Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1974 Dec;71(12):4737–4741. doi: 10.1073/pnas.71.12.4737

Direct Evidence for the Presence of Epstein-Barr Virus DNA and Nuclear Antigen in Malignant Epithelial Cells from Patients with Poorly Differentiated Carcinoma of the Nasopharynx

George Klein *, Beppino C Giovanella , Tomas Lindahl *, Philip J Fialkow , Surjit Singh §, John S Stehlin
PMCID: PMC433971  PMID: 4373728

Abstract

A well-differentiated squamous cell carcinoma and three poorly differentiated carcinomas of the nasopharynx were analyzed for the presence of Epstein-Barr virus DNA by hybridization with radioactive complementary RNA. The well-differentiated carcinoma contained no detectable Epstein-Barr virus DNA, whereas the three anaplastic carcinomas contained 41, 16, and 14 viral genome equivalents per cell. The anaplastic carcinomas were heavily infiltrated with lymphocytes and other non-neoplastic cells. All four tumors were successfully passaged in nude (thymusless) mice. Mouse cell admixture in the heterotransplanted tumors was estimated by analysis of glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and varied between 25% and 80%. After two passages in nude mice, the carcinoma that was negative for Epstein-Barr virus DNA remained negative, while the three anaplastic carcinomas retained their viral DNA. After correction for mouse cell admixture, the latter tumors were found to contain about 80, 55, and 160 Epstein-Barr virus genome equivalents per human cell. The virus-determined nuclear antigen was localized in the large carcinoma cell clusters in all three mouse-passaged tumors positive for the viral DNA, but other virus-determined antigens were not detected, indicating the absence of virus induction. In contrast to the original tumor biopsies, the nude-mouse-passaged tumors showed virtually no lymphocytic infiltration. It is concluded that the Epstein-Barr virus DNA found in biopsies of human nasopharyngeal carcinomas is localized in the neoplastic cells.

Keywords: nude mice, transplantation, nucleic acid hybridization, isozyme

Full text

PDF
4740

Images in this article

Selected References

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

  1. De-Thé G., Ho H. C., Kwan H. C., Desgranges C., Favre M. C. Nasopharyngeal carcinoma (NPC). I. Types of cultures derived from tumour biopsies and non-tumorous tissues of Chinese patients with special reference to lymphoblastoid transformation. Int J Cancer. 1970 Sep 15;6(2):189–206. doi: 10.1002/ijc.2910060206. [DOI] [PubMed] [Google Scholar]
  2. Diehl V., Henle G., Henle W., Kohn G. Demonstration of a herpes group virus in cultures of peripheral leukocytes from patients with infectious mononucleosis. J Virol. 1968 Jul;2(7):663–669. doi: 10.1128/jvi.2.7.663-669.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. FILDES R. A., PARR C. W. HUMAN RED-CELL PHOSPHOGLUCONATE DEHYDROGENASES. Nature. 1963 Nov 30;200:890–891. doi: 10.1038/200890a0. [DOI] [PubMed] [Google Scholar]
  4. Fialkow P. J., Klein E., Klein G., Clifford P., Singh S. Immunoglobulin and glucose-6-phosphate dehydrogenase as markers of cellular origin in Burkitt lymphoma. J Exp Med. 1973 Jul 1;138(1):89–102. doi: 10.1084/jem.138.1.89. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fialkow P. J., Klein G., Gartler S. M., Clifford P. Clonal origin for individual Burkitt tumours. Lancet. 1970 Feb 21;1(7643):384–386. doi: 10.1016/s0140-6736(70)91517-5. [DOI] [PubMed] [Google Scholar]
  6. Fialkow P. J., Klein G., Giblett E. R., Gothoskar B., Clifford P. Foreign-cell contamination in Burkitt tumours. Lancet. 1971 May 1;1(7705):883–886. doi: 10.1016/s0140-6736(71)92443-3. [DOI] [PubMed] [Google Scholar]
  7. Gerber P., Lucas S., Nonoyama M., Perlin E., Goldstein L. I. Oral excretion of Epstein-Barr virus by healthy subjects and patients with infectious mononucleosis. Lancet. 1972 Nov 11;2(7785):988–989. doi: 10.1016/s0140-6736(72)92402-6. [DOI] [PubMed] [Google Scholar]
  8. Gergely L., Klein G., Ernberg I. The action of DNA antagonists on Epstein-Barr virus (EBV)-associated early antigen (EA) in Burkitt lymphoma lines. Int J Cancer. 1971 Mar 15;7(2):293–302. doi: 10.1002/ijc.2910070214. [DOI] [PubMed] [Google Scholar]
  9. Giovanella B. C., Stehlin J. S. Heterotransplantation of human malignant tumors in "nude" thymusless mice. I. Breeding and maintenance of "nude" mice. J Natl Cancer Inst. 1973 Aug;51(2):615–619. [PubMed] [Google Scholar]
  10. Giovanella B. C., Stehlin J. S., Williams L. J., Jr Heterotransplantation of human malignant tumors in "nude" thymusless mice. II. Malignant tumors induced by injection of cell cultures derived from human solid tumors. J Natl Cancer Inst. 1974 Mar;52(3):921–930. doi: 10.1093/jnci/52.3.921. [DOI] [PubMed] [Google Scholar]
  11. Henle W., Henle G., Ho H. C., Burtin P., Cachin Y., Clifford P., de Schryver A., de-Thé G., Diehl V., Klein G. Antibodies to Epstein-Barr virus in nasopharyngeal carcinoma, other head and neck neoplasms, and control groups. J Natl Cancer Inst. 1970 Jan;44(1):225–231. [PubMed] [Google Scholar]
  12. Ho J. H. Nasopharyngeal carcinoma (NPC). Adv Cancer Res. 1972;15:57–92. doi: 10.1016/s0065-230x(08)60372-3. [DOI] [PubMed] [Google Scholar]
  13. Kawai Y., Nonoyama M., Pagano J. S. Reassociation kinetics for Epstein-Barr virus DNA: nonhomology to mammalian DNA and homology of viral DNA in various diseases. J Virol. 1973 Nov;12(5):1006–1012. doi: 10.1128/jvi.12.5.1006-1012.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Klein G., Gergely L., Goldstein G. Two-colour immunofluorescence studies on EBV-determined antigens. Clin Exp Immunol. 1971 Apr;8(4):593–602. [PMC free article] [PubMed] [Google Scholar]
  15. Klein G. Immunological studies on a human tumor. Dilemmas of the experimentalist. Isr J Med Sci. 1971 Jan;7(1):111–131. [PubMed] [Google Scholar]
  16. Lindahl T., Klein G., Reedman B. M., Johansson B., Singh S. Relationship between Epstein-Barr virus (EBV) DNA and the EBV-determined nuclear antigen (EBNA) in Burkitt lymphoma biopsies and other lymphoproliferative malignancies. Int J Cancer. 1974 Jun 15;13(6):764–772. doi: 10.1002/ijc.2910130605. [DOI] [PubMed] [Google Scholar]
  17. Miller G., Niederman J. C., Andrews L. L. Prolonged oropharyngeal excretion of Epstein-Barr virus after infectious mononucleosis. N Engl J Med. 1973 Feb 1;288(5):229–232. doi: 10.1056/NEJM197302012880503. [DOI] [PubMed] [Google Scholar]
  18. Nilsson K. Characteristics of established myeloma and lymphoblastoid cell lines derived from an E myeloma patient: a comparative study. Int J Cancer. 1971 May 15;7(3):380–396. doi: 10.1002/ijc.2910070303. [DOI] [PubMed] [Google Scholar]
  19. Nilsson K., Klein G., Henle W., Henle G. The establishment of lymphoblastoid lines from adult and fetal human lymphoid tissue and its dependence on EBV. Int J Cancer. 1971 Nov 15;8(3):443–450. doi: 10.1002/ijc.2910080312. [DOI] [PubMed] [Google Scholar]
  20. Pagano J. S., Huang C. H., Levine P. Absence of Epstein-Barr viral DNA in Amercian Burkitt's lymphoma. N Engl J Med. 1973 Dec 27;289(26):1395–1399. doi: 10.1056/NEJM197312272892604. [DOI] [PubMed] [Google Scholar]
  21. Reedman B. M., Klein G. Cellular localization of an Epstein-Barr virus (EBV)-associated complement-fixing antigen in producer and non-producer lymphoblastoid cell lines. Int J Cancer. 1973 May;11(3):499–520. doi: 10.1002/ijc.2910110302. [DOI] [PubMed] [Google Scholar]
  22. Reedman B. M., Klein G., Pope J. H., Walters M. K., Hilgers J., Singh S., Johansson B. Epstein-Barr virus-associated complement-fixing and nuclear antigens in Burkitt lymphoma biopsies. Int J Cancer. 1974 Jun 15;13(6):755–763. doi: 10.1002/ijc.2910130604. [DOI] [PubMed] [Google Scholar]
  23. Wolf H., zur Hausen H., Becker V. EB viral genomes in epithelial nasopharyngeal carcinoma cells. Nat New Biol. 1973 Aug 22;244(138):245–247. doi: 10.1038/newbio244245a0. [DOI] [PubMed] [Google Scholar]
  24. de Schryver A., Friberg S., Jr, Klein G., Henle W., Henle G., De-Thé G., Clifford P., Ho H. C. Epstein-Barr virus-associated antibody patterns in carcinoma of the post-nasal space. Clin Exp Immunol. 1969 Nov;5(5):443–459. [PMC free article] [PubMed] [Google Scholar]
  25. zur Hausen H., Schulte-Holthausen H., Klein G., Henle W., Henle G., Clifford P., Santesson L. EBV DNA in biopsies of Burkitt tumours and anaplastic carcinomas of the nasopharynx. Nature. 1970 Dec 12;228(5276):1056–1058. doi: 10.1038/2281056a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES