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. 1991 Jun 15;88(12):5096–5100. doi: 10.1073/pnas.88.12.5096

Direct derivation of conditionally immortal cell lines from an H-2Kb-tsA58 transgenic mouse.

P S Jat 1, M D Noble 1, P Ataliotis 1, Y Tanaka 1, N Yannoutsos 1, L Larsen 1, D Kioussis 1
PMCID: PMC51818  PMID: 1711218

Abstract

Studies on cell lines have greatly improved our understanding of many important biological questions. Generation of cell lines is facilitated by the introduction of immortalizing oncogenes into cell types of interest. One gene known to immortalize many different cell types in vitro encodes the simian virus 40 (SV40) large tumor (T) antigen (TAg). To circumvent the need for gene insertion in vitro to generate cell lines, we created transgenic mice harboring the SV40 TAg gene. Since previous studies have shown that TAg expression in transgenic mice is associated with tumorigenesis and aberrant development, we utilized a thermolabile TAg [from a SV40 strain, tsA58, temperature sensitive (ts) for transformation] to reduce the levels of functional TAg present in vivo. To direct expression to a broad range of tissues, we used the mouse major histocompatibility complex H-2Kb promoter, which is both widely active and can be further induced by interferons. tsA58 TAg mRNA was expressed in tissues of all animals harboring the hybrid construct. Development of all tissues was macroscopically normal except for thymus, which consistently showed hyperplasia. Fibroblast and cytokeratin+ thymic epithelial cultures from these mice were readily established without undergoing crisis and were conditionally immortal in their growth; the degree of conditionality was correlated with the levels of tsA58 TAg detected. One strain of H-2Kb-tsA58 mice has been bred through several generations to homozygosity and transmits a functional copy of the transgene.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baldwin A. S., Jr, Sharp P. A. Binding of a nuclear factor to a regulatory sequence in the promoter of the mouse H-2Kb class I major histocompatibility gene. Mol Cell Biol. 1987 Jan;7(1):305–313. doi: 10.1128/mcb.7.1.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bayley S. A., Stones A. J., Smith C. G. Immortalization of rat keratinocytes by transfection with polyomavirus large T gene. Exp Cell Res. 1988 Jul;177(1):232–236. doi: 10.1016/0014-4827(88)90042-0. [DOI] [PubMed] [Google Scholar]
  3. Birren S. J., Anderson D. J. A v-myc-immortalized sympathoadrenal progenitor cell line in which neuronal differentiation is initiated by FGF but not NGF. Neuron. 1990 Feb;4(2):189–201. doi: 10.1016/0896-6273(90)90094-v. [DOI] [PubMed] [Google Scholar]
  4. Botteri F. M., van der Putten H., Wong D. F., Sauvage C. A., Evans R. M. Unexpected thymic hyperplasia in transgenic mice harboring a neuronal promoter fused with simian virus 40 large T antigen. Mol Cell Biol. 1987 Sep;7(9):3178–3184. doi: 10.1128/mcb.7.9.3178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brinster R. L., Chen H. Y., Messing A., van Dyke T., Levine A. J., Palmiter R. D. Transgenic mice harboring SV40 T-antigen genes develop characteristic brain tumors. Cell. 1984 Jun;37(2):367–379. doi: 10.1016/0092-8674(84)90367-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burns J. S., Lemoine L., Lemoine N. R., Williams E. D., Wynford-Thomas D. Thyroid epithelial cell transformation by a retroviral vector expressing SV40 large T. Br J Cancer. 1989 May;59(5):755–760. doi: 10.1038/bjc.1989.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. David-Watine B., Israël A., Kourilsky P. The regulation and expression of MHC class I genes. Immunol Today. 1990 Aug;11(8):286–292. doi: 10.1016/0167-5699(90)90114-o. [DOI] [PubMed] [Google Scholar]
  8. Efrat S., Linde S., Kofod H., Spector D., Delannoy M., Grant S., Hanahan D., Baekkeskov S. Beta-cell lines derived from transgenic mice expressing a hybrid insulin gene-oncogene. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9037–9041. doi: 10.1073/pnas.85.23.9037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Evrard C., Galiana E., Rouget P. Immortalization of bipotential glial progenitors and generation of permanent "blue" cell lines. J Neurosci Res. 1988 Sep;21(1):80–87. doi: 10.1002/jnr.490210112. [DOI] [PubMed] [Google Scholar]
  10. Frederiksen K., Jat P. S., Valtz N., Levy D., McKay R. Immortalization of precursor cells from the mammalian CNS. Neuron. 1988 Aug;1(6):439–448. doi: 10.1016/0896-6273(88)90175-4. [DOI] [PubMed] [Google Scholar]
  11. Hammang J. P., Baetge E. E., Behringer R. R., Brinster R. L., Palmiter R. D., Messing A. Immortalized retinal neurons derived from SV40 T-antigen-induced tumors in transgenic mice. Neuron. 1990 May;4(5):775–782. doi: 10.1016/0896-6273(90)90204-s. [DOI] [PubMed] [Google Scholar]
  12. Harlow E., Crawford L. V., Pim D. C., Williamson N. M. Monoclonal antibodies specific for simian virus 40 tumor antigens. J Virol. 1981 Sep;39(3):861–869. doi: 10.1128/jvi.39.3.861-869.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Israel A., Kimura A., Fournier A., Fellous M., Kourilsky P. Interferon response sequence potentiates activity of an enhancer in the promoter region of a mouse H-2 gene. Nature. 1986 Aug 21;322(6081):743–746. doi: 10.1038/322743a0. [DOI] [PubMed] [Google Scholar]
