Genotypic analysis on the ORF-K1 gene of human herpesvirus 8 from patients with Kaposi's sarcoma in Xinjiang,China

Human herpesvirus 8 （HHV-8） is thought to be essential for the development of all forms of Kaposi＇s sarcoma （KS）. HHV-8 DNA is present virtually in all KS tumor biopsy samples. Genes at both ends of the HHV-8 genome have been shown to vary considerably. Seven major molecular subtypes of HHV-8 were defined based on the amino acid sequence of the open reading frame K1 （ORF-K1）, generally known as A, B, C, D, E, F, and Z. Most strains collected worldwide were clustered into two subtypes （A and C）. Here, the K1/VRI region of HHV-8 was amplified by nested PCR in 22 （81.48%） of 27 cases from Xinjiang Uygur Autonomous Region, a province in northwestern China. Phylogenetic analysis on the basis of the K1/VR1 amino acid sequence indicated that the majority of these KS patients were infected by subtype C HHV-8 （n = 18, including 15 belonging to the C2 group）, and several by subtype A （n = 4, including 3 being the A1 group）. This is the first report of subtype A HHV-8 in China. Furthermore, the correlations between different forms and lesions of KS and different subtypes of HHV-8 were analyzed. The findings showed that subtype A HHV-8 resulted in significantly more frequent mucosal KS lesions than subtype C. However, there was no obvious correlation between different forms of KS and different subtypes of HHV-8.

Human herpesvirus-8 positive iatrogenic Kaposi's sarcoma in the setting of refractory ulcerative colitis

Although Kaposi sarcoma(KS) has been more traditionally considered an AIDS-defining illness,it may also be seen in individuals on immunosuppresive therapy.We report a case of a patient who presented to the hospital in the setting of increasingly refractory ulcerative colitis.Computed tomography scan of the abdomen was consistent with sigmoid diverticulititis and blood cultures were positive for Klebsiella.After a course of antibiotics with resolution of infection,a colonoscopy was performed to evaluate his diverticulitis and incidentally revealed a new rectal tumor.Immunohistochemistry showed the tumor was consistent with KS,with cells staining strongly positive for human herpesvirus-8.This case not only illustrates a rare case of KS found in an HIV-negative individual,but it also highlights the importance of considering an alternative diagnosis in a patient refractory to medical treatment.We discuss the management and care of an ulcerative colitis patient diagnosed with KS on immunosuppressive therapy.

The Role of Human Herpesvirus 8 Molecular Characterization in the Management of HIV Infected Patients Diagnosed with Malignancies Associated with Its Infection

Despite the progress has been reached with Human herpesvirus 8 (HHV-8) research, there are gaps in the knowledge of viral induced oncogenesis. The aim of the present study was to identify possible associations between HHV-8 subtypes, HHV-8 loads and clinical manifestations of HIV infected patients diagnosed with different malignancies associated with HHV-8 infection. Forty six HIV-1 infected individuals diagnosed with different HHV-8 associated diseases were studied [37 epidemic Kaposi’s sarcoma (KS), 3 pleural effusion lymphoma (PEL);5 peripheral lymphadenopathies (PL);1 Hodgkin’s lymphoma (HL);1 non Hodgkin’s lymphoma (NHL)]. HHV-8 loads were determined by quantitative real time PCR (qRT-PCR) whilst HHV-8 subtypes were determined by open-reading frame (ORF)-K1 gen genotyping. HHV-8 subtypes B, A, C, A5 and E were exhibited by 31.8%, 23.4%, 19.1%, 17% and 8.5% of the studied patients, respectively. The median HHV-8 viral load did not differ between subtypes (p > 0.05) but HHV-8 viral loads were significantly higher in PEL than in epidemic KS lesion or lymph nodes (p = 0.04). Subtype B was detected in 60% of patients with B cell lymphoma (NHL, PEL and HL) whereas subtype E was only detected in patients with epidemic KS diagnosis. Our data suggest that HHV-8 DNA quantification instead of subtype identification could be used as a surrogate marker for monitoring its infection, not only in epidemic KS patients but also in HIV infected individuals with lymphoproliferative disorders.

