인체 기관지 상피세포에서 Mycoplasma pneumoniae 감염에 의한 천식 매개물질의 발현

Mycoplasma pneumoniae-induced production of proasthmatic mediators in airway epithelium

  • 김경원 (연세대학교 의과대학 소아과학교실 및 알레르기 연구소) ;
  • 이병철 (국립보건원 세균부 리케치아과) ;
  • 이경은 (연세대학교 의과대학 소아과학교실 및 알레르기 연구소) ;
  • 김은수 (연세대학교 의과대학 소아과학교실 및 알레르기 연구소) ;
  • 송태원 (연세대학교 의과대학 소아과학교실 및 알레르기 연구소) ;
  • 박미연 (국립보건원 세균부 리케치아과) ;
  • 손명현 (연세대학교 의과대학 소아과학교실 및 알레르기 연구소) ;
  • 김규언 (연세대학교 의과대학 소아과학교실 및 알레르기 연구소)
  • Kim, Kyung Won (Department of Pediatrics and Institute of Allergy, Yonsei University College of Medicine and Rickettsial and Zoonotic Diseases Division) ;
  • Lee, Byung Chul (Department of Microbiology, National Institute of Health) ;
  • Lee, Kyung Eun (Department of Pediatrics and Institute of Allergy, Yonsei University College of Medicine and Rickettsial and Zoonotic Diseases Division) ;
  • Kim, Eun Soo (Department of Pediatrics and Institute of Allergy, Yonsei University College of Medicine and Rickettsial and Zoonotic Diseases Division) ;
  • Song, Tae Won (Department of Pediatrics and Institute of Allergy, Yonsei University College of Medicine and Rickettsial and Zoonotic Diseases Division) ;
  • Park, Mi Yeoun (Department of Microbiology, National Institute of Health) ;
  • Sohn, Myung Hyun (Department of Pediatrics and Institute of Allergy, Yonsei University College of Medicine and Rickettsial and Zoonotic Diseases Division) ;
  • Kim, Kyu-Earn (Department of Pediatrics and Institute of Allergy, Yonsei University College of Medicine and Rickettsial and Zoonotic Diseases Division)
  • 투고 : 2006.06.02
  • 심사 : 2006.07.10
  • 발행 : 2006.09.15

초록

목 적 : M. pneumoniae는 기도 점막의 상피세포에서 증식하면서 호흡기 질환을 일으키며 기관지 천식의 발생이나 악화와 관계된다고 알려져 있다. IL-6은 급성 염증을 유도하는 사이토카인으로 B 세포의 분화에 관여하며 Th2 염증반응을 촉진시키며, IL-8은 기도에서 염증부위로의 세포이동을 매개하는 케모카인으로 알레르기 염증 반응에 밀접한 관련을 가진다. 기도 상피세포에서 생성되는 NO는 기도 염증과 기도과민성의 조절에 중요하다. VEGF는 천식에서 기도개형에 주된 역할을 담당한다. 본 연구에서는 기관지 상피세포에 M. pneumoniae를 감염시켰을 때 IL-6, IL-8, NO 및 VEGF의 발현을 살펴보았다. 방 법 : M. pneumoniae를 기관지 상피 세포주인 A549 세포에 감염시킨 후 여러 시간 간격으로 배양하여 세포와 배양 상층액을 수거하였다. 면역 형광 염색과 M. pneumoniae 16S ribosomal RNA gene의 oligonucleotide primers를 이용한 중합효소 연쇄반응을 시행하여 감염을 확인하였다. IL-6, IL-8, VEGF 단백질 생성은 ELISA kit를 이용하여 정량하였고, NO는 Griess 반응을 이용하여 측정하였다. 역전사 중합효소 연쇄반응을 이용하여 IL-6, IL-8, VEGF의 mRNA 발현을 관찰하였다. 결 과 : 기관지 상피세포에 M. pneumoniae를 감염시킨 후 시간이 지남에 따라 IL-6, IL-8, NO, VEGF의 생성이 증가하였고, IL-6, IL-8, VEGF mRNA 발현이 증가됨을 관찰하였다. 결 론 : M. pneumoniae 감염은 IL-6, IL-8, NO, VEGF 등의 매개물질의 발현을 증가시켜 기관지 천식의 알레르기 염증반응에 관여할 것으로 사료된다.

