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. 1982 Aug 1;156(2):454–464. doi: 10.1084/jem.156.2.454

Purification of human interleukin 2 to apparent homogeneity and its molecular heterogeneity

PMCID: PMC2186775  PMID: 6980256

Abstract

Interleukin 2 (IL-2), produced with and without co-stimulation by the Burkitt's lymphoma line Daudi, was purified 37,000-fold to apparent homogeneity from lymphocyte conditioned medium by (NH4)2SO4 precipitation, DEAE-cellulose ion-exchange chromatography, gel filtration, and chromatography on blue agarose and on Procion-red agarose. The purified IL-2 showed a 10(6) U/mg protein sp act. IL-2 produced in the absence of Daudi cells had a mol wt of 26,000 as measured by gel filtration and an isoelectric point of 6.7. This IL-2 showed a 16,000 and 17,000 mol wt in sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE). IL-2, produced in the presence of Daudi cells (10(6)/ml), showed a mol wt of approximately 14,000, as measured by both gel filtration and SDS-PAGE, and an isoelectric point of 8.1. The purified IL-2 lacked detectable interferon (alpha and gamma), granulocyte-macrophage colony-stimulating factor, B cell growth factor, T cell-replacing factor, and thymocyte- differentiating activity and was free of any contaminating proteins as judged by silver staining in SDS-PAGE. All three molecular forms of IL- 2 were biologically active at concentrations of 10(-11) - 10(-10) M, supporting the growth of human and murine cytotoxic T cell lines.

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Selected References

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  1. Burgess A. W., Metcalf D. The nature and action of granulocyte-macrophage colony stimulating factors. Blood. 1980 Dec;56(6):947–958. [PubMed] [Google Scholar]
  2. Caplan B., Gibbs C., Paetkau V. Properties of sodium dodecyl sulfate-denatured Interleukin 2. J Immunol. 1981 Apr;126(4):1351–1354. [PubMed] [Google Scholar]
  3. Clark-Lewis I., Schrader J. W. Biochemical characterization of regulatory factors derived from T cell hybridomas and spleen cells. II. Evidence for glycosylation of T cell growth factor, T cell replacing factor, and granulocyte-macrophage colony-stimulating factor. J Immunol. 1982 Jan;128(1):175–180. [PubMed] [Google Scholar]
  4. Friedman S. M., Thompson G., Halper J. P., Knowles D. M. OT-CLL: a human T cell chronic lymphocytic leukemia that produces IL 2 in high titer. J Immunol. 1982 Feb;128(2):935–940. [PubMed] [Google Scholar]
  5. Gillis S., Ferm M. M., Ou W., Smith K. A. T cell growth factor: parameters of production and a quantitative microassay for activity. J Immunol. 1978 Jun;120(6):2027–2032. [PubMed] [Google Scholar]
  6. Gillis S., Smith K. A., Watson J. Biochemical characterization of lymphocyte regulatory molecules. II. Purification of a class of rat and human lymphokines. J Immunol. 1980 Apr;124(4):1954–1962. [PubMed] [Google Scholar]
  7. Granelli-Piperno A., Vassalli J. D., Reich E. Purification of murine T cell growth factor. A lymphocyte mitogen with helper activity. J Exp Med. 1981 Aug 1;154(2):422–431. doi: 10.1084/jem.154.2.422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. 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]
  10. Merril C. R., Switzer R. C., Van Keuren M. L. Trace polypeptides in cellular extracts and human body fluids detected by two-dimensional electrophoresis and a highly sensitive silver stain. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4335–4339. doi: 10.1073/pnas.76.9.4335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mier J. W., Gallo R. C. Purification and some characteristics of human T-cell growth factor from phytohemagglutinin-stimulated lymphocyte-conditioned media. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6134–6138. doi: 10.1073/pnas.77.10.6134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mizel S. B., Mizel D. Purification to apparent homogeneity of murine interleukin 1. J Immunol. 1981 Mar;126(3):834–837. [PubMed] [Google Scholar]
  13. Morgan D. A., Ruscetti F. W., Gallo R. Selective in vitro growth of T lymphocytes from normal human bone marrows. Science. 1976 Sep 10;193(4257):1007–1008. doi: 10.1126/science.181845. [DOI] [PubMed] [Google Scholar]
  14. Paetkau V. Lymphokines on the move. Nature. 1981 Dec 24;294(5843):689–690. doi: 10.1038/294689a0. [DOI] [PubMed] [Google Scholar]
  15. Palacios R., Möller G. HLA-DR antigens render resting T cells sensitive to interleukin-2 and induce production of the growth factor in the autologous mixed lymphocyte reaction. Cell Immunol. 1981 Sep 1;63(1):143–153. doi: 10.1016/0008-8749(81)90035-6. [DOI] [PubMed] [Google Scholar]
  16. Robb R. J., Munck A., Smith K. A. T cell growth factor receptors. Quantitation, specificity, and biological relevance. J Exp Med. 1981 Nov 1;154(5):1455–1474. doi: 10.1084/jem.154.5.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Robb R. J., Smith K. A. Heterogeneity of human T-cell growth factor(s) due to variable glycosylation. Mol Immunol. 1981 Dec;18(12):1087–1094. doi: 10.1016/0161-5890(81)90024-9. [DOI] [PubMed] [Google Scholar]
  18. Ruscetti F. W., Gallo R. C. Human T-lymphocyte growth factor: regulation of growth and function of T lymphocytes. Blood. 1981 Mar;57(3):379–394. [PubMed] [Google Scholar]
  19. Shimizu S., Smith R. T., Norcross M. A., Maino V. C. Mechanisms controlling TCGF production by cloned sublines of EL-4 azgr in response to stimulation by anti-Thy-1 antibody. J Immunol. 1982 Jan;128(1):296–301. [PubMed] [Google Scholar]
  20. Smith K. A., Baker P. E., Gillis S., Ruscetti F. W. Functional and molecular characteristics of T-cell growth factor. Mol Immunol. 1980 May;17(5):579–589. doi: 10.1016/0161-5890(80)90156-x. [DOI] [PubMed] [Google Scholar]
  21. Watson J., Gillis S., Marbrook J., Mochizuki D., Smith K. A. Biochemical and biological characterization of lymphocyte regulatory molecules. I. Purification of a class of murine lymphokines. J Exp Med. 1979 Oct 1;150(4):849–861. doi: 10.1084/jem.150.4.849. [DOI] [PMC free article] [PubMed] [Google Scholar]

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