Regulation of Wnt/β-catenin signaling by herpesviruses

The Wnt/β-catenin signaling pathway is instrumental in successful differentiation and proliferation of mammalian cells. It is therefore not surprising that the herpesvirus family has developed mechanisms to interact with and manipulate this pathway. Successful coexistence with the host requires that herpesviruses establish a lifelong infection that includes periods of latency and reactivation or persistence. Many herpesviruses establish latency in progenitor cells and viral reactivation is linked to host-cell proliferation and differentiation status. Importantly, Wnt/β-catenin is tightly connected to stem/progenitor cell maintenance and differentiation. Numerous studies have linked Wnt/β-catenin signaling to a variety of cancers, emphasizing the importance of Wnt/β-catenin pathways in development, tissue homeostasis and disease. This review details how the alpha-, beta-, and gammaherpesviruses interact and manipulate the Wnt/β-catenin pathway to promote a virus-centric agenda.

The replication and transcription activator (RTA) of Kaposi's sarcoma-associated herpesvirus/human herpesvirus-8

Kaposi’s sarcoma-associated herpesvirus(KSHV)is γ-2 herpesvirus with latency and lytic replication stages in its life-cycle.The viral replication and transcription activator(RTA)is the key protein for triggering KSHV lytic gene expression and replication from latency.In this review,we will discuss the gene expression program in KSHV lytic replication and latency,the regulation of the RTA expression,the RTA protein and the mechanisms that RTA utilizes to transactivate its target genes.We will focus on the RTA-mediated transactivation mechanisms,including DNA-binding,interacting with cellular co-factors and promoting repressor degradation.

新疆Kaposi肉瘤组织内EBV、HHV-8双重感染的调查

应用PCR方法，对20例新疆Kaposi肉瘤病理组织进行了EBV和HHV－8双重感染的调查。结果：20例Kaposi肉瘤病理组织中14例检出HHV－8DNA（70％），EBV均为阴性。正常皮肤对照：10例这两种疱疹类病毒均为阴性。作者认为新疆Kaposi肉瘤的发生与EBV的相关性很小，但明显与HHV－8感染有关，但是否HHV－8感染就是新疆Kaposi肉瘤发生的决定因素，仍需进一步研究。

人类疱疹病毒8型k12/T0.7基因亚型与Kaposi肉瘤的相关性研究

目的:了解Kaposi肉瘤患者所感染的疱疹病毒8型(HHV-8)k12/T0.7基因亚型分类,初步探索k12/T0.7基因亚型与Kaposi肉瘤不同临床分型的相关性。方法:对30例病理诊断明确的Kaposi肉瘤石蜡包埋组织进行HHV-8 DNA提取、k12/T0.7基因扩增、双向测序。使用DNAMAN软件、ClustalW软件和PHYLIP软件包对测序结果进行系统发生学分析,确定HHV-8 k12/T0.7基因亚型。最后用Fisher确切概率法对结果进行统计学分析。结果:30例Kaposi肉瘤中有22例HHV-8阳性,阳性率为73.33%,其中7例艾滋病相关型患者HHV-8均阳性。30例HHV-8阳性患者中,10例为HHV-8 k12 A亚型,12例为A/C亚型。Kaposi肉瘤患者不同亚型间有无黏膜损害及临床分型的分布差异均无统计学意义(P>0.05)。结论:Kaposi肉瘤患者感染HHV-8 k12亚型以A亚型和A/C亚型为主,不同亚型与临床分型无关。

Kaposi肉瘤组织内人疱疹病毒8型的K14.1基因型研究

目的:确定人疱疹病毒8型K14.1基因等位基因型在Kaposi肉瘤中的分布,及与Kaposi肉瘤临床分型的关系。方法:使用酚-氯仿-异戊醇法对32例Kaposi肉瘤石蜡包埋组织进行病毒DNA提取,使用PCR法扩增K14.1基因片段,然后确定其等位基因型。结果:32例中有27例人疱疹病毒8型感染为阳性,阳性率为84.38%,其中5例艾滋病相关型KS患者HHV-8感染均为阳性,感染率100%;在分析的27例HHV-8病毒株中,24例为P等位基因型(包括5例艾滋病相关型KS患者皮损),3例为M等位基因型(均来自经典型KS)。结论:本研究中,Kaposi肉瘤感染的人疱疹病毒8型的K14.1等位基因型以P等位基因型为主。

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.

新疆Kaposi肉瘤患者感染的人类疱疹病毒8型ORF75基因多态性研究

目的通过对人类疱疹病毒8型(HHV-8)ORF75基因多态性研究,初步探讨其与新疆Kaposi肉瘤的相关性。方法对25例新疆Kaposi肉瘤患者石蜡包埋组织进行HHV-8 DNA抽提、扩增及双向测序,使用CLUSTALW软件和PHYLIP软件包对ORF75基因进行核苷酸多态性分析。结果①25例新疆Kaposi肉瘤患者有21例HHV-8感染阳性,阳性率为84%,其中7例艾滋病相关型Kaposi肉瘤(AIDS-KS)患者HHV-8感染均为阳性。②新疆Kaposi肉瘤患者HHV-8 ORF75基因核苷酸具有多态性,存在462、618、647三个主要突变位点。结论人类疱疹病毒8型ORF75基因多态性与新疆Kaposi肉瘤具有相关性。

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.

Lipids, lipid metabolism and Kaposi's sarcoma-associated herpesvirus pathogenesis

Lipids are essential for mammalian cells to maintain many physiological functions. Emerging evidence has shown that cancer cells can develop specific alterations in lipid biosynthesis and metabolism to facilitate their survival and various malignant behaviors. To date, the precise role of cellular lipids and lipid metabolism in viral oncogenesis is still largely unclear with only a handful of literature covering this topic to implicate lipid metabolism in oncogenic virus associated pathogenesis. In this review, we focus on the role of lipid biosynthesis and metabolism in the pathogenesis of the Kaposi's sarcoma-associated herpesvirus, a common causative factor for cancers arising in the immunocompromised settings.

Prevalence of Kaposi's sarcoma-associated herpesvirus among intravenous drug users： a systematic review and meta-analysis

Intravenous drug users(IDUs) have been demonstrated to be highly vulnerable to HIV/AIDS.Nevertheless, the prevalence of Kaposi's sarcoma associated herpesvirus(KSHV), an important co-infected agent with HIV, among this population remained obscure. We conducted a systematic review on the epidemiological features of KSHV among IDUs worldwide. Eligible studies were retrieved from 6 electronic databases(Pub Med, EMBASE, Web of Science, CBM, CNKI and Wanfang).We calculated the pooled prevalence and 95% confidence interval(CI) overall and among subgroups using either random-effects model or fixed-effects model depending on between-study heterogeneity. The potential publication bias was assessed by the Egger's test. A meta-regression analysis was performed to explore the sources of heterogeneity. Finally, twenty-two studies with a total sample of 7881 IDUs were included in the analysis. The pooled prevalence of KSHV was14.71%(95% CI 11.12%–19.46%) among IDUs. Specifically, KSHV prevalence was 10.86%(95% CI6.95%–16.96%) in HIV-negative IDUs, and 13.56%(95% CI 10.57%–17.38%) in HIV-positive IDUs.Moreover, prevalence among IDUs from the three continents involved in the current study was similar:16.10%(95%CI 7.73%–33.54%) in Asia; 14.22%(95%CI 8.96%–22.57%) in Europe and 14.06%(95%CI11.38%–17.37%) in America. Globally, IDUs are at higher risk of the KSHV infection when compared with the general population, regardless of geographical region or HIV-infection status.

