Advanced Glycated End Products Alter Neutrophil Effect on Regulation of CD4+ T Cell Differentiation Through Induction of Myeloperoxidase and Neutrophil Elastase Activities
CD4+ T cell subset imbalance plays an important role in the development of diabetic complications. Neutrophils have recently been known as the regulator of CD4+ T cell differentiation. However, whether neutrophils affect CD4+ T cell population in diabetes is still elusive. In this study, we investigated the effect of neutrophils stimulated with advanced glycated end products (AGEs), the marker of diabetes, on CD4+ T cell differentiation and its underlying mechanism. Our data showed that the cultural medium of healthy adult neutrophils treated with AGEs increased expressions of both Th1 (IFN-γ) and Th17 (IL-17) phenotypes and the transcription factors of Th1 (Tbet) and Th17 (RORγt) in naive CD4+T cells and CD4+CD25+FoxP3+ (Treg) T cells in vitro. Next, we found that AGEs induced the generations of myeloperoxidase (MPO) and neutrophil elastase (NE) in neutrophils; inhibition of MPO or NE attenuated the effect of AGE-stimulated neutrophils on CD4+ T cell bias. Furthermore, receptor for AGEs (RAGE) inhibitor interrupted AGE-induced MPO and NE expressions, but MPO and NE inhibitions did not change AGE-increased RAGE gene expression. These results suggested that AGEs drive the effect of neutrophils on CD4+ T cell differentiation into pro-inflammatory program through inducing MPO and NE productions in neutrophils, which is mediated by AGE–RAGE interaction.
KEY WORDSadvanced glycated end products neutrophils CD4+ T cell subsets myeloperoxidase neutrophil elastase
Advanced glycated end products
Receptor for AGEs
Regulatory T cells
Neutrophil extracellular traps
Retinoic orphan receptor
Bovine serum albumin
Intracellular cytokine staining
We are grateful for the participants who volunteered for this study and the physicians and nurses in the Department of Neurology for their assistances during the execution of the work.
Role of Funding Source
The study was supported by Talent Initial funding YY2017-002 sponsored by Guangdong Second Provincial General Hospital, Grant 201707010436, from Guangzhou city scientific and technological research program and research program, Grant 20173024, sponsored by Chinese Traditional Medicine Bureau of Guangdong Province.
Compliance with Ethical Standards
This study was approved by the Ethical Review Committee of Guangdong Second Provincial General Hospital. Informed consent of blood donation was given by healthy adult volunteers who had no record of food allergy or sensitization.
Conflict of Interest
The authors declare that they have no conflict of interest.
- 4.Soboleva, A., R. Schmidt, M. Vikhnina, T. Grishina, and A. Frolov. 2017. Maillard proteomics: opening new pages. International Journal of Molecular Sciences 18.Google Scholar
- 9.Drew, W., D.V. Wilson, and E. Sapey. 2017. Inflammation and neutrophil immunosenescence in health and disease: targeted treatments to improve clinical outcomes in the elderly. Experimental Gerontology.Google Scholar
- 15.Thomson, J.T., M.B. Brand, and P. FruFonteh. 2017. The role of IL17-A in the second hit of acute pancreatitis. South African Journal of Surgery 55: 51.Google Scholar
- 16.Balato, A., E. Scala, N. Balato, G. Caiazzo, R. Di Caprio, G. Monfrecola, A. Raimondo, S. Lembo, and F. Ayala. 2017. Biologics that inhibit the Th17 pathway and related cytokines to treat inflammatory disorders. Expert Opinion on Biological Therapy 17: 1363–1374.Google Scholar
- 17.Kitz, A., E. Singer, and D. Hafler. 2018. Regulatory T cells: from discovery to autoimmunity. Cold Spring Harbor Perspectives in Medicine.Google Scholar
