Detection of Th17/Treg cells and related factors in gingival tissues and peripheral blood of rats with experimental periodontitis

Document Type: Original Article


1 Department of Periodontology, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat -sen University, Guangzhou 510055,China

2 Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China


Objective(s): This study aimed to investigate the role and the possible mechanisms involved in the immunoregulation of experimental periodontitis by Th17/Treg.
Materials and Methods: Experimental periodontitis was established by silk thread ligation with Porphyromonasgingivalis daubing in the bilateral maxillary second molar of Male Sprague-Dawley (SD) rats. Alveolar bones were scanned by Micro-CT. Histological examination was stained with H&E. The proportions of Th17 and Treg cells in peripheral blood were detected by flow cytometry. RT-PCR was used to measure the expression of RORγt, Foxp3 mRNA in the gingival tissues. The concentrations of IL-17, IL-10, and TGF-β in peripheral blood and gingival crevicular fluid were measured by ELISA.
Results: Experimental rats showed profound bone resorption and inflammatory cell infiltration. The percentages of Th17 significantly increased in the peripheral blood, which was consistent with gingival tissues study that Th17 cells related transcription factor RORγt mRNA and IL-17 increased in the course of periodontitis. The percentages of CD25+Foxp3+ Treg significantly increased in the peripheral blood, which was consistent with gingival tissues study that Treg cells related transcription factor Foxp3 mRNA and cytokines IL-10 and TGF-β increased in the course of periodontitis. The ratio of Th17/Treg cells was significantly increased in the peripheral circulation, however, the Th17/Treg balance is in wave motion in inflamed gingival tissues in the different stages of periodontitis.
Conclusion: Th17/Treg balance may be associated with the progression of periodontitis and pathological tissue destruction. Moreover, local inflammation would result in the up-regulation ratio of Th17/Treg in peripheral blood, which may influence some periodontally involved systemic diseases.


1. Lavu V, Venkatesan V, Rao SR.The epigenetic paradigm in periodontitis pathogenesis. J Indian Soc Periodontol 2015;19:142-149.

2. Hajishengallis G. Periodontitis: from microbial immune subversion to systemic inflammation. Nat Rev Immunol 2014;15:30-44.

3. Llambés F, Arias-Herrera S, Caffesse R. Relationship between diabetes and periodontal infection.World J Diabetes 2015;6:927-935.

4. Kholy KE, Genco RJ, Van Dyke TE. Oral infections and cardiovascular disease. Trends Endocrinol Metab 2015; 26:315-321.

5. Hussain M, Stover CM, Dupont A. P.gingivalis in Periodontal Disease and Atherosclerosis-Scenes of Action for Antimicrobial Peptides and Complement. Front Immunol 2015; 6: 45.

6. Hernández M, Dutzan N, García-Sesnich J, Abusleme L, Dezerega A, Silva N, et al. Host-pathogen interactions in progressive chronic periodontitis. J Dent Res 2011; 90:1164-1170.

7. Teng YT. The role of acquired immunity and periodontal disease progression. Crit Rev Oral Biol Med 2003;14:237-252.

8. Gemmell E, Yamazaki K, Seymour GJ. The role of T cells in periodontal disease: homeostasis and autoimmunity. Periodontol 2007;43:14-40.

9. Graves D. Cytokines that promote periodontal tissue destruction. J Periodontol 2008; 79:1585-1591.

10. Peters A, Lee Y, Kuchroo VK. The many faces of Th17 cells. Curr Opin Immunol 2011; 23:702-706.

11. Isailovic N, Daigo K, Mantovani A, Selmi C. Interleukin-17 and innate immunity in infections and chronic inflammation. J Autoimmun 2015; 60:1-11.

12. Miossec P, Kolls JK. Targeting IL-17 and TH17 cells in chronic inflammation. Nat Rev Drug Discov 2012; 11:763-776.

13. Hienz SA, Paliwal S, Ivanovski S. Mechanisms of Bone Resorption in Periodontitis. J Immunol Res 2015; 2015:615486.

14. Du Y, Chen X, Huang ZM, Ye XH, Niu Q. Increased frequency of Foxp3+ regulatory T cells in mice with hepatocellular carcinoma. Asian Pac J Cancer Prev 2013;13:3815-3819.

15. Haque M, Fino K, Lei F, Xiong X, Song J. Utilizing regulatory T cells against rheumatoid arthritis. Front Oncol 2014; 4:209.

16. Sakaguchi S. Naturally arising CD4+ regulatory T-cells for immunologic self-tolerance and negative control of immune responses. Annu Rev Immunol 2004; 22:531-562.

17. Garlet GP, Cardoso CR, Mariano FS, Claudino M, de Assis GF, Campanelli AP, et al. Regulatory T cells attenuate experimental periodontitis progression in mice. J Clin Periodontol 2010; 37:591-600.

18. Lee SY, Lee SH, Yang EJ, Kim EK, Kim JK, Shin DY, Cho ML. Metformin Ameliorates Inflammatory Bowel Disease by Suppression of the STAT3 Signaling Pathway and Regulation of the between Th17/Treg Balance. PLoS One 2015;10: e0135858.

19. Wei Y, Luo QL, Sun J, Chen MX, Liu F, Dong JC. Bu-Shen-Yi-Qi formulae suppress chronic airway inflammation and regulate Th17/Treg imbalance in the murine ovalbumin asthma model. J Ethnopharmacol 2015;164:368-377.

20. Duan MC, Zhong XN, Liu GN, Wei JR. The Treg/Th17 paradigm in lung cancer. J Immunol Res 2014; 2014:730380.

21. Li XS, Li S, Kellermann G. An integrated liquid chromatography-tandem mass spectrometry approach for the ultra-sensitive determination of catecholamines in human peripheral blood mononuclear cells to assess neural-immune communication.J Chromatogr A 2016;1449:54-61.

22. Bleda S,de Haro J,Varela C,Ferruelo A,Acin F. Elevated levels of triglycerides and vldl-cholesterol provoke activation of nlrp1 inflammasome in endothelial cells. Int J Cardiol 2016;220:52-55.

23. Imai K, Victoriano AF, Ochiai K, Okamoto T. Microbial interaction of periodontopathic bacterium Porphyromonas gingivalis and HIV-possible causal link of periodontal diseases to AIDS progression. Curr HIV Res 2012;10:238-244.

24. Norowski PA Jr, Bumgardner JD. Biomaterial and antibiotic strategies for peri-implantitis: a review. J Biomed Mater Res B Appl Biomater 2009; 88:530-543.

25. Wang L, Wang J, Jin Y, Gao H, Lin X. Oral administration of all-trans retinoic acid suppresses experimental periodontitis by modulating the Th17/Treg imbalance. J Periodontol 2014; 85:740-750.

26. Sato K, Suematsu A, Okamoto K, Yamaguchi A, Morishita Y, Kadono Y, et al. Th17 functions as an osteoclastogenic helper T cell subset that links T cell activation and bone destruction. J Exp Med 2006; 203:2673-2682.

27. Vernal R, Dutzan N, Chaparro A, Puente J, Antonieta Valenzuela M, Gamonal J. Levels of
interleukin-17 in gingival crevicular fluid and in supernatants of cellular cultures of gingival tissue from patients with chronic periodontitis. J Clin Periodontol 2005; 32:383-389.

28. Cardoso CR, Garlet GP, Crippa GE, Rosa AL, Júnior WM, Rossi MA, et al. Evidence of the presence of T helper type 17 cells in chronic lesions of human periodontal disease. Oral Microbiol Immunol 2009; 24:1-6.

29. Yu JJ, Ruddy MJ, Wong GC, Sfintescu C, Baker PJ, Smith JB, et al. An essential role for IL-17 in preventing pathogen-initiated bone destruction: recruitment of neutrophils to inflamed bone requires IL-17 receptor-dependent signals. Blood 2007;109:3794-3802.

30. Kobayashi R, Kono T, Bolerjack BA, Fukuyama Y, Gilbert RS, Fujihashi K, et al. Induction of IL-10-producing CD4+ T-cells in chronic periodontitis. J Dent Res 2011; 90:653-658.

31. Okui T, Aoki Y, Ito H, Honda T, Yamazaki K. The presence of IL-17+/FOXP3+ double-positive cells in periodontitis. J Dent Res 2012:91:574-579.

32. Jin Y, Wang LY , Liu DX, Lin XP. Tamibarotene modulates the local immune response in experimental periodontitis. Int Immunopharmacol 2014; 23:537-545.