研究者詳細 - 木村　宏

Single-Cell Transcriptomic Analysis of Epstein-Barr Virus-Associated Hemophagocytic Lymphohistiocytosis

特集小児臨床検査2024 XIII.感染症検査 4.ウイルス感染症　2)ヘルペスウイルス感染症招待有り

鈴木高子, 木村宏

小児内科 56 巻 ( 13 ) 頁： 620 - 624 2024年10月

Direct evidence of abortive lytic infection-mediated establishment of Epstein-Barr virus latency during B-cell infection (vol 11 , 575255, 2021) 国際共著 Open Access

Inagaki, T; Sato, Y; Ito, J; Takaki, M; Okuno, Y; Yaguchi, M; Al Masud, HMA; Watanabe, T; Sato, K; Iwami, S; Murata, T; Kimura, H

FRONTIERS IN MICROBIOLOGY 15 巻頁： 1426311 2024年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Frontiers in Microbiology

In the published article, there was an error in Table 2 as published. We have carelessly swapped the rows and columns of Table 2. The corrected Table 2 and its caption appear below. Estimated parameters by fitting the mathematical model to experimental data. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.

DOI： 10.3389/fmicb.2024.1426311

Suzuki, T; Sato, Y; Okuno, Y; Torii, Y; Fukuda, Y; Haruta, K; Yamaguchi, M; Kawamura, Y; Hama, A; Narita, A; Muramatsu, H; Yoshikawa, T; Takahashi, Y; Kimura, H; Ito, Y; Kawada, JI

JOURNAL OF CLINICAL IMMUNOLOGY 44 巻 ( 4 ) 頁： 103 2024年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Clinical Immunology

Epstein-Barr virus (EBV) infection can lead to infectious mononucleosis (EBV-IM) and, more rarely, EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH), which is characterized by a life-threatening hyperinflammatory cytokine storm with immune dysregulation. Interferon-gamma (IFNγ) has been identified as a critical mediator for primary HLH; however, the detailed role of IFNγ and other cytokines in EBV-HLH is not fully understood. In this study, we used single-cell RNA sequencing to characterize the immune landscape of EBV-HLH and compared it with EBV-IM. Three pediatric patients with EBV-HLH with different backgrounds, one with X-linked lymphoproliferative syndrome type 1 (XLP1), two with chronic active EBV disease (CAEBV), and two patients with EBV-IM were enrolled. The TUBA1B + STMN1 + CD8 + T cell cluster, a responsive proliferating cluster with rich mRNA detection, was explicitly observed in EBV-IM, and the upregulation of SH2D1A—the gene responsible for XLP1—was localized in this cluster. This proliferative cluster was scarcely observed in EBV-HLH cases. In EBV-HLH cases with CAEBV, upregulation of LAG3 was observed in EBV-infected cells, which may be associated with an impaired response by CD8 + T cells. Additionally, genes involved in type I interferon (IFN) signaling were commonly upregulated in each cell fraction of EBV-HLH, and activation of type II IFN signaling was observed in CD4 + T cells, natural killer cells, and monocytes but not in CD8 + T cells in EBV-HLH. In conclusion, impaired responsive proliferation of CD8 + T cells and upregulation of type I IFN signaling were commonly observed in EBV-HLH cases, regardless of the patients’ background, indicating the key features of EBV-HLH.

DOI： 10.1007/s10875-024-01701-0

Diagnostic and disease severity determination criteria for hydroa vacciniforme lymphoproliferative disorders and severe mosquito bite allergy Open Access

Epstein-Barr virus lytic gene BNRF1 promotes B-cell lymphomagenesis via IFI27 upregulation Open Access

Sagou, K; Sato, Y; Okuno, Y; Watanabe, T; Inagaki, T; Motooka, Y; Toyokuni, S; Murata, T; Kiyoi, H; Kimura, H

PLOS PATHOGENS 20 巻 ( 2 ) 頁： e1011954 2024年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PLoS Pathogens

Epstein-Barr virus (EBV) is a ubiquitous human lymphotropic herpesvirus that is causally associated with several malignancies. In addition to latent factors, lytic replication contributes to cancer development. In this study, we examined whether the lytic gene BNRF1, which is conserved among gamma-herpesviruses, has an important role in lymphomagenesis. We found that lymphoblastoid cell lines (LCLs) established by BNRF1-knockout EBV exhibited remarkably lower pathogenicity in a mice xenograft model than LCLs produced by wild-type EBV (LCLs-WT). RNA-seq analyses revealed that BNRF1 elicited the expression of interferon-inducible protein 27 (IFI27), which promotes cell proliferation. IFI27 knockdown in LCLs-WT resulted in excessive production of reactive oxygen species, leading to cell death and significantly decreased their pathogenicity in vivo. We also confirmed that IFI27 was upregulated during primary infection in B-cells. Our findings revealed that BNRF1 promoted robust proliferation of the B-cells that were transformed by EBV latent infection via IFI27 upregulation both in vitro and in vivo.

DOI： 10.1371/journal.ppat.1011954

Updated guidelines for chronic active Epstein-Barr virus disease Open Access

Kawada, JI; Ito, Y; Ohshima, K; Yamada, M; Kataoka, S; Muramatsu, H; Sawada, A; Wada, T; Imadome, KI; Arai, A; Iwatsuki, K; Ohga, S; Kimura, H

INTERNATIONAL JOURNAL OF HEMATOLOGY 118 巻 ( 5 ) 頁： 568 - 576 2023年11月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：International Journal of Hematology

Chronic active Epstein–Barr virus disease (CAEBV), formerly named chronic active Epstein–Barr virus infection, is characterized by systemic inflammation and clonal proliferation of Epstein–Barr virus (EBV)-infected T or NK cells. As CAEBV is a potentially life-threatening illness, appropriate diagnosis and therapeutic interventions are necessary for favorable clinical outcomes. Substantial evidence regarding the pathogenesis and treatment of CAEBV has been accumulated since previous guidelines for the diagnosis of CAEBV were proposed. To reflect this evidence, we updated the guidelines for the diagnosis and treatment of CAEBV to improve clinical management of the disease. The details of the updated guidelines are presented in this report. Diagnosis of CAEBV now requires confirmation of a high copy number of EBV genome and EBV-infected T or NK cells. An EBV DNA load ≥ 10,000 IU/mL in whole blood is proposed as the diagnostic cutoff value for CAEBV in this updated guideline. A standard treatment approach for CAEBV has not been established, and hematopoietic stem cell transplantation (HSCT) is considered the only curative treatment. Chemotherapy can be administered to control disease activity before HSCT.

