GTS‑21, an α7‑nicotinic acetylcholine receptor agonist, modulates Th1 differentiation in CD4+ T cells from patients with rheumatoid arthritis

Wu,Shiyao; Zhao,Hongjun; Luo,Hui; Xiao,Xianzhong; Zhang,Huali; Li,Tong; Zuo,Xiaoxia

doi:10.3892/etm.2014.1754

GTS‑21, an α7‑nicotinic acetylcholine receptor agonist, modulates Th1 differentiation in CD4+ T cells from patients with rheumatoid arthritis

Authors:
- Shiyao Wu
- Hongjun Zhao
- Hui Luo
- Xianzhong Xiao
- Huali Zhang
- Tong Li
- Xiaoxia Zuo
View Affiliations

Affiliations: Department of Rheumatology, Xiangya Hospital, Central South University, Changsha, Hunan 410008, P.R. China, Department of Pathophysiology, Xiangya School of Medicine, Central South University, Changsha, Hunan 410008, P.R. China
Published online on: June 3, 2014 https://doi.org/10.3892/etm.2014.1754
Pages: 557-562

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

GTS‑21 (also known as DMBX‑anabaseine), a selective α7 nicotinic acetylcholine receptor (α7nAChR) agonist, has previously been found to inhibit the inflammation associated with rheumatoid arthritis (RA). RA is an autoimmune disease, where an abnormal immune system plays a critical role in the occurrence and development of synovium inflammation and bone damage. However, prior to this study, the immunological mechanism by which GTS‑21 protects against RA had not been elucidated. In the present study, the effects of GTS‑21 on T helper 1 (Th1) cells, which have an important role in the inflammation associated with RA, were investigated. Peripheral blood mononuclear cells (PBMCs) and cluster of differentiation (CD)4+ T cells were separated from patients with RA, and the effects of GTS‑21 on PBMCs stimulated with anti‑CD3/‑CD28 antibodies and CD4+ T cells were investigated in the context of Th1‑cell differentiation. ELISA was used to analyze interferon (IFN)‑γ expression and flow cytometric analysis was used to detect the percentage of IFN‑γ+ CD3+CD8‑ T cells. In addition, western blotting was employed to detect the levels of the T‑box transcription factor TBX21, which is a Th1 cell‑specific transcription factor. The present study showed that GTS‑21 reduced IFN‑γ production in PBMCs from patients with RA. Under conditions of Th1‑cell differentiation, GTS‑21 reduced the percentage of IFNγ+CD3+CD8‑ T cells and IFN‑γ production in the culture supernatant and also inhibited the expression of the Th1 cell‑specific transcription factor TBX21. The effects of GTS‑21 were blocked by the α7nAchR antagonist α‑bungarotoxin, which increased the expression of IFN‑γ and TBX21. This study demonstrated that GTS‑21 is able to inhibit RA Th1‑cell differentiation through activation of the α7nAchR.

