艾滋病—1（HIV—1）的性传播

艾滋病—１（ＨＩＶ—１）的传播途径有性接触、血液接触和母婴传播,性传播是ＨＩＶ—１流行性传播的主要途径。ＨＩＶ—１经性接触传播占全球３３４０万ＨＩＶ—感染者的７５％以上。对ＨＩＶ—１的性传播的研究,为预防ＨＩＶ—１经性接触传播有重要意义。本文分析了Ｈ...     

我国首次发现的HIV—1和HIV—2双重感染样品中病毒部分基因的序列特 …

上海市卫生检疫局送检了一例ＨＩＶ－１和ＨＩＶ－２抗体检测均呈阳性的双重感染样品,对其感染的ＨＩＶ前病毒的ｇａｇ和ｅｎｖ基因区进行了序列分析,首次阐明我国发现的ＨＩＶ双重感染样品的ＨＩＶ部分基因特征。从ＨＩＶ感染者淋巴细胞（ｐｅｒｉｐｈｅｒａｌ ｂｌｏｏｄ ｍｏｎｏｎｕｃｌｅａｒ ｃｅｌｌｓ,ＰＢＭＣ）中提取前病毒ＤＮＡ,分别使用ＨＩＶ－１和ＨＩＶ－２特异性引特用套式ＰＣＲ扩增ＨＩＶ－１和ＨＩＶ－２     

HIV－1反式激活因子（Tat）作用机理的研究进展

ＨＩＶ－１反式激活因子（Ｔａｔ）作用机理的研究进展梁臣,耿运琪（南开大学生命科学学院,天津３０００７１）关键词ＨＩＶ－１,Ｔａｔ,ＴＡＲ人免疫缺陷病毒－１（ＨＩＶ－１）属反转录病毒科,慢病毒亚科,为艾滋病的重要病原体。该病毒可长期潜伏于人体内,但在某...     

人I型免疫缺陷病毒gag—env嵌合基因在重组痘苗中的表达

Ｇａｇ和Ｅｎｖ蛋白是人Ｉ型免疫缺陷病毒（Ｈｕｍａｎ ｉｍｍｕｎｏｄｅｔｉｃｉｅｎｃｙ ｖｉｒｕｓ ｔｙｐｅ １,ＨＩＶ－１）的结构蛋白,是ＨＩＶ－１诱导机体产生体液免疫和细胞免疫的主要抗原。本实验通过多交亚克隆,将ｅｎｖ基因以正确的三联密码读框插入ｇａｇ基因的下游制备了ＨＩＶ－１ ｇａｇ－ｅｎｖ嵌合基因,并将嵌合基因分别置于痘苗病毒ｐ７．５启动子和牛痘病毒Ａ型包涵体（ＡＴＩ）启动子的下游,经过同源     

HIV共受体—CC类趋化因子受体—5

ＨＩＶ共受体——ＣＣ类趋化因子受体┐５黄仕和秦椿华（卫生部武汉生物制品研究所,武昌４３００６０）（美国得克萨斯Ａ＆Ｍ大学毒理学系）关键词ＨＩＶ共受体ＣＣ类趋化因子受体－５继冯愈等发现融合素是嗜Ｔ细胞ＨＩＶ－１的融合辅助因子后,邓洪魁及其同事发现了初始...     

人疱疹病毒6型在艾滋病发病中的作用

许多临床和实验研究结果表明人疱疹病毒６型（ＨＨＶ－６）在人类免疫缺陷病毒（ＨＩＶ）自然感染过程中起着促进因子的作用。ＨＨＶ－６与ＨＩＶ有着共同的Ｔ细胞嗜性,除此以外,ＨＨＶ－６还能感染杀伤ＣＤ８＾＋Ｔ细胞,ＮＫ细胞单核－巨噬细胞,故认为免疫系统潜在影响更大。弄清ＨＨＶ－６在ＨＩＶ感染过程中对免疫系统的损伤作用,将有助于阐明艾滋病发病的复杂机制。     

