알림·소식
알림·소식
[2019 Pasteur Colloquium] Daniel Todt 박사, 독일 보훔 루드 대학교
2019-10-08
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[한국파스퇴르연구소] 파스퇴르 콜로퀴엄
일시 : 2019년 10월 10일 (목), 오전 11시
장소 : 모노드 회의실 (1층), 한국파스퇴르연구소
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연자
Daniel Todt 박사
독일 보훔 루드 대학교 분자의학바이러스학 박사후연구원
강연주제
Robust hepatitis E virus infection and transcriptional response in human hepatocytes
*파스퇴르 콜로퀴엄 관련 문의사항은 sukyon.yang@ip-korea.org 로 이메일 송부 부탁드립니다. 연구소 관련 추가 내용은 홈페이지 (www.ip-korea.org) 를 통해 확인하실 수 있습니다.
** 한국파스퇴르연구소 약도: http://www.ip-korea.org/about_us/location.php
[한국파스퇴르연구소] 파스퇴르 콜로퀴엄
일시 : 2019년 10월 10일 (목), 오전 11시
장소 : 모노드 회의실 (1층), 한국파스퇴르연구소
______________________________________________________________________________________________________
연자
Daniel Todt 박사
독일 보훔 루드 대학교 분자의학바이러스학 박사후연구원
강연주제
Robust hepatitis E virus infection and transcriptional response in human hepatocytes
초록
Hepatitis E virus (HEV) is the causative agent of hepatitis E in humans and the leading cause for acute viral hepatitis worldwide. The virus is classified as a member of the genus Orthohepevirus within the Hepeviridae family. Due to the absence of a robust cell culture model for HEV infection, the analysis of the viral life cycle and development of effective antivirals and vaccines is severely limited. In this study, we established a protocol based on the HEV genotype 3 p6 (Kernow C-1) and the human hepatoma cell lines HepG2 and HepG2/C3A with different media conditions to produce HEV cell culture-derived particles (HEVcc) with viral titers between 105 to 106 FFU/mL. Viral titers could be further enhanced by an HEV variant harboring a mutation in the RNA-dependent RNA polymerase. These HEVcc particles were characterized in density gradients and allowed the trans-complementation of subgenomic reporter HEV replicons. In addition, in vitro produced particles were infectious in liver-humanized mice with high RNA copy numbers detectable in serum and feces. Efficient infection of primary human and swine hepatocytes using the developed protocol could be observed and was inhibited by the off-label drug ribavirin. Finally, RNA sequencing studies of HEV infected primary human hepatocytes demonstrated a temporally structured transcriptional defense response. In conclusion, this robust cell culture model of HEV infection provides a powerful tool for studying viral-host interactions that should facilitate the discovery of antiviral drugs and vaccines for this important zoonotic pathogen.
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Hepatitis E virus (HEV) is the causative agent of hepatitis E in humans and the leading cause for acute viral hepatitis worldwide. The virus is classified as a member of the genus Orthohepevirus within the Hepeviridae family. Due to the absence of a robust cell culture model for HEV infection, the analysis of the viral life cycle and development of effective antivirals and vaccines is severely limited. In this study, we established a protocol based on the HEV genotype 3 p6 (Kernow C-1) and the human hepatoma cell lines HepG2 and HepG2/C3A with different media conditions to produce HEV cell culture-derived particles (HEVcc) with viral titers between 105 to 106 FFU/mL. Viral titers could be further enhanced by an HEV variant harboring a mutation in the RNA-dependent RNA polymerase. These HEVcc particles were characterized in density gradients and allowed the trans-complementation of subgenomic reporter HEV replicons. In addition, in vitro produced particles were infectious in liver-humanized mice with high RNA copy numbers detectable in serum and feces. Efficient infection of primary human and swine hepatocytes using the developed protocol could be observed and was inhibited by the off-label drug ribavirin. Finally, RNA sequencing studies of HEV infected primary human hepatocytes demonstrated a temporally structured transcriptional defense response. In conclusion, this robust cell culture model of HEV infection provides a powerful tool for studying viral-host interactions that should facilitate the discovery of antiviral drugs and vaccines for this important zoonotic pathogen.
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*파스퇴르 콜로퀴엄 관련 문의사항은 sukyon.yang@ip-korea.org 로 이메일 송부 부탁드립니다. 연구소 관련 추가 내용은 홈페이지 (www.ip-korea.org) 를 통해 확인하실 수 있습니다.
** 한국파스퇴르연구소 약도: http://www.ip-korea.org/about_us/location.php