Establishment of method for seroepidermiological detection of human herpes virus 8 infection by using the fusion protein in the prokaryotic expression system as antigen for testing

To establish a sensitive and specific method for seroepidermiological detection of human her- pes virus 8(HHV-8)infection,three potent antigenic proteins encoded by open reading frames(ORFs) K8.1,65 and 73C in genome of HHV-8 were produced as glutathione S-transferase fusion protein in the prokaryotic expression system and was used as antigen for testing.The recombinant fusion protein ex- pressed in the prokaryotic expression vector E.coli BL21 was purified by glutathione Sepharose 4B affin- ity chromatography and was quantitated with SDS-PAGE.All these 3 fusion proteins produced in the pro- karyotic expression system showed good immunogenicity as demonstrated by Western blotting and could be recognized by mixed sera of patients with Kaposi′s sarcoma(KS).The immuno-reactivities of the single or compound fusion protein were determined by means of ELISA and compared with the traditional immu- nofluorescence assay(IFA)to determine their sensitivity and specificity of the test.It was demonstrated that the sensitivity of mixed-antigen ELISA method was significantly higher than that of IFA(81.8% vs 34.4%),while the specificity of the former was demonstrated to be 97.9%.The coincidence of the de- tection rate between these two methods was considerably high,approaching up to 90.0%.These results suggest that the mixed antigen ELISA assay appears to be a sensitive and specific method for sero-epide- miological detection of human herpesvirus 8 infection.

Diagnostic value of endothelial markers and HHV-8 staining in gastrointestinal Kaposi sarcoma and its difference in endoscopic tumor staging

AIM: To clarify the diagnostic values of hematoxylin and eosin (HE), D2-40, CD31, CD34, and HHV-8 immunohistochemical (IHC) staining in gastrointestinal Kaposi's sarcoma (GI-KS) in relation to endoscopic tumor staging. METHODS: Biopsy samples (n = 133) from 41 human immunodeficiency virus-infected patients were reviewed. GI-KS was defined as histologically negative for other GI diseases and as a positive clinical response to KS therapy. The receiver operating characteristic area under the curve (ROC-AUC) was compared in relation to lesion size, GI location, and macroscopic appearances on endoscopy. RESULTS: GI-KS was confirmed in 84 lesions (81.6%). Other endoscopic findings were polyps (n = 9), inflammation (n = 4), malignant lymphoma (n = 4), and condyloma (n = 2), which mimicked GI-KS on endoscopy. ROC-AUC of HE, D2-40, blood vessel markers, and HHV-8 showed results of 0.83, 0.89, 0.80, and 0.82, respectively. For IHC staining, the ROC-AUC of D2-40 was significantly higher (P < 0.05) than that of HE staining only. In the analysis of endoscopic appearance, the ROC-AUC of HE and IHC showed a tendency toward an increase in tumor staging (e.g. , small to large, patches, and polypoid to SMT appearance). D2-40 was significantly (P < 0.05) advantageous in the upper GI tract and for polypoid appearance compared with HE staining. CONCLUSION: The diagnostic value of endothelial markers and HHV-8 staining was found to be high, and its accuracy tended to increase with endoscopic tumor staging. D2-40 will be useful for complementing HE staining in the diagnosis of GI-KS, especially in the upper GI tract and for polypoid appearance.

