Comprehensive Comparison of Novel Bovine Leukemia Virus (BLV) Integration Sites between B-Cell Lymphoma Lines BLSC-KU1 and BLSC-KU17 Using the Viral DNA Capture High-Throughput Sequencing Method
Bovine leukemia virus (BLV) infects cattle and integrates into host DNA, causing enzootic bovine leukosis (EBL), an aggressive B-cell lymphoma. Here, we developed a novel proviral DNA-capture sequencing (proviral DNA-capture-seq) method investigating BLV proviral integration in two B-cell lymphoma l...
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MDPI AG
2022-05-01
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| Online Access: | https://www.mdpi.com/1999-4915/14/5/995 |
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| author | Meripet Polat Yamanaka Susumu Saito Kazuyoshi Hosomichi Yoko Aida |
| author_facet | Meripet Polat Yamanaka Susumu Saito Kazuyoshi Hosomichi Yoko Aida |
| author_sort | Meripet Polat Yamanaka |
| collection | DOAJ |
| description | Bovine leukemia virus (BLV) infects cattle and integrates into host DNA, causing enzootic bovine leukosis (EBL), an aggressive B-cell lymphoma. Here, we developed a novel proviral DNA-capture sequencing (proviral DNA-capture-seq) method investigating BLV proviral integration in two B-cell lymphoma lines, BLSC-KU1 and BLSC-KU17, derived from BLV-infected cattle with EBL. We designed BLV-specific biotinylated probes to capture the provirus genome and enrich libraries for next-generation sequencing. Validation showed high specificity and efficient enrichment of target sequence reads as well as identification of three BLV proviral integration sites on BLV persistently infected FLK-BLV cells as a positive control. We successfully detected a single BLV proviral integration site on chromosome 19 of BLSC-KU1 and chromosome 9 of BLSC-KU17, which were confirmed by standard PCR and Sanger sequencing. Further, a defective provirus in BLSC-KU1 and complete BLV proviral sequence in BLSC-KU17 were confirmed using long PCR and sequencing. This is the first study to provide comprehensive information on BLV proviral structure and viral integration in BLSC-KU1 and BLSC-KU17. Moreover, the proposed method can facilitate understanding of the detailed mechanisms underlying BLV-induced leukemogenesis and may be used as an innovative tool to screen BLV-infected cattle at risk at an earlier stage than those that have already developed lymphoma. |
| first_indexed | 2024-03-10T01:36:23Z |
| format | Article |
| id | doaj.art-54675f9ad57248d382c1dc75c3789aeb |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T01:36:23Z |
| publishDate | 2022-05-01 |
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| record_format | Article |
| series | Viruses |
| spelling | doaj.art-54675f9ad57248d382c1dc75c3789aeb2023-11-23T13:31:32ZengMDPI AGViruses1999-49152022-05-0114599510.3390/v14050995Comprehensive Comparison of Novel Bovine Leukemia Virus (BLV) Integration Sites between B-Cell Lymphoma Lines BLSC-KU1 and BLSC-KU17 Using the Viral DNA Capture High-Throughput Sequencing MethodMeripet Polat Yamanaka0Susumu Saito1Kazuyoshi Hosomichi2Yoko Aida3Laboratory of Global Infectious Diseases Control Science, Department of Global Agricultural Science, Graduate School of Agriculture and Life Science, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, JapanViral Infectious Diseases Unit, RIKEN, 2-1 Hirosawa, Wako 351-0198, JapanDepartment of Bioinformatics and Genomics, Graduate School of Medical Sciences, Kanazawa University, Takara-machi 13-1, Kanazawa 920-8640, JapanLaboratory of Global Infectious Diseases Control Science, Department of Global Agricultural Science, Graduate School of Agriculture and Life Science, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, JapanBovine leukemia virus (BLV) infects cattle and integrates into host DNA, causing enzootic bovine leukosis (EBL), an aggressive B-cell lymphoma. Here, we developed a novel proviral DNA-capture sequencing (proviral DNA-capture-seq) method investigating BLV proviral integration in two B-cell lymphoma lines, BLSC-KU1 and BLSC-KU17, derived from BLV-infected cattle with EBL. We designed BLV-specific biotinylated probes to capture the provirus genome and enrich libraries for next-generation sequencing. Validation showed high specificity and efficient enrichment of target sequence reads as well as identification of three BLV proviral integration sites on BLV persistently infected FLK-BLV cells as a positive control. We successfully detected a single BLV proviral integration site on chromosome 19 of BLSC-KU1 and chromosome 9 of BLSC-KU17, which were confirmed by standard PCR and Sanger sequencing. Further, a defective provirus in BLSC-KU1 and complete BLV proviral sequence in BLSC-KU17 were confirmed using long PCR and sequencing. This is the first study to provide comprehensive information on BLV proviral structure and viral integration in BLSC-KU1 and BLSC-KU17. Moreover, the proposed method can facilitate understanding of the detailed mechanisms underlying BLV-induced leukemogenesis and may be used as an innovative tool to screen BLV-infected cattle at risk at an earlier stage than those that have already developed lymphoma.https://www.mdpi.com/1999-4915/14/5/995bovine leukemia virus (BLV)B-cell lymphoma lineintegration siteproviral DNA capture sequencingnext-generation sequencing (NGS)PCR |
| spellingShingle | Meripet Polat Yamanaka Susumu Saito Kazuyoshi Hosomichi Yoko Aida Comprehensive Comparison of Novel Bovine Leukemia Virus (BLV) Integration Sites between B-Cell Lymphoma Lines BLSC-KU1 and BLSC-KU17 Using the Viral DNA Capture High-Throughput Sequencing Method Viruses bovine leukemia virus (BLV) B-cell lymphoma line integration site proviral DNA capture sequencing next-generation sequencing (NGS) PCR |
| title | Comprehensive Comparison of Novel Bovine Leukemia Virus (BLV) Integration Sites between B-Cell Lymphoma Lines BLSC-KU1 and BLSC-KU17 Using the Viral DNA Capture High-Throughput Sequencing Method |
| title_full | Comprehensive Comparison of Novel Bovine Leukemia Virus (BLV) Integration Sites between B-Cell Lymphoma Lines BLSC-KU1 and BLSC-KU17 Using the Viral DNA Capture High-Throughput Sequencing Method |
| title_fullStr | Comprehensive Comparison of Novel Bovine Leukemia Virus (BLV) Integration Sites between B-Cell Lymphoma Lines BLSC-KU1 and BLSC-KU17 Using the Viral DNA Capture High-Throughput Sequencing Method |
| title_full_unstemmed | Comprehensive Comparison of Novel Bovine Leukemia Virus (BLV) Integration Sites between B-Cell Lymphoma Lines BLSC-KU1 and BLSC-KU17 Using the Viral DNA Capture High-Throughput Sequencing Method |
| title_short | Comprehensive Comparison of Novel Bovine Leukemia Virus (BLV) Integration Sites between B-Cell Lymphoma Lines BLSC-KU1 and BLSC-KU17 Using the Viral DNA Capture High-Throughput Sequencing Method |
| title_sort | comprehensive comparison of novel bovine leukemia virus blv integration sites between b cell lymphoma lines blsc ku1 and blsc ku17 using the viral dna capture high throughput sequencing method |
| topic | bovine leukemia virus (BLV) B-cell lymphoma line integration site proviral DNA capture sequencing next-generation sequencing (NGS) PCR |
| url | https://www.mdpi.com/1999-4915/14/5/995 |
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