Clinical and Experimental Pediatrics

The Korean Pediatric Society (대한소아청소년과학회)
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Toll-like receptor 2 expression on monocytes from patients with BCG vaccine-associated suppurative lymphadenitis

BCG 접종 후 발생한 화농성 림프절염 환자의 단핵구에서 Toll-like receptor 2의 발현

  • Oh, Hyun Joo (Department of Pediatrics, Cheju National University School of Medicine) ;
  • Shin, Kyung-Sue (Department of Pediatrics, Cheju National University School of Medicine, Institute of Medical Science, Cheju National University)
  • 오현주 (제주대학교 의학전문대학원 소아과학교실) ;
  • 신경수 (제주대학교 의학전문대학원 소아과학교실, 제주대학교 의과학연구소)
  • Received : 2008.12.05
  • Accepted : 2009.05.13
  • Published : 2009.06.15
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Abstract

Purpose : Toll-like receptor 2 (TLR2) is critical in the immune response to mycobacterial infections. The purpose of this study was to analyze TLR2 surface expressions and TLR2-mediated tumor necrosis factor-alpha ($TNF-{\alpha}$) and interleukin-6 (IL-6) production in patients with BCG vaccine-associated suppurative lymphadenitis. Methods : Peripheral monocytes were separated from 16 patients with BCG vaccine-associated suppurative lymphadenitis and 10 healthy controls using a magnet cell isolation kit. Monocytes ($1{\times}10^6$ cells/well) were incubated with a constant amount of $Pam_3CSK_4$ ($100{\mu}g/mL$) for 24 hours. TLR2 surface expression on monocytes was analyzed by FACS analysis and TLR-2 mRNA expression was determined by RT-PCR. TLR2-mediated $TNF-{\alpha}$ and IL-6 production were measured by ELISA. Results : In patients with BCG vaccine-associated suppurative lymphadenitis, low TLR2 expression on monocytes ($3.39{\pm}$1.2% versus $4.64{\pm}2.6%$) together with significantly lower TLR2 mRNA expression than in the healthy controls was seen after $Pam_3CSK_4$ stimulation. TLR2-mediated $TNF-{\alpha}$ and IL-6 production in patients with BCG vaccine-associated suppurative lymphadenitis ($TNF-{\alpha}$, $775.5{\pm}60.8pg/mL$; IL-6, $4,645.8{\pm}583.9pg/mL$) were also lesser than that in healthy controls ($TNF-{\alpha}$, $1,098.5{\pm}94.3pg/mL$; IL-6, $6,696.3{\pm}544.3pg/mL$). Conclusion : These findings suggest that low TLR2 expression on monocytes might be associated with increased susceptibility to BCG vaccine-associated suppurative lymphadenitis.

목 적 : TLR2는 숙주의 항결핵 방어면역의 중요한 역할을 하는 것으로 알려져 있다. 본 연구는 BCG 접종 후 발생한 화농성 림프절염 환자 단핵구에서 TLR2의 발현과 TLR2 리간드 자극에 의한 $TNF-{\alpha}$와 IL-6의 생성을 조사하여 화농성 림프절염 발병과 TLR2의 연관성을 알아보고자 하였다. 방 법 : BCG 접종 후 발생한 화농성 림프절염 환자 16명과 건강 대조군 10명의 말초 혈액에서 단핵구를 분리하고, TLR2 리간드인 Pam3CSK4로 자극한 후 유세포분석과 역전사중합효소반응을 이용하여 TLR2의 발현을 측정하였고, 자극 후 $TNF-{\alpha}$와 IL-6의 생성을 측정하여 TLR2의 발현 정도를 간접적으로 조사하였다. 결 과 : BCG 접종 후 발생한 화농성 림프절염 환자 단핵구의 TLR2 발현 정도($3.39{\pm}1.2%$)는 대조군($4.64{\pm}2.6%$)에 비하여 유의하게 감소하였고, 단핵구 자극에 의한 $TNF-{\alpha}$와 IL-6의 생성도 대조군($TNF-{\alpha}$, $1,098.5{\pm}94.3pg/mL$; IL-6, $6,696.3{\pm}544.3pg/mL$)에 비하여 환자군($TNF-{\alpha}$, $775.5{\pm}60.8pg/mL$; IL-6, $4,645.8{\pm}583.9pg/mL$)에서 유의하게 감소하였다. 그리고 자극 시간에 따른 TLR2 발현 정도와 $TNF-{\alpha}$와 IL-6의 생성 증가가 유사한 양상을 나타내었다. 결 론 : 본 연구의 결과에서 BCG 접종 후 발생한 화농성 림프절염 환자군 단핵구의 TLR2 발현 감소가 연관되어 있고, M. bovis BCG의 리간드 인식에 TLR2가 관여함을 추정할 수 있다.

