Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1982 Apr 24;10(8):2487–2501. doi: 10.1093/nar/10.8.2487

Molecular cloning of human immune interferon cDNA and its expression in eukaryotic cells.

R Devos, H Cheroutre, Y Taya, W Degrave, H Van Heuverswyn, W Fiers
PMCID: PMC320629  PMID: 6176945

Abstract

Starting with mRNA derived from Staphylococcal enterotoxin A induced human splenocytes, dsDNA was synthesized and inserted into unique BamHI site of the eukaryotic expression vector pSV529 (1). A recombinant plasmid containing human immune interferon (IFN-gamma) cDNA was identified by hybridization of plasmid inserted DNA bound onto nitrocellulose filters with mRNA derived from SEA-induced splenocytes, translation of the eluted RNA in Xenopus laevis oocytes and assaying for IFN activity. Plasmids containing the entire human IFN-gamma cDNA sequence were identified by colony hybridization and were sequenced. A unique coding region was identified which predicted a protein of 166 amino acids, the 20 N-terminal amino acids of which presumably represent a signal peptide. After transfection of monkey cells with plasmid DNA isolated from one of the recombinant clones (pHIIF-SV-gamma 1), IFN was excreted into the culture medium. This IFN was not distinguishable from human IFN-gamma by serological criteria or by cell target species specificity.

