1887

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Volume 31, Issue 3

Use of a Transfection Method to Demonstrate a Monolayer Cell Transforming Agent from the EB3 Line of Burkitt's Lymphoma Cells Free

Abstract

SUMMARY

Primary human amnion cell monolayers which had been treated with DEAE-dextran, washed, and then inoculated with sonicated cells of the EB3 line of Burkitt's lymphoma cells developed foci of transformed amnion cells 7 to 14 days later. When either the DEAE-dextran or the sonicate was omitted, no significant transformation was found. The foci consisted of enlarging mounds of rapidly dividing cells, which upon subculturing continued their high mitotic activity; and strains or lines of the transformed amnion cells were thus readily established. The modal number of chromosomes in such lines was 65 instead of the normal 46. Not all human amnions yielded cells transformable by EB3 cell sonicate, as determined by direct comparisons using the same cultural conditions and testing with the same fresh sonicate preparation in the same experiment. Overall, it appeared that only about 40 to 50% of the amnions yielded transformable cell monolayers; the rest gave monolayers apparently completely refractory to the transformation. The transformed amnion cells contained nuclear and cytoplasmic Epstein-Barr virus (EBV) antigen(s), as revealed by indirect immunofluorescence tests.

EB3 cell sonicate also caused the appearance of rapidly growing transformed cell foci on secondary rat embryo cell monolayers which had been sensitized with DEAE-dextran.

Calcium in the cell maintenance medium decreased the number of transformed foci found, both on the human and on the rat cell monolayers.

Sonicates of cultured normal human leucocytes had no such transforming activity for either the human or the rat cells.

The transforming agent in EB3 cell sonicate was completely destructible by either deoxyribonuclease or trypsin, but not by ribonuclease, and was not neutralizable by anti-EBV serum. The simplest interpretation of these results is that the transforming agent is part or all of the EBV DNA plus some necessary protein, with both the DNA and the protein accessible to hydrolytic enzyme action.

  • Received:
  • Accepted:
  • Published Online:
© Society for General Microbiology 1976
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1976-06-01
2024-05-05
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References

