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. 1992 Feb;75(2):286–292.

Biochemical and immunological properties of rat recombinant interleukin-2 and interleukin-4.

A J McKnight 1, B J Classon 1
PMCID: PMC1384708  PMID: 1551691

Abstract

We have previously described the isolation and sequencing of cDNA clones encoding rat interleukin-2 (IL-2) and interleukin-4 (IL-4). In the present study, we report the generation of stably transfected Chinese hamster ovary (CHO) cell lines which constitutively synthesize and secrete high levels of rat recombinant IL-2 (rIL-2) and IL-4 (rIL-4). The expression of the cytokine cDNA sequences is driven by the human cytomegalovirus promoter/enhancer within the respective pEE6. HCMV-GS vector constructs, following the successful transfection and isolation of methionine sulphoximine (MSX)-resistant CHO cell lines. Analyses of metabolically labelled CHO.rIL-2 and CHO.rIL-4 have been performed, in addition to studies which demonstrate certain biological properties of these recombinant cytokines including T-cell growth factor activity (rIL-2) and the ability to enhance expression of class II major histocompatibility complex (MHC) molecules on spleen cells (rIL-4). The availability of large quantities of these rat recombinant cytokines, conveniently produced by a mammalian cell line, will prove invaluable in future studies into the induction and regulation of immune responses in this species.

