Sage Journals: Discover world-class research

Abstract

Interleukin (IL)-21 has been intensively studied for use in therapy of autoimmune diseases, cancers, and chronic viruses due to its immunomodulatory properties, especially on CD4⁺ and CD8⁺ T cells and natural killer (NK) cells. The objective of this study was to produce an optimized form of IL-21 with improved stability. Plasmids encoding the murine IL-21 alone (pIL-21) or IL-21 genetically fused to portions from mouse IgG3 (pIL-21/Ig) were constructed, and the efficiency of expression, protein kinetics, biodisponibility, and function were analyzed. The genetic constructions of pIL-21 and pIL-21/Ig were transfected into HEK 293 cells, and significant levels of functional IL-21 were obtained. The amino acid of murine IL-21 and IgG3 cloned showed 100% identity with correspondent published sequences. At 24 h of incubation, increased levels of IL-21 were detected in the supernatants of pIL-21. At 72 h of culture, the levels of IL-21 in the supernatant of cells transfected with pIL-21/Ig were significantly higher than those secreted by pIL-21-transfected cells. Furthermore, the data showed that our chimeric IL-21/Ig present improved systemic disponibility in BALB/c mice and conserved the intrinsic ability to increase the frequency of CD4⁺ T cells, NKT cells, and CD8⁺ T cells.

Get full access to this article

View all access options for this article.

References

Barker

B.R.

, Gladstone

M.N.

, et al. (2010). Critical role for IL-21 in both primary and memory anti-viral CD8+ T-cell responses. Eur J Immunol, 40, 3085–3096.

Barouch

D.H.

, Truitt

D.M.

, et al. (2004). Expression kinetics of the interleukin-2/immunoglobulin (IL-2/Ig) plasmid cytokine adjuvant. Vaccine, 22, 3092–3097.

Butler

M.O.

, Imataki

, et al. (2012). Ex vivo expansion of human CD8+ T cells using autologous CD4+ T cell help. PLoS One, 7, e30229.

Calame

K.L.

, Lin

K.I.

, et al. (2003). Regulatory mechanisms that determine the development and function of plasma cells. Annu Rev Immunol, 21, 205–230.

Caprioli

, Sarra

, et al. (2008). Autocrine regulation of IL-21 production in human T lymphocytes. J Immunol, 180, 1800–1807.

Carbo

, Olivares-Villagomez

, et al. (2014). Systems modeling of the role of interleukin-21 in the maintenance of effector CD4+ T cell responses during chronic Helicobacter pylori infection. mBio, 5, e01243–e01214.

Craiu

, Barouch

D.H.

, et al. (2001). An IL-2/Ig fusion protein influences CD4+ T lymphocytes in naive and simian immunodeficiency virus-infected Rhesus monkeys. AIDS Res Hum Retroviruses, 17, 873–886.

Derer

, Kellner

, et al. (2012). Fc engineering: design, expression, and functional characterization of antibody variants with improved effector function. Methods Mol Biol, 907, 519–536.

Dou

, Wang

, et al. (2009). Antitumor efficacy induced by human ovarian cancer cells secreting IL-21 alone or combination with GM-CSF cytokines in nude mice model. Immunobiology, 214, 483–492.

10.

Hamming

O.J.

, Kang

, et al. (2012). Crystal structure of interleukin-21 receptor (IL-21R) bound to IL-21 reveals that sugar chain interacting with WSXWS motif is integral part of IL-21R. J Biol Chem, 287, 9454–9460.

11.

Iannello

, Samarani

, et al. (2010). Comment on “HIV-specific IL-21 producing CD4+ T cells are induced in acute and chronic progressive HIV infection and are associated with relative viral control.”. J Immunol, 185, 5675; author reply 5675.

12.

Iannello

, Tremblay

, et al. (2008). Decreased levels of circulating IL-21 in HIV-infected AIDS patients: correlation with CD4+ T-cell counts. Viral Immunol, 21, 385–388.

13.

Khattar

, Miyahara

, et al. (2014). Interleukin-21 is a critical regulator of CD4 and CD8 T cell survival during priming under interleukin-2 deprivation conditions. PLoS One, 9, e85882.

14.

Lim

D.P.

, Jang

Y.Y.