树Ju免疫细胞体外感染Ⅰ型人免疫缺陷病毒的实验研究

至今可以感染Ⅰ型人免疫缺陷病毒（ＨＩＶ－１）的动物只有黑猩猩和长臂猿,这严重阻碍了ＨＩＶ－１的疫苗研究和治疗研究。因此,寻找新的可以感染ＨＩＶ－１的动物模型成为十分迫切的课题。已知树Ｊu对许多重要的医学病毒易感,为了探讨树Ｊu是否可以感染ＨＩＶ－１,利用不同辅助受体的５种ＨＩＶ－１病毒株。体外感染云南野生成年树Ju的淋巴细胞和单核／巨噬细胞;同时还用这些病毒感染人外周血淋巴细胞或单核细胞。然后用ＲＴ－ＰＣＲ、ＰＣＲ和流式细胞术分别进行了检测,用ＲＴ－ＰＣＲ方法未检测到感染上清中有病毒粒子的存在,用ＰＣＲ法未能发现树Ｊu的这些免疫细胞中有前病毒ＤＮＡ,用流式细胞术也未能在这些感染ＨＩＶ－１的树Ｊu细胞的表面检测到特异抗原;而感染ＨＩＶ－１的人免疫细胞均为阳性结果。实验结果表明树Ju的这些免疫细胞在体外未能感染上ＨＩＶ－１,可能的原因是树Ｊu的这些免疫细胞的ＨＩＶ－１受体（ＣＤ４）和辅助受体（ＣＣＲ５或ＣＸＣＲ４）与人的免疫细胞胞差别较大。     

广西壮族自治区HIV—1流行毒株的基因序列测定和亚型分析

使用ＰＣＲ技术对１４份广西ＨＩＶ－１阳性感染者外周血单核细胞（ＰＢＭＣｓ）样品进行扩增,获得ＨＩＶ－１膜蛋白（ｅｎｖ）基因的核酸片段,并对其Ｃ２－Ｖ３及邻区３５０￣４５０个核苷酸序列进行了测定和分析。结果表明,１４份样品中９份为泰国Ｂ（Ｂ＇）亚型,５份为Ｅ亚型毒株。其中Ｂ＇亚型毒株的基因离散率为４．２％,与Ａ－Ｅ参考亚型及部分Ｂ亚型代表株序列相比较,与包括泰国、缅甸及云南德宏在内的Ｂ亚型毒株序列十     

ＣＣＲ５与ＨＩＶ—１感染的分子机制研究进展

趋化因子受体５（ＣＣＲ５）作为一个辅助受体在ＨＩＶ－１进入巨噬细胞的过程中发挥重要作用。在不同人种中，ＣＣＲ５存在天然的突变基因型，其中包括纯合子和杂合子基因型，它们对ＨＩＶ－１的易感性也各不相同，实验室中膜外袢的突变、缺失突变、插入及膜内基元的信号转导研究从分子水平揭示了一些氨基酸和空间结构对辅助受体的功能起关键性的作用，这对深入了解ＨＩＶ－１与受体作用的分子机制，为寻找有效的阻断途径无疑是必需的和重要的。     

人免疫缺陷病毒1型Rev单链抗体细胞内免疫抗病毒基因治疗研究

应用抗ＨＩＶ－１Ｒｅｖ单链抗体细胞内免疫方法,研究在人Ｔ细胞和周围血淋巴单核细胞内抗病毒复制的效果,探讨细胞内免疫抗ＨＩＶ－１基因治疗的可行性。克隆抗ＨＩＶ－１Ｒｅｖ单链抗抗体（ｓＦｖ）基因,以逆转录病毒为基因载体,将名装后的含靶基因的逆转录病毒转导至人ＣＤ４阳性Ｔ－细胞株ＣＥＭ和ＳｕｐＴ１,以及ＨＩＶ－１阴性自愿者的周围血淋巴单核细胞（ＰＢＭＣ）,再分别用不同剂量（ＭＯＩ）的ＨＩＶ－１病毒株ＰＮ     

Transactivation of the human immunodeficiency virus promoter by human herpesvirus 6 (HHV-6) strains GS and Z-29 in primary human T lymphocytes and identification of transactivating HHV-6(GS) gene fragments.

Human herpesvirus 6 (HHV-6) can activate the human immunodeficiency virus (HIV) promoter and accelerate cytopathic effects in HIV-infected human T cells. This study examines the regions of the HIV promoter required for HHV-6 transactivation in a heterogeneous population of primary human T lymphocytes with or without antigenic stimulation. Two different strains of HHV-6, GS and Z29, transactivated the HIV promoter. The GS strain transactivated the promoter in both stimulated and resting T cells, while the Z29 strain increased HIV promoter activity only in stimulated T cells. Three DNA clones containing HHV-6(GS) genomic fragments transactivated the HIV promoter in cotransfected T cells. A 21.4-kb DNA clone, pZVB70, showed the highest transactivating ability, while two other DNA fragments, pZVB10 (6.2 kb) and pZVH14 (8.7 kb), showed lower activity. One of these clones, pZVH14, activated the HIV promoter construct containing a mutation in the NF kappa B site. However, this mutated NF kappa B promoter was not transactivated during HHV-6(GS) infection or after cotransfection with pZVB70 or pZVB10. These data indicate that the NF kappa B sites of the HIV promoter are essential for its transactivation during HHV-6(GS) infection. By increasing HIV promoter activity in primary T lymphocytes, HHV-6 may consequently increase HIV replication, leading to an increase in the cytopathic effect on coinfected human T cells.     

