Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1998 Jan 15;329(Pt 2):295–302. doi: 10.1042/bj3290295

Identification of nine interferon-alpha subtypes produced by Sendai virus-induced human peripheral blood leucocytes.

T A Nyman 1, H Tölö 1, J Parkkinen 1, N Kalkkinen 1
PMCID: PMC1219044  PMID: 9425112

Abstract

The human interferon-alpha (IFN-alpha) family is encoded by 13 different functional genes, and including all cloned sequence variants there are 28 potential IFN-alpha proteins. To find out which of the described sequences are expressed in normal human leucocytes, we have isolated and partly characterized the components of a highly purified IFN-alpha preparation produced by Sendai virus-induced human peripheral blood leucocytes. The identification protocol consisted of N-terminal sequencing and mass mapping of the proteins separated by reverse-phase HPLC and/or SDS/PAGE. The highly purified leucocyte IFN-alpha preparation was found to contain at least nine different IFN-alpha species: IFN-alpha1a, IFN-alpha2b, IFN-alpha4b, IFN-alpha7a, IFN-alpha8b, IFN-alpha10a, IFN-alpha14c, IFN-alpha17b and IFN-alpha21b. IFN-alpha1a was the major subtype, comprising approx. 30% of total leucocyte IFN-alpha. IFN-alpha14c, the only subtype containing potential N-glycosylation sites, was shown to be glycosylated at Asn-72. Molecular mass determination of the intact proteins by electrospray ionization MS showed that there are no other post-translational modifications in the IFN-alpha subtypes than the glycosylation of IFN-alpha2b and IFN-alpha14c. Only one sequence variant was found for each subtype, suggesting that the other described gene sequences represent allelic variants or mutations that are more rarely found in the general population.

Full Text

The Full Text of this article is available as a PDF (290.0 KB).

