Comparison of Human Sera Reactivities in Immunoblots with Recombinant Human Herpesvirus (HHV)-8 Proteins Associated with the Latent (ORF73) and Lytic (ORFs 65, K8.1A, and K8.1B) Replicative Cycles and in Immunofluorescence Assays with HHV-8-Infected BCBL-1 Cells

Zhu, Liangjin; Wang, Rong; Sweat, Alisa; Goldstein, Elliot; Horvat, Rebecca T.; Chandran, Bala

doi:10.1006/viro.1999.9674

Cited by 70 publications

(68 citation statements)

References 25 publications

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“…The genesis of KSHV seroepidemiology came from studies employing easily cultured, uniformly KSHV-infected B-lymphoma cell lines (PEL cell lines) as the source of viral latent antigen (176,250,355). PELs have since served as the primary target for indirect immunofluorescent-antibody (IFA) approaches to measure levels of antibody to both latent and lytic KSHV proteins in clinical serum samples and have been successfully used as a source of protein extract in confirmatory immunoblots (93,175,176,250,290,343,355,474,545). When clinical sera are incubated with latently infected PEL cells or nuclei, they are scored positive for previous viral exposure if a characteristic punctate nuclear stain is observed (176,250,355).…”

Section: Clinical Detection Of Kshv and Estimates Of Infection In Thmentioning

confidence: 99%

Molecular Genetics of Kaposi's Sarcoma-Associated Herpesvirus (Human Herpesvirus 8) Epidemiology and Pathogenesis

Dourmishev

Palmeri

et al. 2003

Microbiol Mol Biol Rev

309

367

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Kaposi's sarcoma had been recognized as unique human cancer for a century before it manifested as an AIDS-defining illness with a suspected infectious etiology. The discovery of Kaposi's sarcoma-associated herpesvirus (KSHV), also known as human herpesvirus-8, in 1994 by using representational difference analysis, a subtractive method previously employed for cloning differences in human genomic DNA, was a fitting harbinger for the powerful bioinformatic approaches since employed to understand its pathogenesis in KS. Indeed, the discovery of KSHV was rapidly followed by publication of its complete sequence, which revealed that the virus had coopted a wide armamentarium of human genes; in the short time since then, the functions of many of these viral gene variants in cell growth control, signaling apoptosis, angiogenesis, and immunomodulation have been characterized. This critical literature review explores the pathogenic potential of these genes within the framework of current knowledge of the basic herpesvirology of KSHV, including the relationships between viral genotypic variation and the four clinicoepidemiologic forms of Kaposi's sarcoma, current viral detection methods and their utility, primary infection by KSHV, tissue culture and animal models of latent- and lytic-cycle gene expression and pathogenesis, and viral reactivation from latency. Recent advances in models of de novo endothelial infection, microarray analyses of the host response to infection, receptor identification, and cloning of full-length, infectious KSHV genomic DNA promise to reveal key molecular mechanisms of the candidate pathogeneic genes when expressed in the context of viral infection

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Section: Clinical Detection Of Kshv and Estimates Of Infection In Thmentioning

confidence: 99%

Molecular Genetics of Kaposi's Sarcoma-Associated Herpesvirus (Human Herpesvirus 8) Epidemiology and Pathogenesis

Dourmishev

Palmeri

et al. 2003

Microbiol Mol Biol Rev

309

367

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“…It has been shown that the transmembrane glycoprotein K8.1 is part of the viral envelope (27). K8.1 is highly immunogenic in the natural host (40) and is frequently used in HHV-8 serologic assays (26,49,60). The physiological function of K8.1 or the other rhadinoviral glycoproteins encoded at comparable genomic positions has not been identified so far.…”

mentioning

confidence: 99%

Cell Surface Heparan Sulfate Is a Receptor for Human Herpesvirus 8 and Interacts with Envelope Glycoprotein K8.1

Birkmann

Mahr

Ensser

et al. 2001

J Virol

158

159

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An immunodominant envelope glycoprotein is encoded by the human herpesvirus 8 (HHV-8) (also termedKaposi's sarcoma-associated herpesvirus) K8.1 gene. The functional role of glycoprotein K8.1 is unknown, and recognizable sequence homology to K8.1 is not detectable in the genomes of most other closely related gammaherpesviruses, such as herpesvirus saimiri or Epstein-Barr virus. In search for a possible function for K8.1, we expressed the ectodomain of K8.1 fused to the Fc part of human immunoglobulin G1 (K8.1⌬TMFc). K8.1⌬TMFc specifically bound to the surface of cells expressing glycosaminoglycans but not to mutant cell lines negative for the expression of heparan sulfate proteoglycans. Binding of K8.1⌬TMFc to mammalian cells could be blocked by heparin. Interestingly, the infection of primary human endothelial cells by HHV-8 could also be blocked by similar concentrations of heparin. The specificity and affinity of these interactions were then determined by surface plasmon resonance measurements using immobilized heparin and soluble K8.1. This revealed that K8.1 binds to heparin with an affinity comparable to that of glycoproteins B and C of herpes simplex virus, which are known to be involved in target cell recognition by binding to cell surface proteoglycans, especially heparan sulfate. We conclude that cell surface glycosaminoglycans play a crucial role in HHV-8 target cell recognition and that HHV-8 envelope protein K8.1 is at least one of the proteins involved.