Full text

PDF
2487

Images in this article

2491Image
on p.2491

Selected References

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

  1. Ankel H., Krishnamurti C., Besançon F., Stefanos S., Falcoff E. Mouse fibroblast (type I) and immune (type II) interferons: pronounced differences in affinity for gangliosides and in antiviral and antigrowth effects on mouse leukemia L-1210R cells. Proc Natl Acad Sci U S A. 1980 May;77(5):2528–2532. doi: 10.1073/pnas.77.5.2528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blalock J. E., Georgiades J. A., Langford M. P., Johnson H. M. Purified human immune interferon has more potent anticellular activity than fibroblast or leukocyte interferon. Cell Immunol. 1980 Feb;49(2):390–394. doi: 10.1016/0008-8749(80)90041-6. [DOI] [PubMed] [Google Scholar]
  3. Branca A. A., Baglioni C. Evidence that types I and II interferons have different receptors. Nature. 1981 Dec 24;294(5843):768–770. doi: 10.1038/294768a0. [DOI] [PubMed] [Google Scholar]
  4. Catalona W. J., Ratliff T. L., McCool R. E. gamma-Interferon induced by S. aureus protein A augments natural killing and ADCC. Nature. 1981 May 7;291(5810):77–79. doi: 10.1038/291077a0. [DOI] [PubMed] [Google Scholar]
  5. Claeys H., Van Damme J., De Ley M., Vermylen C., Billiau A. Activation of natural cytotoxicity of human peripheral blood mononuclear cells by interferon: a kinetic study and comparison of different interferon types. Br J Haematol. 1982 Jan;50(1):85–94. doi: 10.1111/j.1365-2141.1982.tb01893.x. [DOI] [PubMed] [Google Scholar]
  6. Crane J. L., Jr, Glasgow L. A., Kern E. R., Youngner J. S. Inhibition of murine osteogenic sarcomas by treatment with type I or type II interferon. J Natl Cancer Inst. 1978 Sep;61(3):871–874. [PubMed] [Google Scholar]
  7. Davis B. D., Tai P. C. The mechanism of protein secretion across membranes. Nature. 1980 Jan 31;283(5746):433–438. doi: 10.1038/283433a0. [DOI] [PubMed] [Google Scholar]
  8. Derynck R., Content J., DeClercq E., Volckaert G., Tavernier J., Devos R., Fiers W. Isolation and structure of a human fibroblast interferon gene. Nature. 1980 Jun 19;285(5766):542–547. doi: 10.1038/285542a0. [DOI] [PubMed] [Google Scholar]
  9. Devos R., van Emmelo J., Contreras R., Fiers W. Construction and characterization of a plasmid containing a nearly full-size DNA copy of bacteriophage MS2 RNA. J Mol Biol. 1979 Mar 15;128(4):595–619. doi: 10.1016/0022-2836(79)90295-x. [DOI] [PubMed] [Google Scholar]
  10. Dianzani F., Monahan T. M., Georgiades J., Alperin J. B. Human immune interferon: induction in lymphoid cells by a calcium ionophore. Infect Immun. 1980 Aug;29(2):561–563. doi: 10.1128/iai.29.2.561-563.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dianzani F., Monahan T. M., Scupham A., Zucca M. Enzymatic induction of interferon production by galactose oxidase treatment of human lymphoid cells. Infect Immun. 1979 Dec;26(3):879–882. doi: 10.1128/iai.26.3.879-882.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dianzani F., Zucca M., Scupham A., Georgiades J. A. Immune and virus-induced interferons may activate cells by different derepressional mechanisms. Nature. 1980 Jan 24;283(5745):400–402. doi: 10.1038/283400a0. [DOI] [PubMed] [Google Scholar]
  13. Epstein L. B., Ammann A. J. Evaluation of T lymphocyte effector function in immunodeficiency diseases: abnormality in mitogen-stimulated interferon in patients with selective IgA deficiency. J Immunol. 1974 Feb;112(2):617–626. [PubMed] [Google Scholar]
  14. Epstein L. B. Interferon-gamma: success, structure and speculation. Nature. 1982 Feb 11;295(5849):453–454. doi: 10.1038/295453a0. [DOI] [PubMed] [Google Scholar]
  15. Farrar W. L., Johnson H. M., Farrar J. J. Regulation of the production of immune interferon and cytotoxic T lymphocytes by interleukin 2. J Immunol. 1981 Mar;126(3):1120–1125. [PubMed] [Google Scholar]
  16. Fleischmann W. R., Jr, Georgiades J. A., Osborne L. C., Johnson H. M. Potentiation of interferon activity by mixed preparations of fibroblast and immune interferon. Infect Immun. 1979 Oct;26(1):248–253. doi: 10.1128/iai.26.1.248-253.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fleischmann W. R., Jr, Kleyn K. M., Baron S. Potentiation of antitumor effect of virus-induced interferon by mouse immune interferon preparations. J Natl Cancer Inst. 1980 Nov;65(5):963–966. [PubMed] [Google Scholar]
  18. Gray P. W., Leung D. W., Pennica D., Yelverton E., Najarian R., Simonsen C. C., Derynck R., Sherwood P. J., Wallace D. M., Berger S. L. Expression of human immune interferon cDNA in E. coli and monkey cells. Nature. 1982 Feb 11;295(5849):503–508. doi: 10.1038/295503a0. [DOI] [PubMed] [Google Scholar]
  19. Green J. A., Cooperband S. R., Kibrick S. Immune specific induction of interferon production in cultures of human blood lymphocytes. Science. 1969 Jun 20;164(3886):1415–1417. doi: 10.1126/science.164.3886.1415. [DOI] [PubMed] [Google Scholar]
  20. Gresser I., Bandu M. T., Brouty-Boyé D. Interferon and cell division. IX. Interferon-resistant L1210 cells: characteristics and origin. J Natl Cancer Inst. 1974 Feb;52(2):553–559. doi: 10.1093/jnci/52.2.553. [DOI] [PubMed] [Google Scholar]
  21. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hanahan D., Meselson M. Plasmid screening at high colony density. Gene. 1980 Jun;10(1):63–67. doi: 10.1016/0378-1119(80)90144-4. [DOI] [PubMed] [Google Scholar]
