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. 1991 Sep 1;174(3):673–681. doi: 10.1084/jem.174.3.673

Evidence that natural murine soluble interleukin 4 receptors may act as transport proteins

PMCID: PMC2118951  PMID: 1875167

Abstract

The present studies were undertaken to determine whether the interleukin 4 binding proteins (IL-4BPs) previously identified in the biological fluids of mice are soluble forms of IL-4Rs. We also studied the binding properties of IL-4BPs in order to gain insight into their physiological role in vivo. Affinity-purified IL-4BPs and recombinant soluble IL-4Rs generated similar one-dimensional (Cleveland) peptide maps after digestion with either Staphylococcus aureus V8 protease or trypsin, indicating structural similarities. Furthermore, a rat mAb directed against the murine IL-4Rs immunoprecipitated the IL-4BPs and completely inhibited binding of 125I-IL-4 to a purified preparation of IL-4BPs. Taken together these data indicate that the IL-4BPs are soluble IL-4Rs. At 4 degrees C the IL-4BPs competitively inhibited the binding of IL-4 to membrane IL-4Rs but their ability to prevent binding of IL-4 to cells at 37 degrees C, at the same concentrations, was significantly reduced. Kinetic binding studies of soluble IL-4BPs vs. membrane IL-4Rs disclosed important differences in their rates of dissociation from IL-4. Whereas dissociation at 4 degrees C was slow for both, dissociation of IL-4 from IL-BPs at 37 degrees C was considerably faster (t 1/2 of 2 min) than dissociation of IL-4 from membrane IL-4Rs (t 1/2 of approximately 69 min). Temperature-dependent changes in dissociation kinetics were reversible, and could not be accounted for by either inactivation of the IL-4BPs at 37 degrees C or receptor internalization. Additional experiments also demonstrated that when IL-4BPs bind to IL-4 at 37 degrees C, the IL-4/IL-4BPs complex can rapidly dissociate, allowing IL-4 to bind to membrane IL-4Rs. In addition, binding of IL-4 by the IL-4BPs protects IL-4 from proteolytic degradation. Taken together, these results suggest that the IL-4BPs are naturally occurring forms of soluble IL-4Rs and that some of their properties (fast dissociation kinetics and protection of IL-4 from proteolysis) are consistent with a potential role as carrier proteins for IL-4 in the circulation.