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“…The cDNAs encoding ORF65 and ORF73 were previously identified by screening a cDNA library from TPA-induced BCBL-1 cells with serum from a patient positive for human immunodeficiency virus and KSHV (1,30). They were then expressed as a glutathione S-transferase (GST) fusion protein in a baculovirus system, purified, and injected into rabbits.…”

Section: Methodsmentioning

confidence: 99%

Three-Dimensional Localization of pORF65 in Kaposi's Sarcoma-Associated Herpesvirus Capsid

Atanasov

et al. 2003

J Virol

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Of the six herpesvirus capsid proteins, the smallest capsid proteins (SCPs) share the least sequence homology among herpesvirus family members and have been implicated in virus specificity during infection. The herpes simplex virus-1 (HSV-1) SCP was shown to be horn shaped and to specifically bind the upper domain of each major capsid protein in hexons but not in pentons. In Kaposi's sarcoma-associated herpesvirus (KSHV), the protein encoded by the ORF65 gene (pORF65) is the putative SCP but its location remains controversial due to the absence of such horn-shaped densities from both the pentons and hexons of the KSHV capsid reconstructions. To directly locate the KSHV SCP, we have used electron cryomicroscopy and threedimensional reconstruction techniques to compare the three-dimensional structure of KSHV capsids to that of anti-pORF65 antibody-labeled capsids. Our difference map shows prominent antibody densities bound to the tips of the hexons but not to pentons, indicating that KSHV SCP is attached to the upper domain of the major capsid protein in hexons but not to that in pentons, similar to HSV-1 SCP. The lack of horn-shaped densities on the hexons indicates that KSHV SCP exhibits structural features that are substantially different from those of HSV-1 SCP. The location of SCP at the outermost regions of the capsid suggests a possible role in mediating capsid interactions with the tegument and cytoskeletal proteins during infection.Herpesvirus virions share a characteristic architecture in which the double-stranded DNA genome is surrounded by an icosahedral protein capsid, a thick tegument layer, and a lipid bilayer envelope (15). The capsid, approximately 1,300 Å in diameter, is a T ϭ 16 icosahedron with 12 pentons forming the vertices, 150 hexons forming the faces and edges, and 320 triplexes interconnecting the pentons and hexons (15,17). These structural features of the capsid are built from four of the six capsid proteins. In Kaposi's sarcoma-associated herpesvirus (KSHV), a gammaherpesvirus, the pentons and hexons are composed of five and six copies, respectively, of the major capsid protein (MCP), pORF25. The triplexes are heterotrimers containing a monomer of the pORF62 protein and a dimer of the pORF26 protein (20, 23). The fourth and smallest capsid protein (SCP), pORF65, is homologous to the SCPs in other herpesviruses, including the structurally well-characterized herpes simplex type 1 (HSV-1) and cytomegalovirus (CMV), representative members of the alpha-and betaherpesvirus subfamilies, respectively. However, of the six capsid proteins, SCPs share the lowest sequence homology between HSV-1, human CMV (HCMV), and KSHV (23). This low sequence homology of SCPs may be related to their virus-specific functional roles. It has been recently shown that the HCMV UL48.5-encoded SCP is essential for HCMV infection in vivo (3), but its HSV-1 counterpart, VP26, is dispensable for HSV-1 infection (5,8). While other proteins making up the capsid shell have been shown to have very similar structures located ...

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Comparison of Human Sera Reactivities in Immunoblots with Recombinant Human Herpesvirus (HHV)-8 Proteins Associated with the Latent (ORF73) and Lytic (ORFs 65, K8.1A, and K8.1B) Replicative Cycles and in Immunofluorescence Assays with HHV-8-Infected BCBL-1 Cells

Cited by 70 publications

References 25 publications

Molecular Genetics of Kaposi's Sarcoma-Associated Herpesvirus (Human Herpesvirus 8) Epidemiology and Pathogenesis

Molecular Genetics of Kaposi's Sarcoma-Associated Herpesvirus (Human Herpesvirus 8) Epidemiology and Pathogenesis

Cell Surface Heparan Sulfate Is a Receptor for Human Herpesvirus 8 and Interacts with Envelope Glycoprotein K8.1

Three-Dimensional Localization of pORF65 in Kaposi's Sarcoma-Associated Herpesvirus Capsid

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