Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1984 Sep 1;160(3):772–787. doi: 10.1084/jem.160.3.772

Purification and partial biochemical characterization of normal human interleukin 1

PMCID: PMC2187399  PMID: 6332170

Abstract

A protocol for the rapid, efficient purification of the major charged species of human interleukin 1 (IL-1) has been developed using high performance anion exchange and size exclusion chromatography. The isolated material is pure as determined by sodium dodecyl sulfate (SDS) gradient polyacrylamide gel electrophoresis (PAGE) and analytical isoelectric focusing (IEF). The molecular weight of the purified material is 15,000 and the isoelectric point (pI) is 6.8, values that are in good agreement with those previously reported for human IL-1. 10(-10) M concentrations of the purified material give half-maximal stimulation in the thymocyte proliferation assay. Amounts of IL-1 sufficient for receptor studies and detailed biochemical analysis can now be produced on a regular basis.

Full Text

The Full Text of this article is available as a PDF (983.2 KB).

Selected References

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

  1. Anzano M. A., Roberts A. B., Smith J. M., Lamb L. C., Sporn M. B. Purification by reverse-phase high-performance liquid chromatography of an epidermal growth factor-dependent transforming growth factor. Anal Biochem. 1982 Sep 1;125(1):217–224. doi: 10.1016/0003-2697(82)90405-5. [DOI] [PubMed] [Google Scholar]
  2. Baumann G., Chrambach A. Lack of interaction between hormonal proteins and synthetic carrier ampholytes (Ampholine). Anal Biochem. 1975 Apr;64(2):530–536. doi: 10.1016/0003-2697(75)90464-9. [DOI] [PubMed] [Google Scholar]
  3. Bloomster T. G., Watson D. W. Effects of carrier ampholyte contamination on the biological and biochemical properties of streptococcal pyrogenic exotoxin type C. Infect Immun. 1983 Jan;39(1):311–314. doi: 10.1128/iai.39.1.311-314.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blyden G., Handschumacher R. E. Purification and properties of human lymphocyte activating factor (LAF). J Immunol. 1977 May;118(5):1631–1638. [PubMed] [Google Scholar]
  5. Bonnard G. D., Yasaka K., Jacobson D. Ligand-activated T cell growth factor-induced proliferation: absorption of T cell growth factor by activated T cells. J Immunol. 1979 Dec;123(6):2704–2708. [PubMed] [Google Scholar]
  6. Coutinho A., Larsson E. L., Grönvik K. O., Andersson J. Studies on T lymphocyte activation II. The target cells for concanavalin A-induced growth factors. Eur J Immunol. 1979 Aug;9(8):587–592. doi: 10.1002/eji.1830090803. [DOI] [PubMed] [Google Scholar]
  7. Cuatrecasas P., Hollenberg M. D. Membrane receptors and hormone action. Adv Protein Chem. 1976;30:251–451. doi: 10.1016/s0065-3233(08)60481-7. [DOI] [PubMed] [Google Scholar]
  8. Dinarello C. A., Renfer L., Wolff S. M. Human leukocytic pyrogen: purification and development of a radioimmunoassay. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4624–4627. doi: 10.1073/pnas.74.10.4624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Economou J. S., Shin H. S. Lymphocyte-activating factor. I. Generation and physicochemical characterization. J Immunol. 1978 Oct;121(4):1446–1452. [PubMed] [Google Scholar]
  10. Farr A. G., Dorf M. E., Unanue E. R. Secretion of mediators following T lymphocyte-macrophage interaction is regulated by the major histocompatibility complex. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3542–3546. doi: 10.1073/pnas.74.8.3542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gery I., Gershon R. K., Waksman B. H. Potentiation of the T-lymphocyte response to mitogens. I. The responding cell. J Exp Med. 1972 Jul 1;136(1):128–142. doi: 10.1084/jem.136.1.128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gierthy J. F., Ellem K. A., Kongsvik J. R. Mitogenic and inhibitory effects of carrier ampholytes on quiescent and stimulated human diploid lung fibroblasts. Anal Biochem. 1979 Sep 15;98(1):27–35. doi: 10.1016/0003-2697(79)90701-2. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Henderson L. E., Hewetson J. F., Hopkins R. F., 3rd, Sowder R. C., Neubauer R. H., Rabin H. A rapid, large scale purification procedure for gibbon interleukin 2. J Immunol. 1983 Aug;131(2):810–815. [PubMed] [Google Scholar]
  15. Howard M., Mizel S. B., Lachman L., Ansel J., Johnson B., Paul W. E. Role of interleukin 1 in anti-immunoglobulin-induced B cell proliferation. J Exp Med. 1983 May 1;157(5):1529–1543. doi: 10.1084/jem.157.5.1529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Koopman W. J., Farrar J. J., Oppenheim J. J., Fuller-Bonar J., Dougherty S. Association of a low molecular weight helper factor(s) with thymocyte proliferative activity. J Immunol. 1977 Jul;119(1):55–60. [PubMed] [Google Scholar]
  17. Krakauer T., Oppenheim J. J. Interleukin 1 production by a human acute monocytic leukemia cell line. Cell Immunol. 1983 Sep;80(2):223–229. doi: 10.1016/0008-8749(83)90111-9. [DOI] [PubMed] [Google Scholar]