Identification and characterization of a human herpesvirus 6 gene segment that trans activates the human immunodeficiency virus type 1 promoter.

Human herpesvirus 6 (HHV-6) is a lymphotropic herpesvirus, and in vitro, HHV-6 can productively infect many of the same cell types as can human immunodeficiency virus (HIV). Coinfection by both viruses in vitro can lead to both activation of the HIV promoter and acceleration of cytopathic effects. We have previously demonstrated that a large, 22.25-kb cloned HHV-6 fragment, pZVB70, can trans activate HIV promoter expression in vitro. In this study, we show that the pZVB70 fragment can trans activate the HIV promoter in human T-cell lines as well as in the monkey kidney cell line CV-1. The pZVB70 insert was digested with various restriction enzymes, and individual fragments were transfected into cells to test for their ability to trans activate the HIV promoter. By this method, we have identified a 1.8-kb subfragment, B701, that is involved in trans activation. Sequence analyses show that B701 potentially encodes a 143-amino-acid protein. This protein shares no homology with other herpesvirus proteins, such as ICP0 and ICP4, that have been shown to trans activate the HIV promoter. However, it shows weak sequence homology with the gene products encoded by the cytomegalovirus early US22 gene family, suggesting that the putative B701 protein may be an HHV-6 early regulatory protein. The 143-amino-acid coding sequence of B701 was cloned by polymerase chain reaction, and transfection of this construct into cells activated HIV promoter expression. The target site on the HIV promoter for the putative B701 protein is mapped to the NF-kappa B binding site. Our results suggest that the putative B701 protein may function by directly binding to the NF-kappa B site or may involve cellular factors, such as NF-kappa B, either directly or indirectly.     

Transactivation of human immunodeficiency virus promoter by human herpesvirus 6.

Patients with acquired immunodeficiency syndrome (AIDS) are often infected with a number of other heterologous viruses in addition to the initial human immunodeficiency virus (HIV) infection, and these agents could act as potential reactivating agents of latent HIV. A new antigenically distinct herpesvirus, designated human herpesvirus 6 (HHV-6), has recently been isolated from patients with AIDS and has been shown to infect a number of different human cells, specifically human T cells, B cells, and glial cells. Since these are some of the same cells that harbor the AIDS virus, it is quite important to determine any interaction between this new herpesvirus and HIV. In this report, we demonstrate that HHV-6 can trans-activate the HIV promoter in human T-cell lines as measured by the expression of the bacterial gene chloramphenicol acetyltransferase. This indicates that stimulation of HIV gene expression by HHV-6 could play a role in HIV pathogenesis.     

Prevalence of Human Herpesvirus 6 Variants A and B in Patients with Chronic Fatigue Syndrome

Peripheral blood mononuclear cells collected from 13 patients with chronic fatigue syndrome and 13 healthy controls were analyzed for the presence of human herpesvirus 6 (HHV-6) DNA by variant-specific polymerase chain reaction and dot blot hybridization. HHV-6 DNA was detected in 7 of 13 (53%) patients, and of those 7 patients, 4 were positive for HHV-6 variant A DNA and 3 were for variant B. No HHV-6 DNA was detected in the controls. Serum antibody titers to the late antigen and antibody prevalence to the early antigen of HHV-6 were significantly higher in the patient group. These results suggest active replication of HHV-6 in patients with chronic fatigue syndrome.     

Exogenous human herpesvirus 6 reinfection after tumor-infiltrating T-lymphocyte therapy

Exogenous human herpes virus 6 (HHV-6) reinfection has never been reported in patients receiving tumor-infiltrating T lymphocytes therapy. We report an unusual case of HHV-6 infection following infusion of HHV-6 infected autologous T lymphocytes. HHV-6 infection could interfere with the tumor antigen immune recognition and the efficacy of immunotherapy.     