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Selected References

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  1. Akira S., Hirano T., Taga T., Kishimoto T. Biology of multifunctional cytokines: IL 6 and related molecules (IL 1 and TNF). FASEB J. 1990 Aug;4(11):2860–2867. [PubMed] [Google Scholar]
  2. Beckmann M. P., Schooley K. A., Gallis B., Vanden Bos T., Friend D., Alpert A. R., Raunio R., Prickett K. S., Baker P. E., Park L. S. Monoclonal antibodies block murine IL-4 receptor function. J Immunol. 1990 Jun 1;144(11):4212–4217. [PubMed] [Google Scholar]
  3. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  4. Dukovich M., Wano Y., Le thi Bich Thuy, Katz P., Cullen B. R., Kehrl J. H., Greene W. C. A second human interleukin-2 binding protein that may be a component of high-affinity interleukin-2 receptors. Nature. 1987 Jun 11;327(6122):518–522. doi: 10.1038/327518a0. [DOI] [PubMed] [Google Scholar]
  5. Engelmann H., Aderka D., Rubinstein M., Rotman D., Wallach D. A tumor necrosis factor-binding protein purified to homogeneity from human urine protects cells from tumor necrosis factor toxicity. J Biol Chem. 1989 Jul 15;264(20):11974–11980. [PubMed] [Google Scholar]
  6. Fanslow W. C., Clifford K., VandenBos T., Teel A., Armitage R. J., Beckmann M. P. A soluble form of the interleukin 4 receptor in biological fluids. Cytokine. 1990 Nov;2(6):398–401. doi: 10.1016/1043-4666(90)90047-w. [DOI] [PubMed] [Google Scholar]
  7. Fanslow W. C., Sims J. E., Sassenfeld H., Morrissey P. J., Gillis S., Dower S. K., Widmer M. B. Regulation of alloreactivity in vivo by a soluble form of the interleukin-1 receptor. Science. 1990 May 11;248(4956):739–742. doi: 10.1126/science.2139736. [DOI] [PubMed] [Google Scholar]
  8. Fernandez-Botran R., Sanders V. M., Oliver K. G., Chen Y. W., Krammer P. H., Uhr J. W., Vitetta E. S. Interleukin 4 mediates autocrine growth of helper T cells after antigenic stimulation. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9689–9693. doi: 10.1073/pnas.83.24.9689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fernandez-Botran R., Sanders V. M., Vitetta E. S. Interactions between receptors for interleukin 2 and interleukin 4 on lines of helper T cells (HT-2) and B lymphoma cells (BCL1). J Exp Med. 1989 Feb 1;169(2):379–391. doi: 10.1084/jem.169.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fernandez-Botran R., Vitetta E. S. A soluble, high-affinity, interleukin-4-binding protein is present in the biological fluids of mice. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4202–4206. doi: 10.1073/pnas.87.11.4202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goodwin R. G., Friend D., Ziegler S. F., Jerzy R., Falk B. A., Gimpel S., Cosman D., Dower S. K., March C. J., Namen A. E. Cloning of the human and murine interleukin-7 receptors: demonstration of a soluble form and homology to a new receptor superfamily. Cell. 1990 Mar 23;60(6):941–951. doi: 10.1016/0092-8674(90)90342-c. [DOI] [PubMed] [Google Scholar]
  12. Harada N., Castle B. E., Gorman D. M., Itoh N., Schreurs J., Barrett R. L., Howard M., Miyajima A. Expression cloning of a cDNA encoding the murine interleukin 4 receptor based on ligand binding. Proc Natl Acad Sci U S A. 1990 Feb;87(3):857–861. doi: 10.1073/pnas.87.3.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Josimovic-Alasevic O., Herrmann T., Diamantstein T. Demonstration of two distinct forms of released low-affinity type interleukin 2 receptors. Eur J Immunol. 1988 Nov;18(11):1855–1857. doi: 10.1002/eji.1830181133. [DOI] [PubMed] [Google Scholar]
  14. Kondo N., Kondo S., Shimizu A., Honjo T., Hamuro J. A soluble 'anchorminus' interleukin 2 receptor suppresses in vitro interleukin 2-mediated immune responses. Immunol Lett. 1988 Dec;19(4):299–307. doi: 10.1016/0165-2478(88)90159-9. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Lowenthal J. W., Castle B. E., Christiansen J., Schreurs J., Rennick D., Arai N., Hoy P., Takebe Y., Howard M. Expression of high affinity receptors for murine interleukin 4 (BSF-1) on hemopoietic and nonhemopoietic cells. J Immunol. 1988 Jan 15;140(2):456–464. [PubMed] [Google Scholar]
  17. Matsuda T., Hirano T., Nagasawa S., Kishimoto T. Identification of alpha 2-macroglobulin as a carrier protein for IL-6. J Immunol. 1989 Jan 1;142(1):148–152. [PubMed] [Google Scholar]
  18. Mosley B., Beckmann M. P., March C. J., Idzerda R. L., Gimpel S. D., VandenBos T., Friend D., Alpert A., Anderson D., Jackson J. The murine interleukin-4 receptor: molecular cloning and characterization of secreted and membrane bound forms. Cell. 1989 Oct 20;59(2):335–348. doi: 10.1016/0092-8674(89)90295-x. [DOI] [PubMed] [Google Scholar]
  19. Novick D., Engelmann H., Wallach D., Rubinstein M. Soluble cytokine receptors are present in normal human urine. J Exp Med. 1989 Oct 1;170(4):1409–1414. doi: 10.1084/jem.170.4.1409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ohara J., Paul W. E. Production of a monoclonal antibody to and molecular characterization of B-cell stimulatory factor-1. Nature. 1985 May 23;315(6017):333–336. doi: 10.1038/315333a0. [DOI] [PubMed] [Google Scholar]
  21. Ohara J., Paul W. E. Receptors for B-cell stimulatory factor-1 expressed on cells of haematopoietic lineage. Nature. 1987 Feb 5;325(6104):537–540. doi: 10.1038/325537a0. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Robb R. J., Rusk C. M., Yodoi J., Greene W. C. Interleukin 2 binding molecule distinct from the Tac protein: analysis of its role in formation of high-affinity receptors. Proc Natl Acad Sci U S A. 1987 Apr;84(7):2002–2006. doi: 10.1073/pnas.84.7.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rubin L. A., Kurman C. C., Fritz M. E., Biddison W. E., Boutin B., Yarchoan R., Nelson D. L. Soluble interleukin 2 receptors are released from activated human lymphoid cells in vitro. J Immunol. 1985 Nov;135(5):3172–3177. [PubMed] [Google Scholar]
  25. Schall T. J., Lewis M., Koller K. J., Lee A., Rice G. C., Wong G. H., Gatanaga T., Granger G. A., Lentz R., Raab H. Molecular cloning and expression of a receptor for human tumor necrosis factor. Cell. 1990 Apr 20;61(2):361–370. doi: 10.1016/0092-8674(90)90816-w. [DOI] [PubMed] [Google Scholar]
  26. Teshigawara K., Wang H. M., Kato K., Smith K. A. Interleukin 2 high-affinity receptor expression requires two distinct binding proteins. J Exp Med. 1987 Jan 1;165(1):223–238. doi: 10.1084/jem.165.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wagner D. K., York-Jolley J., Malek T. R., Berzofsky J. A., Nelson D. L. Antigen-specific murine T cell clones produce soluble interleukin 2 receptor on stimulation with specific antigens. J Immunol. 1986 Jul 15;137(2):592–596. [PubMed] [Google Scholar]
  28. Wang H. M., Smith K. A. The interleukin 2 receptor. Functional consequences of its bimolecular structure. J Exp Med. 1987 Oct 1;166(4):1055–1069. doi: 10.1084/jem.166.4.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Watson J. Continuous proliferation of murine antigen-specific helper T lymphocytes in culture. J Exp Med. 1979 Dec 1;150(6):1510–1519. doi: 10.1084/jem.150.6.1510. [DOI] [PMC free article] [PubMed] [Google Scholar]

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