Purpose : There has been an increasing amount of literature concerning the association between Mycoplasma pneumoniae and asthma pathogenesis. Interleukin(IL)-6 stimulates the differentiation of monocytes, and can promote Th2 differentiation and simultaneously inhibit Th1 polarization. IL-8 is a potent chemoattractant and, it has been suggested, has a role in asthma pathogenesis. Nitric oxide (NO) synthesized by airway epithelium may be important in the regulation of airway inflammation and reactivity. Vascular endothelial growth factor(VEGF) has been reported to be a mediator of airway remodeling in asthma. We investigated the effects of M. pneumoniae on IL-6, IL-8, NO and VEGF production in human respiratory epithelial cells. Methods : A549 cells were cultured and inoculated with M. pneumoniae at a dose of 20 cfu/cell. After infection, the presence of M. pneumoniae in epithelial cell cultures was monitored by immunofluorescence and confirmed by polymerase chain reaction(PCR) detection. IL-6, IL-8 and VEGF were determined by an enzyme-linked immunosorbent assay and reverse transcriptase-polymerase chain reaction. NO was measured using the standard Griess reaction. Results : In A549 cells, M. pneumoniaeinduced IL-6, IL-8, NO and VEGF release in time-dependent manners. It also induced mRNA expression of IL-6, IL-8 and VEGF in similar manners. Conclusion : These observations suggest that M. pneumoniae might have a role in the pathogenesis of the allergic inflammation of bronchial asthma.