DOI： 10.1007/s12185-023-03660-5

The 5th edition of The World Health Organization Classification of Haematolymphoid Tumours: Lymphoid Neoplasms (vol 36, pg 1720, 2022) 国際共著 Open Access

Alaggio, R; Amador, C; Anagnostopoulos, I; Attygalle, AD; Araujo, IBD; Berti, E; Bhagat, G; Borges, AM; Boyer, D; Calaminici, M; Chadburn, A; Chan, JKC; Cheuk, W; Chng, WJ; Choi, JK; Chuang, SS; Coupland, SE; Czader, M; Dave, SS; de Jong, D; Di Napoli, A; Du, MQ; Elenitoba-Johnson, KS; Ferry, J; Geyer, J; Gratzinger, D; Guitart, J; Gujral, S; Harris, M; Harrison, CJ; Hartmann, S; Hochhaus, A; Jansen, PM; Karube, K; Kempf, W; Khoury, J; Kimura, H; Klapper, W; Kovach, AE; Kumar, S; Lazar, AJ; Lazzi, S; Leoncini, L; Leung, N; Leventaki, V; Li, XQ; Lim, MS; Liu, WP; Louissaint, A ; Marcogliese, A; Medeiros, LJ; Michal, M; Miranda, RN; Mitteldorf, C; Montes-Moreno, S; Morice, W; Nardi, V; Naresh, KN; Natkunam, Y; Ng, SB; Oschlies, I; Ott, G; Parrens, M; Pulitzer, M; Rajkumar, SV; Rawstron, AC; Rech, K; Rosenwald, A; Said, J; Sarkozy, C; Sayed, S; Saygin, C; Schuh, A; Sewell, W; Siebert, R; Sohani, AR; Suzuki, R; Tooze, R; Traverse-Glehen, A; Vega, F; Vergier, B; Wechalekar, AD; Wood, B; Xerri, L; Xiao, WB; Akinola, NO; Akkari, Y; Allende, LM; Aozasa, K; Araujo, I; Arcaini, L; Ardeshna, KM; Asano, N; Attarbaschi, A; Bacon, CM; Barrans, SL; Batchelor, T; Battistella, M; Baughn, LB; Behdad, A; Berentsen, S; Bianchi, G; Bledsoe, J; Borchmann, P; Bower, M; Buldini, B; Burger, JA; Burkhardt, B; Cassaday, RD; Cazzaniga, G; Cerf-Bensussan, N; Cesarman, E; Chandy, M; Chapman, JR; Chapuy, B; Chen, XY; Cheng, CL; Chiattone, C; Chiorazzi, N; Cook, LB; Cooper, WA; Corboy, GP; Cowan, AJ; Cozzolino, I; Cree, IA; d'Amore, ESG; Davies, AJ; Deckert, M; Delabie, J; Demicco, EG; Deshpande, V; Diepstra, A; Dierickx, D; Dunleavy, K; Eichhorst, B; Ennishi, D; Fajgenbaum, DC; Farinha, P; de Larrea, CF; Fisher, KE; Fitzgibbon, J; Flanagan, M; Fromm, J; Garcia, JF; Geddie, WR; Gertz, M; Gopal, A; Gopal, S; Greipp, PT; Gru, A; Gupta, R; Hansmann, ML; Hebeda, KM; Herfarth, K; Herling, M; Hermine, O; Khe, HX; Hodge, J; Hu, SM; Huang, YH; Hung, YP; Hunger, S; Inaba, H; Inagaki, H; Iqbal, J; Ishitsuka, K; Iwaki, N; Iwatsuki, K; Jain, N; Jeon, YK; Kadin, M; Kaji, S; Kakkar, A; Karadimitris, A; Kataoka, K; Kato, S; Kersten, MJ; Ketterling, RP; Kim, JE; Kratz, CP; Kridel, R; Kristinsson, S; Küppers, R; Kuzu, I; Kwong, YL; Lacasce, A; Lamant-Rochaix, L; Lamy, T; Landgren, O; Laskar, S; Laskin, WB; Lenz, G; Li, SY; Di Li, G; Lin, P; Locatelli, F; Lorsbach, RB; Lossos, I; Loughran, TP Jr; Macon, WR; Maleszewski, JJ; Malhotra, P; Marafioti, T; Maruyama, D; Marx, A; Mbulaiteye, SM; Meignin, V; Mejstrikova, E; Michelow, P; Miettinen, M; Miles, RR; Miyoshi, H; Molina, TJ; Mollejo, M; Momose, S; Mori, T; Morice, WG; Nadel, B; Nagai, H; Nagane, M; Nair, R; Nakamura, N; Nakazawa, A; Nasr, S; Nicholson, AG; Nicolae, A; Ohgami, RS; Oishi, N; Olson, TS; Ortonne, N; Paiva, B; Pan-Hammarström, Q; Parihar, M; Paulli, M; Petersen, A; Picarsic, J; Pileri, A; Pimpinelli, N; Plaza, JA; Rabin, KR; Raderer, M; Rai, K; Randen, U; Rao, H; Robson, A; Rochford, R; Rosenquist, R; Rossi, D; Rossi, ED; Rule, S; Rymkiewicz, G; Sabattini, E; Saha, V; Sakata-Yanagimoto, M; Sander, CA; Sangueza, JM; Sangüeza, OP; Santucci, M; Sato, Y; Satou, A; Schafernak, KT; Schmitt, F; Semenzato, G; Sengar, M; Shanafelt, T; Shimada, K; Slack, GW; Slager, S; Soffietti, R; Solomon, DA; Stamatopoulos, K; Steidl, C; Stilgenbauer, S; Sukswai, N; Takeuchi, K; Tallini, G; Tamaru, J; Tan, SY; Tembhare, P; Tiacci, E; Tokura, Y; Tournilhac, O; Treon, S; Truemper, L; Tsukasaki, K; van Rhee, F; Varghese, A; Vermeer, MH; Vielh, P; Walker, B; Wang, M; Wang, HY; Wang, Z; Watanabe, T; Weigert, O; Wein-Stock, D; Whittaker, SJ; Willemze, R; Woessmann, W; Wu, CJ; Yamaguchi, M; Yamamoto, H; Yamashita, D; Yang, SM; Yang, DT; Yasuda, T; Yin, WH; Zen, Y; Zhao, S; Zhao, WL

LEUKEMIA 37 巻 ( 9 ) 頁： 1944 - 1951 2023年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Leukemia

Correction to: Leukemia, published online 22 June 2022 At the time of original publication, there was an incomplete listing of contributing authors and their institutions. This revision corrects those omissions.

DOI： 10.1038/s41375-023-01962-5

Growth Transformation of B Cells by Epstein-Barr Virus Requires IMPDH2 Induction and Nucleolar Hypertrophy 国際共著 Open Access

Sugimoto, A; Watanabe, T; Matsuoka, K; Okuno, Y; Yanagi, Y; Narita, Y; Mabuchi, S; Nobusue, H; Sugihara, E; Hirayama, M; Ide, T; Onouchi, T; Sato, Y; Kanda, T; Saya, H; Iwatani, Y; Kimura, H; Murata, T

MICROBIOLOGY SPECTRUM 11 巻 ( 4 ) 頁： e0044023 2023年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Microbiology Spectrum

The in vitro growth transformation of primary B cells by Epstein-Barr virus (EBV) is the initial step in the development of posttransplant lymphoproliferative disorder (PTLD). We performed electron microscopic analysis and immunostaining of primary B cells infected with wild-Type EBV. Interestingly, the nucleolar size was increased by two days after infection. A recent study found that nucleolar hypertrophy, which is caused by the induction of the IMPDH2 gene, is required for the efficient promotion of growth in cancers. In the present study, RNA-seq revealed that the IMPDH2 gene was significantly induced by EBV and that its level peaked at day 2. Even without EBV infection, the activation of primary B cells by the CD40 ligand and interleukin-4 increased IMPDH2 expression and nucleolar hypertrophy. Using EBNA2 or LMP1 knockout viruses, we found that EBNA2 and MYC, but not LMP1, induced the IMPDH2 gene during primary infections. IMPDH2 inhibition by mycophenolic acid (MPA) blocked the growth transformation of primary B cells by EBV, leading to smaller nucleoli, nuclei, and cells. Mycophenolate mofetil (MMF), which is a prodrug of MPA that is approved for use as an immunosuppressant, was tested in a mouse xenograft model. Oral MMF significantly improved the survival of mice and reduced splenomegaly. Taken together, these results indicate that EBV induces IMPDH2 expression through EBNA2-dependent and MYC-dependent mechanisms, leading to the hypertrophy of the nucleoli, nuclei, and cells as well as efficient cell proliferation. Our results provide basic evidence that IMPDH2 induction and nucleolar enlargement are crucial for B cell transformation by EBV. In addition, the use of MMF suppresses PTLD. IMPORTANCE EBV infections cause nucleolar enlargement via the induction of IMPDH2, which are essential for B cell growth transformation by EBV. Although the significance of IMPDH2 induction and nuclear hypertrophy in the tumorigenesis of glioblastoma has been reported, EBV infection brings about the change quickly by using its transcriptional cofactor, EBNA2, and MYC. Moreover, we present here, for the novel, basic evidence that an IMPDH2 inhibitor, namely, MPA or MMF, can be used for EBV-positive posttransplant lymphoproliferative disorder (PTLD).

