The molecular basis of IL-21–mediated proliferation

Rong Zeng

Rosanne Spolski

Esther Casas

Wei Zhu

David Levy

Warren Leonard

^{Laboratory of Molecular Immunology, National Heart, Lung, and Blood Institute (NHLBI), National Institutes of Health (NIH), Bethesda, MD;}

^{Departments of Pathology and Microbiology, New York University (NYU) School of Medicine, New York, NY}

^{Corresponding author.}

Received 2006 Oct 30; Accepted 2007 Jan 3.

Abstract

Interleukin-21 (IL-21) is a type I cytokine that modulates functions of T, B, natural killer (NK), and myeloid cells. The IL-21 receptor (IL-21R) is closely related to the IL-2 receptor β chain and is capable of transducing signals through its dimerization with the common cytokine receptor γ chain (γ_c), the protein whose expression is defective in humans with X-linked severe combined immunodeficiency. To clarify the molecular basis of IL-21 actions, we investigated the role of tyrosine residues in the IL-21R cytoplasmic domain. Simultaneous mutation of all 6 tyrosines greatly diminished IL-21–mediated proliferation, whereas retention of tyrosine 510 (Y510) allowed full proliferation. Y510 efficiently mediated IL-21–induced phosphorylation of Stat1 and Stat3, but not of Stat5, and CD8^{T cells from Stat1/Stat3 double knock-out mice exhibited decreased proliferation in response to IL-21 + IL-15. In addition, IL-21 weakly induced phosphorylation of Shc and Akt, and consistent with this, specific inhibitors of the MAPK and PI3K pathways inhibited IL-21–mediated proliferation. Collectively, these data indicate the involvement of the Jak-STAT, MAPK, and PI3K pathways in IL-21 signaling.}

Abstract

Acknowledgments

This work was supported by the Intramural Research Program, NHLBI, NIH (W.J.L.), and NIH grant AI28900 (D.E.L.).

We thank Drs Jian-Xin Lin, Hai-Hui Xue, Wei Liao, and Keji Zhao for valuable discussions and/or critical comments.

Acknowledgments

Footnotes

The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked “advertisement” in accordance with 18 USC section 1734.

Footnotes

References

1. Asao H, Okuyama C, Kumaki S, et al Cutting edge: the common gamma-chain is an indispensable subunit of the IL-21 receptor complex. J Immunol. 2001;167:1–5.[PubMed][Google Scholar]
2. Habib T, Senadheera S, Weinberg K, Kaushansky KThe common gamma chain (gamma c) is a required signaling component of the IL-21 receptor and supports IL-21-induced cell proliferation via JAK3. Biochemistry. 2002;41:8725–8731.[PubMed][Google Scholar]
3. Noguchi M, Yi H, Rosenblatt HM, et al Interleukin-2 receptor gamma chain mutation results in X-linked severe combined immunodeficiency in humans. Cell. 1993;73:147–157.[PubMed][Google Scholar]
4. Ozaki K, Spolski R, Feng CG, et al A critical role for IL-21 in regulating immunoglobulin production. Science. 2002;298:1630–1634.[PubMed][Google Scholar]
5. Leonard WJ, Spolski RInterleukin-21: a modulator of lymphoid proliferation, apoptosis and differentiation. Nat Rev Immunol. 2005;5:688–698.[PubMed][Google Scholar]
6. Mehta DS, Wurster AL, Whitters MJ, Young DA, Collins M, Grusby MJIL-21 induces the apoptosis of resting and activated primary B cells. J Immunol. 2003;170:4111–4118.[PubMed][Google Scholar]
7. Ozaki K, Spolski R, Ettinger R, et al Regulation of B cell differentiation and plasma cell generation by IL-21, a novel inducer of Blimp-1 and Bcl-6. J Immunol. 2004;173:5361–5371.[PubMed][Google Scholar]
8. Jin H, Carrio R, Yu A, Malek TRDistinct activation signals determine whether IL-21 induces B cell costimulation, growth arrest, or Bim-dependent apoptosis. J Immunol. 2004;173:657–665.[PubMed][Google Scholar]
9. Ettinger R, Sims GP, Fairhurst AM, et al IL-21 induces differentiation of human naive and memory B cells into antibody-secreting plasma cells. J Immunol. 2005;175:7867–7879.[PubMed][Google Scholar]
10. Parrish-Novak J, Dillon SR, Nelson A, et al Interleukin 21 and its receptor are involved in NK cell expansion and regulation of lymphocyte function. Nature. 2000;408:57–63.[PubMed][Google Scholar]
11. Zeng R, Spolski R, Finkelstein SE, et al Synergy of IL-21 and IL-15 in regulating CD8+ T cell expansion and function. J Exp Med. 2005;201:139–148.[Google Scholar]
12. Brady J, Hayakawa Y, Smyth MJ, Nutt SLIL-21 induces the functional maturation of murine NK cells. J Immunol. 2004;172:2048–2058.[PubMed][Google Scholar]
13. Kasaian MT, Whitters MJ, Carter LL, et al IL-21 limits NK cell responses and promotes antigen-specific T cell activation: a mediator of the transition from innate to adaptive immunity. Immunity. 2002;16:559–569.[PubMed][Google Scholar]
14. Smyth MJ, Wallace ME, Nutt SL, Yagita H, Godfrey DI, Hayakawa YSequential activation of NKT cells and NK cells provides effective innate immunotherapy of cancer. J Exp Med. 2005;201:1973–1985.[Google Scholar]
15. Toomey JA, Gays F, Foster D, Brooks CGCytokine requirements for the growth and development of mouse NK cells in vitro. J Leuk Biol. 2003;74:233–242.[PubMed][Google Scholar]
16. Wang G, Tschoi M, Spolski R, et al In vivo antitumor activity of interleukin 21 mediated by natural killer cells. Cancer Res. 2003;63:9016–9022.[PubMed][Google Scholar]
17. Brandt K, Bulfone-Paus S, Foster DC, Ruckert RInterleukin-21 inhibits dendritic cell activation and maturation. Blood. 2003;102:4090–4098.[PubMed][Google Scholar]
18. Pelletier M, Bouchard A, Girard DIn vivo and in vitro roles of IL-21 in inflammation. J Immunol. 2004;173:7521–7530.[PubMed][Google Scholar]
19. Leonard WJCytokines and immunodeficiency diseases. Nat Rev Immunol. 2001;1:200–208.[PubMed][Google Scholar]
20. Ozaki K, Kikly K, Michalovich D, Young PR, Leonard WJCloning of a type I cytokine receptor most related to the IL-2 receptor beta chain. Proc Natl Acad Sci U S A. 2000;97:11439–11444.[Google Scholar]
21. Strengell M, Sareneva T, Foster D, Julkunen I, Matikainen SIL-21 up-regulates the expression of genes associated with innate immunity and Th1 response. J Immunol. 2002;169:3600–3605.[PubMed][Google Scholar]
22. Suto A, Nakajima H, Hirose K, et al Interleukin 21 prevents antigen-induced IgE production by inhibiting germ line C(epsilon) transcription of IL-4-stimulated B cells. Blood. 2002;100:4565–4573.[PubMed][Google Scholar]
23. Hunter TSignaling: 2000 and beyond. Cell. 2000;100:113–127.[PubMed][Google Scholar]
24. Pawson TSpecificity in signal transduction: from phosphotyrosine-SH2 domain interactions to complex cellular systems. Cell. 2004;116:191–203.[PubMed][Google Scholar]
25. Durbin JE, Hackenmiller R, Simon MC, Levy DETargeted disruption of the mouse Stat1 gene results in compromised innate immunity to viral disease. Cell. 1996;84:443–450.[PubMed][Google Scholar]
26. Lee CK, Raz R, Gimeno R, et al STAT3 is a negative regulator of granulopoiesis but is not required for G-CSF-dependent differentiation. Immunity. 2002;17:63–72.[PubMed][Google Scholar]
27. Wolfer A, Bakker T, Wilson A, et al Inactivation of Notch 1 in immature thymocytes does not perturb CD4 or CD8T cell development. Nat Immunol. 2001;2:235–241.[PubMed][Google Scholar]
28. Gimeno R, Lee CK, Schindler C, Levy DEStat1 and Stat2 but not Stat3 arbitrate contradictory growth signals elicited by alpha/beta interferon in T lymphocytes. Mol Cell Biol. 2005;25:5456–5465.[Google Scholar]
29. Itoh T, Muto A, Watanabe S, Miyajima A, Yokota T, Arai KGranulocyte-macrophage colony-stimulating factor provokes RAS activation and transcription of c-fos through different modes of signaling. J Biol Chem. 1996;271:7587–7592.[PubMed][Google Scholar]