- 18.Salehipour, Z., D. Haghmorad, M. Sankian, M. Rastin, R. Nosratabadi, D.M. Soltan, N. Tabasi, M. Khazaee, L.R. Nasiraii, and M. Mahmoudi. 2017. Bifidobacterium animalis in combination with human origin of Lactobacillus plantarum ameliorate neuroinflammation in experimental model of multiple sclerosis by altering CD4+ T cell subset balance. Biomedicine & Pharmacotherapy 95: 1535–1548.CrossRefGoogle Scholar
- 22.Prame, K.K., A.J. Nicholls, and C. Wong. 2018. Partners in crime: neutrophils and monocytes/macrophages in inflammation and disease. Cell and Tissue Research.Google Scholar
- 24.Perobelli, S.M., A.C. Mercadante, R.G. Galvani, T. Goncalves-Silva, A.P. Alves, A. Pereira-Neves, M. Benchimol, A. Nobrega, and A. Bonomo. 2016. G-CSF-induced suppressor IL-10+ neutrophils promote regulatory T cells that inhibit graft-versus-host disease in a long-lasting and specific way. Journal of Immunology 197: 3725–3734.CrossRefGoogle Scholar
- 26.Domingo-Gonzalez, R., G.J. Martinez-Colon, A.J. Smith, C.K. Smith, M.N. Ballinger, M. Xia, S. Murray, M.J. Kaplan, G.A. Yanik, and B.B. Moore. 2016. Inhibition of neutrophil extracellular trap formation after stem cell transplant by prostaglandin E2. American Journal of Respiratory and Critical Care Medicine 193: 186–197.CrossRefGoogle Scholar
- 27.Aarts, C., and T.W. Kuijpers. 2018. Neutrophils as myeloid-derived suppressor cells. European Journal of Clinical Investigation: e12989.Google Scholar
- 29.Miyoshi, A., M. Yamada, H. Shida, D. Nakazawa, Y. Kusunoki, A. Nakamura, H. Miyoshi, U. Tomaru, T. Atsumi, and A. Ishizu. 2016. Circulating neutrophil extracellular trap levels in well-controlled type 2 diabetes and pathway involved in their formation induced by high-dose glucose. Pathobiology 83: 243–251.CrossRefGoogle Scholar
- 32.Lin, J., S. Haridas, S.J. Barenkamp, L.C. Lorenset, A. Lee, B.T. Schroeder, G. Peng, and J.M. Koenig. 2017. Neonatal neutrophils stimulated by group B Streptococcus induce a proinflammatory T-helper cell bias. Pediatric Research.Google Scholar
- 34.Najmeh, S., J. Cools-Lartigue, B. Giannias, J. Spicer, and L.E. Ferri. 2015. Simplified human neutrophil extracellular traps (NETs) isolation and handling. Journal of Visualized Experiments.Google Scholar
- 36.Machado-Lima, A., R.T. Iborra, R.S. Pinto, C.H. Sartori, E.R. Oliveira, E.R. Nakandakare, J.T. Stefano, D. Giannella-Neto, M.L. Correa-Giannella, and M. Passarelli. 2013. Advanced glycated albumin isolated from poorly controlled type 1 diabetes mellitus patients alters macrophage gene expression impairing ABCA-1-mediated reverse cholesterol transport. Diabetes/Metabolism Research and Reviews 29: 66–76.CrossRefGoogle Scholar
- 38.Vlassara, H., J. Uribarri, W. Cai, S. Goodman, R. Pyzik, J. Post, F. Grosjean, M. Woodward, and G.E. Striker. 2012. Effects of sevelamer on HbA1c, inflammation, and advanced glycation end products in diabetic kidney disease. Clinical Journal of the American Society of Nephrology 7: 934–942.CrossRefGoogle Scholar
- 43.Scully, I.L., L.K. McNeil, S. Pathirana, C.L. Singer, Y. Liu, S. Mullen, D. Girgenti, A. Gurtman, M.W. Pride, K.U. Jansen, et al. 2017. Neutrophil killing of Staphylococcus aureus in diabetes, obesity and metabolic syndrome: a prospective cellular surveillance study. Diabetology and Metabolic Syndrome 9: 76.CrossRefGoogle Scholar
- 46.DeLeon-Pennell, K.Y., Y. Tian, B. Zhang, C.A. Cates, R.P. Iyer, P. Cannon, P. Shah, P. Aiyetan, G.V. Halade, Y. Ma, et al. 2016. CD36 is a matrix metalloproteinase-9 substrate that stimulates neutrophil apoptosis and removal during cardiac remodeling. Circulation. Cardiovascular Genetics 9: 14–25.CrossRefGoogle Scholar