Go to ContentTo Startpage
Virus Diagnostics ProjectsVirusdiagnostik

Virus Diagnostics

Figure 1: Western blot analysis of EBV infection status in B cell lineage-derived lines stimulated with phorbol ester TPA detected with ZEBRA antibody.

Immortalized human and animal cells are frequently used for the study of virus infection and replication, and for the production of viral antigenes for vaccines. However, virus-infected producer cell lines may pose health risks.  Whereas some viruses can be easily propagated in continuous cell lines (e.g., human retroviruses), propagation of other viruses depends on the microenvironment or maturation of the otherwise permissive cells. Additionally, some viruses exhibit a latent or cryptic infection cycle, during which no active viruses are produced (e.g., Epstein-Barr virus (EBV), and retroviral proviruses). However, latency can be switched to the productive lytic cycle by certain inducers (see Figure 1).

Viruses are sometimes associated with certain tumor types as has been shown for EBV, hepatitis B virus (HBV), human T lymphotropic viruses (HTLV), human papilloma virus (HPV) and several others. Cell lines can be used to test tumorigenicity. Additionally, replication mechanisms or virus cell interactions in human and animal cells can be elucidated using in vitro cell culture systems. We investigated the infection status of human and animal cell lines contaminated with human pathogenic viruses, e.g. EBV, HBV, HTLV, human herpesvirus type 8 (HHV-8) etc. In the event of contamination the type of infection (latent, lytic, persisting, transforming) may thus be determined. Hence, we recently investigated the prevalence of squirrel monkey retroviruses (SMRV) in a wide range of cell lines, and of xenotropic murine leukemia virus related virus (XMRV) and HPV in prostate cancer and oropharyngeal cancer cell lines, respectively.

Applying the extraordinary potential of "Next Generation Sequencing" (NGS) we are now in the position to detect almost any kind of viruses in human cells. As all virus-infected cells produce viral RNA regardless of the infection status (latent or lytic), RNA sequences expressed by the eukaryotic cells can be used to identify DNA and RNA viruses. We are currently screening RNAseq and Whole Exome Sequencing (WES) data for viral sequences to detect viral contaminations of cell cultures on one hand and to investigate the implication of viral infections on the tumorigenesis of human cells on the other hand. 

The replication of EBV and HBV in cell lines was analyzed with respect to the formation of concatemeric DNA intermediates together with the introduction of terminal repeats into the genomes of EBV replication intermediates, and the production of HBV in a hepatocellular carcinoma cell line.

Mycoplasma detection and elimination remains an enduring field of interest, and the routine assays employed at the cell culture collection have been developed and continuously improved over the years.

Photo of Cord C.  Uphoff
Dr. Uphoff, Cord C.
Phone: +49-531/2616-156

Virus Diagnostics

Selected References

1. Drexler HG, Dirks WG, MacLeod RA, Uphoff CC: False and mycoplasma-contaminated leukemia-lymphoma cell lines: time for a reappraisal. Int J Cancer 140 (5): 1209-1214 (2017).

2. Uphoff CC, Lange S, Denkmann SA, Garritsen HS, Drexler HG: Prevalence and characterization of murine leukemia virus contamination in human cell lines. PLoS One 10 (4): e0125622 (2015).

3. Dirks WG, Uphoff CC: Quality control essentials in human cell culture: Cell line cross-contamination and microbiological infections. Animal Cell Biotechnology in Biologics Production: 102-113 (2014).

4. Uphoff CC, Drexler HG: Detection of Mycoplasma contamination in cell cultures. Curr Protoc Mol Biol 106: 28/4/1-28/4/14 (2014).

5. Uphoff CC, Drexler HG: Eradication of Mycoplasma contaminations from cell cultures. Curr Protoc Mol Biol 106: 28/5/1-28/5/12 (2014).

6. Uphoff CC, Denkmann SA, Drexler HG: Treatment of Mycoplasma contamination in cell cultures with plasmocin. J Biomed Biotechnol 2012: 267678 (2012).

7. Uphoff CC, Denkmann SA, Steube KG, Drexler HG: Detection of EBV, HBV, HCV, HIV-1, HTLV-I and -II and SMRV in human and other primate cell lines. J Biomed Biotechnol, Vol. 2010: Article ID 904767, 23 pages (2010).

8. Markoullis K, Bulian D, Hölzlwimmer G, Quintanilla-Martinez L, Heiliger K-J, Zitzelsberger H, Scherb H, Mysliwietz J, Uphoff CC, Drexler HG, Adler T, Busch DH, Schmidt J, Mahabir E: Mycoplasma contamination of murine embryonic stem cells affects cell parameters, germ line transmission and chimeric progeny. Transgenic Res 18: 71-87 (2008).

9. Burnett TA, Dinkla K, Rohde M, Chhatwal GS, Uphoff C, Srivastava M, Cordwell SJ, Geary S, Liao X, Minion FC, Walker MJ, Djordjevic SP: P159 is a proteolytically processed, surface adhesin of Mycoplasma hyopneumoniae: defined domains of P159 bind heparin and promote adherence to eukaryote cells. Mol Microbiol 60: 669-686 (2006).