  23. Hooks J. J., Moutsopoulos H. M., Geis S. A., Stahl N. I., Decker J. L., Notkins A. L. Immune interferon in the circulation of patients with autoimmune disease. N Engl J Med. 1979 Jul 5;301(1):5–8. doi: 10.1056/NEJM197907053010102. [DOI] [PubMed] [Google Scholar]
  24. Kahn M., Kolter R., Thomas C., Figurski D., Meyer R., Remaut E., Helinski D. R. Plasmid cloning vehicles derived from plasmids ColE1, F, R6K, and RK2. Methods Enzymol. 1979;68:268–280. doi: 10.1016/0076-6879(79)68019-9. [DOI] [PubMed] [Google Scholar]
  25. Kozak M. How do eucaryotic ribosomes select initiation regions in messenger RNA? Cell. 1978 Dec;15(4):1109–1123. doi: 10.1016/0092-8674(78)90039-9. [DOI] [PubMed] [Google Scholar]
  26. Langford M. P., Georgiades J. A., Stanton G. J., Dianzani F., Johnson H. M. Large-scale production and physicochemical characterization of human immune interferon. Infect Immun. 1979 Oct;26(1):36–41. doi: 10.1128/iai.26.1.36-41.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mantei N., Schwarzstein M., Streuli M., Panem S., Nagata S., Weissmann C. The nucleotide sequence of a cloned human leukocyte interferon cDNA. Gene. 1980 Jun;10(1):1–10. doi: 10.1016/0378-1119(80)90137-7. [DOI] [PubMed] [Google Scholar]
  28. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. McCutchan J. H., Pagano J. S. Enchancement of the infectivity of simian virus 40 deoxyribonucleic acid with diethylaminoethyl-dextran. J Natl Cancer Inst. 1968 Aug;41(2):351–357. [PubMed] [Google Scholar]
  30. McReynolds L., O'Malley B. W., Nisbet A. D., Fothergill J. E., Givol D., Fields S., Robertson M., Brownlee G. G. Sequence of chicken ovalbumin mRNA. Nature. 1978 Jun 29;273(5665):723–728. doi: 10.1038/273723a0. [DOI] [PubMed] [Google Scholar]
  31. Mizrahi A., O'Malley J. A., Carter W. A., Takatsuki A., Tamura G., Sulkowski E. Glycosylation of interferons. Effects of tunicamycin on human immune interferon. J Biol Chem. 1978 Nov 10;253(21):7612–7615. [PubMed] [Google Scholar]
  32. Nathan I., Groopman J. E., Quan S. G., Bersch N., Golde D. W. Immune (gamma) interferon produced by a human T-lymphoblast cell line. Nature. 1981 Aug 27;292(5826):842–844. doi: 10.1038/292842a0. [DOI] [PubMed] [Google Scholar]
  33. Parnes J. R., Velan B., Felsenfeld A., Ramanathan L., Ferrini U., Appella E., Seidman J. G. Mouse beta 2-microglobulin cDNA clones: a screening procedure for cDNA clones corresponding to rare mRNAs. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2253–2257. doi: 10.1073/pnas.78.4.2253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Perussia B., Mangoni L., Engers H. D., Trinchieri G. Interferon production by human and murine lymphocytes in response to alloantigens. J Immunol. 1980 Oct;125(4):1589–1595. [PubMed] [Google Scholar]
  35. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  36. Sonnenfeld G., Mandel A. D., Merigan T. C. Time and dosage dependence of immunoenhancement by murine type II interferon preparations. Cell Immunol. 1978 Oct;40(2):285–293. doi: 10.1016/0008-8749(78)90336-2. [DOI] [PubMed] [Google Scholar]
  37. Taniguchi T., Pang R. H., Yip Y. K., Henriksen D., Vilcek J. Partial characterization of gamma (immune) interferon mRNA extracted from human lymphocytes. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3469–3472. doi: 10.1073/pnas.78.6.3469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Volckaert G., Tavernier J., Derynck R., Devos R., Fiers W. Molecular mechanisms of nucleotide-sequence rearrangements in cDNA clones of human fibroblast interferon mRNA. Gene. 1981 Nov;15(2-3):215–223. doi: 10.1016/0378-1119(81)90131-1. [DOI] [PubMed] [Google Scholar]
  39. Wallace D. M., Hitchcock M. J., Reber S. B., Berger S. L. Translation of human immune interferon messenger RNA in Xenopus laevis oocytes. Biochem Biophys Res Commun. 1981 May 29;100(2):865–871. doi: 10.1016/s0006-291x(81)80253-7. [DOI] [PubMed] [Google Scholar]
  40. Wheelock E. F. Interferon-Like Virus-Inhibitor Induced in Human Leukocytes by Phytohemagglutinin. Science. 1965 Jul 16;149(3681):310–311. doi: 10.1126/science.149.3681.310. [DOI] [PubMed] [Google Scholar]
  41. Yip Y. K., Barrowclough B. S., Urban C., Vilcek J. Molecular weight of human gamma interferon is similar to that of other human interferons. Science. 1982 Jan 22;215(4531):411–413. doi: 10.1126/science.6173921. [DOI] [PubMed] [Google Scholar]
  42. Yip Y. K., Pang R. H., Urban C., Vilcek J. Partial purification and characterization of human gamma (immune) interferon. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1601–1605. doi: 10.1073/pnas.78.3.1601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. de Ley M., van Damme J., Claeys H., Weening H., Heine J. W., Billiau A., Vermylen C., de Somer P. Interferon induced in human leukocytes by mitogens: production, partial purification and characterization. Eur J Immunol. 1980 Nov;10(11):877–883. doi: 10.1002/eji.1830101113. [DOI] [PubMed] [Google Scholar]
  44. von Wussow P., Platsoucas C. D., Wiranowska-Stewart M., Stewart W. E., 2nd Human gamma interferon production by leukocytes induced with monoclonal antibodies recognizing T cells. J Immunol. 1981 Sep;127(3):1197–1200. [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES