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In Vitro Infectious Risk Assessment of Heliothis virescens ascovirus 3j (HvAV-3j) toward Non-target Vertebrate Cells

  • RESEARCH ARTICLE
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Virologica Sinica

Abstract

As specific pathogens of noctuid pests, including Spodoptera exigua, S. litura, Helicoverpa armigera, and Mythimna separata, ascoviruses are suitable for the development of bioinsecticides. In this study, the infectivity of Heliothis virescens ascovirus 3j (HvAV-3j) on insect and mammalian cells was evaluated. HvAV-3j infection induced drastic morphological changes in Sf9, HzAM1, SeFB, and HaFB cells, including swelling and detachment. Notably, the latter phenomena did not occur in HvAV-3j-inoculated mammalian cells (HEK293, 7402, HePG2, PK15, ST, and TM3). MTT assays indicated that HvAV-3j inhibited the growth of host insect cells from the 6th hpi, but no effects were detected in the HvAV-3j-inoculated mammalian cells. Furthermore, viral DNA replication, gene transcription, and protein expression were investigated, and the results consistently suggested that HvAV-3j viruses were not able to replicate their genomic DNA, transcribe, or express their proteins in the non-target vertebrate cells. The HvAV-3j genes were only transcribed and expressed in the four insect cell lines. These results indicated that HvAV-3j was infectious to cells derived from S. frugiperda, S. exigua, H. armigera, and H. zea but not to cells derived from human, pig, and mouse, suggesting that ascoviruses are safe to non-target vertebrate cells.

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Acknowledgements

The authors thank Dr. Zhi-Hong Hu (Wuhan Institute of Virology, Chinese Academy of Sciences) for generously providing HzAM1, Sf9 cells and E. coli strain of DH10Bac; Dr. Qi-Lian Qin (Institute of Zoology, Chinese Academy of Sciences) for generously providing SeFB cells; Dr. Jian-Hong Li (College of Plant Science & Technology, Huazhong Agricultural University) for kindly providing HaFB cells; Dr. Si Qin (College of Food Science and Technology, Hunan Agriculture University) kindly giving HePG2 cells; Dr. Yun Liu (Zunyi Medical University) generously providing 7402 cells; Dr. Zhong Ding (College of Plant Protection, Hunan Agriculture University) generously providing HEK293 cells; Dr. Qing Yang (College of Veterinary Medicine, Hunan Agriculture University) generously providing PK15, ST, and TM3 cells. This work was supported by the National Natural Science Foundation of China (31700141, 31872027) and the grant of Chinese Post-doctoral special funding (2018T110832) and Program to supporting research activities of female researchers in Ministry of education, culture, sports, science and technology-Japan.

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Authors and Affiliations

  1. Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Hunan Agricultural University, Changsha, 410128, China

    Huan Yu, Yi-Yi Ou-Yang, Ni Li & Guo-Hua Huang

  2. College of Plant Protection, Hunan Agricultural University, Changsha, 410128, China

    Huan Yu, Yi-Yi Ou-Yang, Ni Li & Guo-Hua Huang

  3. Tokyo University of Agriculture and Technology, Saiwai, Fuchu, Tokyo, 183-8509, Japan

    Madoka Nakai

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  1. Huan Yu
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  2. Yi-Yi Ou-Yang
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  3. Ni Li
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  4. Madoka Nakai
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  5. Guo-Hua Huang
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Contributions

HY, MN and GHH contributed to the study design. HY, YYOY and NL performed the experiments. HY, MN and GHH contributed reagents. HY and YYOY analyzed the data. HY, YYOY, MN and GHH wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Madoka Nakai or Guo-Hua Huang.

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The authors declared no competing financial interests in this paper.

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This article does not contain any studies with human or animal subjects performed by any of the authors.

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Yu, H., Ou-Yang, YY., Li, N. et al. In Vitro Infectious Risk Assessment of Heliothis virescens ascovirus 3j (HvAV-3j) toward Non-target Vertebrate Cells. Virol. Sin. 34, 423–433 (2019). https://doi.org/10.1007/s12250-019-00113-4

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