Inactivated Mycobacterium phlei inhalation ameliorates allergic asthma through modulating the balance of CD4+CD25+ regulatory T and Th17 cells in mice

Document Type: Original Article

Authors

Department of Respiratory Medicine, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China 530021

Abstract

Objective(s): Th2 response is related to the aetiology of asthma, but the underlying mechanism is unclear. To address this point, the effect of nebulized inhalation of inactivated Mycobacterium phlei on modulation of asthmatic  airway  inflammation was investigated.
Materials and Methods: 24 male BALB/c mice were randomly divided into three groups: control group (Group A), asthma model group (Group B), and the medicated asthma model group (Group C). Group B and C were sensitized and challenged with ovalbumin (OVA). Group C was treated with aerosol M. phlei once daily before OVA challenge. Airway responsiveness in each group was assessed. All the animals were killed, and lung tissues and bronchoalveolar lavage fluid (BALF) were harvested. Inflammatory response, proportion of Th17 and CD4+CD25+ Treg cells, and the levels of cytokines were analyzed in lung tissue.
Results: The proportion of Th17 cells and expression level of IL17, IL23, and IL23R were increased, while Foxp3 expression was decreased in Group B. Inhaling inactivated M. phlei inhibited airway inflammation and improved airway hyper-responsiveness, as well as peak expiratory flow (PEF). In addition, it significantly increased Th17 proportion, Foxp3 level, and the proportion of CD4+CD25+ Treg cells in lung tissue in Group C.
Conclusion: Inactivated M. phlei was administered by atomization that suppressed airway inflammation and airway hyper responsiveness partially via modulating the balance of CD4+CD25+ regulatory T and Th17 cells.

Keywords


1. Robinson DS. Regulatory T cells and asthma. Clin Exp Allergy 2009; 39:1314-1323.

2. Louten J, Boniface K, de Waal Malefyt R. Development and function of TH17 cells in health and disease. The J Allergy Clin Immunol 2009; 123:1004-10011.

3. Soroosh P, Doherty TA. Th9 and allergic disease. Immunology 2009; 127:450-458.

4. Hellings PW, Kasran A, Liu Z, Vandekerckhove P, Wuyts A, Overbergh L, et al. Interleukin-17 orchestrates the granulocyte influx into airways after allergen inhalation in a mouse model of allergic asthma. Am J Respir Cell Mol Biol 2003; 28:42-50.

5. Durrant DM, Metzger DW. Emerging roles of T helper subsets in the pathogenesis of asthma. Immunol Invest 2010; 39:526-249.

6. Wing K, Fehervari Z, Sakaguchi S. Emerging possibilities in the development and function of regulatory T cells. Int Immunol 2006; 18:991-1000.

7. Ryanna K, Stratigou V, Safinia N, Hawrylowicz C. Regulatory T cells in bronchial asthma. Allergy 2009; 64:335-347.

8. Akdis CA, Akdis M. Mechanisms and treatment of allergic disease in the big picture of regulatory T cells.  J Allergy Clin Immunol 2009; 123:735-746; quiz 47-48.

9. Palomares O, Yaman G, Azkur AK, Akkoc T, Akdis M, Akdis CA. Role of Treg in immune regulation of allergic diseases. Eur J Immunol 2010; 40:1232-1240.

10. Sakaguchi S, Ono M, Setoguchi R, Yagi H, Hori S, Fehervari Z, et al. Foxp3+ CD25+ CD4+ natural regulatory T cells in dominant self-tolerance and autoimmune disease. Immunol Rev 2006; 212:8-27.

11. Annesi-Maesano I, Mourad C, Daures JP, Kalaboka S, Godard P. Time trends in prevalence and severity of childhood asthma and allergies from 1995 to 2002 in France. Allergy 2009; 64:798-800.

12. Asher MI. Recent perspectives on global epidemiology of asthma in childhood. Allergol Immunopathol 2010; 38:83-87.

13. Afshar R, Medoff BD, Luster AD. Allergic asthma: a tale of many T cells. Clin Expe Allergy  2008; 38:1847-1857.

14.Koh YI, Choi IS, Kim WY. BCG infection in allergen-presensitized rats suppresses Th2 immune response and prevents the development of allergic asthmatic reaction. J Clin Immunol 2001; 21:51-59.