키워드

참고문헌

  1. Foy HM. Infections caused by Mycoplasma pneumoniae and possible carrier state in different populations of patients. Clin Infect Dis 1993;17 Suppl 1:37-46 https://doi.org/10.1093/clinids/17.Supplement_1.S37
  2. Johnston SL, Martin RJ. Chlamydophila pneumoniae and Mycoplasma pneumoniae : a role in asthma pathogenesis? Am J Respir Crit Care Med 2005;172:1078-89 https://doi.org/10.1164/rccm.200412-1743PP
  3. Waites KB, Talkington DF. Mycoplasma pneumoniae and its role as a human pathogen. Clin Microbiol Rev 2004;17:697-728 https://doi.org/10.1128/CMR.17.4.697-728.2004
  4. Busse WW. Respiratory infections: their role in airway responsiveness and the pathogenesis of asthma. J Allergy Clin Immunol 1990;85:671-83 https://doi.org/10.1016/0091-6749(90)90181-3
  5. Esposito S, Blasi F, Arosio C, Fioravanti L, Fagetti L, Droghetti R, et al. Importance of acute Mycoplasma pneumoniae and Chlamydia pneumoniae infections in children with wheezing. Eur Respir J 2000;16:1142-6 https://doi.org/10.1034/j.1399-3003.2000.16f21.x
  6. Gil JC, Cedillo RL, Mayagoitia BG, Paz MD. Isolation of Mycoplasma pneumoniae from asthmatic patients. Ann Allergy 1993;70:23-5
  7. Kraft M, Cassell GH, Henson JE, Watson H, Williamson J, Marmion BP, et al. Detection of Mycoplasma pneumoniae in the airways of adults with chronic asthma. Am J Respir Crit Care Med 1998;158:998-1001 https://doi.org/10.1164/ajrccm.158.3.9711092
  8. Lieberman D, Lieberman D, Printz S, Ben-Yaakov M, Lazarovich Z, Ohana B, et al. Atypical pathogen infection in adults with acute exacerbation of bronchial asthma. Am J Respir Crit Care Med 2003;167:406-10 https://doi.org/10.1164/rccm.200209-996OC
  9. Martin RJ, Kraft M, Chu HW, Berns EA, Cassell GH. A link between chronic asthma and chronic infection. J Allergy Clin Immunol 2001;107:595-601 https://doi.org/10.1067/mai.2001.113563
  10. Chu HW, Honour JM, Rawlinson CA, Harbeck RJ, Martin RJ. Effects of respiratory Mycoplasma pneumoniae infection on allergen-induced bronchial hyperresponsiveness and lung inflammation in mice. Infect Immun 2003;71:1520-6 https://doi.org/10.1128/IAI.71.3.1520-1526.2003
  11. Hardy RD, Jafri HS, Olsen K, Hatfield J, Iglehart J, Rogers BB, et al. Mycoplasma pneumoniae induces chronic respiratory infection, airway hyperreactivity, and pulmonary inflammation : a murine model of infection-associated chronic reactive airway disease. Infect Immun 2002;70:649-54 https://doi.org/10.1128/IAI.70.2.649-654.2002
  12. Martin RJ, Chu HW, Honour JM, Harbeck RJ. Airway inflammation and bronchial hyperresponsiveness after Mycoplasma pneumoniae infection in a murine model. Am J Respir Cell Mol Biol 2001;24:577-82 https://doi.org/10.1165/ajrcmb.24.5.4315
  13. Yang J, Hooper WC, Phillips DJ, Talkington DF. Cytokines in Mycoplasma pneumoniae infections. Cytokine Growth Factor Rev 2004;15:157-68 https://doi.org/10.1016/j.cytogfr.2004.01.001
  14. Sohn MH, Lee KE, Choi SY, Kwon BC, Chang MW, Kim KE. Effect of Mycoplasma pneumoniae lysate on interleukin-8 gene expression in human respiratory epithelial cells. Chest 2005;128:322-6 https://doi.org/10.1378/chest.128.1.322
  15. Yang J, Hooper WC, Phillips DJ, Talkington DF. Regulation of proinflammatory cytokines in human lung epithelial cells infected with Mycoplasma pneumoniae. Infect Immun 2002;70:3649-55 https://doi.org/10.1128/IAI.70.7.3649-3655.2002
  16. Marini M, Vittori E, Hollemborg J, Mattoli S. Expression of the potent inflammatory cytokines, granulocyte-macrophage-colony-stimulating factor and interleukin-6 and interleukin-8, in bronchial epithelial cells of patients with asthma. J Allergy Clin Immunol 1992;89:1001-9 https://doi.org/10.1016/0091-6749(92)90223-O
  17. Fonseca-Aten M, Rios AM, Mejias A, Chavez-Bueno S, Katz K, Gomez AM, et al. Mycoplasma pneumoniae induces host-dependent pulmonary inflammation and airway obstruction in mice. Am J Respir Cell Mol Biol 2005;32:201-10 https://doi.org/10.1165/rcmb.2004-0197OC
  18. Watkins DN, Peroni DJ, Basclain KA, Garlepp MJ, Thompson PJ. Expression and activity of nitric oxide synthases in human airway epithelium. Am J Respir Cell Mol Biol 1997;16:629-39 https://doi.org/10.1165/ajrcmb.16.6.9191464
  19. Kharitonov SA, Yates D, Robbins RA, Logan-Sinclair R, Shinebourne EA, Barnes PJ. Increased nitric oxide in exhaled air of asthmatic patients. Lancet 1994;343:133-5 https://doi.org/10.1016/S0140-6736(94)90931-8
  20. Smith AD, Cowan JO, Brassett KP, Herbison GP, Taylor DR. Use of exhaled nitric oxide measurements to guide treatment in chronic asthma. N Engl J Med 2005;352:2163-73 https://doi.org/10.1056/NEJMoa043596
  21. Zeiger RS, Szefler SJ, Phillips BR, Schatz M, Martinez FD, Chinchilli VM, et al. Response profiles to fluticasone and montelukast in mild-to-moderate persistent childhood asthma. J Allergy Clin Immunol 2006;117:45-52 https://doi.org/10.1016/j.jaci.2005.10.012
  22. Lazaar AL, Panettieri RA. Airway smooth muscle : a modulator of airway remodeling in asthma. J Allergy Clin Immunol 2005;116:488-95 https://doi.org/10.1016/j.jaci.2005.06.030
  23. Lee CG, Link H, Baluk P, Homer RJ, Chapoval S, Bhandari V, et al. Vascular endothelial growth factor(VEGF) induces remodeling and enhances Th2-mediated sensitization and inflammation in the lung. Nat Med 2004;10:1095-103 https://doi.org/10.1038/nm1105
  24. Asai K, Kanazawa H, Kamoi H, Shiraishi S, Hirata K, Yoshikawa J. Increased levels of vascular endothelial growth factor in induced sputum in asthmatic patients. Clin Exp Allergy 2003;33:595-9 https://doi.org/10.1046/j.1365-2222.2003.01576.x
  25. Hoshino M, Nakamura Y, Hamid QA. Gene expression of vascular endothelial growth factor and its receptors and angiogenesis in bronchial asthma. J Allergy Clin Immunol 2001;107:1034-8 https://doi.org/10.1067/mai.2001.115626
  26. Dakhama A, Kraft M, Martin RJ, Gelfand EW. Induction of regulated upon activation, normal T cells expressed and secreted(RANTES) and transforming growth factor-beta 1 in airway epithelial cells by Mycoplasma pneumoniae. Am J Respir Cell Mol Biol 2003;29:344-51 https://doi.org/10.1165/rcmb.2002-0291OC
  27. Yu J, Yoo Y, Kim DK, Kang H, Koh YY. Distributions of antibody titers to Mycoplasma pneumoniae in Korean children in 2000-2003. J Korean Med Sci 2005;20:542-7 https://doi.org/10.3346/jkms.2005.20.4.542
  28. Wongtim S MS. Methacholine inhalation challenge in patients with post-Mycoplasma pneumoniae pneumonia. Asian Pac J Allergy Immunol 1995;13:5-10
  29. Abdel-Rahman AM, el-Sahrigy SA, Bakr SI. A comparative study of two angiogenic factors : vascular endothelial growth factor and angiogenin in induced sputum from asthmatic children in acute attack. Chest 2006;129:266-71 https://doi.org/10.1378/chest.129.2.266