Detection of EBV, HBV, HCV, HIV-1, HTLV-I and -II, and SMRV in Human and Other Primate Cell Lines

Cord C Uphoff,Sabine A Denkmann,Klaus G Steube,Hans G Drexler

doi:10.1155/2010/904767

Abstract

The high prevalence of contaminated cell cultures suggests that viral contaminations might be distributed among cultures. We investigated more than 460 primate cell lines for Epstein-Barr (EBV), hepatitis B (HBV), hepatitis C (HCV), human immunodeficiency virus type 1 (HIV-1), human T-cell leukemia/lymphoma virus I and II (HTLV-I/-II), and squirrel monkey retrovirus (SMRV) infections for risk assessment. None of the cell lines were infected with HCV, HIV-1, or HTLV-I/-II. However, one cell line displayed reverse transcriptase activity. Thirty-nine cell lines harbored EBV DNA sequences. Studies on the lytic phase of EBV revealed that five cell lines produce EBV particles and six further cell lines produced EBV upon stimulation. One cell line contained an integrated HBV genome fragment but showed no virus production. Six cell lines were SMRV-infected. Newly established cell lines should be tested for EBV infections to detect B-lymphoblastoid cell lines (B-LCL). B-LCLs established with EBV from cell line B95-8 should be tested for SMRV infections.

Highlights

Human primary cell cultures and cell lines have become fundamental tools for basic research in numerous life science faculties as well as for the production of bioactive reagents in biomedicine and biotechnology
Cell lines were usually established from patient material which might be infected with those viruses or perhaps with viruses linked to specific tumors, for example, human herpes virus type 8 (HHV-8) or novel types of papilloma viruses [2]
We describe the use of polymerase chain reaction (PCR) assays, enzyme-linked immunosorbent assay (ELISA), Southern and Western blotting for the detection of latent and active forms of the human pathogenic viruses (EBV, hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus type 1 (HIV-1), HTLV-I and -II), squirrel monkey retrovirus (SMRV), and retroviruses in general in a large panel of continuous primate cell lines to determine the potential risk for handling these cell lines

Summary

Introduction

Human primary cell cultures and cell lines have become fundamental tools for basic research in numerous life science faculties as well as for the production of bioactive reagents in biomedicine and biotechnology. They are already used for several decades and frozen cell cultures or blood and tissue samples obtained many years ago can be found in numerous laboratories. Cell lines were usually established from patient material which might be infected with those viruses or perhaps with viruses linked to specific tumors, for example, human herpes virus type 8 (HHV-8) or novel types of papilloma viruses [2]. The risk of emerging pathogens has to be kept under constant review [3]

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