Stability and inhibitory function of Treg cells under inflammatory conditions in vitro

Guo,Huifang; Xun,Liru; Zhang,Ruisan; Hu,Fengrui; Luan,Jing; Lao,Kejing; Wang,Xiaolong; Gou,Xingchun

doi:10.3892/etm.2019.7873

Stability and inhibitory function of Treg cells under inflammatory conditions in vitro

Authors:
- Huifang Guo
- Liru Xun
- Ruisan Zhang
- Fengrui Hu
- Jing Luan
- Kejing Lao
- Xiaolong Wang
- Xingchun Gou
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Affiliations: Shaanxi Key Laboratory of Brain Disorders and School of Basic Medical Science, Xi'an Medical University, Xi'an, Shaanxi 710021, P.R. China, Nephrology Department of Shaanxi Provincial People's Hospital Affiliated to Xi'an Medical University, Xi'an, Shaanxi 710068, P.R. China
Published online on: August 8, 2019 https://doi.org/10.3892/etm.2019.7873
Pages: 2443-2450
Copyright: © Guo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Immunotherapy with transplanted T‑regulatory (Treg) cells is currently in use. However, patients have complex internal environments with confounding factors, including the presence of inflammatory cytokines. The present study aimed to detect Treg cell function under simulated inflammatory conditions to provide a foundation for Treg cell‑based immunotherapy. CD4+CD25high Treg cells were sorted from peripheral blood mononuclear cells and cultured for 14 days in the presence of recombinant human interleukin‑2 (rhIL‑2) and anti‑CD3/CD28 beads, with or without 25 ng/ml rhIL‑6. Next, the absolute count of Treg cells was determined, the stability and activity were detected by measuring the expression levels of forkhead box (Fox)P3 and CD39, and the suppressive function of Treg cells was investigated by assessing the suppression of T‑effector cell proliferation by Treg cells after co‑culture for 5 days. The number of Treg cells cultured in the presence of 25 ng/ml rhIL‑6 for 14 days was reduced by 49.7% when compared with that of cells cultured without rhIL‑6. Of the Treg cells continually cultured for 14 days without or with 25 ng/ml rhIL‑6, 56.15 and 24.7% expressed FoxP3, respectively. There was no difference in the activity of the FoxP3+ Treg cells after culture for 14 days without or with 25 ng/ml rhIL‑6. The suppressive function of Treg cells tended to deteriorate in the presence of rhIL‑6. In conclusion, IL‑6 inhibited the proliferation and stability of Treg cells, suggesting that administration of increased numbers of Treg cells may be required during Treg cell‑based immunotherapy.