Keywords

Acknowledgement

Supported by : Wyeth

References

  1. Baek HS, Chang JY, Moon SJ, Oh SH. Lymphadenitis following intradermal BCG vaccination. Korean J Pediatr 2006;49:46-50 https://doi.org/10.3345/kjp.2006.49.1.46
  2. Teo S, Smeulders N, Shingadia DV. BCG vaccine-associated suppurative lymphadenitis. Vaccine 2005;23:2676-9 https://doi.org/10.1016/j.vaccine.2004.07.052
  3. Stenger S, Modlin RL. Control of Mycobacterium tuberculosis through mammalian Toll-like receptors. Curr Opin Immunol 2002;14:452-7 https://doi.org/10.1016/S0952-7915(02)00355-2
  4. Heldwein KA, Liang MD, Andressen TK, Thomas KE, Marthy AM, Cuesta N, et al. TLR2 and TLR4 serve distinct roles in the host immune response against Mycobacterium bovis BCG. J Leukoc Biol 2003;74:277-86 https://doi.org/10.1189/jlb.0103026
  5. Nicolle D, Fremond C, Pichon X, Bouchot A, Maillet I, Ryffel B, et al. Long-term control of Mycobacterium bovis BCG infection in the absence of Toll-like receptors: investigation of TLR2-, TLR6-, or TLR2-TLR4-deficient mice. Infect Immun 2004;72:6994-7004 https://doi.org/10.1128/IAI.72.12.6994-7004.2004
  6. Gilleron M, Nigou J, Nicolle D, Quesniaux V, Puzo G. The acylation state of mycobacterial lipomannans modulates innate immunity response through toll-like receptor 2. Chem Biol 2006;13:39-47 https://doi.org/10.1016/j.chembiol.2005.10.013
  7. Underhill DM, Ozinsky A, Smith KD, Aderem A. Toll-like receptor-2 mediates mycobacteria-induced proinflammatory signaling in macrophages. PNAS 1999;96:14459-63 https://doi.org/10.1073/pnas.96.25.14459
  8. Leal IS, Florido M, Andersen P, Appelberg R. Interlukin-6 regulates the phenotype of the immune response to a tuberculosis subunit vaccine. Immunology 2001;103:375-81 https://doi.org/10.1046/j.1365-2567.2001.01244.x
  9. Singla A, Singh S, Goraya JS, Radhika S, Sharma M. The natural course of nonsuppurative Calmette-Guerin bacillus lymphadenitis. Pediatr Infect Dis J 2002;21:446-8 https://doi.org/10.1097/00006454-200205000-00026
  10. Flowaczny M, Glas J, Torok HP, Limbersky O, Flowaczny C. Toll-like receptor 2 and 4 mutations in peiodontal disease. Clin Exp Immunol 2004;135:330-5 https://doi.org/10.1111/j.1365-2249.2004.02383.x
  11. Lotte A, Wasz-Hockert O, Poisson N, Dumitrescu N, Verron M, Couvet E. BCG complications. Estimates of the risks among vaccinated subjects and statistical analysis of their main chracteristics. Adv Tuberc Res 1984;21:107-93
  12. Behr MA. BCG-different strains, different vaccines? Lancet Infect Dis 2002;2:86-92 https://doi.org/10.1016/S1473-3099(02)00182-2
  13. Na KH, Rim SS, Kim EY, Kim KS, Kim YW. Factors affecting clinical course of BCG lymphadenitis. Korean J Pediatr Infect Dis 2001;8:181-90
  14. Kim MS, Jo DS, Kang MK, Kim SJ, Kim JS. The effect of local rifampicin instillation on the treatment of suppurative BCG lymphadenitis. Korean J Pediatr 2006;49:40-5 https://doi.org/10.3345/kjp.2006.49.1.40
  15. Hwang DH, Han JW, Choi KM, Shin KM, Kim DS. Expression of Toll-like receptor-2 on the peripheral monocytes in Kawasaki disease patients. Korean J Pediatr 2005;48:315-20
  16. Wikén M, Grunewald J, Eklund A, Wahlström J. Higher monocyte expression of TLR2 and TLR4, and enhanced pro- inflammatory synergy of TLR2 with NOD2 stimualtion in Sarcoidosis. J Clin Immunol 2009;29:78-89 https://doi.org/10.1007/s10875-008-9225-0
  17. Han YP, Li J, Wan YF, Kong LH, Cai J, Dong L, et al. The influence of hepatitis B e antigen on the expression of toll- like receptor 2 on peripheral monocytes [abstract]. Zhonghua Gan Zang Bing Za Zhi 2008;16:739A