DOI： 10.1128/spectrum.00440-23

Hirai, Y; Asada, H; Hamada, T; Kawada, J; Kimura, H; Arai, A; Ohshima, K; Ohga, S; Iwatsuki, K

JOURNAL OF DERMATOLOGY 50 巻 ( 7 ) 頁： E198 - E205 2023年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Dermatology

Hydroa vacciniforme lymphoproliferative disorder (HV-LPD) and severe mosquito bite allergy (SMBA) are both cutaneous forms of Epstein–Barr virus (EBV)-associated T/natural killer (NK) cell LPDs and are closely related to chronic active EBV disease (CAEBV) and EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH). HV-LPD is further divided into classic HV, a benign subtype mediated by EBV-positive γδT cells, and systemic HV, another life-threatening subtype mainly associated with EBV-positive αβT or γδT cells. The vast majority of patients with SMBA have increased numbers of EBV-infected NK cells in the blood. Clinical symptoms of HV-LPD and SMBA often overlap in the same patient and may progress to more serious disease conditions equivalent to the systemic form of CAEBV. To define the disease spectrum of HV-LPD and SMBA, we propose the diagnostic criteria and the determination criteria for disease severity. The proposed diagnostic criteria are consistent with those for CAEBV and EBV-HLH in the guidelines for the management for CAEBV and related disorders 2023.

DOI： 10.1111/1346-8138.16842

Epstein-Barr Virus BBLF1 Mediates Secretory Vesicle Transport to Facilitate Mature Virion Release Open Access

Uddin, MK; Watanabe, T; Arata, M; Sato, Y; Kimura, H; Murata, T

JOURNAL OF VIROLOGY 97 巻 ( 6 ) 頁： e0043723 2023年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Virology

Enveloped viruses undergo a complex multistep process of assembly, maturation, and release into the extracellular space utilizing host secretory machinery. Several studies of the herpesvirus subfamily have shown that secretory vesicles derived from the trans-Golgi network (TGN) or endosomes transport virions into the extracellular space. However, the regulatory mechanism underlying the release of Epstein-Barr virus, a human oncovirus, remains unclear. We demonstrate that disruption of BBLF1, a tegument component, suppressed viral release and resulted in the accumulation of viral particles on the inner side of the vesicular membrane. Organelle separation revealed the accumulation of infectious viruses in fractions containing vesicles derived from the TGN and late endosomes. Deficiency of an acidic amino acid cluster in BBLF1 reduced viral secretion. Moreover, truncational deletion of the C-terminal region of BBLF1 increased infectious virus production. These findings suggest that BBLF1 regulates the viral release pathway and reveal a new aspect of tegument protein function.

DOI： 10.1128/jvi.00437-23

特集新しい時代の小児感染症各論:原因微生物別ウイルスヘルペスウイルス科招待有り

川田潤一, 木村宏

小児内科 55 巻 ( 4 ) 頁： 623 - 628 2023年4月

Salvage Human Leukocyte Antigen-Haploidentical Hematopoietic Cell Transplant With Posttransplant Cyclophosphamide for Graft Failure in a Patient With Chronic Active Epstein-Barr Virus Infection

Nakaya, Y; Koh, H; Harada, N; Okamura, H; Misugi, T; Suekane, T; Tachibana, D; Kimura, H; Hino, M; Nakamae, H

EXPERIMENTAL AND CLINICAL TRANSPLANTATION 21 巻 ( 2 ) 頁： 184 - 188 2023年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Experimental and Clinical Transplantation

Salvage human leukocyte antigen-haploidentical hematopoietic cell transplant using posttransplant cyclophosphamide has shown promising results for graft failure in various hematological disorders. However, to our knowledge, no such findings have been reported for a case of chronic active Epstein-Barr virus infection, although graft failure is relatively common in patients with chronic active Epstein-Barr virus infection. We report a case of a 32-year-old woman with natural killer-cell type chronic active Epstein-Barr virus infection who experienced graft failure after a first allogeneic hematopoietic cell transplant from an unrelated human leukocyte antigen-matched donor. The patient received a second allogeneic hematopoietic cell transplant with human leukocyte antigen-haploidentical hematopoietic cell transplant using posttransplant cyclophosphamide (cyclophosphamide, 50 mg/kg, on day 3 and day 4) following reduced-intensity conditioning as rescue therapy. Neutrophils successfully engrafted on day 19, and the patient sustained remission without severe transplant-related complication 10 months after salvage human leukocyte antigen-haploidentical hematopoietic cell transplant using posttransplant cyclophosphamide. This report suggests that salvage human leukocyte antigen-haploidentical hematopoietic cell transplant using posttransplant cyclophosphamide may be a feasible therapeutic option for graft failure in patients with chronic active Epstein-Barr virus infection.

DOI： 10.6002/ect.2022.0322

症例 CD8＋αβT細胞優位型種痘様水疱症リンパ増殖異常症の1例

大林三裕佳, 中村賢人, 古橋卓也, 岩月啓氏, 三宅智子, 平井陽至, 谷田けい, 金兼弘和, 西田徹也, 木村宏, 森田明理

皮膚科の臨床 65 巻 ( 2 ) 頁： 273 - 278 2023年2月

N6-methyladenosine Modification of Hepatitis B Virus RNA in the Coding Region of HBx Open Access

Murata, T; Iwahori, S; Okuno, Y; Nishitsuji, H; Yanagi, Y; Watashi, K; Wakita, T; Kimura, H; Shimotohno, K

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 24 巻 ( 3 ) 2023年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：International Journal of Molecular Sciences

N6-methyladenosine (m6A) is a post-transcriptional modification of RNA involved in transcript transport, degradation, translation, and splicing. We found that HBV RNA is modified by m6A predominantly in the coding region of HBx. The mutagenesis of methylation sites reduced the HBV mRNA and HBs protein levels. The suppression of m6A by an inhibitor or knockdown in primary hepatocytes decreased the viral RNA and HBs protein levels in the medium. These results suggest that the m6A modification of HBV RNA is needed for the efficient replication of HBV in hepatocytes.

DOI： 10.3390/ijms24032265

The 100th anniversary of the Nagoya Journal of Medical Science: a message from the Dean

Kimura, H

NAGOYA JOURNAL OF MEDICAL SCIENCE 85 巻 ( 1 ) 頁： 5 - 6 2023年2月

The Epstein-Barr Virus Enhancer Interaction Landscapes in Virus-Associated Cancer Cell Lines 国際共著 Open Access

Ding, WY; Wang, C; Narita, Y; Wang, HB; Leong, MML; Huang, A; Liao, YF; Liu, XF; Okuno, Y; Kimura, H; Gewurz, B; Teng, MX; Jin, SL; Sato, Y; Zhao, B

JOURNAL OF VIROLOGY 96 巻 ( 18 ) 頁： e0073922 2022年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Virology

Epstein-Barr virus (EBV) persists in human cells as episomes. EBV episomes are chromatinized and their 3D conformation varies greatly in cells expressing different latency genes. We used HiChIP, an assay which combines genome-wide chromatin conformation capture followed by deep sequencing (Hi-C) and chromatin immunoprecipitation (ChIP), to interrogate the EBV episome 3D conformation in different cancer cell lines. In an EBV-transformed lymphoblastoid cell line (LCL) GM12878 expressing type III EBV latency genes, abundant genomic interactions were identified by H3K27ac HiChIP. A strong enhancer was located near the BILF2 gene and looped to multiple genes around BALFs loci. Perturbation of the BILF2 enhancer by CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa) altered the expression of BILF2 enhancer-linked genes, including BARF0 and BALF2, suggesting that this enhancer regulates the expression of linked genes. H3K27ac ChIP followed by deep sequencing (ChIP-seq) identified several strong EBV enhancers in T/NK (natural killer) lymphoma cells that express type II EBV latency genes. Extensive intragenomic interactions were also found which linked enhancers to target genes. A strong enhancer at BILF2 also looped to the BALF loci. CRISPRi also validated the functional connection between BILF2 enhancer and BARF1 gene. In contrast, H3K27ac HiChIP found significantly fewer intragenomic interactions in type I EBV latency gene-expressing primary effusion lymphoma (PEL) cell lines. These data provided new insight into the regulation of EBV latency gene expression in different EBV-associated tumors.

DOI： 10.1128/jvi.00739-22

EBV-associated NK and T-cell lymphoid neoplasms 国際共著 Open Access

Kimura, H; de Leval, L; Cai, QQ; Kim, WS

CURRENT OPINION IN ONCOLOGY 34 巻 ( 5 ) 頁： 422 - 431 2022年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Current Opinion in Oncology

Purpose of reviewEpstein-Barr virus (EBV)-associated neoplasms derived from natural killer (NK) or T cells comprise a group of clinically and biologically heterogenous disorders affecting children and adults, which are overall rare but more prevalent in Asia and South America. This review focuses on neoplasms presenting in the adulthood, addressing recent genomic discoveries as well as therapeutic developments in these highly aggressive disorders.Recent findingsDistinct molecular subtypes of extranodal NK/T-cell lymphomas (ENKTCLs) have been described, with differences in cell of origin, EBV pattern, genomic alterations, clinical characteristics, response to asparaginase-based therapies and to more recent approaches targeting molecular aberrations of the lymphoma. For the last two decades, progress in the clinical management of ENKTCL was based on L-asapraginase containing combinations and the incoroperation of radiotherapy. A subset of cases with PDL1-2 structural alterations may be more responsive to treatment with immune checkpoint inhibitors. Primary nodal EBV+ lymphomas derived from T or NK cells have distinctive features separating them from both peripheral T-cell lymphoma not otherwise specified and ENKTCL. Treatment algorithms correspond to those for advanced ENKTCL.SummaryWith better understanding of lymphomagenesis, genomic landscape and immunologic aspects of the diseases, future treatment options will include targeted therapies including immune checkpoint inhibitors and novel antibodies.

DOI： 10.1097/CCO.0000000000000889

Comprehensive Analyses of Intraviral Epstein-Barr Virus Protein-Protein Interactions Hint Central Role of BLRF2 in the Tegument Network 国際共著 Open Access

Hara, Y; Watanabe, T; Yoshida, M; Al Masud, HMA; Kato, H; Kondo, T; Suzuki, R; Kurose, S; Uddin, MK; Arata, M; Miyagi, S; Yanagi, Y; Sato, Y; Kimura, H; Murata, T

JOURNAL OF VIROLOGY 96 巻 ( 14 ) 頁： e0051822 2022年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Virology

Protein-protein interactions (PPIs) are crucial for various biological processes. Epstein-Barr virus (EBV) proteins typically form complexes, regulating the replication and persistence of the viral genome in human cells. However, the role of EBV protein complexes under physiological conditions remains unclear. In this study, we performed comprehensive analyses of EBV PPIs in living cells using the NanoBiT system. We identified 195 PPIs, many of which have not previously been reported. Computational analyses of these PPIs revealed that BLRF2, which is only found in gammaherpesviruses, is a central protein in the structural network of EBV tegument proteins. To characterize the role of BLRF2, we generated two BLRF2 knockout EBV clones using CRISPR/Cas9. BLRF2 knockout significantly decreased the production of infectious virus particles, which was partially restored by exogenous BLRF2 expression. In addition, self-association of BLRF2 protein was found, and mutation of the residues crucial for the self-association affected stability of the protein. Our data imply that BLRF2 is a tegument network hub that plays important roles in progeny virion maturation.

DOI： 10.1128/jvi.00518-22

EBV genome variations enhance clinicopathological features of nasopharyngeal carcinoma in a non-endemic region Open Access

Genome-wide CRISPR screen for HSV-1 host factors reveals PAPSS1 contributes to heparan sulfate synthesis Open Access

Suzuki, T; Sato, Y; Okuno, Y; Goshima, F; Mikami, T; Umeda, M; Murata, T; Watanabe, T; Watashi, K; Wakita, T; Kitagawa, H; Kimura, H

COMMUNICATIONS BIOLOGY 5 巻 ( 1 ) 頁： 694 2022年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Communications Biology

Herpes simplex virus type 1 (HSV-1) is a ubiquitous pathogen that causes various diseases in humans, ranging from common mucocutaneous lesions to severe life-threatening encephalitis. However, our understanding of the interaction between HSV-1 and human host factors remains incomplete. Here, to identify the host factors for HSV-1 infection, we performed a human genome-wide CRISPR screen using near-haploid HAP1 cells, in which gene knockout (KO) could be efficiently achieved. Along with several already known host factors, we identified 3′-phosphoadenosine 5′-phosphosulfate synthase 1 (PAPSS1) as a host factor for HSV-1 infection. The KO of PAPSS1 in HAP1 cells reduced heparan sulfate (HepS) expression, consequently diminishing the binding of HSV-1 and several other HepS-dependent viruses (such as HSV-2, hepatitis B virus, and a human seasonal coronavirus). Hence, our findings provide further insights into the host factor requirements for HSV-1 infection and HepS biosynthesis.

DOI： 10.1038/s42003-022-03581-9

The 5th edition of the World Health Organization Classification of Haematolymphoid Tumours: Lymphoid Neoplasms 国際共著 Open Access

LEUKEMIA 36 巻 ( 7 ) 頁： 1720 - 1748 2022年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Leukemia

We herein present an overview of the upcoming 5th edition of the World Health Organization Classification of Haematolymphoid Tumours focussing on lymphoid neoplasms. Myeloid and histiocytic neoplasms will be presented in a separate accompanying article. Besides listing the entities of the classification, we highlight and explain changes from the revised 4th edition. These include reorganization of entities by a hierarchical system as is adopted throughout the 5th edition of the WHO classification of tumours of all organ systems, modification of nomenclature for some entities, revision of diagnostic criteria or subtypes, deletion of certain entities, and introduction of new entities, as well as inclusion of tumour-like lesions, mesenchymal lesions specific to lymph node and spleen, and germline predisposition syndromes associated with the lymphoid neoplasms.

