人类8型疱疹病毒的研究进展

人类8型疱疹病毒（human herpesvirus-8,HHV-8）又称卡波氏肉瘤相关疱疹病毒（Kapo- si's sarcoma- associated herpesvirus,KSHV）,是一种新的肿瘤病毒,目前被认为是卡波氏肉瘤（Kaposi's sarcoma,KS）致病因子,并且与primary effusion lymphoma （PEL）和multicentric Castleman's disease（MCD）相关。该病毒编码许多蛋白,包括潜伏感染相关蛋白,裂解感染相关蛋白和HHV-8特有基因表达蛋白,在KS和HHV-8相关疾病的发病中起到关键作用。     

卡波济肉瘤相关疱疹病毒／人类疱疹病毒8型研究进展

本文综述了１９９６年以来对卡波济肉瘤（ＫＳ）相关疱疹病毒（ＫＳＨＶ）／人类疱疹病毒８型（ＨＨＶ－８）的流行病学,人体内病毒分布,细胞模型及溶细胞型生长培养系统的建立,电镜超微结构,血清学检测及传播途径等方面的研究成果。     

人类疱疹病毒6型

<正>人类疱疹病毒6型(HHV-6)最先是从淋巴细胞增生性疾病患者中分离出来的一种新的疱疹病毒。其抗原性和遗传学特性与疱疹病毒1型,疱疹病毒2型,巨细胞病毒(CMV),水痘一带状疱疹病毒(VZV)及EBV均不相同。后来发现HHV-6是幼儿急疹(ES)的病原体。本文就HHV-6的血清流行病学,从ES患儿中的分离检测,与HHV— 6感染有关的疾病及其病毒学特性等综述如下。     

人类疱疹病毒8型K1和K15基因致病机制的研究进展

人类疱疹病毒8型K1和K15是HHV-8基因组所特有的基因,K1和K15基因结构的独特性决定了其在疾病的致病机制中发挥着一定的作用;近年来有关K1和K15基因致病机制的研究较多,本文结合近几年的文献将时K1和K15基因致病机制做一综述.     

人疱疹病毒8型KS330基因片段的检出与Kaposi肉瘤的关系

HHV-8 sequences were recently identified in 100% of the amplifiable samples from AIDS patients with Kaposi's sarcoma(KS)and in 15% of the non-KS tissue samples from AIDS patients, so there is a strong correlation of Kaposi's sarcoma with HHV-8. Serum and DNA samples from a clinically diagnosed Kaposi's sarcoma Chinese patient were tested. HHV-8 antibody was tested positive by IFA and HIV-I antibody was negative by Western blot. The KS330 PCR product was found both in peripheral blood mononuclear cells and in KS tumor cells from this Chinese patient. This supports the hypothesis that Kaposi's sarcoma results from infection of HHV-8.     

HIV-1 Tat蛋白对人类疱疹病毒8型复制的影响

用HindⅢ将HIV-1Tat101蛋白编码基因从pEV质粒中切出,BamHI、NotⅠ将绿色荧光蛋白(GFP)编码基因从表达质粒pcDNA3．1 ／GFP中切出,分别插入到质粒LZRSpBMN-Z中,构建成重组反转录病毒表达质粒LZRS—Tat101和LZRS—GFP。采用磷酸钙转染法将两重组质粒转染到含反转录病毒env,gal和pol编码基因的包装细胞Phoenix(φNX)中,嘌呤霉素筛选获得稳定细胞系。分别收集稳定细胞系分泌的病毒上清,并感染体外培养的原发性渗出性淋巴瘤(PEL)BC2BL-1细胞。收集LZRS—GFP重组病毒感染的BCBL-1细胞进行流式细胞计数,检测GFP表达水平。收集LZRS—Tat101重组病毒感染的BCBL-1细胞,提取蛋白作Western blot,检测Tat蛋白表达状况;取细胞总RNA作Northem blot和定量PCR,检查HHV-8次要衣壳蛋白ORF26 mRNA转录水平。重组LZRS—Tat101病毒进一步感染HL3T1细胞(HeLa细胞包含HIV-1-LTR／CAT报告基因),收集感染细胞提取蛋白,检测CAT活性,评价Tat生物学功能。PCR扩增HHV-8复制和转录激活蛋白Rta启动子区上游序列,并克隆至pGL-3载体中,构建Rta启动子 虫荧光素酶(Luciferase)报告基因重组质粒。此重组质粒进一步电转染预先感染了LZRS—Tat101病毒的BC-3细胞,TPA刺激后收集细胞,检测Luciferase活性。结果显示：①重组反转录病毒感染BCBL-1细胞,一次感染效率达56％;②重组LZRS—Tat101毒能够在其感染的BCBL-1细胞中表达Tat蛋白,且表达蛋白具有转录激活功能;③Tat蛋白不能有效上调HHV-8Rta启动子活性;④细胞内HIV-1Tat蛋白诱导HHV-8可溶性周期复制的能力较弱。提示,单纯HIV-1Tat蛋白并不能激活潜伏感染的HHV-8。     

