Clinical and Experimental Pediatrics

The Korean Pediatric Society (대한소아청소년과학회)
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Toll-like receptor 9 expression and interferon-α secretion upon CpG-ODN stimulation in allergic subjects

알레르기 환자에서 TLR9 ligand인 CpG-ODN 자극에 의한 IFN-α 분비와 TLR9 발현

  • Han, Man Yong (Departments of Pediatrics, CHA University School of Medicine) ;
  • Jee, Hye Mi (Departments of Pediatrics, CHA University School of Medicine) ;
  • Kim, Hyeong Yoon (Departments of Pediatrics, CHA University School of Medicine) ;
  • Lee, Cho Ae (Departments of Pediatrics, CHA University School of Medicine) ;
  • Cho, Hyo-Jin (Departments of Internal Medicine, CHA University School of Medicine) ;
  • Hwang, Seong-Gyu (Departments of Internal Medicine, CHA University School of Medicine) ;
  • Kim, Kyu-Earn (Department of Pediatrics, Yonsei University College of Medicine)
  • 한만용 (차 의과학대학교 소아과학교실) ;
  • 지혜미 (차 의과학대학교 소아과학교실) ;
  • 김형윤 (차 의과학대학교 소아과학교실) ;
  • 이초애 (차 의과학대학교 소아과학교실) ;
  • 조효진 (차 의과학대학교 내과학교실) ;
  • 황성규 (차 의과학대학교 내과학교실) ;
  • 김규언 (연세대학교 의과대학 소아과학교실)
  • Received : 2009.06.22
  • Accepted : 2009.08.22
  • Published : 2009.09.15
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Abstract

Purpose:The aim of this study is to explore the effect of the Toll-like receptor 9 (TLR9) expressed in plasmacytoid dendritic cells (pDCs) that respond to antigen to Th2 immune deviation in allergic patients. Methods:Subjects consisted of 19 allergic patients and 17 healthy volunteers. Skin prick tests and nasal provocation tests were performed for the two groups. Peripheral blood mononuclear cells (PBMCs) were collected from subjects and analyzed for the Lineage Cocktail (CD3, CD14, CD16, CD19, CD20, CD56) (-), HLA-DR (+), and CD123 (+) using flow cytometry. In addition, we analyzed TLR9 mRNA by reverse transcriptase-polymerase chain reaction. The level of $interferon-{\alpha}$ ($IFN-{\alpha}$) of the PBMCs following stimulation with the TLR9 ligand CpG-ODN 2216 was also evaluated. Results:Analyses of CD123 (+) revealed a nearly similar distribution for the classical pDC markers in the allergic group ($0.1%{\pm}0.04%$) and in the controls ($0.25%{\pm}0.23%$). The mRNA levels of TLR9 on PBMCs were not different between the allergic group and the controls ($1.29{\pm}0.41$ vs. $1.25{\pm}0.23$, respectively). Additionally, the level of $IFN-{\alpha}$ in PBMCs exposed to stimuli of the TLR9 ligand CpG-ODN 2216 was not significantly different between the two groups ($911{\pm}829$ vs. $1,095{\pm}888pg/mL$, respectively). Conclusion:We found no evidence that TLR9-dependent immune responses in human pDCs are associated with allergic status.

목 적 : 알레르기 환자와 정상인에서 형질세포양 수지상세포의 분포, TLR9 mRNA 양과 $IFN-{\alpha}$의 분비능에 차이가 있는지 알아보고자 하였다. 방 법 : 19명의 알레르기 환자와 17명의 건강한 성인을 대상으로 하였다. 말초혈액단핵세포를 채취하여 Lineage Cocktail(CD3, CD14, CD16, CD19, CD20, CD56)음성, HLA-DR 양성이면서 CD123양성을 유세포 분석기로 분석하였다. 말초혈액단핵세포에 TLR9 작용제(agonists)인 CpG-ODN 2216과 음성 대조를 위해 CpG-ODN 2206으로 자극하고 24시간 후 상청액을 추출하여 $IFN-{\alpha}$의 농도를 측정하였다. 또한 real time RT-PCR을 이용하여 TLR9 mRNA 정량분석을 시행하였다. 결 과 : 말초혈액단핵세포에서 형질세포양 수지상세포의 분포는 알레르기 환자가 평균 $0.1{\pm}0.04%$, 대조군이 평균 $0.25{\pm}0.23%$이었다. TLR9 mRNA 상대적인 양을 나타내는 ${\Delta}{\Delta}Ct$는 알레르기 환자에서 $1.29{\pm}0.41$이었고 대조군은 $1.25{\pm}0.23$이었다. TLR9 리간드인 CpG-ODN 2216 자극에 따른 $IFN-{\alpha}$의 분비능은 알레르기 환자에서 $911{\pm}829pg/mL$ 이었고 대조군에서 $1,095{\pm}888pg/mL$ 이었다. 이 세 결과에서 통계적인 차이는 없었다. 결 론 : TLR9을 통한 신호전달이 알레르기 환자의 면역반응을 대표하지는 않는 것으로 보이며, 향후 더 자세한 TLR9의 역할에 대한 연구가 필요하다.

Keywords

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