  18. Link A, Selejan S, Maack C, Lenz M, Böhm M. Phosphodiesterase 4 inhibition but not beta-adrenergic stimulation suppresses tumor necrosis factor-alpha release in peripheral blood mononuclear cells in septic shock. Critical Care 2008; 12:R159 https://doi.org/10.1186/cc7158
  19. Quesniaux VJ, Nicolle DM, Torres D, Kremmer L, Guerardel Y, Nigou J, et al. Toll-like receptor 2-dependent-positive and TLR2-independent-negative regulation of proinflammatory cytokines by mycobacterial lipomannans. J Immunol 2004; 172:4425-34 https://doi.org/10.4049/jimmunol.172.7.4425
  20. Gilleron M, Quesniaux VF, Puzo G. Acylation state of the phosphatidylinositol hexamannosides from Mycobacterium bovis bacillus Calmette-Gurin and Mycobacterium tuberculosis H37Rv and its implication in Toll-like receptor response. J Biol Chem 2003;278:29880-9 https://doi.org/10.1074/jbc.M303446200
  21. Droemann D, Goldmann T, Tiedje T, Zabel P, Dalhoff K, Schaaf B. Toll-like receptor 2 expression is decreased on alveolar macrophages in cigarette smokers and COPD patients. Respir Res 2005;6:68 https://doi.org/10.1186/1465-9921-6-68
  22. Bosisio D, Polentarutti N, Sironi M, Bernasconi S, Miyake K, Webb GR, et al. Stimulation of toll-like receptor 4 expression in human mononuclear phagocytes by interferon-gamma: a molecular basis for priming and synergism with bacterial lipopolysaccharide. Blood 2002;99:3427-31 https://doi.org/10.1182/blood.V99.9.3427
  23. Nomura F, Akashi S, Sakao Y, Sato S, Kawai T, Matsumoto M, et al. Endotoxin tolerance in mouse peritoneal macrophages correlates with down-regulation of surface Toll-like receptor 4 expression. J Immunol 2000;164:3476-9 https://doi.org/10.4049/jimmunol.164.7.3476
  24. Iwahashi M, Tamamura M, Aita T, Okamoto A, Ueno A, Ogawa N, et al. Expression of Toll-like receptor 2 on CD16+ blood monocytes and synovial tissue macrophages in rheumatoid arthritis. Arthritis Rheum 2004;50:1457-67 https://doi.org/10.1002/art.20219
  25. Krause SW, Kreutz M, Andreesen R. Differential effcts of cell adherence on LPS-stimulated cytokine production by human monocytes and macrophages. Immunobiology 1996; 196:522-34
  26. Underhill DM, Ozinsky A, Hajjar AM, Stevens A, Wilson CB, Bassetti M, et al. The Toll-like receptor 2 is recruited to macrophage phagosomes and discriminates between pathogens. Nature 1999;401:811-5 https://doi.org/10.1038/44605
  27. Kang TJ, Chae GT. Detection of Toll-like receptor2 (TLR2) mutation in the lepromatous leprosy patients. FEMS Immunol Med Microbiol 2001;31:53-8 https://doi.org/10.1111/j.1574-695X.2001.tb01586.x
  28. Schr\ddot{o}der NWJ, Hermann C, Hamann L, Gobel UB, Hartung T, Schumann RR. High frequency of polymorphism Arg753Gln of the Toll-like receptor-2 gene detected by a novel allele-specific PCR. J Mol Med 2003;81:368-72 https://doi.org/10.1007/s00109-003-0443-x
  29. Ben-Ali M, Barbouche MR, Bousnina S, Chabbou A, Dellagi K. Toll-like receptor 2 Arg677Trp polymorphism is associated with susceptibility to tuberculosis in Tunisian patients. Clin Diagn Lab Immunol 2004;11:625-6
  30. Lorenz E, Mira JP, Cornish KL, Arbour NC, Schwartz DA. A novel polymorphism in the toll-like receptor 2 gene and its potential association with staphylococcal infection. Infect Immun 2000;68:6398-401 https://doi.org/10.1128/IAI.68.11.6398-6401.2000
  31. Yoon HJ, Choi JY, Kim CO, Park YS, Kim MS, Kim YK, et al. Lack of Toll-like receptor 4 and 2 polymorphism in Korean patients with bacteremia. J Korean Med Sci 2006;21: 979-82 https://doi.org/10.3346/jkms.2006.21.6.979