Identification of human herpesvirus-8 in Kaposi＆#39;s sarcoma with bullous pemphigoid

Kaposi＆#39;s sarcoma （KS） is a rare,malignant vascular.tumor of the skin,mucosa,and viscera with an increased incidence in human immunodeficiency virus （HIV）-positive patients.According to the clinical characteristics and the affected population,four clinical types of KS have been described successively：classic,endemic to Africa,acquired immunodeficiency syndrome （AIDS）-associated,and immunosuppression associated type.Bullous pemphigoid （BP） is an acquired autoimmune bullous skin disease that frequently occurs in the elderly and is characterized by intraepidermal blisters and immunoglobulin G （IgG） deposits on the basement membrane.A few cases of KS have occurred in immunosuppressed patients with BP.Here,we report a Chinese female patient diagnosed with immunosuppression associated KS that was induced by the short-term use of immunosuppressive agents for BP.We also present a review of the literature on immunosuppressed KS with BP and discuss the potential role of human herpesvirus-8 （HHV-8） in the pathogenic mechanism.

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。１ＫＳＨＶ的发现Ｋａｐｏ...

福建地区经典型Kaposi肉瘤感染病毒的亚型研究

目的:研究福建地区经典型Kaposi肉瘤感染的病毒亚型。方法:收集经典型Kaposi肉瘤患者临床资料及石蜡包埋组织标本24例,利用QIAamp® DNA FFPE Tissue试剂盒提取患者皮损蜡块组织中的DNA,设计扩增人类疱疹病毒(HHV)-8(ORF-K1、T0.7/K12、ORF-K14.1/K15、ORF26及ORF75)基因的引物,巢式PCR扩增提取的DNA,然后进行双向测序,再采用SnapGene软件对测序结果进行确认,从而确定HHV-8各病毒亚型的分布情况。结果:24例福建地区经典型Kaposi肉瘤均来源于汉族人,皮损好发于老年人双下肢,以结节为主,患者感染HHV-8不同基因扩增阳性率:ORF26 100.00%、ORF-K133.33%、T0.7/K12 29.17%、ORF-K14.1/K15 33.33%、ORF75 25.00%。序列比对分析显示24例经典型Kaposi肉瘤患者感染HHV-8 ORF26的病毒亚型均为C亚型。结论:福建地区经典型Kaposi肉瘤感染HHV-8的基因型主要为ORF26,且均为C亚型。

Manipulation of the host cell membrane by humanγ-herpesviruses EBV and KSHV for pathogenesis

The cell membrane regulates many physiological processes including cellular communication,homing and metabolism. It is therefore not surprising that the composition of the host cell membrane is manipulated by intracellular pathogens. Among these, the human oncogenic herpesviruses Epstein–Barr virus(EBV) and Kaposi's sarcoma-associated herpesvirus(KSHV)exploit the host cell membrane to avoid immune surveillance and promote viral replication.Accumulating evidence has shown that both EBV and KSHV directly encode several similar membrane-associated proteins, including receptors and receptor-specific ligands(cytokines and chemokines), to increase virus fitness in spite of host antiviral immune responses. These proteins are expressed individually at different phases of the EBV/KSHV life cycle and employ various mechanisms to manipulate the host cell membrane. In recent decades, much effort has been made to address how these membrane-based signals contribute to viral tumorigenesis. In this review, we summarize and highlight the recent understanding of how EBV and KSHV similarly manipulate host cell membrane signals, particularly how remodeling of the cell membrane allows EBV and KSHV to avoid host antiviral immune responses and favors their latent and lytic infection.

Hepatic Kaposi sarcoma: A case report and review of the literature

Kaposi sarcoma(KS) is an aggressive cancer caused by human herpesvirus-8, primarily seen in immunocompromised patients. As opposed to the well-described cutaneous manifestations and pulmonary complications of KS, hepatic KS is rarely reported before death as most patients with hepatic KS do not manifest symptoms or evidence of liver injury. In patients with acquired immune deficiency syndrome, hepatic involvement of KS is present in 12%-24% of the population on incidental imaging and in approximately 35% of patients with cutaneous KS if an autopsy was completed after their death. Patients with clinically significant hepatic injury due to hepatic KS usually have an aggressive course of disease with hepatic failure often progressing to multi-organ failure and death. Here we report an unusual presentation of acute liver injury due to hepatic KS and briefly review the published literature on hepatic KS.