Presence of lytic Epstein‐Barr virus infection in nasopharyngeal carcinoma

Yu, F.; Lu, Yanan; Petersson, Fredrik; Wang, Deyun; Loh, Kwok Seng

doi:10.1002/hed.25131

Cited by 14 publications

(19 citation statements)

References 23 publications

(38 reference statements)

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“…Further analysis by cell lineage markers revealed that the majority of cells expressed CD20 (>85%), which is indicative of B cell lineage ( Figure 1A ), with smaller populations of cells that expressed pan-cytokeratin (<5%) or CD3 (<10%) ( Figure 1B ). To establish the presence of EBV and to characterize its replication cycle in the CD20+ subpopulation, we exploited chromogenic EBER-ISH as well as fluorescent EBER-ISH and BRLF-ISH assays, which detect gene products specific to latency and lytic transcriptional programs[17,18]. These assays revealed that although latent infection was predominant, the virus also underwent spontaneous lytic reactivation in B cells at unusually high levels ( Figures 1A and 1C ).…”

Section: Resultsmentioning

confidence: 99%

“…Fourthly, we also re-examined sections of the NPC biopsy using RNAscope in situ hybridization assays with single-molecule sensitivity for detecting EBV-specific gene products [17–19], and observed that EBER-ISH signals were exclusively detected on pan-cytokeratin cells but absent on CD45+ lymphocytes. Finally, by parsing the whole-genomes of EBV isolates from the original NPC biopsy, xenograft, and cell line, we were able to ascertain that these three EBV isolates exhibit homologous sequences at highly discriminative genomic loci such as the genes encoding the nuclear antigens (EBNA-2 , −3, −4, −6) and LMP1 .…”

Section: Discussionmentioning

confidence: 99%

“…For Giemsa staining, dissociated cells were cytospun onto slides was fixed in absolute methanol for 30 seconds, and then submerged in 10% Giemsa solution for 30 minutes. Fluorescent in situ hybridization of EBV viral mRNA with immunohistochemical (IHC) staining was performed as described in our previous study[17]. All images were captured using an Olympus fluorescent microscope equipped with appropriate filters, and image analysis was conducted using the in-built Cellsens software.…”

Section: Methodsmentioning

confidence: 99%

“…Two milliliters of cell-free supernatant was admixed with 2 × 10 5 primary nasopharyngeal epithelial cells or Ramos cells in six-well plates in 2 ml of medium (~7 × 10 7 DNA copies/ml). Cells were fixed after 24 h and cell-free infection was assessed by EBERISH and BRLF1-ISH as previously described [17].…”

Section: Methodsmentioning

confidence: 99%

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Characterization and establishment of a novel EBV strain simultaneously associated with nasopharyngeal carcinoma and B-cell lymphoma

Syn

et al. 2020

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41Epstein-Barr virus (EBV) -the prototypical human tumor virus -is responsible for 1-2% of the 42 global cancer burden, but divergent strains seem to exist in different geographical regions with 43 distinct predilections for causing lymphoid or epithelial malignancies. Here we report the 44 establishment and characterization of Yu103, an Asia Pacific EBV strain with a highly remarkable 45 provenance of being derived from nasopharyngeal carcinoma biopsy but subsequently 46 propagated in human B-lymphoma cells and xenograft models. Unlike previously characterized 47 EBV strains which are either predominantly B-lymphotropic or epitheliotropic, Yu103 evinces an 48 uncanny capacity to infect and transform both B-lymphocytes and nasopharyngeal epithelial cells. 49 Genomic and phylogenetic analyses indicated that Yu103 EBV lies midway along the spectrum 50 of EBV strains known to drive lymphomagenesis or carcinogenesis, and harbors molecular 51 features which likely account for its unusual properties. To our knowledge, Yu103 EBV is currently 52 the only EBV isolate shown to drive human nasopharyngeal carcinoma and B-lymphoma, and 53 should therefore provide a powerful novel platform for research on EBV-driven hematological and 54 epithelial malignancies. PLoS Pathogens 55

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Characterization and establishment of a novel EBV strain simultaneously associated with nasopharyngeal carcinoma and B-cell lymphoma

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et al. 2020

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“…PBMC and tumor samples were obtained from a 69 years old Asian woman (patient A311), nonsmoker, with a lung cancer. Immunohistochemistry for H&E, PD-L1 and EBERish were performed on PFA fixed tissue sections [45]. Tumor single cell suspensions were prepared as previously described [46].…”

Section: Human Samples Immunohistochemistry and Cells Isolationmentioning

confidence: 99%

Partial absence of PD-1 expression by tumor-specific CD8⁺T cells in EBV-driven lymphoepithelioma-like carcinoma: a case report

Simoni

Becht

et al. 2019

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Lymphoepithelioma-like carcinoma (LELC) is an uncommon lung cancer, typically observed in young, non-smoking Asian populations. LELC is associated with Epstein-Barr virus (EBV) infection of lung tumor cells of epithelial origin, suggesting a carcinogenic role of EBV as observed in nasopharyngeal carcinoma (NPC). Here, we studied the antigen specificity and phenotype of CD8 + tumor infiltrating lymphocytes (TILs) in one LELC patient positive for EBV infection in lung tumor cells. Using MHC class I tetramers, we detected two populations of EBV-specific CD8 + TILs, which can be considered as tumor-specific CD8 + T cells, in the tumor of this patient. Transcriptomic analyses of these two populations reveal their distinct exhausted profiles and polyclonal TCR repertoire. High dimensional analyses at single cell level using mass cytometry showed showed that populations of tumor specific CD8 + TILs are phenotypically heterogeneous, although they consistently express CD39. Unexpectedly, although the LELC tumor cells expressed abundant PD-L1, these tumor-specific CD8 + TILs mostly did not express PD-1, suggesting that anti-PD1/PD-L1 immunotherapy may not be an appropriate strategy for disinhibiting EBV-specific cells for the treatment of LELC patients. These results might also help to explain low rates of checkpoint blockade immunotherapy response for NPC, despite the antigenicity of EBV for both tumor types.

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Partial absence of PD‐1 expression by tumor‐infiltrating EBV‐specific CD8⁺ T cells in EBV‐driven lymphoepithelioma‐like carcinoma

Simoni

Becht

et al. 2020

Clin & Trans Imm

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Objectives. Lymphoepithelioma-like carcinoma (LELC) is an uncommon lung cancer, typically observed in young, non-smoking Asian populations. LELC is associated with Epstein-Barr virus (EBV) infection of lung tumor cells of epithelial origin, suggesting a carcinogenic role of EBV as observed in nasopharyngeal carcinoma (NPC). Here, we studied the antigen specificity and phenotype of EBV-specific CD8 + T cells in blood and tumor of one LELC patient positive for EBV infection in lung tumor cells. Methods. Using multiplex MHC class I tetramers, mass cytometry and mRNA sequencing, we studied EBV-specific CD8 + T cells at the transcriptomic and phenotypic levels in blood and tumor tissues of the LELC patient. Results. Lymphoepithelioma-like carcinoma lung tumor cells were positive for EBV infection. In both blood and tumor tissues, we detected two populations of EBV-specific CD8 + T cells targeting the EBV lytic cycle proteins: BRLF1 and BMLF1. Transcriptomic analyses of these two populations in the tumor, which can be considered as tumor-specific, revealed their distinct exhausted profile and polyclonal TCR repertoire. High-dimensional phenotypical analysis revealed the distinct phenotype of these cells between blood and tumor tissues. In tumor tissue, EBVspecific CD8 + TILs were phenotypically heterogeneous, but consistently expressed CD39. Unexpectedly, although the LELC tumor cells expressed abundant PD-L1, these tumor-specific CD8 + tumor-infiltrating lymphocytes (TILs) mostly did not express PD-1. Conclusion. Epstein-Barr virus-specific CD8 + TILs in EBV-driven tumor are heterogeneous and partially lack PD-1 expression, suggesting that anti-PD1/PD-L1 immunotherapy may not be an appropriate strategy for disinhibiting EBV-specific cells in the treatment of LELC patients.

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Presence of lytic Epstein‐Barr virus infection in nasopharyngeal carcinoma

Cited by 14 publications

References 23 publications

Characterization and establishment of a novel EBV strain simultaneously associated with nasopharyngeal carcinoma and B-cell lymphoma

Characterization and establishment of a novel EBV strain simultaneously associated with nasopharyngeal carcinoma and B-cell lymphoma

Partial absence of PD-1 expression by tumor-specific CD8⁺T cells in EBV-driven lymphoepithelioma-like carcinoma: a case report

Partial absence of PD‐1 expression by tumor‐infiltrating EBV‐specific CD8⁺ T cells in EBV‐driven lymphoepithelioma‐like carcinoma

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Presence of lytic Epstein‐Barr virus infection in nasopharyngeal carcinoma

Cited by 14 publications

References 23 publications

Characterization and establishment of a novel EBV strain simultaneously associated with nasopharyngeal carcinoma and B-cell lymphoma

Characterization and establishment of a novel EBV strain simultaneously associated with nasopharyngeal carcinoma and B-cell lymphoma

Partial absence of PD-1 expression by tumor-specific CD8+T cells in EBV-driven lymphoepithelioma-like carcinoma: a case report

Partial absence of PD‐1 expression by tumor‐infiltrating EBV‐specific CD8+ T cells in EBV‐driven lymphoepithelioma‐like carcinoma

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Partial absence of PD-1 expression by tumor-specific CD8⁺T cells in EBV-driven lymphoepithelioma-like carcinoma: a case report

Partial absence of PD‐1 expression by tumor‐infiltrating EBV‐specific CD8⁺ T cells in EBV‐driven lymphoepithelioma‐like carcinoma