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초록

영어

Mx is regulated by type I interferons and contains a typical GTP-binding motif like other members of the GTPase dynamin family. However, the functions and working mechanisms of the Mx protein in chicken spermatogonial stem cells (SSCs) are not well documented. In the present study, Mx-overexpressing chicken SSCs (chMx-SSCs) were established and the antiviral activity of chMx-SSCs against Newcastle disease virus, avian influenza viruses was investigated in vitro. For chicken SSCs isolation, day 20 fetal males derived testes were initially subjected to digestion by collagenase IV followed by 0.25% trypsin–EDTA. After discarding the supernatant, the cells were cultured in SSC medium. SSC colonies expressed pluripotent markers such as stagespecific embryonic antigen-1, Oct-4, Nanog, and Sox-2. Chicken Mx gene was constructed in plasmid DNA vector (pcDNA3.1/V5-His A-chMX) and ChMx-SSCs lines were established with chMX constructs. The antiviral activity of ChMx-SSCs was determined by real-time RT-PCR, flow cytometry, and western blot analyses after infection with Newcastle disease virus-green fluorescent protein (GFP) and avian influenza viruses (H9N2 and H1N1). ChMx-SSCs inhibited recombinant Newcastle disease virus (rNDV)- GFP replication as determined by the calculation of the proportion GFP signal- positive cells by FACS analysis. When SSCs showed 100% GFP expression, chMx- SSCs had only 3.6% GFP expression. At 24 h after avian influenza virus infection, chMx-SSCs had a lower hemagglutinin protein level and a higher level of Mx protein. When the number of released virion particles was estimated by plaque-formation assay, chMx-SSCs had significantly fewer stained visible plaques in the MDCK layer than SSCs. Our results suggest that overexpression of chicken Mx directly stimulates antiviral activity resulting in downregulation of viral progeny release. Chicken Mx overexpression in chicken SSCs can be applied for the production of virus resistant transgenic chicken via direct transplantation of chMx-SSCs into the testis.

저자

• Ju-Young Ji [ Animal Biotechnology Division, National Institute of Animal Science (NIAS), Rural Development Administration (RDA), College of Veterinary Medicine, Konkuk University ]
• Kuppannan Gobianand [ Animal Biotechnology Division, National Institute of Animal Science (NIAS) ]
• Jong-Ju Park [ Animal Biotechnology Division, National Institute of Animal Science (NIAS), Rural Development Administration (RDA) ]
• Jin-Gu No [ Animal Biotechnology Division, National Institute of Animal Science (NIAS), Rural Development Administration (RDA) ]
• Ju Sung Yang [ Department of Vaccine Development, Bioneer, Daejeon, Republic of Korea ]
• Man Sung Park [ College of Medicine, Hanlim University ]
• Dong Kee Jeong [ Department of Animal Biotechnology, Jeju National University ]
• Dong-Hoon Kim [ Animal Biotechnology Division, National Institute of Animal Science (NIAS), Rural Development Administration (RDA) ]
• Sung-June Byun [ Animal Biotechnology Division, National Institute of Animal Science (NIAS), Rural Development Administration (RDA) ]
• Jin Ki Park [ Animal Biotechnology Division, National Institute of Animal Science (NIAS), Rural Development Administration (RDA) ]
• Chang Sun Song [ College of Veterinary Medicine, Konkuk University ]
• Jae Gyu Yoo [ Animal Biotechnology Division, National Institute of Animal Science (NIAS), Rural Development Administration (RDA) ]

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발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  Reproductive & Developmental Biology(Supplement)

• 간기

  연간

• 수록기간

  2001~2017

• 십진분류

  KDC 527 DDC 636