DOI： 10.1038/s41375-022-01620-2

Kondo, S; Okuno, Y; Murata, T; Dochi, H; Wakisaka, N; Mizokami, H; Moriyama-Kita, M; Kobayashi, E; Kano, M; Komori, T; Hirai, N; Ueno, T; Nakanishi, Y; Endo, K; Sugimoto, H; Kimura, H; Yoshizaki, T

CANCER SCIENCE 113 巻 ( 7 ) 頁： 2446 - 2456 2022年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Cancer Science

Nasopharyngeal carcinoma (NPC) is caused by infection with Epstein–Barr virus (EBV) and endemic in certain geographic regions. EBV lytic gene, BALF2, closely associates with viral reactivation and BALF2 gene variation, the H-H-H strain, causes NPC in endemic region, southern China. Here, we investigate whether such EBV variations also affect NPC in a non-endemic region, Japan. Viral genome sequencing with 47 EBV isolates of Japanese NPC were performed and compared with those of other EBV-associated diseases from Japan or NPC in Southern China. EBV genomes of Japanese NPC are different from those of other diseases in Japan or endemic NPC; Japanese NPC was not affected by the endemic strain (the BALF2 H-H-H) but frequently carried the type 2 EBV or the strain with intermediate risk of endemic NPC (the BALF2 H-H-L). Seven single nucleotide variations were specifically associated with Japanese NPC, of which six were present in both type 1 and 2 EBV genomes, suggesting the contribution of the type 2 EBV-derived haplotype. This observation was supported by a higher viral titer and stronger viral reactivation in NPC with either type 2 or H-H-L strains. Our results highlight the importance of viral strains and viral reactivation in the pathogenesis of non-endemic NPC.

DOI： 10.1111/cas.15381

Elderly-onset systemic Epstein-Barr virus-positive T-cell lymphoma of childhood Open Access

Nishikubo, M; Hiramoto, N; Yamashita, D; Imoto, H; Sato, Y; Okuno, Y; Haga, H; Nannya, Y; Ogawa, S; Kimura, H; Ishikawa, T

PATHOLOGY INTERNATIONAL 72 巻 ( 7 ) 頁： 376 - 378 2022年7月

Epstein-Barr virus tegument protein BGLF2 in exosomes released from virus-producing cells facilitates de novo infection 国際共著 Open Access

Sato, Y; Yaguchi, M; Okuno, Y; Ishimaru, H; Sagou, K; Ozaki, S; Suzuki, T; Inagaki, T; Umeda, M; Watanabe, T; Fujimuro, M; Murata, T; Kimura, H

CELL COMMUNICATION AND SIGNALING 20 巻 ( 1 ) 頁： 95 2022年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Cell Communication and Signaling

Background: Viruses must adapt to the environment of their host cells to establish infection and persist. Diverse mammalian cells, including virus-infected cells, release extracellular vesicles such as exosomes containing proteins and miRNAs, and use these vesicles to mediate intercellular communication. However, the roles of exosomes in viral infection remain unclear. Results: We screened viral proteins to identify those responsible for the exosome-mediated enhancement of Epstein–Barr virus (EBV) infection. We identified BGLF2 protein encapsulated in exosomes, which were released by EBV-infected cells. BGLF2 protein is a tegument protein that exists in the space between the envelope and nucleocapsid, and it is released into the cytoplasm shortly after infection. BGLF2 protein-containing exosomes enhanced viral gene expression and repressed innate immunity, thereby supporting the EBV infection. Conclusions: The EBV tegument protein BGLF2 is encapsulated in exosomes and released by infected cells to facilitate the establishment of EBV infection. These findings suggest that tegument proteins support viral infection not only between the envelope and nucleocapsid, as well as in extraviral particles such as exosomes. Graphical abstract: [Figure not available: see fulltext.] [MediaObject not available: see fulltext.].

DOI： 10.1186/s12964-022-00902-7

EBV Exploits RNA m6A Modification to Promote Cell Survival and Progeny Virus Production During Lytic Cycle Open Access

Yanagi, Y; Watanabe, T; Hara, Y; Sato, Y; Kimura, H; Murata, T

FRONTIERS IN MICROBIOLOGY 13 巻頁： 870816 2022年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Frontiers in Microbiology

N6-methyladenosine (m6A) mediates various biological processes by affecting RNA stability, splicing, and translational efficiency. The roles of m6A modification in Epstein-Barr virus (EBV) infection in the lytic phase are unclear. Here, knockout of the m6A methyltransferase, N6-methyladenosine methyltransferase-like 3 (METTL3), or inhibition of methylation by DAA or UZH1a decreased the expression of viral lytic proteins and reduced progeny virion production. Interestingly, cell growth and viability were decreased by induction of the lytic cycle in METTL3-knockout or inhibitor-treated cells. Apoptosis was induced in those conditions possibly because of a decreased level of the anti-apoptotic viral protein, BHRF1. Therefore, m6A shows potential as a target of lytic induction therapy for EBV-associated cancers, including Burkitt lymphoma.

DOI： 10.3389/fmicb.2022.870816

PD-L1 upregulation by lytic induction of Epstein-Barr Virus Open Access

Yanagi, Y; Hara, Y; Mabuchi, S; Watanabe, T; Sato, Y; Kimura, H; Murata, T

VIROLOGY 568 巻頁： 31 - 40 2022年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Virology

Epstein-Barr virus (EBV) is an etiologic agent of infectious mononucleosis and several malignancies. Here, we found that reactivation of EBV resulted in increased programmed cell death-ligand 1 (PD-L1) expression in a cell type-dependent manner. Lytic induction in EBV-positive Akata, AGS, MutuI, and Jijoye cell lines increased PD-L1 levels, but cells such as EBV-negative Akata, MutuIII, and P3HR1 did not have increased PD-L1. EBV in the P3HR1 cell line has a deletion in the EBNA2 gene, while EBV in its parental cell line, Jijoye, has the complete EBNA2 gene. PD-L1 expression by lytic induction was reduced when EBNA2 was knocked down. In addition, pharmacological inhibition indicated involvement of nuclear factor kappa B, mitogen-activated protein kinase, and AKT signaling. These results suggest that EBV likely evades immunity by inducing PD-L1 upon reactivation, through the increased expression of EBNA2 and activation of signaling pathways.

DOI： 10.1016/j.virol.2022.01.006

In Silico Analysis and Synthesis of Nafamostat Derivatives and Evaluation of Their Anti-SARS-CoV-2 Activity Open Access

Fujimoto, KJ; Hobbs, DCF; Umeda, M; Nagata, A; Yamaguchi, R; Sato, Y; Sato, A; Ohmatsu, K; Ooi, T; Yanai, T; Kimura, H; Murata, T

VIRUSES-BASEL 14 巻 ( 2 ) 2022年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Viruses

Inhibition of transmembrane serine protease 2 (TMPRSS2) is expected to block the spike protein-mediated fusion of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Nafamostat, a potent TMPRSS2 inhibitor as well as a candidate for anti-SARS-CoV-2 drug, possesses the same acyl substructure as camostat, but is known to have a greater antiviral effect. A unique aspect of the molecular binding of nafamostat has been recently reported to be the formation of a covalent bond between its acyl substructure and Ser441 in TMPRSS2. In this study, we investigated crucial elements that cause the difference in anti-SARS-CoV-2 activity of nafamostat and camostat. In silico analysis showed that Asp435 significantly contributes to the binding of nafamostat and camostat to TMPRSS2, while Glu299 interacts strongly only with nafamostat. The estimated binding affinity for each compound with TMPRSS2 was actually consistent with the higher activity of nafamostat; however, the evaluation of the newly synthesized nafamostat derivatives revealed that the predicted binding affinity did not correlate with their anti-SARS-CoV-2 activity measured by the cytopathic effect (CPE) inhibition assay. It was further shown that the substitution of the ester bond with amide bond in nafamostat resulted in significantly weakened anti-SARS-CoV-2 activity. These results strongly indicate that the ease of covalent bond formation with Ser441 in TMPRSS2 possibly plays a major role in the anti-SARS-CoV-2 effect of nafamostat and its derivatives.

DOI： 10.3390/v14020389

Molecular Basis of Epstein-Barr Virus Latency Establishment and Lytic Reactivation 国際共著 Open Access

Murata, T; Sugimoto, A; Inagaki, T; Yanagi, Y; Watanabe, T; Sato, Y; Kimura, H

VIRUSES-BASEL 13 巻 ( 12 ) 2021年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Viruses

Epstein–Barr virus (EBV) is a causative agent of infectious mononucleosis and several types of cancer. Like other herpesviruses, it establishes an asymptomatic, life-long latent infection, with occasional reactivation and shedding of progeny viruses. During latency, EBV expresses a small number of viral genes, and exists as an episome in the host–cell nucleus. Expression patterns of latency genes are dependent on the cell type, time after infection, and milieu of the cell (e.g., germinal center or peripheral blood). Upon lytic induction, expression of the viral immediate-early genes, BZLF1 and BRLF1, are induced, followed by early gene expression, viral DNA replication, late gene expression, and maturation and egress of progeny virions. Furthermore, EBV reactivation involves more than just progeny production. The EBV life cycle is regulated by signal transduction, transcription factors, promoter sequences, epigenetics, and the 3D structure of the genome. In this article, the molecular basis of EBV latency establishment and reactivation is summarized.

DOI： 10.3390/v13122344

A STING inhibitor suppresses EBV-induced B cell transformation and lymphomagenesis 国際共著 Open Access

Miyagi, S; Watanabe, T; Hara, Y; Arata, M; Uddin, MK; Mantoku, K; Sago, K; Yanagi, Y; Suzuki, T; Al Masud, HMA; Kawada, J; Nakamura, S; Miyake, Y; Sato, Y; Murata, T; Kimura, H

CANCER SCIENCE 112 巻 ( 12 ) 頁： 5088 - 5099 2021年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Cancer Science

Epstein-Barr virus-associated lymphoproliferative disease (EBV-LPD) is frequently fatal. Innate immunity plays a key role in protecting against pathogens and cancers. The stimulator of interferon genes (STING) is regarded as a key adaptor protein allowing DNA sensors recognizing exogenous cytosolic DNA to activate the type I interferon signaling cascade. In terms of EBV tumorigenicity, the role of STING remains elusive. Here we showed that treatment with the STING inhibitor, C-176, suppressed EBV-induced transformation in peripheral blood mononuclear cells. In an EBV-LPD mouse model, C-176 treatment also inhibited tumor formation and prolonged survival. Treatment with B cells alone did not affect EBV transformation, but suppression of EBV-induced transformation was observed in the presence of T cells. Even without direct B cell-T cell contact in a transwell system, the inhibitor reduced the transformation activity, indicating that intercellular communication by humoral factors was critical to prevent EBV-induced transformation. These findings suggest that inhibition of STING signaling pathway with C-176 could be a new therapeutic target of EBV-LPD.

DOI： 10.1111/cas.15152

Editorial: Epstein-Barr Virus-Associated T/NK-Cell Lymphoproliferative Diseases

Deletion of Viral microRNAs in the Oncogenesis of Epstein-Barr Virus-Associated Lymphoma Open Access

Kimura, H; Okuno, Y; Sato, Y; Watanabe, T; Murata, T

FRONTIERS IN MICROBIOLOGY 12 巻頁： 667968 2021年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Frontiers in Microbiology

Epstein–Barr virus (EBV), which encodes >80 genes and nearly 50 non-coding RNAs, is a double-stranded DNA virus. EBV is associated with various types of lymphomas and lymphoproliferative disorders not only of B-cell but also T/NK-cell origin. However, the oncogenic mechanism remains poorly understood, including the EBV receptors expressed on T/NK cells, relationship of EBV with host genes, and epigenetic regulation of EBV and host genes. The roles of host and viral non-coding RNAs during tumorigenesis have been elucidated. EBV encodes at least 49 mature microRNAs (miRNAs), of which 44 are located in BamHI-A rightward transcripts (BARTs) region, and the remaining five are located in BamHI-H rightward fragment 1. BART miRNAs modulate cell differentiation, proliferation, apoptosis, and the cell cycle, and they are considered positive regulators of oncogenesis. We and others have recently reported that EBV-positive lymphomas frequently possess large deletions in BART miRNA clusters, suggesting that some viral miRNAs have suppressive effects on oncogenesis, and that deletion of these miRNAs may aid lymphoma formation.

DOI： 10.3389/fmicb.2021.667968

RNAseq analysis identifies involvement of EBNA2 in PD-L1 induction during Epstein-Barr virus infection of primary B cells 査読有り国際共著

Yusuke Yanagi, Yusuke Okuno, Yohei Narita, H M Abdullah Al Masud , Takahiro Watanabe , Yoshitaka Sato, Teru Kanda, Hiroshi Kimura, Takayuki Murata

Virology 557 巻頁： 44 - 54 2021年5月

Oncolytic activity of naturally attenuated herpes-simplex virus HF10 against an immunocompetent model of oral carcinoma. 査読有り

Takano G, Esaki S, Goshima F, Enomoto A, Hatano Y, Ozaki H, Watanabe T, Sato Y, Kawakita D, Murakami S, Murata T, Nishiyama Y, Iwasaki S, Kimura H.

Molecular Therapy – Oncolytics 20 巻頁： 220 - 227 2021年3月

Role of Epstein-Barr Virus C Promoter Deletion in Diffuse Large B Cell Lymphoma 査読有り国際共著

Mabuchi Seiyo, Hijioka Fumiya, Watanabe Takahiro, Yanagi Yusuke, Okuno Yusuke, Al Masud H. M. Abdullah, Sato Yoshitaka, Murata Takayuki, Kimura Hiroshi

CANCERS 13 巻 ( 3 ) 2021年2月

Direct Evidence of Abortive Lytic Infection-Mediated Establishment of Epstein-Barr Virus Latency During B-Cell Infection 査読有り国際共著

Inagaki Tomoki, Sato Yoshitaka, Ito Jumpei, Takaki Mitsuaki, Okuno Yusuke, Yaguchi Masahiro, Al Masud H. M. Abdullah, Watanabe Takahiro, Sato Kei, Iwami Shingo, Murata Takayuki, Kimura Hiroshi

FRONTIERS IN MICROBIOLOGY 11 巻 2021年1月

慢性活動性EBウイルス感染症

木村宏

日本造血細胞移植学会雑誌 10 巻 ( 2 ) 頁： 87 - 93 2021年

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：一般社団法人日本造血・免疫細胞療法学会

慢性活動性EBウイルス感染症は，EBVがT細胞あるいはNK細胞に潜伏感染し，クローナリティを持って増殖，臓器に浸潤し，多彩な症状を呈する難治性疾患である。予後は，概して不良で，無治療の場合には，臓器合併症・急性転化により死に至ることが多い。筆者らは，慢性活動性EBウイルス感染症患者に対して大規模な網羅的遺伝子解析を行い，1）生殖細胞列の変異はほとんど認められないため原発性免疫不全症が潜んでいる可能性は低いこと，2）EBV感染細胞では，DDX3X，KMT2Dなどのドライバー遺伝子変異が認められ，経時的に観察できた例では，EBV感染細胞のクローン進化が確認されること，3）高率に欠失ウイルスが認められ発症に関与している可能性があること，を明らかにした。本稿では，これら遺伝子解析によって解明されつつある慢性活動性EBウイルス感染症の発症病理と，診断・治療に関わる最新の知見について概説する。