人类疱疹病毒7型的研究进展

人类疱疹病毒７型（ＨＨＶ－７）是近期发现的又一种新型人类疱疹病毒。本文介绍该病毒的分离,病毒对ＣＤ４＾＋Ｔ细胞免疫的影响,病毒和人类免疫缺陷病毒（ＨＩＶ）之间的干扰现象及致病性方面的研究进展。     

一种新的人类疱疹病毒:Kaposi's肉瘤相关疱疹病毒(KSHV)

1994年Y.Chang等人从AIDS患者的Kaposi's肉瘤(KS)组织中分离到了两种独有的序列[1],并由KS建立了基因组文库,对其中外源序列进行克隆和序列分析[2],初步确认一种新的人类疱疹病毒--KS相关疱疹病毒.     

人类疱疹病毒8型ORF75基因亚型在鳞癌中的分布

目的:明确鳞癌感染人类疱疹病毒8型(HV-8)ORF75基因亚型分类并初步探讨其与鳞癌的相关性。方法:对40例皮肤鳞癌、46例食管鳞癌石蜡包埋组织进行HHV-8 DNA抽提、扩增、双向测序,使用DNASTAR软件、Crustal W软件和PHYLIP软件包对测序结果进行系统发生学分析,从而确定HHV-8 ORF75基因亚型,最后运用Fisher确切概率法对结果进行统计学分析。结果:①40例皮肤鳞癌有10例HHV-8感染为阳性,阳性率为25.0%;46例食管鳞癌有12例为阳性,阳性率为26.1%.②皮肤鳞癌感染HHV-8 ORF75亚型6例为A亚型,4例为C亚型;食管鳞癌感染HHV-8 ORF75亚型8例为A亚型,4例为C亚型③鳞癌感染HHV-8 ORF75基因亚型与鳞癌患者感染部位及病理分级之间没有统计学意义(P>0.05)。结论:鳞癌患者感染HHV-8 ORF75亚型属于A亚型和C亚型;鳞癌感染HHV-8 ORF75基因亚型与鳞癌患者感染部位及病理分级无关。     

Reciprocal regulatory interaction between human herpesvirus 8 and human immunodeficiency virus type 1

Human herpesvirus 8 (HHV8) is the primary viral etiologic agent in Kaposi's sarcoma (KS). However, individuals dually infected with both HHV8 and human immunodeficiency virus type 1 (HIV-1) show an enhanced prevalence of KS when compared with those singularly infected with HHV8. Host immune suppression conferred by HIV infection cannot wholly explain this increased presentation of KS. To better understand how HHV8 and HIV-1 might interact directly in the pathogenesis of KS, we queried for potential regulatory interactions between the two viruses. Here, we report that HHV8 and HIV-1 reciprocally up-regulate the gene expression of each other. We found that the KIE2 immediate-early gene product of HHV8 interacted synergistically with Tat in activating expression from the HIV-1 long terminal repeat. On the other hand, HIV-1 encoded Tat and Vpr proteins increased intracellular HHV8-specific expression. These results provide molecular insights correlating coinfection with HHV8 and HIV-1 with an unusually high incidence of KS.     

The relationship between tumor promoter binding and epstein-barr virus induction in human lymphoblastoid cell lines

The tumor promoter TPA (12-0-tetradecanoyl-phorbol-13-acetate) efficiently induces the synthesis of Epstein-Barr virus (EBV) antigens in EBV-genome-harboring human lymphoblastoid cells. We attempted to obtain information on the binding of TPA to cells and on the relationship between TPA-binding and EBV induction by the use of tritiated TPA (³H-TPA). Our data show: (1) In the absence of cells TPA can bind to serum completely within 60 minutes. (2) Cells can compete for a proportion of serum-bound TPA. (3) Binding of TPA to cells reaches equilibrium within 60 minutes and is higher at 37° than 0°C. In the absence of serum, the rate of binding is about twice as high as in the presence of serum. (4) Dissociation of TPA from cells also seems to be rapid. When the cells are incubated with cold TPA after prior treatment with ³H-TPA, followed by washing, a much higher rate of release of labeled TPA is observed than in cultures to which fresh medium is added exclusively. Dissociation is higher in the presence of serum than in the absence of it. If radiolabeled cells are analyzed after serial washing, the rate of cell-associated/noncell-associated radioactivity indicates that the proportion of molecules required for EBV induction is dissociated rapidly.     

