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. 1997 Jul 1;325(Pt 1):147–153. doi: 10.1042/bj3250147

Responsiveness of human neutrophils to interleukin-4: induction of cytoskeletal rearrangements, de novo protein synthesis and delay of apoptosis.

D Girard 1, R Paquin 1, A D Beaulieu 1
PMCID: PMC1218539  PMID: 9224640

Abstract

Interleukin-4 (IL-4) and IL-13 are cytokines that share many biological activities. We have previously demonstrated that IL-13 affects a number of neutrophil responses, and here we extend our observations to IL-4. We present, for the first time, direct evidence for the presence of functional IL-4 receptors on human neutrophils. We report that IL-4 induces RNA synthesis in a concentration-dependent manner and, based on observations of the induction of morphological cell shape changes and spreading onto glass, we demonstrate that IL-4 activates neutrophil cytoskeletal rearrangements. We further show that IL-4 is a potent activator of de novo protein synthesis in neutrophils, and we identify by microsequencing one of these proteins as the cytoskeletal protein actin. We were also able to demonstrate for the first time that actin is cleaved into at least two fragments of approximately 30 kDa (pI 5.4) and approximately 25 kDa (pI 5.0) in neutrophils. Finally, we report that IL-4 delays neutrophil apoptosis, as assessed by morphological observations from cytocentrifuge preparations, as well as by measurement of differences in staining by flow cytometry with both propidium iodide and Hoechst reagent. Taken together, we conclude that IL-4 is a more potent neutrophil agonist than previously believed. We discuss the possibility that the induction of the de novo synthesis of actin by IL-4 is related to the mechanism by which this cytokine delays apoptosis; in addition, the cleavage of this protein is likely to contribute to the apoptotic process.

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Selected References

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  1. Atkins M. B., Vachino G., Tilg H. J., Karp D. D., Robert N. J., Kappler K., Mier J. W. Phase I evaluation of thrice-daily intravenous bolus interleukin-4 in patients with refractory malignancy. J Clin Oncol. 1992 Nov;10(11):1802–1809. doi: 10.1200/JCO.1992.10.11.1802. [DOI] [PubMed] [Google Scholar]
  2. Beaulieu A. D., Paquin R., Rathanaswami P., McColl S. R. Nuclear signaling in human neutrophils. Stimulation of RNA synthesis is a response to a limited number of proinflammatory agonists. J Biol Chem. 1992 Jan 5;267(1):426–432. [PubMed] [Google Scholar]
  3. Biffl W. L., Moore E. E., Moore F. A., Barnett C. C., Jr Interleukin-6 suppression of neutrophil apoptosis is neutrophil concentration dependent. J Leukoc Biol. 1995 Nov;58(5):582–584. doi: 10.1002/jlb.58.5.582. [DOI] [PubMed] [Google Scholar]
  4. Bober L. A., Waters T. A., Pugliese-Sivo C. C., Sullivan L. M., Narula S. K., Grace M. J. IL-4 induces neutrophilic maturation of HL-60 cells and activation of human peripheral blood neutrophils. Clin Exp Immunol. 1995 Jan;99(1):129–136. doi: 10.1111/j.1365-2249.1995.tb03483.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boey H., Rosenbaum R., Castracane J., Borish L. Interleukin-4 is a neutrophil activator. J Allergy Clin Immunol. 1989 May;83(5):978–984. doi: 10.1016/0091-6749(89)90115-2. [DOI] [PubMed] [Google Scholar]
  6. Bosco M. C., Pulkki K., Rowe T. K., Zea A. H., Musso T., Longo D. L., Varesio L., Espinoza-Delgado I. IL-4 inhibits IL-2-induced tumoricidal activity and secretory functions of human monocytes. Modulation of IL-2 binding and IL-2 receptor beta gamma chain expression. J Immunol. 1995 Aug 1;155(3):1411–1419. [PubMed] [Google Scholar]
  7. Brach M. A., deVos S., Gruss H. J., Herrmann F. Prolongation of survival of human polymorphonuclear neutrophils by granulocyte-macrophage colony-stimulating factor is caused by inhibition of programmed cell death. Blood. 1992 Dec 1;80(11):2920–2924. [PubMed] [Google Scholar]
  8. Brizzi M. F., Aronica M. G., Rosso A., Bagnara G. P., Yarden Y., Pegoraro L. Granulocyte-macrophage colony-stimulating factor stimulates JAK2 signaling pathway and rapidly activates p93fes, STAT1 p91, and STAT3 p92 in polymorphonuclear leukocytes. J Biol Chem. 1996 Feb 16;271(7):3562–3567. doi: 10.1074/jbc.271.7.3562. [DOI] [PubMed] [Google Scholar]
  9. Colotta F., Re F., Muzio M., Bertini R., Polentarutti N., Sironi M., Giri J. G., Dower S. K., Sims J. E., Mantovani A. Interleukin-1 type II receptor: a decoy target for IL-1 that is regulated by IL-4. Science. 1993 Jul 23;261(5120):472–475. doi: 10.1126/science.8332913. [DOI] [PubMed] [Google Scholar]
  10. Darzynkiewicz Z., Bruno S., Del Bino G., Gorczyca W., Hotz M. A., Lassota P., Traganos F. Features of apoptotic cells measured by flow cytometry. Cytometry. 1992;13(8):795–808. doi: 10.1002/cyto.990130802. [DOI] [PubMed] [Google Scholar]
  11. Favre C., Saeland S., Caux C., Duvert V., De Vries J. E. Interleukin-4 has basophilic and eosinophilic cell growth-promoting activity on cord blood cells. Blood. 1990 Jan 1;75(1):67–73. [PubMed] [Google Scholar]
  12. Finney M., Guy G. R., Michell R. H., Gordon J., Dugas B., Rigley K. P., Callard R. E. Interleukin 4 activates human B lymphocytes via transient inositol lipid hydrolysis and delayed cyclic adenosine monophosphate generation. Eur J Immunol. 1990 Jan;20(1):151–156. doi: 10.1002/eji.1830200122. [DOI] [PubMed] [Google Scholar]
  13. Gilleece M. H., Scarffe J. H., Ghosh A., Heyworth C. M., Bonnem E., Testa N., Stern P., Dexter T. M. Recombinant human interleukin 4 (IL-4) given as daily subcutaneous injections--a phase I dose toxicity trial. Br J Cancer. 1992 Jul;66(1):204–210. doi: 10.1038/bjc.1992.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Girard D., Gosselin J., Heitz D., Paquin R., Beaulieu A. D. Effects of interleukin-2 on gene expression in human neutrophils. Blood. 1995 Aug 1;86(3):1170–1176. [PubMed] [Google Scholar]
  15. Girard D., Paquet M. E., Paquin R., Beaulieu A. D. Differential effects of interleukin-15 (IL-15) and IL-2 on human neutrophils: modulation of phagocytosis, cytoskeleton rearrangement, gene expression, and apoptosis by IL-15. Blood. 1996 Oct 15;88(8):3176–3184. [PubMed] [Google Scholar]
  16. Girard D., Paquin R., Naccache P. H., Beaulieu A. D. Effects of interleukin-13 on human neutrophil functions. J Leukoc Biol. 1996 Mar;59(3):412–419. doi: 10.1002/jlb.59.3.412. [DOI] [PubMed] [Google Scholar]
  17. Ishii N., Takeshita T., Kimura Y., Tada K., Kondo M., Nakamura M., Sugamura K. Expression of the IL-2 receptor gamma chain on various populations in human peripheral blood. Int Immunol. 1994 Aug;6(8):1273–1277. doi: 10.1093/intimm/6.8.1273. [DOI] [PubMed] [Google Scholar]
  18. Kayalar C., Ord T., Testa M. P., Zhong L. T., Bredesen D. E. Cleavage of actin by interleukin 1 beta-converting enzyme to reverse DNase I inhibition. Proc Natl Acad Sci U S A. 1996 Mar 5;93(5):2234–2238. doi: 10.1073/pnas.93.5.2234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Keegan A. D., Nelms K., Wang L. M., Pierce J. H., Paul W. E. Interleukin 4 receptor: signaling mechanisms. Immunol Today. 1994 Sep;15(9):423–432. doi: 10.1016/0167-5699(94)90272-0. [DOI] [PubMed] [Google Scholar]
  20. Komai-Koma M., Liew F. Y., Wilkinson P. C. Interactions between IL-4, anti-CD40, and anti-immunoglobulin as activators of locomotion of human B cells. J Immunol. 1995 Aug 1;155(3):1110–1116. [PubMed] [Google Scholar]
  21. Kondo M., Takeshita T., Ishii N., Nakamura M., Watanabe S., Arai K., Sugamura K. Sharing of the interleukin-2 (IL-2) receptor gamma chain between receptors for IL-2 and IL-4. Science. 1993 Dec 17;262(5141):1874–1877. doi: 10.1126/science.8266076. [DOI] [PubMed] [Google Scholar]
  22. Lee A., Whyte M. K., Haslett C. Inhibition of apoptosis and prolongation of neutrophil functional longevity by inflammatory mediators. J Leukoc Biol. 1993 Oct;54(4):283–288. [PubMed] [Google Scholar]
  23. Liu J. H., Wei S., Ussery D., Epling-Burnette P. K., Leonard W. J., Djeu J. Y. Expression of interleukin-2 receptor gamma chain on human neutrophils. Blood. 1994 Dec 1;84(11):3870–3875. [PubMed] [Google Scholar]
  24. Marcelletti J. F., Ohara J., Katz D. H. Collagen-induced arthritis in mice. Relationship of collagen-specific and total IgE synthesis to disease. J Immunol. 1991 Dec 15;147(12):4185–4191. [PubMed] [Google Scholar]
  25. McColl S. R., Paquin R., Ménard C., Beaulieu A. D. Human neutrophils produce high levels of the interleukin 1 receptor antagonist in response to granulocyte/macrophage colony-stimulating factor and tumor necrosis factor alpha. J Exp Med. 1992 Aug 1;176(2):593–598. doi: 10.1084/jem.176.2.593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Miwa T., Manabe Y., Kurokawa K., Kamada S., Kanda N., Bruns G., Ueyama H., Kakunaga T. Structure, chromosome location, and expression of the human smooth muscle (enteric type) gamma-actin gene: evolution of six human actin genes. Mol Cell Biol. 1991 Jun;11(6):3296–3306. doi: 10.1128/mcb.11.6.3296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Müller K. M., Jaunin F., Masouyé I., Piguet P. F., Saurat J. H., Hauser C. Involvement of granulocytes and the adhesion receptors intercellular adhesion molecule-1 and lymphocyte function-associated antigen-1 in tissue inflammation induced by Th2-type helper cells. J Invest Dermatol. 1995 Mar;104(3):350–354. doi: 10.1111/1523-1747.ep12665378. [DOI] [PubMed] [Google Scholar]
  28. Müller K. M., Jaunin F., Masouyé I., Saurat J. H., Hauser C. Th2 cells mediate IL-4-dependent local tissue inflammation. J Immunol. 1993 Jun 15;150(12):5576–5584. [PubMed] [Google Scholar]
  29. Nakarai T., Robertson M. J., Streuli M., Wu Z., Ciardelli T. L., Smith K. A., Ritz J. Interleukin 2 receptor gamma chain expression on resting and activated lymphoid cells. J Exp Med. 1994 Jul 1;180(1):241–251. doi: 10.1084/jem.180.1.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. 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]
  33. Paul W. E. Interleukin-4: a prototypic immunoregulatory lymphokine. Blood. 1991 May 1;77(9):1859–1870. [PubMed] [Google Scholar]
  34. Perez V. L., Lederer J. A., Lichtman A. H., Abbas A. K. Stability of Th1 and Th2 populations. Int Immunol. 1995 May;7(5):869–875. doi: 10.1093/intimm/7.5.869. [DOI] [PubMed] [Google Scholar]
  35. Punnonen J., Aversa G., Cocks B. G., de Vries J. E. Role of interleukin-4 and interleukin-13 in synthesis of IgE and expression of CD23 by human B cells. Allergy. 1994 Sep;49(8):576–586. doi: 10.1111/j.1398-9995.1994.tb00122.x. [DOI] [PubMed] [Google Scholar]
  36. Racke M. K., Bonomo A., Scott D. E., Cannella B., Levine A., Raine C. S., Shevach E. M., Röcken M. Cytokine-induced immune deviation as a therapy for inflammatory autoimmune disease. J Exp Med. 1994 Nov 1;180(5):1961–1966. doi: 10.1084/jem.180.5.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Romagnani S. Regulatory role of IL4 and other cytokines in the function and development of human T-cell clones. Res Immunol. 1993 Oct;144(8):625–628. doi: 10.1016/s0923-2494(05)80015-x. [DOI] [PubMed] [Google Scholar]
  38. Schlossman S. F., Boumsell L., Gilks W., Harlan J. M., Kishimoto T., Morimoto C., Ritz J., Shaw S., Silverstein R. L., Springer T. A. CD antigens 1993. Blood. 1994 Feb 15;83(4):879–880. [PubMed] [Google Scholar]
  39. Taniguchi T. Cytokine signaling through nonreceptor protein tyrosine kinases. Science. 1995 Apr 14;268(5208):251–255. doi: 10.1126/science.7716517. [DOI] [PubMed] [Google Scholar]
  40. Tepper R. I., Coffman R. L., Leder P. An eosinophil-dependent mechanism for the antitumor effect of interleukin-4. Science. 1992 Jul 24;257(5069):548–551. doi: 10.1126/science.1636093. [DOI] [PubMed] [Google Scholar]
  41. Vercelli D., Jabara H. H., Arai K., Geha R. S. Induction of human IgE synthesis requires interleukin 4 and T/B cell interactions involving the T cell receptor/CD3 complex and MHC class II antigens. J Exp Med. 1989 Apr 1;169(4):1295–1307. doi: 10.1084/jem.169.4.1295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Whyte M. K., Meagher L. C., MacDermot J., Haslett C. Impairment of function in aging neutrophils is associated with apoptosis. J Immunol. 1993 Jun 1;150(11):5124–5134. [PubMed] [Google Scholar]
  43. Zurawski S. M., Vega F., Jr, Huyghe B., Zurawski G. Receptors for interleukin-13 and interleukin-4 are complex and share a novel component that functions in signal transduction. EMBO J. 1993 Jul;12(7):2663–2670. doi: 10.1002/j.1460-2075.1993.tb05927.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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