Skip to main content
Cytotechnology logoLink to Cytotechnology
. 2001 Oct;37(2):83–92. doi: 10.1023/A:1019945416505

Endogenous retrovirus particles and their repercussion effects on the growth behaviour of continuous hybridoma cultivation processes

Andrea Hawerkamp 1, Frank Gudermann 1, Anna Falkenhain 1, Dirk Luetkemeyer 1, Uwe Kahmann 2, Jürgen Lehmann 1
PMCID: PMC3449696  PMID: 19002905

Abstract

Hybridoma cells used for the production of monoclonal antibodies are also known to form growth inhibitory substances. Growth inhibitors already described in the literature belong to the class of peptides and proteins likeTGF-ß (Transforming Growth Factor-ß). The endogenous retrovirus particles - a further potential substance producing this kind of effect - are described here. To examine whether the retrovirus particles participated in growth inhibitory effects hybridoma cells were cultivated in continuous perfusion mode by using a special reactor set-up. A rapid increase of the signal in the supernatant which coincided with a decrease of viability could be observed by monitoring the reverse transcriptase-activity during this type of fermentation process. The examination of concentrated and fractionated supernatant from this period showed growth inhibitory effects in the biological assay (MTT-assay). Investigations of respective fractions demonstrated retrovirus particles with reverse transcriptase-activity. Based on RT-PCR data it was shown that only inhibitory fractions contain retrovirus particles which were of E-MuLV and MCF origin.

Keywords: Growth inhibition effects, Hybridoma cells, Retrovirus particles, Reverse transcriptase-activity

