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Medycyna Doświadczalna i Mikrobiologia 2012, 64(3): 239-244

Replikacja wirusa HCV w linii komórkowej Huh–7.5
[HCV replication in Huh–7.5 cell line]

Marcin Komorowski, Agnieszka Kołakowska, Paulina Godzik, Kazimierz Madaliński

Streszczenie

Zakażenie wirusem zapalenia wątroby C (HCV) stanowi globalny problem zdrowotny. Prowadzone są intensywne badania nad stworzeniem komórko­wego modelu replikacji wirusa. Opracowany konstrukt HCV o genotypie 2a (JFH1 – Japanese Fulminant Hepatitis) badano in vitro początkowo w linii komórkowej Huh–7. Ze względu na niską infekcyjność JFH1 w stosunku do linii Huh–7 zastąpiono ją linią komórkową Huh–7.5. Celem bieżącej pracy było zbadanie dynamiki zmian ilości wirusowego RNA w hodowli Huh–7.5.

Abstract

Introduction. Infection with hepatitis C virus is a serious worldwide health problem. Since its discovery in 1989, the development of a cell culture system for HCV has been a major goal for scientists worldwide. In 2005 the first tissue culture that led to the production of HCV particles (2a genotype) has been created. The aim of the study was to determine viral RNA level in an infectious HCV cell culture system.Methods. Huh-7.5 cell line was infected with the JFH1 (Japanese Fulminant Hepatitis) RNA by lipofection. The level of HCV RNA was measured in supernatant of the cell culture by Real-Time PCR method.Results. HCV RNA was detected in the supernatant of Huh-7.5 cell line infected with the use of JFH1 RNA and lipofectamin. The maximal level of HCV RNA (3.69x109 copies/ml) was detected 96h after transfection. After about 30 days of transfection, HCV RNA was on the stable level (107–108).Conclusions. Huh-7.5 cell line infected with HCV form a robust cell culture model of HCV infection. HCV replicates on high levels in Huh-7.5, which is the crucial event for the investigation of new antivirals in in vitro model.

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