  1. AL-MOSLIH M. I., DUBES G. R. 1973a; The kinetics of DEAE-dextran-induced œil sensitization to trans-fection. Journal of General Virology 18:189–193
     [Google Scholar]
  2. AL-MOSLIH M. I., DUBES G. R. 1973b; Orthophosphate and amino acids as nutritional factors stimulating diethylaminoethyl-dextran-induced cell sensitization to transfection. Archiv für die gesamte Virusforschung 41:71–79
     [Google Scholar]
  3. AL-MOSLIH M. I., DUBES G. R., CRONN J. C., RITCHIE A. E. 1975; Studies of production, assay, and properties of a deoxyribonuclease-labile infective unit from adenovirus. Abstracts of the Annual Meeting of the American Society for Microbiology256
     [Google Scholar]
  4. AL-MOSLIH M. I., WHITE R. J. 1974; Transformation of diethylaminoethyl-dextran pretreated human amnion and rat embryo cells by sonicated EB3 cells. Abstracts of the Annual Meeting of the American Society for Microbiology223
     [Google Scholar]
  5. BOYD V. A. L., BUTEL J. S. 1972; Demonstration of infectious deoxyribonucleic acid in transformed cells. I. Recovery of simian virus 40 from yielder and non-yielder transformed cells. Journal of Virology 10:399–409
     [Google Scholar]
  6. DIEHL V., HENLE G., HENLE W., KOHN G. 1969; Effect of a herpes group virus (EBV) on growth of peripheral leukocyte cultures. In Vitro 4:92–99
     [Google Scholar]
  7. DUBES G. R., ROEGNER K. R., CRONN J. C. 1968; Deoxyribonuclease-labile infective units from adenovirus. Experientia 24:401–403
     [Google Scholar]
  8. DULBECCO R., VOGT M. 1954; Plaque formation and isolation of pure lines with poliomyelitis viruses. Journal of Experimental Medicine 99:167–182
     [Google Scholar]
  9. EPSTEIN M. A., ACHONG B. G., BARR Y. M. 1964; Virus particles in cultured lymphoblasts from Burkitt’s lymphoma. Lancet i:702–703
     [Google Scholar]
  10. EPSTEIN M. A., BARR Y. M. 1964; Cultivation in vitro of human lymphoblasts from Burkitt’s malignant lymphoma. Lancet i:252–253
     [Google Scholar]
  11. EPSTEIN M. A., HENLE G., ACHONG B. G., BARR Y. M. 1965; Morphological and biological studies on a virus in cultured lymphoblasts from Burkitt’s lymphoma. Journal of Experimental Medicine 121:761–770
     [Google Scholar]
  12. FREEMAN A. E., BLACK P. H., WOLFORD R., HUEBNER R. J. 1967; Adenovirus type 12-rat embryo transformation system. Journal of Virology 1:362–367
     [Google Scholar]
  13. FREEMAN A. E., CALISHER C. H., PRICE P. J., TURNER H. C., HUEBNER R. J. 1966; Calcium sensitivity of cell cultures derived from adenovirus-induced tumors. Proceedings of the Society of Experimental Biology and Medicine 122:835–840
     [Google Scholar]
  14. FREEMAN A. E., HOLLINGER S., PRICE P. J., CALISHER C. H. 1965; The effect of calcium on cell lines derived from adenovirus type 12-induced hamster tumors. Experimental Cell Research 39:259–264
     [Google Scholar]
  15. GERBER P., WHANG-PENG J., MONROE J. H. 1969; Transformation and chromosome changes induced by Epstein-Barr virus in normal human leukocyte cultures. Proceedings of the National Academy of Sciences of the United States of America 63:740–747
     [Google Scholar]
  16. GLASER R., RAPP F. 1972; Rescue of Epstein-Barr virus from somatic cell hybrids of Burkitt lympho-blastoid cells. Journal of Virology 10:288–296
     [Google Scholar]
  17. HENLE G., HENLE W. 1966; Immunofluorescence in cells derived from Burkitt’s lymphoma. Journal of Bacteriology 91:1248–1256
     [Google Scholar]
  18. HENLE W., DIEHL V., KOHN G., ZUR HAUSEN H., HENLE G. 1967; Herpes-type virus and chromosome marker in normal leukocytes after growth with irradiated Burkitt cells. Science, New York 157:1064–1065
     [Google Scholar]
  19. HENLE W., HENLE G. 1968; Effect of arginine-deficient media on the herpes-type virus associated with cultured Burkitt tumor cells. Journal of Virology 2:182–191
     [Google Scholar]
  20. HENLE W., HENLE G., ZAJAC B. A., PEARSON G., WAUBKE R., SCRIBA M. 1970; Differential reactivity of human serums with early antigens induced by Epstein-Barr virus. Science, New York 169:188–190
     [Google Scholar]
  21. HENLE W., HUMMELER K., HENLE G. 1966; Antibody coating and agglutination of virus particles separated from the EB3 line of Burkit lymphoma cells. Journal of Bacteriology 92:269–271
     [Google Scholar]
  22. JENKINS M. E., EISEN J., SEQUIN F. 1971; Congenital asymmetry and diploid-triploid mosaicism. American Journal of Diseases of Children 122:80–84
     [Google Scholar]
  23. KOHN G., MELLMAN W. J., MOORHEAD P. S., LOFTUS J., HENLE G. 1967; Involvement of C group chromosomes in five Burkitt lymphoma cell lines. Journal of the National Cancer Institute 38:209–222
     [Google Scholar]
  24. MAURER B. A., IMAMURA T., WILBERT S. M. 1970; Incidence of EB virus-containing cells in primary and secondary clones of several Burkitt lymphoma cell lines. Cancer Research 30:2870–2875
     [Google Scholar]
  25. MILES C. P., O’NEILL F. 1967; Chromosome studies of 8 in vitro lines of Burkitt’s lymphoma. Cancer Research 27:392–402
     [Google Scholar]
  26. MILLER G., ENDERS J. F. 1968; Vaccinia virus replication and cytopathic effect in cultures of phytohemag-glutinin-treated human peripheral blood leukocytes. Journal of Virology 2:787–792
     [Google Scholar]
  27. PAOANO J. S., VAHERI A. 1965; Enhancement of infectivity of poliovirus RNA with diethylaminoethyl-dextran (DEAE-D). Archiv für die gesamte Virusforschung 17:456–464
     [Google Scholar]
  28. POPE J. H., HÖRNE M. K., SCOTT W. 1968; Transformation of foetal human leukocytes in vitro by filtrates of a human leukaemic cell line containing herpes-like virus. International journal of Cancer 3:857–866
     [Google Scholar]
  29. POPE J. H., HÖRNE M. K., SCOTT W. 1969; Identification of the filterable leukocyte-transforming factor of QIMR-WIL cells as herpes-like virus. International Journal of Cancer 4:255–260
     [Google Scholar]
  30. PULVERTAFT R. J. V. 1964; Cytology of Burkitt’s tumour (African lymphoma). Lancet i:238–240
     [Google Scholar]
  31. ROTHFELS K. H., SIMINOVITCH L. 1958; An air-drying technique for flattening chromosomes in mammalian cells grown in vitro. Stain Technology 33:73–77
     [Google Scholar]
  32. WILLIAMS J. F., USTACELEBI S. 1971 Strategy of the Viral Genome275–290 Edited by Wolstenholme G. E. W., O’Connor M. Edinburgh and London: Churchill Livingstone;
     [Google Scholar]
  33. ZAJAC B. A., KOHN G. 1970; Epstein-Barr antigens, marker chromosome, and interferon production in clones derived from cultured Burkitt tumor cells. Journal of the National Cancer Institute 45:399–406
     [Google Scholar]
  34. ZUR HAUSEN H., SCHULTE-HOLTHAUSEN H. 1970; Presence of EB virus nucleic acid homology in a ‘virus-free’ line of Burkitt tumour cells. Nature, London 227:245–248
     [Google Scholar]
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Use of a Transfection Method to Demonstrate a Monolayer Cell Transforming Agent from the EB3 Line of Burkitt's Lymphoma Cells
J. Gen. Virol. 31, 331 (1976); https://doi.org/10.1099/0022-1317-31-3-331
/content/journal/jgv/10.1099/0022-1317-31-3-331
/content/journal/jgv/10.1099/0022-1317-31-3-331
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