View Figures

View References

1	Cope AP, Schulze-Koops H and Aringer M: The central role of T cells in rheumatoid arthritis. Clin Exp Rheumatol. 25(5 Suppl 46): S4–S11. 2007.PubMed/NCBI
2	Yamada H, Nakashima Y, Okazaki K, et al: Th1 but not Th17 cells predominate in the joints of patients with rheumatoid arthritis. Ann Rheum Dis. 67:1299–1304. 2008. View Article : Google Scholar : PubMed/NCBI
3	Cañete JD, Martinez SE, Farrés J, et al: Differential Th1/Th2 cytokine patterns in chronic arthritis: interferon gamma is highly expressed in synovium of rheumatoid arthritis compared with seronegative spondyloarthropathies. Ann Rheum Dis. 59:263–268. 2000.
4	Nissinen R, Leirisalo-Repo M, Tiittanen M, et al: CCR3, CCR5, interleukin 4, and interferon-gamma expression on synovial and peripheral T cells and monocytes in patients with rheumatoid arthritis. J Rheumatol. 30:1928–1934. 2003.PubMed/NCBI
5	Szabo SJ, Sullivan BM, Stemmann C, Satoskar AR, Sleckman BP and Glimcher LH: Distinct effects of T-bet in TH1 lineage commitment and IFN-gamma production in CD4 and CD8 T cells. Science. 295:338–342. 2002. View Article : Google Scholar : PubMed/NCBI
6	Yang DD, Conze D, Whitmarsh AJ, et al: Differentiation of CD4⁺ T cells to Th1 cells requires MAP kinase JNK2. Immunity. 9:575–585. 1998.
7	Pavlov VA, Ochani M, Yang LH, et al: Selective alpha7-nicotinic acetylcholine receptor agonist GTS-21 improves survival in murine endotoxemia and severe sepsis. Crit Care Med. 35:1139–1144. 2007. View Article : Google Scholar : PubMed/NCBI
8	Wang H, Liao H, Ochani M, et al: Cholinergic agonists inhibit HMGB1 release and improve survival in experimental sepsis. Nat Med. 10:1216–1221. 2004. View Article : Google Scholar : PubMed/NCBI
9	Wang H, Yu M, Ochani M, et al: Nicotinic acetylcholine receptor alpha7 subunit is an essential regulator of inflammation. Nature. 421:384–388. 2003. View Article : Google Scholar : PubMed/NCBI
10	Bruchfeld A, Goldstein RS, Chavan S, et al: Whole blood cytokine attenuation by cholinergic agonists ex vivo and relationship to vagus nerve activity in rheumatoid arthritis. J Intern Med. 268:94–101. 2010.PubMed/NCBI
11	de Jonge WJ and Ulloa L: The alpha7 nicotinic acetylcholine receptor as a pharmacological target for inflammation. Br J Pharmacol. 151:915–929. 2007.PubMed/NCBI
12	Kitagawa H, Takenouchi T, Azuma R, et al: Safety, pharmacokinetics, and effects on cognitive function of multiple doses of GTS-21 in healthy, male volunteers. Neuropsychopharmacol. 28:542–551. 2003. View Article : Google Scholar : PubMed/NCBI
13	Olincy A, Harris JG, Johnson LL, et al: Proof-of-concept trial of an alpha7 nicotinic agonist in schizophrenia. Arch Gen Psychiatry. 63:630–638. 2006. View Article : Google Scholar : PubMed/NCBI
14	Conti-Fine BM, Navaneetham D, Lei S and Maus AD: Neuronal nicotinic receptors in non-neuronal cells: new mediators of tobacco toxicity? Eur J Pharmacol. 393:279–294. 2000. View Article : Google Scholar : PubMed/NCBI
15	Ahmed SA, Gogal RJ Jr and Walsh JE: A new rapid and simple non-radioactive assay to monitor and determine the proliferation of lymphocytes: an alternative to [3H]thymidine incorporation assay. J Immunol Methods. 170:211–224. 1994.PubMed/NCBI
16	O’Brien J, Wilson I, Orton T and Pognan F: Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem. 267:5421–5426. 2000.PubMed/NCBI
17	Petersen CM, Christensen EI, Andresen BS and Møller BK: Internalization, lysosomal degradation and new synthesis of surface membrane CD4 in phorbol ester-activated T-lymphocytes and U-937 cells. Exp Cell Res. 201:160–173. 1992. View Article : Google Scholar : PubMed/NCBI
18	Mascher B, Schlenke P and Seyfarth M: Expression and kinetics of cytokines determined by intracellular staining using flow cytometry. J Immunol Methods. 223:115–121. 1999. View Article : Google Scholar : PubMed/NCBI
19	Kox M, van Velzen JF, Pompe JC, Hoedemaekers CW, van der Hoeven JG and Pickkers P: GTS-21 inhibits pro-inflammatory cytokine release independent of the Toll-like receptor stimulated via a transcriptional mechanism involving JAK2 activation. Biochem Pharmacol. 78:863–872. 2009. View Article : Google Scholar : PubMed/NCBI
20	Nizri E, Hamra-Amitay Y, Sicsic C, Lavon I and Brenner T: Anti-inflammatory properties of cholinergic up-regulation: A new role for acetylcholinesterase inhibitors. Neuropharmacology. 50:540–547. 2006. View Article : Google Scholar : PubMed/NCBI
21	Sato KZ, Fujii T, Watanabe Y, et al: Diversity of mRNA expression for muscarinic acetylcholine receptor subtypes and neuronal nicotinic acetylcholine receptor subunits in human mononuclear leukocytes and leukemic cell lines. Neurosci Lett. 266:17–20. 1999. View Article : Google Scholar
22	McGrath J, McDonald JW and Macdonald JK: Transdermal nicotine for induction of remission in ulcerative colitis. Cochrane Database Syst Rev. 18:CD0047222004.PubMed/NCBI
23	van Westerloo DJ, Giebelen IA, Florquin S, et al: The cholinergic anti-inflammatory pathway regulates the host response during septic peritonitis. J Infect Dis. 191:2138–2148. 2005.PubMed/NCBI
24	van Maanen MA, Lebre MC, van der Poll T, et al: Stimulation of nicotinic acetylcholine receptors attenuates collagen-induced arthritis in mice. Arthritis Rheum. 60:114–122. 2009.PubMed/NCBI
25	Li T, Zuo X, Zhou Y, et al: The vagus nerve and nicotinic receptors involve inhibition of HMGB1 release and early pro-inflammatory cytokines function in collagen-induced arthritis. J Clin Immunol. 30:213–220. 2010. View Article : Google Scholar : PubMed/NCBI