Comparison of Humoral Immune Responses to Epstein-Barr Virus and Kaposi’s Sarcoma–Associated Herpesvirus Using a Viral Proteome Microarray

Zheng, Dasheng; Wan, Jun; Cho, Yong Gu; Wang, Leyao; Chiou, Chuang Jiun; Pai, Shweta; Woodard, Crystal; Zhu, Jian; Liao, Gangling; Martı́nez-Maza, Otoniel; Jiang, Qian; Zhu, Heng; Hayward, Gary S.; Ambinder, Richard F.; Hayward, S. Diane

doi:10.1093/infdis/jir645

Cited by 32 publications

(41 citation statements)

References 51 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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“…The results with the 19 and 12 kD antigens need to be confirmed with an independent method, such as ELISA with recombinant protein. The search for the most relevant EBV antigen for MS should also be extended using additional broad and unbiased methods, such as the recently developed EBV protein microarray (Zheng et al, 2011). This will lead to a better understanding of the role of EBV in MS. 6.…”

Section: Discussionmentioning

confidence: 99%

“…Although measuring responses to EBNA, VCA, and EA is useful for the diagnosis and staging of EBV infection, it is not clear whether these are the main targets of the antibody response for either normal subjects or MS patients. A recent study using a protein microarray covering the entire EBV proteome suggests that healthy subjects have strong antibody responses to multiple EBV proteins (Zheng et al, 2011), including many which have received little attention in research or clinical use.…”

Section: Introductionmentioning

confidence: 99%

The antibody response to Epstein–Barr virions is altered in multiple sclerosis

Lindsey

Khan

Ansari

et al. 2013

Journal of Neuroimmunology

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“…Discordance between these assays and the lack of "gold standard" reference samples have hindered the understanding of the prevalence, transmission, and natural history of KSHV infection (8). Furthermore, studies have shown that the humoral response to viruses such as KSHV is complex (15), and appropriate serological assays may require approaches that detect multiple antigens simultaneously. Other proteins, such as the complement inhibitory protein (KCP) and viral cyclin, have been shown to be useful targets for serological screening of KSHV infections (14,41).…”

Section: Figmentioning

confidence: 99%

“…Low viral loads in blood or saliva limit the ability of even sensitive PCR-based approaches to be used for diagnosis (11,12). Several multiantigen tests have been recently developed in order to have a wide-based screen for serological evidence of virus infection (13)(14)(15).…”

mentioning

confidence: 99%

Development of Whole-Virus Multiplex Luminex-Based Serological Assays for Diagnosis of Infections with Kaposi's Sarcoma-Associated Herpesvirus/Human Herpesvirus 8 Homologs in Macaques

Ryan¹,

Rose

2013

Clin. Vaccine Immunol.

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bKaposi's sarcoma-associated herpesvirus (KSHV)/human herpesvirus 8 is a tumorigenic rhadinovirus that is associated with all forms of Kaposi's sarcoma. Current serological detection of KSHV is based on enzyme-linked immunosorbent or immunofluorescence assays that suffer from a variety of problems, including the lack of defined standards for test comparison. While KSHV is the only known human rhadinovirus, two lineages of KSHV-like rhadinoviruses are found in Old World primates: the RV1 lineage includes KSHV and retroperitoneal fibromatosis herpesvirus (RFHV) in macaques, and the RV2 lineage includes RRV and MneRV2 from different macaque species. To develop animal models of KSHV-associated diseases, we developed quantitative multiplex bead-based serological assays to detect antibodies against rhadinovirus antigens. Proteins from KSHV (RV1) and MneRV2 (RV2) virions were coupled to spectrally distinct fluorescent beads and used in Luminex flow cytometry-based assays to detect immune responses in macaques. Both assays showed large dynamic ranges with high levels of seroreactivity to both KSHV and MneRV2 proteins. A large set of macaque serum samples from the Washington National Primate Research Center was screened, and most of the samples (82%) were positive in both assays, consistent with the high level of RV1-RV2 coinfection detected by PCR. The macaque sera showed broad, variable, and unique serological responses to the different viral antigens, allowing an initial seroprevalence to be determined for the macaque viruses. The Luminex assays offer a novel multiplexed approach to assess rhadinovirus infection patterns in both humans and nonhuman primates. This will help advance our understanding of rhadinovirus biology and associated host immunological responses.

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Comparison of Humoral Immune Responses to Epstein-Barr Virus and Kaposi’s Sarcoma–Associated Herpesvirus Using a Viral Proteome Microarray

Abstract: The protein array provided a sensitive platform for global screening; identified new, frequently recognized viral antigens; and revealed a broader humoral response to EBV compared with KSHV in the same patients.

Cited by 32 publications

References 51 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

The antibody response to Epstein–Barr virions is altered in multiple sclerosis

Development of Whole-Virus Multiplex Luminex-Based Serological Assays for Diagnosis of Infections with Kaposi's Sarcoma-Associated Herpesvirus/Human Herpesvirus 8 Homologs in Macaques

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