Flow cytometric evaluation of antiviral agents against human herpesvirus 6

Antiviral activities of acyclovir (9-[(2-hydroxyethoxy) methyl] guanine, ACV), penciclovir (9-[4-hydroxy-3-(hydroxymethyl) butyl] guanine, PCV), ganciclovir ([9-(1,3-dihydroxy-2-propoxy) methyl] guanine, GCV), and foscarnet (phosphonoformic acid, PFA) were determined against Human Herpesvirus 6 (HHV-6) by flow cytometric technique. The technique is based on the detection of gp116 antigen expression in virus infected cells. Susceptibility was defined in terms of drug concentration which reduced the number of cells expressing HHV-6 gp116 antigen with a mean fluorescent intensity (MFI) by 50% as compared to virus infected untreated cells. GCV was found to be most effective against HHV-6 followed by PFA, PCV and ACV. For HHV-6A, the mean 50% inhibitory concentrations (IC50) of GCV and PFA were found to be 3.4 microM and 34.7 microM respectively, whereas the IC50 of ACV and PCV were found to be 53.7 microM and 37.9 microM respectively. For HHV-6B, the IC50 of GCV and PFA were found to be 5.7 microM and 71.4 microM respectively, whereas the IC50 of ACV and PCV were found to be 119.0 microM and 77.8 microM respectively. Flow cytometry is a valuable technique for the evaluation of antiviral compounds against viruses including HHV-6.     

Nucleotide sequence analysis of a 38.5-kilobase-pair region of the genome of human herpesvirus 6 encoding human cytomegalovirus immediate-early gene homologs and transactivating functions.

Human herpesvirus 6 (HHV-6) is prevalent in the human population, with primary infection occurring early in life. Its predominant CD4+ T-lymphocyte tropism, its ability to activate human immunodeficiency virus type 1 (HIV-1) gene expression in vitro, and its upregulation of CD4 expression has led to speculation that HHV-6 may act as a positive cofactor in the progression of HIV infection to AIDS in individuals infected with both viruses. Previous sequencing studies of restricted regions of the 161.5-kbp genome of HHV-6 have demonstrated unequivocally that it is a member of the betaherpesvirus subgroup and have indicated that the HHV-6 genome is generally collinear with the unique long (UL) component of human cytomegalovirus (HCMV). In the work described in this report we have extended these sequencing studies by determining the primary structure of 38.5-kbp of the HHV-6 genome (genomic position 21.0 to 59.5 kbp). Within the sequenced region lie 31 open reading frames, 20 of which are homologous to positional counterparts in HCMV. Of particular significance is the identification of homologs of the HCMV UL36-38 and US22-type genes, which have been shown to encode transactivating proteins. We show that DNA sequences encoding these HHV-6 homologs were able to transactivate HIV-1 long terminal repeat-directed chloramphenicol acetyltransferase expression in cotransfection assays, thus demonstrating functional as well as structural conservation of these betaherpesvirus-specific gene products. Our data therefore confirm the close relationship between HHV-6 and HCMV and identify putative immediate-early regulatory genes of HHV-6 likely to play key roles in lytic replication and possibly also in the interactions between HHV-6 and HIV in dually infected cells.     

脂筏在人类疱疹病毒6型装配中的作用

为了探讨脂筏在人类疱疹病毒6型(HHV-6)装配中的作用,用HHV-6 GS株感染HSB2细胞,用非离子去污剂Triton X-100提取脂筏成分,利用Western blot分析HHV-6包膜糖蛋白与脂筏的相关性.并用免疫荧光双标记的方法,从分子共定位的角度研究HHV-6糖蛋白B(gB)与GPI(glycosyl-phosphatidyl inosital)锚固蛋白CD59分子以及神经节苷脂GMI(monosialotetrahexosyl ganglioside)分子之间的表达与分布关系.结果发现HHV-6包膜糖蛋白B、H、L、Q1和Q2(gB、gH、gL、gQ1和gQ2)分布在脂筏部位.激光共聚焦显微镜可观察到CD59分子及GM1均与HHV-6包膜糖蛋白B有着相同的分布,即脂筏提供HHV-6装配的平台.关于脂筏在人类疱疹病毒6型装配中的作用,这是第一次报道.     

Estimating immunoregulatory gene networks in human herpesvirus type 6-infected T cells

The immune response to viral infection involves complex network of dynamic gene and protein interactions. We present here the dynamic gene network of the host immune response during human herpesvirus type 6 (HHV-6) infection in an adult T-cell leukemia cell line. Using a pathway-focused oligonucleotide DNA microarray, we found a possible association between chemokine genes regulating Th1/Th2 balance and genes regulating T-cell proliferation during HHV-6B infection. Gene network analysis using an integrated comprehensive workbench, VoyaGene, revealed that a gene encoding a TEC-family kinase, ITK, might be a putative modulator in the host immune response against HHV-6B infection. We conclude that Th2-dominated inflammatory reaction in host cells may play an important role in HHV-6B-infected T cells, thereby suggesting the possibility that ITK might be a therapeutic target in diseases related to dysregulation of Th1/Th2 balance. This study describes a novel approach to find genes related with the complex host-virus interaction using microarray data employing the Bayesian statistical framework.