  14. Jat P. S., Sharp P. A. Cell lines established by a temperature-sensitive simian virus 40 large-T-antigen gene are growth restricted at the nonpermissive temperature. Mol Cell Biol. 1989 Apr;9(4):1672–1681. doi: 10.1128/mcb.9.4.1672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jat P. S., Sharp P. A. Large T antigens of simian virus 40 and polyomavirus efficiently establish primary fibroblasts. J Virol. 1986 Sep;59(3):746–750. doi: 10.1128/jvi.59.3.746-750.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kimura A., Israël A., Le Bail O., Kourilsky P. Detailed analysis of the mouse H-2Kb promoter: enhancer-like sequences and their role in the regulation of class I gene expression. Cell. 1986 Jan 31;44(2):261–272. doi: 10.1016/0092-8674(86)90760-9. [DOI] [PubMed] [Google Scholar]
  17. Kollias G., Spanopoulou E., Grosveld F., Ritter M., Beech J., Morris R. Differential regulation of a Thy-1 gene in transgenic mice. Proc Natl Acad Sci U S A. 1987 Mar;84(6):1492–1496. doi: 10.1073/pnas.84.6.1492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lane E. B. Monoclonal antibodies provide specific intramolecular markers for the study of epithelial tonofilament organization. J Cell Biol. 1982 Mar;92(3):665–673. doi: 10.1083/jcb.92.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lemoine N. R., Mayall E. S., Jones T., Sheer D., McDermid S., Kendall-Taylor P., Wynford-Thomas D. Characterisation of human thyroid epithelial cells immortalised in vitro by simian virus 40 DNA transfection. Br J Cancer. 1989 Dec;60(6):897–903. doi: 10.1038/bjc.1989.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mellon P. L., Windle J. J., Goldsmith P. C., Padula C. A., Roberts J. L., Weiner R. I. Immortalization of hypothalamic GnRH neurons by genetically targeted tumorigenesis. Neuron. 1990 Jul;5(1):1–10. doi: 10.1016/0896-6273(90)90028-e. [DOI] [PubMed] [Google Scholar]
  21. Noble M., Fok-Seang J., Cohen J. Glia are a unique substrate for the in vitro growth of central nervous system neurons. J Neurosci. 1984 Jul;4(7):1892–1903. doi: 10.1523/JNEUROSCI.04-07-01892.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Petit C. A., Gardes M., Feunteun J. Immortalization of rodent embryo fibroblasts by SV40 is maintained by the A gene. Virology. 1983 May;127(1):74–82. doi: 10.1016/0042-6822(83)90372-0. [DOI] [PubMed] [Google Scholar]
  23. Radna R. L., Caton Y., Jha K. K., Kaplan P., Li G., Traganos F., Ozer H. L. Growth of immortal simian virus 40 tsA-transformed human fibroblasts is temperature dependent. Mol Cell Biol. 1989 Jul;9(7):3093–3096. doi: 10.1128/mcb.9.7.3093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ridley A. J., Paterson H. F., Noble M., Land H. Ras-mediated cell cycle arrest is altered by nuclear oncogenes to induce Schwann cell transformation. EMBO J. 1988 Jun;7(6):1635–1645. doi: 10.1002/j.1460-2075.1988.tb02990.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rüther U., Müller W., Sumida T., Tokuhisa T., Rajewsky K., Wagner E. F. c-fos expression interferes with thymus development in transgenic mice. Cell. 1988 Jun 17;53(6):847–856. doi: 10.1016/s0092-8674(88)90289-9. [DOI] [PubMed] [Google Scholar]
  26. Santerre R. F., Cook R. A., Crisel R. M., Sharp J. D., Schmidt R. J., Williams D. C., Wilson C. P. Insulin synthesis in a clonal cell line of simian virus 40-transformed hamster pancreatic beta cells. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4339–4343. doi: 10.1073/pnas.78.7.4339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Spanopoulou E., Early A., Elliott J., Crispe N., Ladyman H., Ritter M., Watt S., Grosveld F., Kioussis D. Complex lymphoid and epithelial thymic tumours in Thy1-myc transgenic mice. Nature. 1989 Nov 9;342(6246):185–189. doi: 10.1038/342185a0. [DOI] [PubMed] [Google Scholar]
  28. Tegtmeyer P. Function of simian virus 40 gene A in transforming infection. J Virol. 1975 Mar;15(3):613–618. doi: 10.1128/jvi.15.3.613-618.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wallach D., Fellous M., Revel M. Preferential effect of gamma interferon on the synthesis of HLA antigens and their mRNAs in human cells. Nature. 1982 Oct 28;299(5886):833–836. doi: 10.1038/299833a0. [DOI] [PubMed] [Google Scholar]
  30. Weiss E. H., Mellor A., Golden L., Fahrner K., Simpson E., Hurst J., Flavell R. A. The structure of a mutant H-2 gene suggests that the generation of polymorphism in H-2 genes may occur by gene conversion-like events. Nature. 1983 Feb 24;301(5902):671–674. doi: 10.1038/301671a0. [DOI] [PubMed] [Google Scholar]
  31. Williams D. A., Rosenblatt M. F., Beier D. R., Cone R. D. Generation of murine stromal cell lines supporting hematopoietic stem cell proliferation by use of recombinant retrovirus vectors encoding simian virus 40 large T antigen. Mol Cell Biol. 1988 Sep;8(9):3864–3871. doi: 10.1128/mcb.8.9.3864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zaret K. S., DiPersio C. M., Jackson D. A., Montigny W. J., Weinstat D. L. Conditional enhancement of liver-specific gene transcription. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9076–9080. doi: 10.1073/pnas.85.23.9076. [DOI] [PMC free article] [PubMed] [Google Scholar]

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