DOI： 10.7889/hct-20-026

CiNii Research

Oncolytic activity of HF10 in head and neck squamous cell carcinomas. 査読有り

Esaki S, Goshima F, Ozaki H, Takano G, Hatano Y, Kawakita D, Ijichi K, Watanabe T, Sato Y, Murata T, Iwata H, Shibamoto Y, Murakami S, Nishiyama Y, Kimura H

Cancer gene therapy 27 巻 ( 7-8 ) 頁： 585 - 598 2020年8月

Number of concomitant drugs with thrombocytopenic adverse effects and the extent inflammatory response resolution are risk factors for thrombocytopenia in patients treated with linezolid for more than 14 days 査読有り

Sato Yoshitaka, Iguchi Mitsutaka, Kato Yoshiaki, Morioka Hiroshi, Hirabayashi Aki, Tetsuka Nobuyuki, Tomita Yuka, Kato Daizo, Yamada Kiyofumi, Kimura Hiroshi, Yagi Tetsuya

NAGOYA JOURNAL OF MEDICAL SCIENCE 82 巻 ( 3 ) 頁： 407-414 2020年8月

Nationwide survey of systemic chronic active EBV infection in Japan in accordance with the new WHO classification 査読有り

Yonese Ichiro, Sakashita Chizuko, Imadome Ken-Ichi, Kobayashi Tohru, Yamamoto Masahide, Sawada Akihisa, Ito Yoshinori, Fukuhara Noriko, Hirose Asao, Takeda Yusuke, Makita Masanori, Endo Tomoyuki, Kimura Shun-Ichi, Ishimura Masataka, Miura Osamu, Ohga Shouichi, Kimura Hiroshi, Fujiwara Shigeyoshi, Arai Ayako

BLOOD ADVANCES 4 巻 ( 13 ) 頁： 2918-2926 2020年7月

A systemic form chronic active Epstein-Barr virus infection diagnosed from erythema nodosum-like skin lesions. 査読有り

Shibata A, Ishiguro Y, Makita S, Yamaga Y, Kimura H, Akiyama M

European journal of dermatology : EJD 30 巻 ( 3 ) 頁： 314-316 2020年6月

Epstein-Barr virus NK and T cell lymphoproliferative disease: report of a 2018 international meeting 査読有り国際共著

Cohena Jeffrey I., Iwatsuki Keiji, Ko Young-Hyeh, Kimura Hiroshi, Manoli Irini, Ohshima Koichi, Pittaluga Stefania, Quintanilla-Martinez Leticia, Jaffe Elaine S.

LEUKEMIA & LYMPHOMA 61 巻 ( 4 ) 頁： 808-819 2020年3月

Oncogenesis of CAEBV revealed: Intragenic deletions in the viral genome and leaky expression of lytic genes 査読有り

Murata Takayuki, Okuno Yusuke, Sato Yoshitaka, Watanabe Takahiro, Kimura Hiroshi

REVIEWS IN MEDICAL VIROLOGY 30 巻 ( 2 ) 頁： e2095 2020年3月

Antitumor activity of CDK inhibitor alsterpaullone in Epstein-Barr virus-associated lymphoproliferative disorders. 査読有り国際共著

Watanabe T, Sato Y, Masud HMAA, Takayama M, Matsuda H, Hara Y, Yanagi Y, Yoshida M, Goshima F, Murata T, Kimura H

Cancer science 2020年1月

Epstein-Barr Virus BBRF2 Is Required for Maximum Infectivity Open Access

Al Masud, HMA; Yanagi, Y; Watanabe, T; Sato, Y; Kimura, H; Murata, T

MICROORGANISMS 7 巻 ( 12 ) 2019年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Microorganisms

Epstein-Barr virus (EBV) is a member of the gammaherpesvirinae, which causes infectious mononucleosis and several types of cancer. BBRF2 is an uncharacterized gene of EBV and is expressed during the lytic phase. To evaluate its function, BBRF2-knockout EBV was prepared using bacterial artificial chromosome (BAC) technology and the CRISPR/Cas9 system. Although viral gene expression, DNA synthesis, and progeny secretion were not affected, the infectivity of progeny viruses was significantly reduced by the disruption of BBRF2. When expressed alone, BBRF2 protein localized to the nucleus and cytoplasm, while the coexpression of an interacting partner, BSRF1, resulted in its relocalization to the cytoplasm. Interestingly, the coexpression of BBRF2 protected BSRF1 from proteasome/ubiquitin-dependent degradation. Therefore, BBRF2, together with BSRF1, augments viral infectivity.

DOI： 10.3390/microorganisms7120705

NK-cell post-transplant lymphoproliferative disease with chronic active Epstein-Barr virus infection-like clinical findings.

Iemura T, Kondo T, Hishizawa M, Yamashita K, Kimura H, Takaori-Kondo A

International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases 2019年8月

Fujiwara Shigeyoshi, Kimura Hiroshi

FRONTIERS IN PEDIATRICS 7 巻 ( JULY ) 頁： 285 2019年7月

Viral DNA loads in various blood components of patients with Epstein-Barr virus-positive T/NK cell lymphoproliferative diseases.

Kawada JI, Kamiya Y, Sawada A, Iwatsuki K, Izutsu K, Torii Y, Kimura H, Ito Y

The Journal of infectious diseases 2019年6月

The BOLF1 gene is necessary for effective Epstein-Barr viral infectivity

Al Masud H. M. Abdullah, Watanabe Takahiro, Sato Yoshitaka, Goshima Fumi, Kimura Hiroshi, Murata Takayuki

VIROLOGY 531 巻頁： 114-125 2019年5月

S-Like-Phase Cyclin-Dependent Kinases Stabilize the Epstein-Barr Virus BDLF4 Protein To Temporally Control Late Gene Transcription Open Access

Sato, Y; Watanabe, T; Suzuki, C; Abe, Y; Al Masud, HMA; Inagaki, T; Yoshida, M; Suzuki, T; Goshima, F; Adachi, J; Tomonaga, T; Murata, T; Kimura, H

JOURNAL OF VIROLOGY 93 巻 ( 8 ) 2019年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Virology