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1	Togashi Y and Nishikawa H: Regulatory T cells: Molecular and cellular basis for immunoregulation. Curr Top Microbiol Immunol. 410:3–27. 2017.PubMed/NCBI
2	Vignali DA, Collison LW and Workman CJ: How regulatory T cells work. Nat Rev Immunol. 8:523–532. 2008. View Article : Google Scholar : PubMed/NCBI
3	Collison LW, Chaturvedi V, Henderson AL, Giacomin PR, Guy C, Bankoti J, Finkelstein D, Forbes K, Workman CJ, Brown SA, et al: IL-35-mediated induction of a potent regulatory T cell population. Nat Immunol. 11:1093–1101. 2010. View Article : Google Scholar : PubMed/NCBI
4	Guo H, Xun L, Zhang R and Gou X: Ratio of CD147^high/CD147^low in CD4⁺CD25⁺ T cells: A potential biomarker for early diagnosis and prediction of response to therapy for autoimmune diseases. Med Hypotheses. 115:1–4. 2018. View Article : Google Scholar : PubMed/NCBI
5	Talaat RM, Mohamed SF, Bassyouni IH and Raouf AA: Th1/Th2/Th17/Treg cytokine imbalance in systemic lupus erythematosus (SLE) patients: Correlation with disease activity. Cytokine. 72:146–153. 2015. View Article : Google Scholar : PubMed/NCBI
6	Bluestone JA, Buckner JH, Fitch M, Gitelman SE, Gupta S, Hellerstein MK, Herold KC, Lares A, Lee MR, Li K, et al: Type 1 diabetes immunotherapy using polyclonal regulatory T cells. Sci Transl Med. 7:315ra1892015. View Article : Google Scholar : PubMed/NCBI
7	Ou HX, Guo BB, Liu Q, Li YK, Yang Z, Feng WJ and Mo ZC: Regulatory T cells as a new therapeutic target for atherosclerosis. Acta Pharmacol Sin. 39:1249–1258. 2018. View Article : Google Scholar : PubMed/NCBI
8	Martin-Moreno PL, Tripathi S and Chandraker A: Regulatory T cells and kidney transplantation. Clin J Am Soc Nephrol. 13:1760–1764. 2018. View Article : Google Scholar : PubMed/NCBI
9	Qiu R, Zhou L, Ma Y, Zhou L, Liang T, Shi L, Long J and Yuan D: Regulatory T cell plasticity and stability and autoimmune diseases. Clin Rev Allergy Immunol. Nov 17–2018.(Epub ahead of print). View Article : Google Scholar : PubMed/NCBI
10	Alvarez C, Rojas C, Rojas L, Cafferata EA, Monasterio G and Vernal R: Regulatory T lymphocytes in periodontitis: A translational view. Mediators Inflamm 2018. 78069122018.
11	Sakaguchi S, Sakaguchi N, Asano M, Itoh M and Toda M: Pillars article: Immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor α-chains (CD25). Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases. J. Immunol. 1995. J Immunol. 186:3808–3821. 2011.PubMed/NCBI
12	Jeffery HC, Braitch MK, Brown S and Oo YH: Clinical potential of regulatory T cell therapy in liver diseases: An overview and current perspectives. Front Immunol. 7:3342016. View Article : Google Scholar : PubMed/NCBI
13	Lam AJ, Hoeppli RE and Levings MK: Harnessing advances in T regulatory cell biology for cellular therapy in transplantation. Transplantation. 101:2277–2287. 2017. View Article : Google Scholar : PubMed/NCBI
14	Ait-Oufella H, Taleb S, Mallat Z and Tedgui A: Recent advances on the role of cytokines in atherosclerosis. Arterioscler Thromb Vasc Biol. 31:969–979. 2011. View Article : Google Scholar : PubMed/NCBI
15	Fatkhullina AR, Peshkova IO and Koltsova EK: The role of cytokines in the development of atherosclerosis. Biochemistry (Mosc). 81:1358–1370. 2016. View Article : Google Scholar : PubMed/NCBI
16	Badal D, Kumar R, Paul M, Dayal D, Bhansali A, Bhadada SK, Kumar R and Sachdeva N: Peripheral blood mononuclear cells of patients with latent autoimmune diabetes secrete higher levels of pro- & anti-inflammatory cytokines compared to those with type-1 diabetes mellitus following in vitro stimulation with β-cell autoantigens. Indian J Med Res. 145:767–776. 2017. View Article : Google Scholar : PubMed/NCBI
17	Yao Y, Wang JB, Xin MM, Li H, Liu B, Wang LL, Wang LQ and Zhao L: Balance between inflammatory and regulatory cytokines in systemic lupus erythematosus. Genet Mol Res. 15:2016. View Article : Google Scholar
18	He X, Smeets RL, van Rijssen E, Boots AM, Joosten I and Koenen HJ: Single CD28 stimulation induces stable and polyclonal expansion of human regulatory T cells. Sci Rep. 7:430032017. View Article : Google Scholar : PubMed/NCBI