Selected References

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

  1. Adolf G. R., Kalsner I., Ahorn H., Maurer-Fogy I., Cantell K. Natural human interferon-alpha 2 is O-glycosylated. Biochem J. 1991 Jun 1;276(Pt 2):511–518. doi: 10.1042/bj2760511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adolf G. R. Monoclonal antibodies and enzyme immunoassays specific for human interferon (IFN) omega 1: evidence that IFN-omega 1 is a component of human leukocyte IFN. Virology. 1990 Apr;175(2):410–417. doi: 10.1016/0042-6822(90)90425-q. [DOI] [PubMed] [Google Scholar]
  3. Allen G., Diaz M. O. Nomenclature of the human interferon proteins. J Interferon Cytokine Res. 1996 Feb;16(2):181–184. doi: 10.1089/jir.1996.16.181. [DOI] [PubMed] [Google Scholar]
  4. Antonelli G. Antibodies to interferon in patients undergoing IFN therapy: an update. J Biol Regul Homeost Agents. 1995 Oct-Dec;9(4):123–131. [PubMed] [Google Scholar]
  5. Cantell K., Hirvonen S., Kauppinen H. L., Myllylä G. Production of interferon in human leukocytes from normal donors with the use of Sendai virus. Methods Enzymol. 1981;78(Pt A):29–38. doi: 10.1016/0076-6879(81)78094-7. [DOI] [PubMed] [Google Scholar]
  6. Dipaola M., Smith T., Ferencz-Biro K., Liao M. J., Testa D. Interferon-alpha 2 produced by normal human leukocytes is predominantly interferon-alpha 2b. J Interferon Res. 1994 Dec;14(6):325–332. doi: 10.1089/jir.1994.14.325. [DOI] [PubMed] [Google Scholar]
  7. Familletti P. C., McCandliss R., Pestka S. Production of high levels of human leukocyte interferon from a continuous human myeloblast cell culture. Antimicrob Agents Chemother. 1981 Jul;20(1):5–9. doi: 10.1128/aac.20.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Finter N. B. Large scale production of human interferon from lymphoblastoid cells. Tex Rep Biol Med. 1981;41:175–178. [PubMed] [Google Scholar]
  9. Foster G. R., Rodrigues O., Ghouze F., Schulte-Frohlinde E., Testa D., Liao M. J., Stark G. R., Leadbeater L., Thomas H. C. Different relative activities of human cell-derived interferon-alpha subtypes: IFN-alpha 8 has very high antiviral potency. J Interferon Cytokine Res. 1996 Dec;16(12):1027–1033. doi: 10.1089/jir.1996.16.1027. [DOI] [PubMed] [Google Scholar]
  10. Gharahdaghi F., Kirchner M., Fernandez J., Mische S. M. Peptide-mass profiles of polyvinylidene difluoride-bound proteins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in the presence of nonionic detergents. Anal Biochem. 1996 Jan 1;233(1):94–99. doi: 10.1006/abio.1996.0012. [DOI] [PubMed] [Google Scholar]
  11. Gitlin G., Tsarbopoulos A., Patel S. T., Sydor W., Pramanik B. N., Jacobs S., Westreich L., Mittelman S., Bausch J. N. Isolation and characterization of a monomethioninesulfoxide variant of interferon alpha-2b. Pharm Res. 1996 May;13(5):762–769. doi: 10.1023/a:1016059902645. [DOI] [PubMed] [Google Scholar]
  12. Goeddel D. V., Leung D. W., Dull T. J., Gross M., Lawn R. M., McCandliss R., Seeburg P. H., Ullrich A., Yelverton E., Gray P. W. The structure of eight distinct cloned human leukocyte interferon cDNAs. Nature. 1981 Mar 5;290(5801):20–26. doi: 10.1038/290020a0. [DOI] [PubMed] [Google Scholar]
  13. Golovleva I., Kandefer-Szerszen M., Beckman L., Lundgren E. Polymorphism in the interferon-alpha gene family. Am J Hum Genet. 1996 Sep;59(3):570–578. [PMC free article] [PubMed] [Google Scholar]
  14. Hiscott J., Cantell K., Weissmann C. Differential expression of human interferon genes. Nucleic Acids Res. 1984 May 11;12(9):3727–3746. doi: 10.1093/nar/12.9.3727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hua J., Liao M. J., Rashidbaigi A. Cytokines induced by Sendai virus in human peripheral blood leukocytes. J Leukoc Biol. 1996 Jul;60(1):125–128. doi: 10.1002/jlb.60.1.125. [DOI] [PubMed] [Google Scholar]
  16. Hussain M., Gill D. S., Liao M. J. Identification of interferon-alpha 7, -alpha 14, and -alpha 21 variants in the genome of a large human population. J Interferon Cytokine Res. 1996 Oct;16(10):853–859. doi: 10.1089/jir.1996.16.853. [DOI] [PubMed] [Google Scholar]
  17. Hussain M., Gill D. S., Liao M. J., Testa D. Interferon-alpha 8b is the only variant of interferon-alpha 8 identified in a large human population. J Interferon Cytokine Res. 1996 Jul;16(7):523–529. doi: 10.1089/jir.1996.16.523. [DOI] [PubMed] [Google Scholar]
  18. Jensen O. N., Podtelejnikov A., Mann M. Delayed extraction improves specificity in database searches by matrix-assisted laser desorption/ionization peptide maps. Rapid Commun Mass Spectrom. 1996;10(11):1371–1378. doi: 10.1002/(SICI)1097-0231(199608)10:11<1371::AID-RCM682>3.0.CO;2-5. [DOI] [PubMed] [Google Scholar]
  19. Kontsek P. Human type I interferons: structure and function. Acta Virol. 1994 Dec;38(6):345–360. [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Lee N., Ni D., Brissette R., Chou M., Hussain M., Gill D. S., Liao M. J., Testa D. Interferon-alpha 2 variants in the human genome. J Interferon Cytokine Res. 1995 Apr;15(4):341–349. doi: 10.1089/jir.1995.15.341. [DOI] [PubMed] [Google Scholar]
  22. Liu X., Sosic Z., Krull I. S. Capillary isoelectric focusing as a tool in the examination of antibodies, peptides and proteins of pharmaceutical interest. J Chromatogr A. 1996 May 31;735(1-2):165–190. doi: 10.1016/0021-9673(95)01357-1. [DOI] [PubMed] [Google Scholar]
  23. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  24. McKenna R. M., Oberg K. E. Antibodies to interferon-alpha in treated cancer patients: incidence and significance. J Interferon Cytokine Res. 1997 Mar;17(3):141–143. doi: 10.1089/jir.1997.17.141. [DOI] [PubMed] [Google Scholar]
  25. Nagata S., Mantei N., Weissmann C. The structure of one of the eight or more distinct chromosomal genes for human interferon-alpha. Nature. 1980 Oct 2;287(5781):401–408. doi: 10.1038/287401a0. [DOI] [PubMed] [Google Scholar]
  26. Ortaldo J. R., Herberman R. B., Harvey C., Osheroff P., Pan Y. C., Kelder B., Pestka S. A species of human alpha interferon that lacks the ability to boost human natural killer activity. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4926–4929. doi: 10.1073/pnas.81.15.4926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rosenfeld J., Capdevielle J., Guillemot J. C., Ferrara P. In-gel digestion of proteins for internal sequence analysis after one- or two-dimensional gel electrophoresis. Anal Biochem. 1992 May 15;203(1):173–179. doi: 10.1016/0003-2697(92)90061-b. [DOI] [PubMed] [Google Scholar]
  28. Sperber S. J., Gocke D. J., Haberzettl C., Kuk R., Schwartz B., Pestka S. Anti-HIV-1 activity of recombinant and hybrid species of interferon-alpha. J Interferon Res. 1992 Oct;12(5):363–368. doi: 10.1089/jir.1992.12.363. [DOI] [PubMed] [Google Scholar]
  29. Streuli M., Hall A., Boll W., Stewart W. E., 2nd, Nagata S., Weissmann C. Target cell specificity of two species of human interferon-alpha produced in Escherichia coli and of hybrid molecules derived from them. Proc Natl Acad Sci U S A. 1981 May;78(5):2848–2852. doi: 10.1073/pnas.78.5.2848. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Todokoro K., Kioussis D., Weissmann C. Two non-allelic human interferon alpha genes with identical coding regions. EMBO J. 1984 Aug;3(8):1809–1812. doi: 10.1002/j.1460-2075.1984.tb02050.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weck P. K., Apperson S., May L., Stebbing N. Comparison of the antiviral activities of various cloned human interferon-alpha subtypes in mammalian cell cultures. J Gen Virol. 1981 Nov;57(Pt 1):233–237. doi: 10.1099/0022-1317-57-1-233. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES