Clinical and Experimental Pediatrics

The Korean Pediatric Society (대한소아청소년과학회)
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DNA Methylation Change of IL-4 Gene from T Cell in Allergic Children

영유아기 아토피 환아에서 말초혈액 T 림프구에서 Interleukin-4 유전자의 DNA 메틸화 변화

  • Oh, Jae Won (Department of Pediatrics, College of Medicine, Hanyang University) ;
  • Yum, Myung Gul (Department of Pediatrics, College of Medicine, Hanyang University) ;
  • Kim, Chang Ryul (Department of Pediatrics, College of Medicine, Hanyang University) ;
  • Seol, In-Joon (Department of Pediatrics, College of Medicine, Hanyang University) ;
  • Shin, Su A (Department of Pediatrics, College of Medicine, Hanyang University) ;
  • Lee, Ha Baik (Department of Pediatrics, College of Medicine, Hanyang University) ;
  • Jang, Se Jin (Department of Pathology, Asan Medical Center, University of Ulsan, College of Medicine)
  • 오재원 (한양대학교 의과대학 소아과학교실) ;
  • 염명걸 (한양대학교 의과대학 소아과학교실) ;
  • 김창렬 (한양대학교 의과대학 소아과학교실) ;
  • 설인준 (한양대학교 의과대학 소아과학교실) ;
  • 신수아 (한양대학교 의과대학 소아과학교실) ;
  • 이하백 (한양대학교 의과대학 소아과학교실) ;
  • 장세진 (울산대학교 의과대학 병리학교실)
  • Received : 2004.12.30
  • Accepted : 2005.03.10
  • Published : 2005.06.15
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Abstract

Purpose : An understanding of the immunological process is required if primary prevention of atopic diseases is to be developed in early childhood. But, it is too hard to distinguish atopy from nonatopy under the age of two clinically, because the expression of phenotype and cytokines is vague in early childhood. We evaluated DNA methylation changes at Th2 interleukin-4 gene in peripheral blood from atopic children. Methods : We selected 15 allergic children(mild : eight, moderate to severe : seven) and seven normal controls by using family allergy scores and clinical histories. We measured Total IgE and Der f II specific IgE levels and cultured peripheral blood mononuclear cells with Der f II stimulation and extracted DNA from Der f II specific T cells. We examined the change of CpG methylation in DNA from atopic and nonatopic children. Results : In T cells from normal children, IL-4 DNA were predominantly methylated; otherwise, CpG demethylation occurred in Der f II specific T cells from allergic children. Conclusion : IL-4 DNA methylation changes occurred in T genes from allergic children and DNA methylation assay in early childhood.

목 적 : 2세 미만의 소아에서는 임상적으로 확실한 증상이 있거나 알레르기 가족력이 있는 것을 제외하고는 실제적으로 아토피 유무를 판별하는 것은 성인에 비해 제한점이 많다. 본 연구에서는 영유아기에 알레르기 질환과 관련된 사이토카인이 유전자 배열에서 발현하는 지를 말초혈액의 Th2 림프구에서 interleukin-4 유전자의 DNA 탈메틸화를 통하여 알아보고자 하였다. 방 법 : 대상은 한양대학교 구리병원 소아알레르기클리닉을 내원한 3세 미만 소아에 대해 소아의 병력과 가족력을 조사하여 family allergy score와 개인력을 기준으로 알레르기질환에 대한 고위험군 7명, 저위험군 8명, 정상 대조군 7명이었다. 이들 모두에서 총 IgE치와 Der f II 특이 IgE를 측정하였고 이들의 말초혈액에서 T세포를 Der f II로 자극하여 배양한 후 DNA 추출을 하여 IL-4 유전자 두 번째 인트론의 CpG 섬에 대한 메틸화 분석을 실시하였다. 결 과 : 정상군에 비해 알레르기 환아군에서 IL4 유전자의 두번째 인트론에 위치한 CpG 섬의 두 부위 모두에서 메틸화 특이 PCR 신호강도가 약하게 나타나는 반면 정상아에서는 118 bp PCR 산물과 205 bp PCR 산물이 모두 강하게 나타나서 메틸화된 것으로 보이며, 저위험군 알레르기 환아에서 최소한 한 부위에서 탈메틸화가 일어났으며 고위험군 알레르기 환아에서는 두 부위 모두에서 탈메틸화가 일어났다. 결 론 : 정상 대조군의 T 세포에서는 IL-4 유전자의 메틸화가 일어나고 알레르기 환아에서 고위험군과 저위험군 모두 정도의 차이는 있으나 메틸화의 소실이 관찰되었다. 이를 이용하여 향후 영유아기에서 아토피 유무를 판별할 수 있는 가능성을 제시하였으나 더 많은 연구가 필요할 것으로 사료된다.

Keywords

Acknowledgement

Supported by : 대한소아과학회

References

  1. Hagendorens MM, Van Bever HP, Schuerwegh AJ, De Clerck LS, Bridts CH, Stevens WJ. Determination of T-cell subpopulations and intracellular cytokine production(interleukin-2, interleukin-4, and interferon-$\gamma$) by cord blood Tlymphocytes of neonates from atopic and non-atopic parents. Pediatr Allergy Immunol 2000;11:12-9
  2. Host A. Development of atopy in childhood. Allergy 1997;52:695-7. https://doi.org/10.1111/j.1398-9995.1997.tb01225.x
  3. Ricci M, Rossi O, Bertoni M, Matucci S. The importance of Th2-like cells in the pathogenesis of airway allergic inflammation. Clin Exp Allergy 1993;23:360-9 https://doi.org/10.1111/j.1365-2222.1993.tb00340.x
  4. Mosmann TR, Sad S. The expanding universe of T-cell subsets : Th1, Th2 and more. Immunol Today 1996;17:138-46 https://doi.org/10.1016/0167-5699(96)80606-2
  5. Kopp MV, Zehle C, Pichler J, Szepfalusi Z, Moseler M, Deichmann K, et al. Allergen-specific T cell reactivity in cord blood : the influence of maternal cytokine production. Clin Exp Allergy 2001;31:1536-43. https://doi.org/10.1046/j.1365-2222.2001.01198.x
  6. Kass SU, Pruss D, Wolffe AP. How does DNA methylation repress transcription? Trends Genet 1997;13:444-9 https://doi.org/10.1016/S0168-9525(97)01268-7
  7. Bird JJ, Brown DR, Mullen AC, Moskowitz NH, Mahowald MA, Sider JR, et al. Helper T cell differentiation is controlled by the cell cycle. Immunity 1998;9:229-37 https://doi.org/10.1016/S1074-7613(00)80605-6
  8. Li E. The mojo of methylation. Nat Genet 199;23:5-6 https://doi.org/10.1038/12595
  9. Bullens DMA, Rafiq K, Kasran A, van Gool SW, Ceuppens JL. Naive human T cells can be a source of IL-4 during primary immune responses. Clin Exp Immunol 1999;118: 384-91 https://doi.org/10.1046/j.1365-2249.1999.01072.x
  10. Bhattacharya SK, Ramchandani S, Cervoni N, Szyf M. A mammalian protein with specific demethylase activity for mCpG DNA. Nature 1999;397:579-69 https://doi.org/10.1038/17492
  11. Santangelo S, Cousins DJ, Winkelmann NEE, Staynov DZ. DNA methylation changes at human Th2 cytokine genes coincide with DNase I hypersensitive site formation during CD4＋ T cell differentiation. J Immunol 2002;169:1893-903
  12. Hide DW, Arshad SH, Twiselton R, Stevens M. Cord serum IgE : an insensitive method for prediction of atopy. Clin Exp Allergy 1991;21:739-45 https://doi.org/10.1111/j.1365-2222.1991.tb03204.x
  13. Kjellman NIM, Croner S. cord blood IgE determination for allergy prediction-a follow-up to seven years of age in 1,651 children. Ann Allergy 1984;53:167-74
  14. Kondo N, Kobayashi Y, Shinoda S. Cord blood lymphocyte responses to food antigens for the prediction of allergic disorders. Arch Dis Child 1992;67:1003-7 https://doi.org/10.1136/adc.67.8.1003
  15. Yabuhara A, Macaubas C, Prescott SL, Venaille TJ, Holt BJ, Habre W, et al. Th2-polarized immunological memory to inhalant allergens in atopics is established during infancy and early childhood. Clin Exp Allergy 1997;27:1261-9 https://doi.org/10.1111/j.1365-2222.1997.tb01170.x
  16. Maggi E, Parronchi P, Manetti P. Reciprocal regulatory effects of IFN-gamma and IL-4 on the in vitro development of human Th1 and Th2 clones. J Immunol 1992;148:2142-7
  17. Rengarajan J, Szabo SJ, Glimcher LH. Transcriptional regulation of Th1/Th2 polarization. Immunol Today 2000;21: 479-83 https://doi.org/10.1016/S0167-5699(00)01712-6
  18. Grogan JL, Locksley RM. T helper cell differentiation : on again, off again. Curr Opin Immunol 2002;14:366-72 https://doi.org/10.1016/S0952-7915(02)00340-0
  19. Guo L, Hu-Li J, Zhu J, Watson CJ, Difilippantonia MJ, Pannetier C, et al. In Th2 cells the IL4 gene has a series of accessibility states associated with distinctive probabilities of IL-4 prodution. PNAS 2002;99:10623-8 https://doi.org/10.1073/pnas.162360199
  20. Lee DU, Agarwal S, Rao A. The lineage commitment and efficient IL-4 production involves extended demethylation of the IL-4 gene. Immunity 2002;16:649-60 https://doi.org/10.1016/S1074-7613(02)00314-X
  21. Cedar H, Verdine GL. The amazing demethylase. Nature 1999;397:568-569 https://doi.org/10.1038/17492
  22. Danbara M, Kameyama K, Higashihara M, Takagaki Y. DNA methylation dominates transcriptiona silencing of Pax5 in terminally differentiated B cell lines. Mole Immunol 2001;38:1161-6