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Selected References

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  1. Aguet M., Dembić Z., Merlin G. Molecular cloning and expression of the human interferon-gamma receptor. Cell. 1988 Oct 21;55(2):273–280. doi: 10.1016/0092-8674(88)90050-5. [DOI] [PubMed] [Google Scholar]
  2. Cantrell M. A., Anderson D., Cerretti D. P., Price V., McKereghan K., Tushinski R. J., Mochizuki D. Y., Larsen A., Grabstein K., Gillis S. Cloning, sequence, and expression of a human granulocyte/macrophage colony-stimulating factor. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6250–6254. doi: 10.1073/pnas.82.18.6250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cockett M. I., Bebbington C. R., Yarranton G. T. High level expression of tissue inhibitor of metalloproteinases in Chinese hamster ovary cells using glutamine synthetase gene amplification. Biotechnology (N Y) 1990 Jul;8(7):662–667. doi: 10.1038/nbt0790-662. [DOI] [PubMed] [Google Scholar]
  4. Cohen D. R., Hapel A. J., Young I. G. Cloning and expression of the rat interleukin-3 gene. Nucleic Acids Res. 1986 May 12;14(9):3641–3658. doi: 10.1093/nar/14.9.3641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Conradt H. S., Nimtz M., Dittmar K. E., Lindenmaier W., Hoppe J., Hauser H. Expression of human interleukin-2 in recombinant baby hamster kidney, Ltk-, and Chinese hamster ovary cells. Structure of O-linked carbohydrate chains and their location within the polypeptide. J Biol Chem. 1989 Oct 15;264(29):17368–17373. [PubMed] [Google Scholar]
  6. Crawford R. M., Finbloom D. S., Ohara J., Paul W. E., Meltzer M. S. B cell stimulatory factor-1 (interleukin 4) activates macrophages for increased tumoricidal activity and expression of Ia antigens. J Immunol. 1987 Jul 1;139(1):135–141. [PubMed] [Google Scholar]
  7. Davis S. J., Ward H. A., Puklavec M. J., Willis A. C., Williams A. F., Barclay A. N. High level expression in Chinese hamster ovary cells of soluble forms of CD4 T lymphocyte glycoprotein including glycosylation variants. J Biol Chem. 1990 Jun 25;265(18):10410–10418. [PubMed] [Google Scholar]
  8. Devos R., Plaetinck G., Cheroutre H., Simons G., Degrave W., Tavernier J., Remaut E., Fiers W. Molecular cloning of human interleukin 2 cDNA and its expression in E. coli. Nucleic Acids Res. 1983 Jul 11;11(13):4307–4323. doi: 10.1093/nar/11.13.4307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dijkema R., van der Meide P. H., Pouwels P. H., Caspers M., Dubbeld M., Schellekens H. Cloning and expression of the chromosomal immune interferon gene of the rat. EMBO J. 1985 Mar;4(3):761–767. doi: 10.1002/j.1460-2075.1985.tb03694.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Karasuyama H., Melchers F. Establishment of mouse cell lines which constitutively secrete large quantities of interleukin 2, 3, 4 or 5, using modified cDNA expression vectors. Eur J Immunol. 1988 Jan;18(1):97–104. doi: 10.1002/eji.1830180115. [DOI] [PubMed] [Google Scholar]
  12. Le H. V., Ramanathan L., Labdon J. E., Mays-Ichinco C. A., Syto R., Arai N., Hoy P., Takebe Y., Nagabhushan T. L., Trotta P. P. Isolation and characterization of multiple variants of recombinant human interleukin 4 expressed in mammalian cells. J Biol Chem. 1988 Aug 5;263(22):10817–10823. [PubMed] [Google Scholar]
  13. Leitenberg D., Feldbush T. L. Lymphokine regulation of surface Ia expression on rat B cells. Cell Immunol. 1988 Feb;111(2):451–460. doi: 10.1016/0008-8749(88)90108-6. [DOI] [PubMed] [Google Scholar]
  14. Lomedico P. T., Gubler U., Hellmann C. P., Dukovich M., Giri J. G., Pan Y. C., Collier K., Semionow R., Chua A. O., Mizel S. B. Cloning and expression of murine interleukin-1 cDNA in Escherichia coli. 1984 Nov 29-Dec 5Nature. 312(5993):458–462. doi: 10.1038/312458a0. [DOI] [PubMed] [Google Scholar]
  15. McKnight A. J., Barclay A. N., Mason D. W. Molecular cloning of rat interleukin 4 cDNA and analysis of the cytokine repertoire of subsets of CD4+ T cells. Eur J Immunol. 1991 May;21(5):1187–1194. doi: 10.1002/eji.1830210514. [DOI] [PubMed] [Google Scholar]
  16. McKnight A. J., Mason D. W., Barclay A. N. Sequence of rat interleukin 2 and anomalous binding of a mouse interleukin 2 cDNA probe to rat MHC class II-associated invariant chain mRNA. Immunogenetics. 1989;30(2):145–147. doi: 10.1007/BF02421547. [DOI] [PubMed] [Google Scholar]
  17. Mosmann T. R., Yokota T., Kastelein R., Zurawski S. M., Arai N., Takebe Y. Species-specificity of T cell stimulating activities of IL 2 and BSF-1 (IL 4): comparison of normal and recombinant, mouse and human IL 2 and BSF-1 (IL 4). J Immunol. 1987 Mar 15;138(6):1813–1816. [PubMed] [Google Scholar]
  18. Noelle R., Krammer P. H., Ohara J., Uhr J. W., Vitetta E. S. Increased expression of Ia antigens on resting B cells: an additional role for B-cell growth factor. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6149–6153. doi: 10.1073/pnas.81.19.6149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Northemann W., Braciak T. A., Hattori M., Lee F., Fey G. H. Structure of the rat interleukin 6 gene and its expression in macrophage-derived cells. J Biol Chem. 1989 Sep 25;264(27):16072–16082. [PubMed] [Google Scholar]
  20. Park L. S., Friend D., Grabstein K., Urdal D. L. Characterization of the high-affinity cell-surface receptor for murine B-cell-stimulating factor 1. Proc Natl Acad Sci U S A. 1987 Mar;84(6):1669–1673. doi: 10.1073/pnas.84.6.1669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Park L. S., Friend D., Sassenfeld H. M., Urdal D. L. Characterization of the human B cell stimulatory factor 1 receptor. J Exp Med. 1987 Aug 1;166(2):476–488. doi: 10.1084/jem.166.2.476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Paterson D. J., Jefferies W. A., Green J. R., Brandon M. R., Corthesy P., Puklavec M., Williams A. F. Antigens of activated rat T lymphocytes including a molecule of 50,000 Mr detected only on CD4 positive T blasts. Mol Immunol. 1987 Dec;24(12):1281–1290. doi: 10.1016/0161-5890(87)90122-2. [DOI] [PubMed] [Google Scholar]
  23. Richter G., Blankenstein T., Diamantstein T. Evolutionary aspects, structure, and expression of the rat interleukin 4 gene. Cytokine. 1990 May;2(3):221–228. doi: 10.1016/1043-4666(90)90020-t. [DOI] [PubMed] [Google Scholar]
  24. Robb R. J., Kutny R. M., Panico M., Morris H., DeGrado W. F., Chowdhry V. Posttranslational modification of human T-cell growth factor. Biochem Biophys Res Commun. 1983 Nov 15;116(3):1049–1055. doi: 10.1016/s0006-291x(83)80248-4. [DOI] [PubMed] [Google Scholar]
  25. Rosenberg S. A., Grimm E. A., McGrogan M., Doyle M., Kawasaki E., Koths K., Mark D. F. Biological activity of recombinant human interleukin-2 produced in Escherichia coli. Science. 1984 Mar 30;223(4643):1412–1414. doi: 10.1126/science.6367046. [DOI] [PubMed] [Google Scholar]
  26. Scahill S. J., Devos R., Van der Heyden J., Fiers W. Expression and characterization of the product of a human immune interferon cDNA gene in Chinese hamster ovary cells. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4654–4658. doi: 10.1073/pnas.80.15.4654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Smith G. E., Ju G., Ericson B. L., Moschera J., Lahm H. W., Chizzonite R., Summers M. D. Modification and secretion of human interleukin 2 produced in insect cells by a baculovirus expression vector. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8404–8408. doi: 10.1073/pnas.82.24.8404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Taniguchi T., Matsui H., Fujita T., Takaoka C., Kashima N., Yoshimoto R., Hamuro J. Structure and expression of a cloned cDNA for human interleukin-2. Nature. 1983 Mar 24;302(5906):305–310. doi: 10.1038/302305a0. [DOI] [PubMed] [Google Scholar]
  29. Yokota T., Lee F., Rennick D., Hall C., Arai N., Mosmann T., Nabel G., Cantor H., Arai K. Isolation and characterization of a mouse cDNA clone that expresses mast-cell growth-factor activity in monkey cells. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1070–1074. doi: 10.1073/pnas.81.4.1070. [DOI] [PMC free article] [PubMed] [Google Scholar]

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