  18. Lachman L. B., Hacker M. P., Handschumacher R. E. Partial purification of human lymphocyte-activating factor (LAF) by ultrafiltration and electrophoretic techniques. J Immunol. 1977 Dec;119(6):2019–2023. [PubMed] [Google Scholar]
  19. Lachman L. B., Page S. O., Metzgar R. S. Purification of human interleukin 1. J Supramol Struct. 1980;13(4):457–466. doi: 10.1002/jss.400130405. [DOI] [PubMed] [Google Scholar]
  20. Luger T. A., Stadler B. M., Luger B. M., Mathieson B. J., Mage M., Schmidt J. A., Oppenheim J. J. Murine epidermal cell-derived thymocyte-activating factor resembles murine interleukin 1. J Immunol. 1982 May;128(5):2147–2152. [PubMed] [Google Scholar]
  21. Malamud D., Drysdale J. W. Isoelectric points of proteins: a table. Anal Biochem. 1978 Jun 1;86(2):620–647. doi: 10.1016/0003-2697(78)90790-x. [DOI] [PubMed] [Google Scholar]
  22. Mizel S. B., Dayer J. M., Krane S. M., Mergenhagen S. E. Stimulation of rheumatoid synovial cell collagenase and prostaglandin production by partially purified lymphocyte-activating factor (interleukin 1). Proc Natl Acad Sci U S A. 1981 Apr;78(4):2474–2477. doi: 10.1073/pnas.78.4.2474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mizel S. B., Dukovich M., Rothstein J. Preparation of goat antibodies against interleukin 1: use of an immunoadsorbent to purify interleukin 1. J Immunol. 1983 Oct;131(4):1834–1837. [PubMed] [Google Scholar]
  24. Mizel S. B., Oppenheim J. J., Rosenstreich D. L. Characterization of lymphocyte-activating factor (LAF) produced by the macrophage cell line, P388D1. I. Enhancement of LAF production by activated T lymphocytes. J Immunol. 1978 May;120(5):1497–1503. [PubMed] [Google Scholar]
  25. Mizel S. B. Physicochemical characterization of lymphocyte-activating factor (LAF). J Immunol. 1979 Jun;122(6):2167–2172. [PubMed] [Google Scholar]
  26. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  27. Murphy P. A., Simon P. L., Willoughby W. F. Endogenous pyrogens made by rabbit peritoneal exudate cells are identical with lymphocyte-activating factors made by rabbit alveolar macrophages. J Immunol. 1980 May;124(5):2498–2501. [PubMed] [Google Scholar]
  28. Niedel J. E., Kuhn L. J., Vandenbark G. R. Phorbol diester receptor copurifies with protein kinase C. Proc Natl Acad Sci U S A. 1983 Jan;80(1):36–40. doi: 10.1073/pnas.80.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Postlethwaite A. E., Lachman L. B., Mainardi C. L., Kang A. H. Interleukin 1 stimulation of collagenase production by cultured fibroblasts. J Exp Med. 1983 Feb 1;157(2):801–806. doi: 10.1084/jem.157.2.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Regnier F. E. High-performance liquid chromatography of biopolymers. Science. 1983 Oct 21;222(4621):245–252. doi: 10.1126/science.6353575. [DOI] [PubMed] [Google Scholar]
  31. Rosenstreich D. L., Mizel S. B. Signal requirements for T lymphocyte activation. I. Replacement of macrophage function with phorbol myristic acetate. J Immunol. 1979 Oct;123(4):1749–1754. [PubMed] [Google Scholar]
  32. Rubinstein M., Rubinstein S., Familletti P. C., Miller R. S., Waldman A. A., Pestka S. Human leukocyte interferon: production, purification to homogeneity, and initial characterization. Proc Natl Acad Sci U S A. 1979 Feb;76(2):640–644. doi: 10.1073/pnas.76.2.640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schmidt J. A., Mizel S. B., Cohen D., Green I. Interleukin 1, a potential regulator of fibroblast proliferation. J Immunol. 1982 May;128(5):2177–2182. [PubMed] [Google Scholar]
  34. Schmidt J. A., Oliver C. N., Lepe-Zuniga J. L., Green I., Gery I. Silica-stimulated monocytes release fibroblast proliferation factors identical to interleukin 1. A potential role for interleukin 1 in the pathogenesis of silicosis. J Clin Invest. 1984 May;73(5):1462–1472. doi: 10.1172/JCI111350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Simon P. L., Willoughby W. F. The role of subcellular factors in pulmonary immune function: physicochemical characterization of two distinct species of lymphocyte-activating factor produced by rabbit alveolar macrophages. J Immunol. 1981 Apr;126(4):1534–1541. [PubMed] [Google Scholar]
  36. Smith K. A., Lachman L. B., Oppenheim J. J., Favata M. F. The functional relationship of the interleukins. J Exp Med. 1980 Jun 1;151(6):1551–1556. doi: 10.1084/jem.151.6.1551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Spector T. Refinement of the coomassie blue method of protein quantitation. A simple and linear spectrophotometric assay for less than or equal to 0.5 to 50 microgram of protein. Anal Biochem. 1978 May;86(1):142–146. doi: 10.1016/0003-2697(78)90327-5. [DOI] [PubMed] [Google Scholar]
  38. Sztein M. B., Vogel S. N., Sipe J. D., Murphy P. A., Mizel S. B., Oppenheim J. J., Rosenstreich D. L. The role of macrophages in the acute-phase response: SAA inducer is closely related to lymphocyte activating factor and endogenous pyrogen. Cell Immunol. 1981 Sep 1;63(1):164–176. doi: 10.1016/0008-8749(81)90037-x. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES