LONG TERM PERSISTENCE OF VIRAL CENES IN DIVIDING HOST CELLS IN CULTURE AFTER PRODUCTIVE REPLICATION OF A MUTANT OF INFLUENZA VIRUS A/WSN.

流感病毒 A/WSN 突变体有效复制后，培养物中分裂宿主细胞中病毒场景的长期存留。

• 批准号: 04454203
• 负责人: TOBITA Kiyotake
• 金额: $ 3.07万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454203/
• 关键词: influevza virus; latent infection; polymerase chain reaction; copetitive PCR; viral genes; pUC19ベクター; 遺伝子の塩基配列

By repeated superinfections of MDBK cells with AWBY-140, a weakly cytolytic mutant of influenza virus A/WAN ( HINI ), at a high multiplicity and subsequent cultivations of the surviving cells, we established a latent infection of influenza virus in cell culture ( MDBK-RR ). Within MDBK-RR cells, we detected all the 8 genes of influenza virus by a polymerase chain reaction ( PCR ) using influenza viral gene-specific primer pairs.At least some of them retained their full size, accompanied by a few point mutations.In addition to viral RNA ( vRNA ), comlementary RNA ( cRNA ) and viral messenger RNA ( mRNA )were also detected within the cells.Competitive PCR revesled that the copy number of the persisting viral genes was 10^5 to 10^6 per 8 x 10^6 cells, which did not alter significantry as the passage number of the culture increased.

通过用流感病毒 A/WAN ( HINI ) 的弱溶细胞突变体 AWBY-140 重复感染 MDBK 细胞，并在随后培养存活细胞，我们在细胞培养物 (MDBK-RR ) 中建立了流感病毒的潜伏感染。在MDBK-RR细胞内，我们使用流感病毒基因特异性引物对，通过聚合酶链式反应（PCR）检测到了流感病毒的所有8个基因。至少其中一些基因保留了完整大小，并伴有一些点突变。除了病毒RNA（vRNA）外，还检测到了互补RNA（cRNA）和病毒信使RNA（mRNA）。 竞争性 PCR 表明，每 8 x 10^6 细胞中持续存在的病毒基因的拷贝数为 10^5 至 10^6，并且随着培养物传代次数的增加，该拷贝数不会发生显着变化。

项目成果

Urabe,M.et al.: "Persistence of viral genes in a variant of MDBK cell after productive replication of a mutant of influenza virus A/WSN." Archives of Virology. 128. 97-110 (1993)

Urabe,M.等人：“流感病毒 A/WSN 突变体有效复制后，MDBK 细胞变体中病毒基因的持久性。”

Urabe,M.et al.: "MDBX cells which survived infection with a mutant of influenza virus A/WSN and subsequently received many passages contained viral M and NS genes in full length in the absence of virus production." Archives of Virology. 130. 457-462 (1993

Urabe,M.等人：“MDBX 细胞在流感病毒 A/WSN 突变体感染后存活下来，随后在没有病毒产生的情况下接受了包含全长病毒 M 和 NS 基因的多次传代。”

Urabe, M., Tanaka, T., and Tobita, K.: "MDBK cells which survived infection with a mutant of influenzs virus A/WSN and subsequently received many passages contained viral M and NS genes full length in the absence of virus production." Archives of Virology

Urabe, M.、Tanaka, T. 和 Tobita, K.：“MDBK 细胞在流感病毒 A/WSN 突变体感染后存活下来，随后在没有病毒产生的情况下接受了包含全长病毒 M 和 NS 基因的许多传代

M.Urabe: "MDBK cells which survived infection with a mutant of influenza virus A/WSN and subsequently received many passages contained viral M and NS genes in full length in the absence of virus production." Archives of Virology. (1993)

M.Urabe：“MDBK 细胞在流感病毒 A/WSN 突变体的感染中存活下来，并随后在没有病毒产生的情况下接受了包含全长病毒 M 和 NS 基因的多次传代。”

Urabe, M., Tanaka, T., Odagiri, T., Tashiro, M., and Tobita, K.: "Persistence of viral genes in a variant of MDBK cell after productive replication of a mutant of influenzs virus A/WSN." Archives of Virology. 128. 97-110 (1993)

Urabe, M.、Tanaka, T.、Odagiri, T.、Tashiro, M. 和 Tobita, K.：“流感病毒 A/WSN 突变体有效复制后，病毒基因在 MDBK 细胞变体中持续存在。