30. Naviaux RK, Costanzi E, Haas M, Verma IMThe pCL vector system: rapid production of helper-free, high-titer, recombinant retroviruses. J Virol. 1996;70:5701–5705.[Google Scholar]
31. Xue HH, Kovanen PE, Pise-Masison CA, et al IL-2 negatively regulates IL-7 receptor alpha chain expression in activated T lymphocytes. Proc Natl Acad Sci U S A. 2002;99:13759–13764.[Google Scholar]
32. Friedmann MC, Migone TS, Russell SM, Leonard WJDifferent interleukin 2 receptor beta-chain tyrosines couple to at least two signaling pathways and synergistically mediate interleukin 2-induced proliferation. Proc Natl Acad Sci U S A. 1996;93:2077–2082.[Google Scholar]
33. Goldsmith MA, Lai SY, Xu W, et al Growth signal transduction by the human interleukin-2 receptor requires cytoplasmic tyrosines of the beta chain and non-tyrosine residues of the gamma c chain. J Biol Chem. 1995;270:21729–21737.[PubMed][Google Scholar]
34. Hou J, Schindler U, Henzel WJ, Ho TC, Brasseur M, McKnight SLAn interleukin-4-induced transcription factor: IL-4 Stat. Science. 1994;265:1701–1706.[PubMed][Google Scholar]
35. Lin JX, Migone TS, Tsang M, et al The role of shared receptor motifs and common Stat proteins in the generation of cytokine pleiotropy and redundancy by IL-2, IL-4, IL-7, IL-13, and IL-15. Immunity. 1995;2:331–339.[PubMed][Google Scholar]
36. Yu CL, Meyer DJ, Campbell GS, et al Enhanced DNA-binding activity of a Stat3-related protein in cells transformed by the Src oncoprotein. Science. 1995;269:81–83.[PubMed][Google Scholar]
37. Bromberg JF, Wrzeszczynska MH, Devgan G, et al Stat3 as an oncogene. Cell. 1999;98:295–303.[PubMed][Google Scholar]
38. Kelly JA, Spolski R, Kovanen PE, et al Stat5 synergizes with T cell receptor/antigen stimulation in the development of lymphoblastic lymphoma. J Exp Med. 2003;198:79–89.[Google Scholar]
39. Onishi M, Nosaka T, Misawa K, et al Identification and characterization of a constitutively active STAT5 mutant that promotes cell proliferation. Mol Cell Biol. 1998;18:3871–3879.[Google Scholar]
40. Kovanen PE, Leonard WJCytokines and immunodeficiency diseases: critical roles of the gamma(c)-dependent cytokines interleukins 2, 4, 7, 9, 15, and 21, and their signaling pathways. Immunol Rev. 2004;202:67–83.[PubMed][Google Scholar]
41. Izquierdo Pastor M, Reif K, Cantrell DThe regulation and function of p21ras during T-cell activation and growth. Immunol Today. 1995;16:159–164.[PubMed][Google Scholar]
42. Zhang L, Lorenz U, Ravichandran KSRole of Shc in T-cell development and function. Immunol Rev. 2003;191:183–195.[PubMed][Google Scholar]
43. Brazil DP, Park J, Hemmings BAPKB binding proteins: getting in on the Akt. Cell. 2002;111:293–303.[PubMed][Google Scholar]
44. Brenne AT, Baade Ro T, Waage A, Sundan A, Borset M, Hjorth-Hansen HInterleukin-21 is a growth and survival factor for human myeloma cells. Blood. 2002;99:3756–3762.[PubMed][Google Scholar]
45. Fujii H, Nakagawa Y, Schindler U, et al Activation of Stat5 by interleukin 2 requires a carboxyl-terminal region of the interleukin 2 receptor beta chain but is not essential for the proliferative signal transmission. Proc Natl Acad Sci U S A. 1995;92:5482–5486.[Google Scholar]
46. Migone TS, Lin JX, Cereseto A, et al Constitutively activated Jak-STAT pathway in T cells transformed with HTLV-I. Science. 1995;269:79–81.[PubMed][Google Scholar]
47. Fujitani Y, Hibi M, Fukada T, et al An alternative pathway for STAT activation that is mediated by the direct interaction between JAK and STAT. Oncogene. 1997;14:751–761.[PubMed][Google Scholar]
48. Imada K, Bloom ET, Nakajima H, et al Stat5b is essential for natural killer cell-mediated proliferation and cytolytic activity. J Exp Med. 1998;188:2067–2074.[Google Scholar]
49. Nakajima H, Liu XW, Wynshaw-Boris A, et al An indirect effect of Stat5a in IL-2-induced proliferation: a critical role for Stat5a in IL-2-mediated IL-2 receptor alpha chain induction. Immunity. 1997;7:691–701.[PubMed][Google Scholar]
50. Yao Z, Cui Y, Watford WT, et al Stat5a/b are essential for normal lymphoid development and differentiation. Proc Natl Acad Sci U S A. 2006;103:1000–1005.[Google Scholar]
51. Stahl N, Farruggella TJ, Boulton TG, Zhong Z, Darnell JE, Jr, Yancopoulos GDChoice of STATs and other substrates specified by modular tyrosine-based motifs in cytokine receptors. Science. 1995;267:1349–1353.[PubMed][Google Scholar]
52. Takeda K, Kaisho T, Yoshida N, Takeda J, Kishimoto T, Akira SStat3 activation is responsible for IL-6-dependent T cell proliferation through preventing apoptosis: generation and characterization of T cell-specific Stat3-deficient mice. J Immunol. 1998;161:4652–4660.[PubMed][Google Scholar]
53. Akaishi H, Takeda K, Kaisho T, et al Defective IL-2-mediated IL-2 receptor alpha chain expression in Stat3-deficient T lymphocytes. Int Immunol. 1998;10:1747–1751.[PubMed][Google Scholar]
54. Leonard W. Type I cytokines and interferons and their receptors. In: Paul W, editor. Fundamental Immunology. 5th ed. Philadelphia, PA: Lippincott Williams & Wilkins; 2003. pp. 701–747. [PubMed]
55. O'Shea JJ, Gadina M, Schreiber RDCytokine signaling in 2002: new surprises in the Jak/Stat pathway. Cell. 2002;109(suppl):S121–S131.[PubMed][Google Scholar]
56. Levy DE, Darnell JE., Jr Stats: transcriptional control and biological impact. Nat Rev Mol Cell Biol. 2002;3:651–662.[PubMed]