19	Fan MY, Low JS, Tanimine N, Finn KK, Priyadharshini B, Germana SK, Kaech SM and Turka LA: Differential roles of IL-2 signaling in developing versus mature tregs. Cell Rep. 25:1204–1213.e4. 2018. View Article : Google Scholar : PubMed/NCBI
20	Baecher-Allan C, Brown JA, Freeman GJ and Hafler DA: CD4+CD25high regulatory cells in human peripheral blood. J Immunol. 167:1245–1253. 2001. View Article : Google Scholar : PubMed/NCBI
21	Rubtsov YP, Niec RE, Josefowicz S, Li L, Darce J, Mathis D, Benoist C and Rudensky AY: Stability of the regulatory T cell lineage in vivo. Science. 329:1667–1671. 2010. View Article : Google Scholar : PubMed/NCBI
22	de la Rosa M, Rutz S, Dorninger H and Scheffold A: Interleukin-2 is essential for CD4+CD25+ regulatory T cell function. Eur J Immunol. 34:2480–2488. 2004. View Article : Google Scholar : PubMed/NCBI
23	Levine AG, Arvey A, Jin W and Rudensky AY: Continuous requirement for the TCR in regulatory T cell function. Nat Immunol. 15:1070–1078. 2014. View Article : Google Scholar : PubMed/NCBI
24	Guo H, Zheng M, Zhang K, Yang F, Zhang X, Han Q, Chen ZN and Zhu P: Functional defects in CD4+ CD25high FoxP3+ regulatory cells in ankylosing spondylitis. Sci Rep. 6:375592016. View Article : Google Scholar : PubMed/NCBI
25	Wells AD, Gudmundsdottir H and Turka LA: Following the fate of individual T cells throughout activation and clonal expansion. Signals from T cell receptor and CD28 differentially regulate the induction and duration of a proliferative response. J Clin Invest. 100:3173–3183. 1997. View Article : Google Scholar : PubMed/NCBI
26	Gudmundsdottir H, Wells AD and Turka LA: Dynamics and requirements of T cell clonal expansion in vivo at the single-cell level: Effector function is linked to proliferative capacity. J Immunol. 162:5212–5223. 1999.PubMed/NCBI
27	Borsellino G, Kleinewietfeld M, Di Mitri D, Sternjak A, Diamantini A, Giometto R, Höpner S, Centonze D, Bernardi G, Dell'Acqua ML, et al: Expression of ectonucleotidase CD39 by Foxp3+ Treg cells: Hydrolysis of extracellular ATP and immune suppression. Blood. 110:1225–1232. 2007. View Article : Google Scholar : PubMed/NCBI
28	Fontenot JD, Rasmussen JP, Gavin MA and Rudensky AY: A function for interleukin 2 in Foxp3-expressing regulatory T cells. Nat Immunol. 6:1142–1151. 2005. View Article : Google Scholar : PubMed/NCBI
29	Li X and Zheng Y: Regulatory T cell identity: Formation and maintenance. Trends Immunol. 36:344–353. 2015. View Article : Google Scholar : PubMed/NCBI
30	Passerini L and Bacchetta R: Forkhead-box-P3 gene transfer in human CD4(+) T conventional cells for the generation of stable and efficient regulatory T cells, suitable for immune modulatory therapy. Front Immunol. 8:12822017. View Article : Google Scholar : PubMed/NCBI
31	Ezzelarab MB: Regulatory T cells from allo- to xenotransplantation: Opportunities and challenges. Xenotransplantation. 25:e124152018. View Article : Google Scholar : PubMed/NCBI
32	Min B: Heterogeneity and stability in Foxp3+ regulatory T cells. J Interferon Cytokine Res. 37:386–397. 2017. View Article : Google Scholar : PubMed/NCBI
33	Tanaka T, Narazaki M and Kishimoto T: IL-6 in inflammation, immunity, and disease. Cold Spring Harb Perspect Biol. 6:a0162952014. View Article : Google Scholar : PubMed/NCBI
34	Wiesinger M, Stoica D, Roessner S, Lorenz C, Fischer A, Atreya R, Neufert CF, Atreya I, Scheffold A, Schuler-Thurner B, et al: Good manufacturing practice-compliant production and lot-release of ex vivo expanded regulatory T cells as basis for treatment of patients with autoimmune and inflammatory disorders. Front Immunol. 8:13712017. View Article : Google Scholar : PubMed/NCBI
35	Fletcher JM, Lonergan R, Costelloe L, Kinsella K, Moran B, O'Farrelly C, Tubridy N and Mills KH: CD39+Foxp3+ regulatory T cells suppress pathogenic Th17 cells and are impaired in multiple sclerosis. J Immunol. 183:7602–7610. 2009. View Article : Google Scholar : PubMed/NCBI
36	Gu J, Ni X, Pan X, Lu H, Lu Y, Zhao J, Guo Zheng S, Hippen KL, Wang X and Lu L: Human CD39(hi) regulatory T cells present stronger stability and function under inflammatory conditions. Cell Mol Immunol. 14:521–528. 2017. View Article : Google Scholar : PubMed/NCBI