Temporally controlled gene expression is necessary for the propagation of herpesviruses. To achieve this, herpesviruses encode several transcriptional regulators. In Epstein-Barr virus, BcRF1 associates with five viral proteins (BDLF4, BGLF3, BFRF2, BVLF1, and BDLF3.5) to form the viral late (L) gene regulatory complex, which is called the viral preinitiation complex (vPIC), on TATT-containing promoters. However, regulation of the vPIC has been largely unexplored. In this study, we performed two screens using a kinase inhibitor library and identified a series of cyclin-dependent kinase (CDK) inhibitors that downregulated the expression of L genes without any impact on viral DNA replication through destabilization of the BDLF4 protein. Knockdown of CDK2 by short hairpin RNA (shRNA) and proteasome inhibitor treatment showed that phosphorylation of the BDLF4 protein prevented ubiquitin-mediated degradation. Moreover, we demonstrated that cyclin A- and E-associated CDK2 complexes phosphorylated BDLF4 in vitro, and we identified several serine/threonine phosphorylation sites in BDLF4. Phosphoinactive and phosphomimic mutants revealed that phosphorylation at threonine 91 plays a role in stabilizing BDLF4. Therefore, our findings indicate that S-like-phase CDKs mediate the regulation of L gene expression through stabilization of the BDLF4 protein, which makes the temporal L gene expression system more robust. IMPORTANCE Late (L) genes represent more than one-third of the herpesvirus genome, suggesting that many of these genes are indispensable for the life cycle of the virus. With the exception of BCRF1, BDLF2, and BDLF3, Epstein-Barr virus L genes are transcribed by viral regulators, which are known as the viral preinitiation complex (vPIC) and the host RNA polymerase II complex. Because the vPIC is conserved in beta- and gammaherpesviruses, studying the control of viral L gene expression by the vPIC contributes to the development of drugs that specifically inhibit these processes in beta- and gammaherpesvirus infections/diseases. In this study, we demonstrated that CDK inhibitors induced destabilization of the vPIC component BDLF4, leading to a reduction in L gene expression and subsequent progeny production. Our findings suggest that CDK inhibitors may be a therapeutic option against beta- and gammaherpesviruses in combination with existing inhibitors of herpesvirus lytic replication, such as ganciclovir.

DOI： 10.1128/JVI.01707-18

Initial Characterization of the Epstein-Barr Virus BSRF1 Gene Product

Yanagi Yusuke, Al Masud H. M. Abdullah, Watanabe Takahiro, Sato Yoshitaka, Goshima Fumi, Kimura Hiroshi, Murata Takayuki

VIRUSES-BASEL 11 巻 ( 3 ) 2019年3月

Defective Epstein-Barr virus in chronic active infection and haematological malignancy (vol 4, pg 404, 2019) 国際共著 Open Access

Okuno, Y; Murata, T; Sato, Y; Muramatsu, H; Ito, Y; Watanabe, T; Okuno, T; Murakami, N; Yoshida, K; Sawada, A; Inoue, M; Kawa, K; Seto, M; Ohshima, K; Shiraishi, Y; Chiba, K; Tanaka, H; Miyano, S; Narita, Y; Yoshida, M; Goshima, F; Kawada, JI; Nishida, T; Kiyoi, H; Kato, S; Nakamura, S; Morishima, S; Yoshikawa, T; Fujiwara, S; Shimizu, N; Isobe, Y; Noguchi, M; Kikuta, A; Iwatsuki, K; Takahashi, Y; Kojima, S; Ogawa, S; Kimura, H

NATURE MICROBIOLOGY 4 巻 ( 3 ) 頁： 544 - 544 2019年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Nature Microbiology

In the version of this Letter originally published, in the sentence beginning “The major driver role of DDX3X mutations..”, the citation “Fig. 2a–f” should have been “Fig. 2”. In addition, in the sentence beginning “Another finding of interest was the presence of identical driver mutations..”, the citation “Fig. 3a,b and Fig. 4” should have been “Fig. 3”. This has now been amended in all versions of the Letter.

DOI： 10.1038/s41564-019-0387-8

[Clinical features of adult-onset chronic active Epstein-Barr virus infection].

EBV Viral Loads in Diagnosis, Monitoring, and Response Assessment Open Access

Kimura, H; Kwong, YL

FRONTIERS IN ONCOLOGY 9 巻 ( FEB ) 頁： 62 2019年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Frontiers in Oncology

The quantification of circulating Epstein Barr virus (EBV) DNA loads has played an important role in the diagnosis and management of EBV-associated lymphoid malignancies. Viral load measurement is particularly useful for monitoring EBV-DNA in hematopoietic stem cell transplant patients, and for assessing the prognosis or response to therapy of EBV-associated intractable lymphomas like extranodal NK/T-cell lymphoma, nasal type. Cell-free EBV-DNA in plasma can be used as a biomarker for estimating the severity or prognosis of these lymphomas. In addition to plasma, whole blood has been used for the management of transplant patients. Although measuring EBV-DNA has been useful, there is a lack of standardization and the optimal specimens for measuring viral loads are unknown. This can be attributed to the different forms of EBV-DNA that exist in peripheral blood and the different pathologies that result from diverse EBV disease states. As a result, guidelines for EBV diagnosis or the initiation of treatment are unclear. However, the newly established World Health Organization standard for EBV quantification will encourage collaborative studies across institutions and countries to establish proper guidelines for EBV diagnosis and the initiation of treatment.

DOI： 10.3389/fonc.2019.00062

Overview of EBV-Associated T/NK-Cell Lymphoproliferative Diseases

Kimura Hiroshi, Fujiwara Shigeyoshi

FRONTIERS IN PEDIATRICS 6 巻 2019年1月

Kawamoto K, Miyoshi H, Seto M, Kimura H, Ohshima K

[Rinsho ketsueki] The Japanese journal of clinical hematology 60 巻 ( 8 ) 頁： 944 - 952 2019年

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：一般社団法人日本血液学会

慢性活動性EBウイルス感染症（chronic active Epstein-Barr virus infection, CAEBV）は，Epstein-Barr virus（EBV）の再活性化（感染細胞の再増殖）による慢性持続性もしくは再発性の発熱やリンパ節腫脹，肝脾腫，皮疹などの伝染性単核球症様症状などを呈する疾患である。CAEBVの診断までに明確な免疫異常や既往歴などはなくその発症機序については現在も不明である。CAEBVはEBVに感染したT細胞もしくはNK細胞のclonal expansionによって血球貪食症候群や悪性リンパ腫などの致命的な疾患をきたす予後不良の疾患である。造血幹細胞移植がおこなわれなければ，CAEBVは予後不良の経過をたどる。このentitiyはほとんどが小児発症であるため改定2016WHO分類にはEBV-positive T-cell and NK-cell lymphoproliferative disease of childhoodのうちのchronic active EBV infection of T- and NK-cell type, systemic formとして収載されている。しかし同様の病態が成人にも発症することが知られるようになり，今回我々は成人発症CAEBV（adult-onset CAEBV）の臨床的特徴について多数例において検討した。その検討結果を報告するとともに，近年報告されているCAEBVを含むEBV-NK/T/LPDsにおける遺伝子異常などの結果を踏まえて成人発症のCAEBVの病態について概説する。

DOI： 10.11406/rinketsu.60.944

[EBV and Lymphomagenesis].

CiNii Research

Kimura H, Okuno Y

Uirusu 69 巻 ( 1 ) 頁： 13 - 22 2019年

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：日本ウイルス学会

Epstein-Barr virus (EBV)はヘルペスウイルス科に属する二本鎖DNAウイルスである．腫瘍ウイルスであるEBVは様々なリンパ系腫瘍と関連しており，そのリンパ腫原性については，MYCの転座・活性化など，Burkittリンパ腫を中心に広範な研究がなされてきた．しかしながら，Burkittリンパ腫以外のリンパ系腫瘍については，レセプターや宿主遺伝子の役割を含め，腫瘍化メカニズムについて不明な点が多く，その全貌は明らかとなっていない．近年，我々はEBV関連リンパ腫で，高率に欠損ウイルスが認められること，ウイルス複製に関わる遺伝子を欠損した場合にリンパ腫形成能が増すことをマウスモデルで示し，報告した．これまでも他の腫瘍ウイルスの遺伝子変異について多くの報告があるが，EBVのように70以上の遺伝子を有する大型ウイルスでの遺伝子欠失の成り立ちと意義については，未だ不明な点が多い．本稿では，欠失EBVを含めた最近の知見を中心に，EBVのリンパ腫原性について概説する．

DOI： 10.2222/jsv.69.13