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

References

  1. Bartal A.H., Feit C., Erlandson R.A., Hirshaut Y. Detection of retroviral particles in hybridomas secreting mono-clonal antibodies. Med. Microbiol. Immunol. 1986;174:325–332. doi: 10.1007/BF02123685. [DOI] [PubMed] [Google Scholar]
  2. Büntemeyer H., Lutkemeyer D., Lehmann J. Optimization of serum free fermentation processes for antibody pro-duction. Cytotechnology. 1991;5:57–67. doi: 10.1007/BF00365534. [DOI] [PubMed] [Google Scholar]
  3. Dalton A.J. Micromorphology of murine tumor viruses and of affected cells. Fed. Proc. 1962;21:936–941. [PubMed] [Google Scholar]
  4. Deo Y., Ghebremariam H., Cloyd M. Detection and characterization of murine ecotropic recombinant virus in myeloma and hybridoma cells. Hybridoma. 1994;13:69–76. doi: 10.1089/hyb.1994.13.69. [DOI] [PubMed] [Google Scholar]
  5. Froud S.J., Birch J., McLean C., Shepherd A.J., Smith K.T. Viral contaminants found in mouse cell lines used in the production of biological products. In: Carrondo M.J.T., Griffiths J.B., Moreira J.L.P., editors. Animal Cell Technology: From Vaccines to Genetic Medicine. Dordrecht: Kluwer Academic Publishers; 1997. pp. 681–686. [Google Scholar]
  6. Hansen M.B., Nielsen S.E., Berg K. Re-examination and further development of a precise and rapid dye method for measuring cell growth/ cell kill. J. Immunol. Methods. 1989;119:203–210. doi: 10.1016/0022-1759(89)90397-9. [DOI] [PubMed] [Google Scholar]
  7. Hawerkamp A., Lütkemeyer D., Gudermann F., Falkenhain A., Büntemeyer H., Lehmann J. Effects on growth behaviour in continuous hybridoma cell cultures: The role of viral contamination. Cytotechnology. 1998;28:9–29. doi: 10.1023/A:1008000924426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hawerkamp A. 1999. Zellwachstumsinhibitorische Effekte in Hybridomzellkultivierungen: Die Rolle endogener Retroviren, PhD thesis, Institute of Cell Culture Technology, University of Bielefeld pp. 161.
  9. Heidemann R., Lütkemeyer D., Buntemeyer H., Lehmann J. Effects of dissolved oxygen levels and the role of extra and intracellular amino acid concentrations upon the metabolism of mammalian cell lines during batch and continuous cultures. Cytotechnology. 1998;26:185–197. doi: 10.1023/A:1007917409455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Heinemeyer T., Klingenhoff A., Hansen W., Schumacher L., Hauser H., Wirth M. A sensitive method for the detection of murine C-type retroviruses. J. Virol. Meth. 1997;63:155–165. doi: 10.1016/S0166-0934(96)02124-6. [DOI] [PubMed] [Google Scholar]
  11. Hull R. Purification, biophysical and biochemical characteri-sation of viruses with especial reference to plant viruses. In: Mahy B.W.J., editor. Virology a practical approach. Oxford: IRL Press; 1985. pp. 1–24. [Google Scholar]
  12. Irving J.M., Chang L.W.S., Castillo F.J. A reverse transcriptase-polymerase chain reaction assay for the detection and quantification of murine retroviruses. Bio/Technology. 1993;11:1042–1046. doi: 10.1038/nbt0993-1042. [DOI] [PubMed] [Google Scholar]
  13. Jäger V., Lehmann J., Friedel P. Serum-free growth for the cultivation of a wide spectrum of mammalian cells in stirred bioreactors. Cytotechnology. 1988;1:319–329. doi: 10.1007/BF00365077. [DOI] [PubMed] [Google Scholar]
  14. Johnson L., Bryant J., Barford J.P., Harbour C., Fletcher A. Analysis of endogenous retrovirus production in a murine hybridoma by flow cytometry. In: Carrondo M.J.T., Griffiths J.B., Moreira J.L.P., editors. Animal Cell Technology: From Vaccines to Genetic Medicine. Dordrecht: Kluwer Academic Publishers; 1997. pp. 687–690. [Google Scholar]
  15. Joner E., Christiansen G.D. Hybridoma technology products: required virus testing. Biopharm. 1988;1:50–55. [Google Scholar]
  16. Jurriaans S., Van-Gemen B., Weverling G.J., Van-Strijp D., Nara P., Coutinho R., et al. The natural history of HIV-1 infection: virus load and virus phenotype independent determinants of clinical course? Virology. 1994;204:223–233. doi: 10.1006/viro.1994.1526. [DOI] [PubMed] [Google Scholar]
  17. Kavanaugh M.P., Wang H., Zhang Z., Zhang W., Wu Y.N., Dechant E., et al. Control of cationic amino acid transport and retroviral receptor functions in a membrane protein family. J. Biol. Chem. 1994;269:15445–15450. [PubMed] [Google Scholar]
  18. Lehmann J., Vorlop J., Büntemeyer H. Bubble-free reactors and their development for continuous culture with cell recycle. In: Spier R.E., Griffiths J.B., editors. Animal Cell Biotechnology. London: Academic Press; 1988. pp. 222–237. [Google Scholar]
  19. Mietz J.A., Grossman Z., Lueders K.K., Kuff E.L. Nucleotide sequence of a complete mouse intracisternal A-particle genome: relationship to known aspects of particle assembly and function. J. Virol. 1987;61:3020–3029. doi: 10.1128/jvi.61.10.3020-3029.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mossmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J. Immunol. Methods. 1983;65:55–63. doi: 10.1016/0022-1759(83)90303-4. [DOI] [PubMed] [Google Scholar]
  21. Perk K., Dahlberg J.E. Murine intracisternal A type particles fail to separate from the membrane of the endoplasmatic reticulum. J. Virol. 1974;14:1304–1306. doi: 10.1128/jvi.14.5.1304-1306.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wang H., Dechant E., Kavanaugh M., North A., Kabat D. Effects of ecotropic murine retroviruses on the dual-function cell surface receptor / basic amino acid transporter. J. Biol. Chem. 1992;267:23617–23624. [PubMed] [Google Scholar]
  23. Wang H., Kavanaugh M.P., Kabat D. A critical site in the cell surface receptor for ecotropic murine retroviruses required for amino acid transport but not for viral reception. Virology. 1994;202:1058–1060. doi: 10.1006/viro.1994.1439. [DOI] [PubMed] [Google Scholar]
  24. Weiss R.A. Retroviruses produced by hybridomas. N. Engl. J. Med. 1982;307:1587. doi: 10.1056/NEJM198212163072509. [DOI] [PubMed] [Google Scholar]
  25. Weiss R.A. Hybridomas produce viruses as well as antibodies. Immunol. Today. 1982;3:292–294. doi: 10.1016/0167-5699(82)90028-7. [DOI] [PubMed] [Google Scholar]
  26. Wells V., Mallucci L. Properties of a cell growth inhibitor produced by mouse embryo fibroblasts. J. Cell Physiol. 1983;117:148–154. doi: 10.1002/jcp.1041170203. [DOI] [PubMed] [Google Scholar]
  27. Wivel N.A., Smith G.H. Distribution of intracisternal A-particles in a variety of normal and neoplastic mouse tissues. Int. J. Cancer. 1971;7:167–175. doi: 10.1002/ijc.2910070119. [DOI] [PubMed] [Google Scholar]
  28. Yoshimoto T., Yoshimoto E., Meruelo D. Identification of amino acid residues critical for infection with ecotropic murine leukemia retroviruses. J. Virol. 1993;67:1310–1314. doi: 10.1128/jvi.67.3.1310-1314.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Cytotechnology are provided here courtesy of Springer Science+Business Media B.V.

RESOURCES