作用。  相似文献   

19.
Mycosis fungoides, a rare form of cutaneous T cell leukemia/lymphoma, is suspected of having a viral etiology on the basis of certain similarities to adult T cell leukemia, which is associated with human T cell leukemia/lymphoma virus type I (HTLV-I) infection. Cell lines were established from peripheral blood mononuclear cells (PBMC) of an HTLV-I-seronegative patient with mycosis fungoides. DNA hybridization analysis revealed the presence of HTLV-I-related sequences with unusual restriction endonuclease sites. Sequence analysis of subcloned fragments demonstrated the presence of a monoclonally integrated provirus with a 5.5-kilobase deletion involving large regions of gag and env and all of pol. Additional evidence for the presence of deleted proviruses was found by polymerase chain reaction (PCR) amplification of DNA from cutaneous lesions of five other HTLV-I-seronegative patients. The findings suggest that HTLV-I infection may be involved in the etiology of at least certain cases of mycosis fungoides.  相似文献   

20.
In the polymerase chain reaction (PCR), two specific oligonucleotide primers are used to amplify the sequences between them. However, this technique is not suitable for amplifying genes that encode molecules where the 5' portion of the sequences of interest is not known, such as the T cell receptor (TCR) or immunoglobulins. Because of this limitation, a novel technique, anchored polymerase chain reaction (A-PCR), was devised that requires sequence specificity only on the 3' end of the target fragment. It was used to analyze TCR delta chain mRNA's from human peripheral blood gamma delta T cells. Most of these cells had a V delta gene segment not previously described (V delta 3), and the delta chain junctional sequences formed a discrete subpopulation compared with those previously reported.  相似文献   

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