Heterogeneity and Breadth of Host Antibody Response to KSHV Infection Demonstrated by Systematic Analysis of the KSHV Proteome

Labò, Nazzarena; Miley, Wendell; Marshall, Vickie; Gillette, William; Esposito, Dominic; Bess, Matthew; Turano, Alexandra; Uldrick, Thomas S.; Polizzotto, Mark N.; Wyvill, Kathleen M.; Bagni, Rachel; Yarchoan, Robert; Whitby, Denise

doi:10.1371/journal.ppat.1004046

Cited by 60 publications

(72 citation statements)

References 50 publications

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“…Recent studies revealed generally strong antibody responses to ORF38 among KSHV-infected patients (59,60). In apparent agreement with this high immunogenicity in humans, most hybridoma clones recognized a strong and specific signal of ϳ10 kDa in KSHV-infected cells (Fig.…”

Section: Discussionsupporting

confidence: 74%

ORF33 and ORF38 of Kaposi's Sarcoma-Associated Herpesvirus Interact and Are Required for Optimal Production of Infectious Progeny Viruses

Avey

et al. 2016

J Virol

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We recently showed that the interaction between Kaposi's sarcoma-associated herpesvirus (KSHV) tegument proteins ORF33 and ORF45 is crucial for progeny virion production, but the exact functions of KSHV ORF33 during lytic replication were unknown (J. Gillen, W. Li, Q. Liang, D. Avey, J. Wu, F. Wu, J. Myoung, and F. Zhu, J Virol 89:4918 -4931, 2015, http://dx.doi.org/10 .1128/JVI.02925-14). Therefore, here we investigated the relationship between ORF33 and ORF38, whose counterparts in both alpha-and betaherpesviruses interact with each other. Using specific monoclonal antibodies, we found that both proteins are expressed during the late lytic cycle with similar kinetics and that both are present in mature virions as components of the tegument. Furthermore, we confirmed that ORF33 interacts with ORF38. Interestingly, we observed that ORF33 tightly associates with the capsid, whereas ORF38 associates with the envelope. We generated ORF33-null, ORF38-null, and double-null mutants and found that these mutants apparently have identical phenotypes: the mutations caused no apparent effect on viral gene expression but reduced the yield of progeny virion by about 10-fold. The progeny virions also lack certain virion component proteins, including ORF45. During viral lytic replication, the virions associate with cytoplasmic vesicles. We also observed that ORF38 associates with the membranes of vesicles and colocalizes with the Golgi membrane or early endosome membrane. Further analyses of ORF33/ORF38 mutants revealed the reduced production of virion-containing vesicles, suggesting that ORF33 and ORF38 are involved in the transport of newly assembled viral particles into cytoplasmic vesicles, a process important for viral maturation and egress. IMPORTANCEHerpesvirus assembly is an essential step in virus propagation that leads to the generation of progeny virions. It is a complicated process that depends on the delicate regulation of interactions among virion proteins. We previously revealed an essential role of ORF45-ORF33 binding for virus assembly. Here, we report that ORF33 and its binding partner, ORF38, are required for infectious virus production due to their important role in the tegumentation process. Moreover, we found that both ORF33 and ORF38 are involved in the transportation of virions through vesicles during maturation and egress. Our results provide new insights into the important roles of ORF33 and ORF38 during viral assembly, a process critical for virus propagation that is intimately linked to KSHV pathobiology.

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Section: Discussionsupporting

confidence: 74%

ORF33 and ORF38 of Kaposi's Sarcoma-Associated Herpesvirus Interact and Are Required for Optimal Production of Infectious Progeny Viruses

Avey

et al. 2016

J Virol

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“…If the protein were expressed as predicted, it is not clear what effect a K8.1-deficient virion and/or virion-independent, soluble K8.1 would have in the context of an in vivo infection. K8.1 is highly immunogenic compared to other KSHV proteins (37), and exogenous expression of soluble K8.1 induces an interferon response (38). Together, the ZM124 predicted K8.1 may elicit a modified innate and humoral immune response.…”

Section: Discussionmentioning

confidence: 99%

Whole-Genome Sequencing of Kaposi's Sarcoma-Associated Herpesvirus from Zambian Kaposi's Sarcoma Biopsy Specimens Reveals Unique Viral Diversity