, et al. (2014). Effect of exposure to interleukin-21 at various time points on human natural killer cell culture. Cytotherapy, 16, 1419–1430.

15.

Linterman

M.A.

, Beaton

, et al. (2010). IL-21 acts directly on B cells to regulate Bcl-6 expression and germinal center responses. J Exp Med, 207, 353–363.

16.

Liu

, Yang

, et al. (2011). Interleukin-21 maintains the expression of CD16 on monocytes via the production of IL-10 by human naive CD4+ T cells. Cell Immunol, 267, 102–108.

17.

Marcais

, Viel

, et al. (2013). Regulation of mouse NK cell development and function by cytokines. Front Immunol, 4, 450.

18.

Okuda

, Kawamoto

, et al. (2000). Cytokine and costimulatory factor-encoding plasmids as adjuvants for DNA vaccination. Methods Mol Med, 29, 197–204.

19.

Parrish-Novak

, Dillon

S.R.

, et al. (2000). Interleukin 21 and its receptor are involved in NK cell expansion and regulation of lymphocyte function. Nature, 408, 57–63.

20.

Pouw

, Treffers-Westerlaken

, et al. (2010). Combination of IL-21 and IL-15 enhances tumour-specific cytotoxicity and cytokine production of TCR-transduced primary T cells. Cancer Immunology Immunother, 59, 921–931.

21.

Publicover

, Goodsell

, et al. (2011). IL-21 is pivotal in determining age-dependent effectiveness of immune responses in a mouse model of human hepatitis B. J Clin Invest, 121, 1154–1162.

22.

Raveney

B.J.

, Oki

, et al. (2013). Nuclear receptor NR4A2 orchestrates Th17 cell-mediated autoimmune inflammation via IL-21 signalling. PLoS One, 8, e56595.

23.

Rodrigues

, Nandakumar

, et al. (2009). IL-21 and IL-15 cytokine DNA augments HSV specific effector and memory CD8+ T cell response. Mol Immunol, 46, 1494–1504.

24.

Souza

A.P.

, Bonorino

, et al. (2013). Interleukin-21 expanded NKDC in vitro reduces the B16F10 tumor growth in vivo . Cytokine, 61, 154–160.

25.

Spolski

, and Leonard

W.J.

(2009). Cytokine mediators of Th17 function. Eur J Immunol, 39, 658–661.

26.

Spolski

, and Leonard

W.J.

(2014). Interleukin-21: a double-edged sword with therapeutic potential. Nat Rev Drug Discov, 13, 379–395.

27.

Stolfi

, Rizzo

, et al. (2011). Involvement of interleukin-21 in the regulation of colitis-associated colon cancer. J Exp Med, 208, 2279–2290.

28.

Sutherland

A.P.

, Joller

, et al. (2013). IL-21 promotes CD8+ CTL activity via the transcription factor T-bet. J Immunol, 190, 3977–3984.

29.

Terrier

, Costedoat-Chalumeau

, et al. (2012). Interleukin 21 correlates with T cell and B cell subset alterations in systemic lupus erythematosus. J Rheumatol, 39, 1819–1828.

30.

Vivanco-Cid

, Maldonado-Renteria

M.J.

, et al. (2014). Dynamics of interleukin-21 production during the clinical course of primary and secondary dengue virus infections. Immunol Lett, 161, 89–95.

31.

Williams

, Rafei

, et al. (2010). A fusion of GMCSF and IL-21 initiates hypersignaling through the IL-21Ralpha chain with immune activating and tumoricidal effects in vivo . Mol Ther, 18, 1293–1301.

32.

J.S.

, Ingram

J.T.

, et al. (2010). IL-21 deficiency influences CD8 T cell quality and recall responses following an acute viral infection. J Immunol, 185, 4835–4845.

33.

Zhao

, Dou

, et al. (2010). Enhancing therapy of B16F10 melanoma efficacy through tumor vaccine expressing GPI-anchored IL-21 and secreting GM-CSF in mouse model. Vaccine, 28, 2846–2852.

Supplementary Material

Please find the following supplemental material available below.

For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.

For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.

0.00 MB

0.05 MB

Construction and Functional Characterization of a Fusion Protein Interleukin-21/Immunoglobulin for Long-Term In Vivo Biodisponibility

Abstract

Get full access to this article

References

Supplementary Material