Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang, PR China.
Heilongjiang Provincial Key Laboratory for Infection and Immunity, Department of Immunology, Harbin Medical University, Harbin, PR China.
Virulence. 2024 Dec;15(1):2435372. doi: 10.1080/21505594.2024.2435372. Epub 2024 Nov 29.
Interferon-αs (IFN-αs) are crucial cytokines for inducing protective antiviral responses. The baculovirus-mediated gene transduction of mammalian cells (BacMam) is an efficient delivery tool for recombinant protein expression in mammalian cells. This study focuses on the delivery of bovine IFN-α (BoIFNα) using the BacMam system. A recombinant pACEMam1-BoIFNα bacmid was constructed, and a recombinant BacMam-BoIFNα virus was obtained. After transducing HEK293T cells with this virus, BoIFNα protein was successfully secreted into the cell supernatant, displaying antiviral activities against VSV, BPIV3, BEV, and BVDV in bovine-derived cells. Additionally, the BacMam-BoIFNα virus inhibited the replication of these viruses in MDBK cells, induced the transcription of ISGs, and the expression of M × 1 in both MDBK and BT cells. These induction effects were significantly reduced following treatment with a JAK1 inhibitor, suggesting that the BacMam-BoIFNα virus exerts antiviral activity in MDBK cells through JAK-STAT signaling pathway. Furthermore, the study found that the BacMam-BoIFNα virus significantly inhibited the replication of BPIV3 in the lung and trachea of mice. Overall, this study demonstrates that the BacMam-BoIFNα virus have antivirus activities both and , signaling through JAK-STAT pathway, and provides an efficient interferon gene delivery tool for combating bovine viral infections.
干扰素-αs(IFN-αs)是诱导保护性抗病毒反应的关键细胞因子。杆状病毒介导的哺乳动物细胞基因转导(BacMam)是在哺乳动物细胞中表达重组蛋白的有效传递工具。本研究聚焦于利用 BacMam 系统递送牛干扰素-α(BoIFNα)。构建了重组 pACEMam1-BoIFNα 杆状病毒载体,并获得了重组 BacMam-BoIFNα 病毒。该病毒转导 HEK293T 细胞后,BoIFNα 蛋白成功分泌到细胞上清液中,对牛源细胞中的 VSV、BPIV3、BEV 和 BVDV 表现出抗病毒活性。此外,BacMam-BoIFNα 病毒抑制了 MDBK 细胞中这些病毒的复制,诱导了 ISGs 的转录和 MDBK 及 BT 细胞中 Mx1 的表达。用 JAK1 抑制剂处理后,这些诱导作用显著降低,表明 BacMam-BoIFNα 病毒通过 JAK-STAT 信号通路在 MDBK 细胞中发挥抗病毒活性。此外,研究发现 BacMam-BoIFNα 病毒显著抑制了 BPIV3 在小鼠肺和气管中的复制。综上所述,本研究表明 BacMam-BoIFNα 病毒具有抗病毒活性,通过 JAK-STAT 通路发挥作用,并为防治牛病毒性感染提供了一种有效的干扰素基因传递工具。