The relationship between tumor necrosis factor and human immunodeficiency virus gene expression in lymphoid tissue.

In tissue culture models of chronic human immunodeficiency virus type 1 (HIV-1) infection, cytokines such as tumor necrosis factor alpha (TNF-alpha) activate viral gene expression. We sought evidence that TNF-alpha might similarly regulate viral gene expression in vivo in the major lymphoid tissue (LT) reservoir. We used in situ hybridization, quantitative image analysis, and double-label techniques to compare cytokine and HIV-1 RNA levels in sections of tonsil and lymph node tissues obtained from individuals in early and later stages of HIV-1 infection. The levels of TNF-alpha gene expression in LT from HIV-1-infected an uninfected individuals were indistinguishable, and we found no correlation between TNF-alpha gene expression in LT and the level of HIV-1 gene expression in LT. There is thus little evidence that in vivo TNF-alpha significantly influences HIV production in LT.     

Extensive homology between the herpes simplex virus type 2 glycoprotein F gene and the herpes simplex virus type 1 glycoprotein C gene.

The region of the herpes simplex virus type 2 (HSV-2) genome which maps colinearly with the HSV-1 glycoprotein C (gC) gene has been cloned, and the DNA sequence of a 2.29-kilobase region has been determined. Contained within this sequence is a major open reading frame of 479 amino acids. The carboxyterminal three-fourths of the derived HSV-2 protein sequence showed a high degree of sequence homology to the HSV-1 gC amino acid sequence reported by Frink et al. (J. Virol. 45:634-647, 1983). The amino-terminal region of the HSV-2 sequence, however, showed very little sequence homology to HSV-1 gC. In addition, the HSV-1 gC sequence contained 27 amino acids in the amino-terminal region which were missing from the HSV-2 protein. Computer-assisted analysis of the hydrophilic and hydrophobic properties of the derived HSV-2 sequence demonstrated that the protein contained structures characteristic of membrane-bound glycoproteins, including an amino-terminal signal sequence and carboxy-terminal hydrophobic transmembrane domain and charged cytoplasmic anchor. The HSV-2 protein sequence also contained seven putative N-linked glycosylation sites. These data, in conjunction with mapping studies of Para et al. (J. Virol. 45:1223-1227, 1983) and Zezulak and Spear (J. Virol. 49:741-747, 1984), suggest that the protein sequence derived from the HSV-2 genome corresponds to gF, the HSV-2 homolog of HSV-1 gC.     

Altered cytokine production after human herpes virus type 6 infection.

Several strategies allow viruses to elude the surveillance of the immune system and to establish persistent infection in the host. One of such mechanisms is the immunosuppression caused by the direct infection and functional impairment of immune cells. Human Herpes virus type 6 (HHV-6) is a typical immunosuppressive agent, as suggested by its tropism for both CD4+ and CD8+ T cells, B cells, monocytes/macrophages, megakaryocytes and NK cells. In this study the production of IL-10 and IL-12 by peripheral blood mononuclear cells (PBMC) was evaluated during HHV-6 infection "in vitro". Our results demonstrate that HHV-6 up-regulates IL-10 production by PBMC. Furthermore, our data suggest that rhIFN gamma addition counteracts the effect of HHV-6 in promoting IL-10 release. To gain more insight into the role of IFN gamma, anti-IFN gamma monoclonal antibodies were added to PBMC stimulated with LPS. Neutralization of endogenous IFN gamma upregulated IL-10 release. Furthermore, HHV-6 infection inhibited IFN gamma release induced by LPS in PBMC. No basal production of IL-12 was found in PBMC. Moreover, HHV-6 infection did not induce IL-12 release by PBMC. On the contrary, IL-12 was detected in the supernatants of PBMC treated with LPS with or without rhIFN gamma. In these experimental conditions the further addition of HHV-6 markedly impaired IL-12 production. Moreover, the neutralization of IL-10 resulted in a significant up-regulation of IL-12. Finally our data suggest that the immunodysregulation induced by HHV-6 could be accounted for by a shift from a Th-1 to a Th-2 type cytokine profile.     

Accessory genes define the relationship between the herpes simplex virus and its host

Accessory genes of herpes simplex virus are implicated in the interplay between the virus and host responses to infection, ensuring the survival of the virus in the host and maintaining its transmission cycle in human populations. They will provide vital clues about novel vaccination strategies and gene and oncolytic therapies.