Olp

Jeanniard

Marimo

et al. 2015

J Virol

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Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiological agent for Kaposi's sarcoma (KS). Both KSHV and KS are endemic in sub-Saharan Africa where approximately 84% of global KS cases occur. Nevertheless, whole-genome sequencing of KSHV has only been completed using isolates from Western countries-where KS is not endemic. The lack of whole-genome KSHV sequence data from the most clinically important geographical region, sub-Saharan Africa, represents an important gap since it remains unclear whether genomic diversity has a role on KSHV pathogenesis. We hypothesized that distinct KSHV genotypes might be present in sub-Saharan Africa compared to Western countries. Using a KSHV-targeted enrichment protocol followed by Illumina deep-sequencing, we generated and analyzed 16 unique Zambian, KS-derived, KSHV genomes. We enriched KSHV DNA over cellular DNA 1,851 to 18,235-fold. Enrichment provided coverage levels up to 24,740-fold; therefore, supporting highly confident polymorphism analysis. Multiple alignment of the 16 newly sequenced KSHV genomes showed low level variability across the entire central conserved region. This variability resulted in distinct phylogenetic clustering between Zambian KSHV genomic sequences and those derived from Western countries. Importantly, the phylogenetic segregation of Zambian from Western sequences occurred irrespective of inclusion of the highly variable genes K1 and K15. We also show that four genes within the more conserved region of the KSHV genome contained polymorphisms that partially, but not fully, contributed to the unique Zambian KSHV whole-genome phylogenetic structure. Taken together, our data suggest that the whole KSHV genome should be taken into consideration for accurate viral characterization. IMPORTANCEOur results represent the largest number of KSHV whole-genomic sequences published to date and the first time that multiple genomes have been sequenced from sub-Saharan Africa, a geographic area where KS is highly endemic. Based on our new sequence data, it is apparent that whole-genome KSHV diversity is greater than previously appreciated and differential phylogenetic clustering exists between viral genomes of Zambia and Western countries. Furthermore, individual genes may be insufficient for KSHV genetic characterization. Continued investigation of the KSHV genetic landscape is necessary in order to effectively understand the role of viral evolution and sequence diversity on KSHV gene functions and pathogenesis. K aposi's sarcoma-associated herpesvirus (KSHV), or human herpesvirus 8 (HHV-8), is the etiologic agent for all forms of Kaposi's sarcoma (KS) (1). KS manifests as an endothelial tumor primarily on the skin but can also involve mucosal membranes and visceral organs. Among the HIV-uninfected population, KS is rare worldwide; however, HIV infection and immunosuppression greatly increase the risk of developing KS (2). In sub-Saharan Africa, HIV is epidemic, and KSHV is endemic. Accordingly, KS is one of the most common cancers in sub-Saharan A...

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“…Serum IgG against synthetic peptides covering immunodominant epitopes of the viral capsid antigen P18 (VCA), EBV nuclear antigen 1 (EBNA1), diffuse early antigen complex (EAd) and against the EBV-encoded recombinant glycoprotein 350/220 (gp350) 10 were measured using a multiplexed beadbased assay as previously described, 11 at a 1/200 serum dilution. Background-subtracted median fluorescent intensity levels (MFI) were log10 transformed for subsequent analyses.…”

Section: Serologymentioning

confidence: 99%

Mutual detection of Kaposi's sarcoma‐associated herpesvirus and Epstein–Barr virus in blood and saliva of Cameroonians with and without Kaposi's sarcoma

Labò

Marshall

Miley

et al. 2019

Intl Journal of Cancer

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Kaposi's sarcoma‐associated herpesvirus (KSHV) and Epstein–Barr virus (EBV) are prevalent in sub‐Saharan Africa, together with HIV; the consequent burden of disease is grave. The cofactors driving transmission of the two viruses and pathogenesis of associated malignancies are not well understood. We measured KSHV and EBV DNA in whole blood and saliva as well as serum antibodies levels in 175 Cameroonians with Kaposi's sarcoma and 1,002 age‐ and sex‐matched controls with and without HIV. KSHV seroprevalence was very high (81%) in controls, while EBV seroprevalence was 100% overall. KSHV DNA was detectable in the blood of 36–46% of cases and 6–12% of controls; EBV DNA was detected in most participants (72–89%). In saliva, more cases (50–58%) than controls (25–28%) shed KSHV, regardless of HIV infection. EBV shedding was common (75–100%); more HIV+ than HIV− controls shed EBV. Cases had higher KSHV and EBV VL in blood and saliva then controls, only among HIV+ participants. KSHV and EBV VL were also higher in HIV+ than in HIV− controls. Cases (but not controls) were more likely to have detectable KSHV in blood if they also had EBV, whereas shedding of each virus in saliva was independent. While EBV VL in saliva and blood were modestly correlated, no correlation existed for KSHV. Numerous factors, several related to parasitic coinfections, were associated with detection of either virus or with VL. These findings may help better understand the interplay between the two gammaherpesviruses and generally among copathogens contributing to cancer burden in sub‐Saharan Africa.

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Heterogeneity and Breadth of Host Antibody Response to KSHV Infection Demonstrated by Systematic Analysis of the KSHV Proteome

Cited by 60 publications

References 50 publications

ORF33 and ORF38 of Kaposi's Sarcoma-Associated Herpesvirus Interact and Are Required for Optimal Production of Infectious Progeny Viruses

ORF33 and ORF38 of Kaposi's Sarcoma-Associated Herpesvirus Interact and Are Required for Optimal Production of Infectious Progeny Viruses

Whole-Genome Sequencing of Kaposi's Sarcoma-Associated Herpesvirus from Zambian Kaposi's Sarcoma Biopsy Specimens Reveals Unique Viral Diversity

Mutual detection of Kaposi's sarcoma‐associated herpesvirus and Epstein–Barr virus in blood and saliva of Cameroonians with and without Kaposi's sarcoma

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