15.Erb KJ, Holloway JW, Sobeck A, Moll H, Le Gros G. Infection of mice with Mycobacterium bovis-Bacillus Calmette-Guerin (BCG) suppresses allergen-induced airway eosinophilia. J Exp Med 1998; 187:561-569.

16.Herz U, Gerhold K, Gruber C, Braun A, Wahn U, Renz H, et al. BCG infection suppresses allergic sensitization and development of increased airway reactivity in an animal model. J Allergy Clin Immunol 1998; 102:867-874.

17. Wang CC, Rook GA. Inhibition of an established allergic response to ovalbumin in BALB/c mice by killed Mycobacterium vaccae. Immunology 1998; 93:307-313.

18. Hopfenspirger MT, Agrawal DK. Airway hyper responsiveness, late allergic response, and eosinophilia are reversed with mycobacterial antigens in ovalbumin-presensitized mice. J Immunol 2002; 168:2516-2522.

19. Zuany-Amorim C, Sawicka E, Manlius C, Le Moine A, Brunet LR, Kemeny DM, et al. Suppression of airway eosinophilia by killed Mycobacterium vaccae-induced allergen-specific regulatory T-cells. Nat Med 2002; 8:625-629.

20. Zhang J, Li C, Guo S. Effects of inhaled inactivated Mycobacterium phlei on airway inflammation in mouse asthmatic models. J Aerosol Med Pulm Drug Deliv 2012; 25:96-103.

21. Cheng C, Ho WE, Goh FY, Guan SP, Kong LR, Lai WQ, et al. Anti-malarial drug artesunate attenuates experimental allergic asthma via inhibition of the phosphoinositide 3-kinase/Akt pathway. PloS One 2011; 6:e20932.

22. Qu SY, Ou-Yang HF, He YL, Wan Q, Shi JR, Wu CG.. Der p 2 recombinant bacille Calmette-Guerin targets dendritic cells to inhibit allergic airway inflammation in a mouse model of asthma. Respiration 2013; 85:49-58.

23. Karani L, Tolo F, Karanja S, Khayeka C. Safety and efficacy of Prunus africana and Warburgia ugandensis against induced asthma in BALB/c Mice. Eur J Med Pl 2013; 3:345-368.

24. Tsai YG, Niu DM, Yang KD, Hung CH, Yeh YJ, Lee CY, et al. Functional defects of CD46-induced regulatory T cells to suppress airway inflammation in mite allergic asthma. Lab Invest 2012; 92:1260-1269.

25. Roh GS, Seo S-W, Yeo S, Lee JM, Choi J-W, Kim E, et al. Efficacy of a traditional Korean medicine, Chung-Sang-Bo-Ha-Tang, in a murine model of chronic asthma. Int Immunopharmacol 2005; 5:427-436.

26. Yang X, Wang S, Fan Y, Zhu L. Systemic mycobacterial infection inhibits antigen-specific immunoglobulin E production, bronchial mucus production and eosinophilic inflammation induced by allergen. Immunology 1999; 98:329-337.

27. Cavallo GP, Elia M, Giordano D, Baldi C, Cammarota R. Decrease of specific and total IgE levels in allergic patients after BCG vaccination: preliminary report. Arch Otolaryngol Head Neck Surg 2002; 128:1058-1060.

28.Round JL, Lee SM, Li J, Tran G, Jabri B, Chatila TA, et al. The Toll-like receptor 2 pathway establishes colonization by a commensal of the human microbiota. Science 2011; 332:974-977.

29.Olszak T, An D, Zeissig S, Vera MP, Richter J, Franke A, et al. Microbial exposure during early life has persistent effects on natural killer T cell function. Science 2012; 336:489-493.

30. Navarro S, Cossalter G, Chiavaroli C, Kanda A, Fleury S, Lazzari A, et al. The oral administration of bacterial extracts prevents asthma via the recruitment of regulatory T cells to the airways. Mucosal Immunol 2011; 4:53-65.

31.Strickland DH, Judd S, Thomas JA, Larcombe AN, Sly PD, Holt PG. Boosting airway T-regulatory cells by gastrointestinal stimulation as a strategy for asthma control. Mucosal Immunol 2011; 4:43-52.

32.Han ER, Choi IS, Eom SH, Kim HJ. Preventive effects of mycobacteria and their culture supernatants against asthma development in BALB/c mice. Allergy Asthma Immunol Res 2010; 2:34-40.

33. Choi IS, Lin XH, Koh YA, Koh YI, Lee HC. Strain-dependent suppressive effects of BCG vaccination on asthmatic reactions in BALB/c mice. Ann Allergy Asthma Immunol 2005; 95:571-578.

34.Choi IS, Lin XH, Koh YA, Cui Y. Inoculation route-dependent and allergen-specific suppressive effects of bacille Calmette-Guerin vaccination on asthmatic reactions in BALB/c mice. Lung 2007; 185:179-186.

35.Lagranderie M, Vanoirbeek JAJ, Vargaftig BB, Guyonvarc’h P-M, Marchal G, Roux X. Therapeutic administration of Mycobacterium bovis BCG killed by Extended Freeze-Drying modulates airway inflammation in a chronic murine model of asthma. Open J Respir Dis 2013; 3:79.

36.Deng Y, Li W, Luo Y, Wang LJ, Xie XH, Luo J, et al. Inhibition of IFN-γ promotes anti-asthma effect of Mycobacterium bovis Bacillus Calmette-Guerin neonatal vaccination: A murine asthma model. Vaccine 2014; 32:2070-2078.

37.Arnoldussen DL, Linehan M, Sheikh A. BCG vaccination and allergy: a systematic review and meta-analysis. J Allergy Clin Immunol 2011; 127:246-53, 53.e1-21.

38. Qu SY, Ou-Yang HF, He YL, Li ZK, Shi JR, Song LQ, et al. Der p2 recombinant bacille Calmette-Guerin priming of bone marrow-derived dendritic cells suppresses Der p2-induced T helper17 function in a mouse model of asthma. Respirology  2014; 19:122-131.

39.Ahrens B, Gruber C, Rha RD, Freund T, Quarcoo D, Awagyan A, et al. BCG priming of dendritic cells enhances T regulatory and Th1 function and suppresses allergen-induced Th2 function in vitro and in vivo. Int Arch Allergy Immunol 2009; 150:210-220.

40. Yao B, Li M, Pang Y. [Effect of Mycobacterium phlei F.U.36 on balance of CD4(+)CD25(+) regulatory T cells and Th17 cells in asthmatic mice]. Zhongguo Dang Dai Er Ke Za Zhi = Chinese J Contemporary Pediatr 2013; 15:1018-1022.

41.Xia Y, Zhang JH, Ji ZH, Li XD, Yu ZW, Liu HY. [Effect of bacillus calmette-guerin treatment on airway inflammation and T regulatory cells in mice with asthma]. Zhongguo Dang Dai Er Ke Za Zhi = Chinese J Contemporary Pediatr 2006; 8:413-416.

42.Deng Y, Chen W, Zang N, Li S, Luo Y, Ni K, et al. The antiasthma effect of neonatal BCG vaccination does not depend on the Th17/Th1 but IL-17/IFN-gamma balance in a BALB/c mouse asthma model. J Clin Immunol 2011; 31:419-429.

43.O'Callaghan C, Barry PW. The science of nebulised drug delivery. Thorax 1997; 52:S31-44.

44.Melani AS. Inhalatory therapy training: a priority challenge for the physician. Acta Biomed 2007; 78:233-245.

45.Torchilin VP. Drug targeting. Eur J Pharm Sci 2000; 1:S81-91.

46.Hansel TT, Johnston SL, Openshaw PJ. Microbes and mucosal immune responses in asthma. Lancet  2013; 381:861-873.