KREC/TRECによる先天性免疫不全症スクリーニング法開発と病態解析への応用

2011 Fiscal Year Annual Research Report

• Project/Area Number: 22390213
• Research Institution: National Defense Medical College
• Principal Investigator: 野々山 恵章 防衛医科大学校, 医学教育部・医学科専門課程, 教授 (40280961)
• Co-Investigator(Kenkyū-buntansha): 大嶋 宏一 京都大学, iPS細胞研究所, 特定研究員 (60525377) ; 今井 耕輔 東京医科歯科大学, 大学院・医歯学総合研究科, 准教授 (90332626) ; 小原 收 (財)かずさDNA研究所, ヒトゲノム研究部, 副所長 (20370926)
• Keywords: 免疫学 / 遺伝子 / 移植・再生医療 / 発生・分化 / 感染症

Research Abstract

先天性免疫不全症は診断が困難で、治療の遅れなどにより感染症が重症化し致命的になる事がある。一方で、早期診断により適切な治療を行えば予後が劇的に改善する。また、病態解明は、より適切な治療法の選択につながる。先天性免疫不全症の新生児スクリーニングと病態解析を目的に以下の研究を行った。
a)TREC/KREC測定による免疫不全症新生児スクリーニング法の確立
TREC(T cell receptor recombination circles)とKREC(Kappa-chain recombination excision circles)は、T細胞新生、B細胞新生の良いマーカーである。患者および健常者からえた乾燥濾紙血のTREC/KRECを測定した。その結果、全例で健常新生児ではTREC/KRECともに陽性、T細胞欠損患者ではTRECが陰性、B細胞欠損患者ではKRECが陰性であった。さらに、Ataxia telangiectasiaではKRECが全例陰性であることも見出した。以上、新生児マススクリーニングが可能であることを示した。
b)TREC/KREC測定による免疫不全症の病態解明
先天性免疫不全症の中で最も頻度の高いCommon variable immunodeficiency(CVID,分類不能型免疫不全症)患者43例でTREC,KRECの測定を行った。その結果、A群(TREC(+),KREC(+)),B群(TREC(+),KREC(-))、C群(TREC(-),KREC(+))、D群(TREC(-),KREC(-))の4群に分かれることを見出した。4群で臨床症状、予後が異なることを明らかにした。γグロブリン定期補充で予後が良いA群、造血幹細胞移植が必要なD群が鑑別でき、TREC/KREC測定により免疫不全症の病態に即した治療法が選択できることを示した。さらにTREC/KREC陰性群からGATA2欠損、RAG1欠損,LigIV欠損症を見出し、原因遺伝子同定にも役立つことも示した。

Current Status of Research Progress

1: Research has progressed more than it was originally planned.

Reason

新生児スクリーニングの新規技術を開発ができた。また、TREC/KRECでCVIDが4群に分けられ、重症度と相関すること、γグロブリン補充に加え、移植適応などの治療法の選択に有用であること、GATA2変異患者で世界で初めてTREC/KRECが陰性であることを見出し、病態解明につなげたことなど、計画以上の実績がえられた。

Strategy for Future Research Activity

TREC/KRECにより4群に分けたCVID患者の原因遺伝子を、Exome解析により同定していく。Exome解析で得られた候補遺伝子が真の原因遺伝子であることを、患者同意下で作製してあるiPS細胞に変異のない正常遺伝子を導入し、B細胞、T細胞、好中球、樹状細胞などの免疫担当細胞の分化が正常化することを示す方法で、確定する。
原因遺伝子の機能解析により、CVID患者の病態を明らかにし、より適切な治療法を開発する。将来的な遺伝子治療にも結びつく。

Research Products

• [Journal Article] The kinase Btk negatively regulates the production of reactive oxygen species and stimulation-induced apoptosis in human neutrophils (2012)
  • Author(s): Honda F, Kano H, Kanegane H, Nonoyama S, Kim ES, Lee SK, Takagi M, Mizutani S, Morio T
  • Journal Title: Nat Immunol Volume: 26 Pages: 369-378
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• [Journal Article] GATA-2 anomaly and clinical phenotype of a sporadic case of lymphedema, dendritic cell, monocyte, B- and NK-cell (DCML) deficiency, and myelodysplasia (2012)
  • Author(s): Ishida H, Imai K, Homma K, Tamura S, Imamura T, Itoh M, Nonoyama S
  • Journal Title: Eur J Pediatr Volume: 171 Pages: 1273-1276
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• [Journal Article] Hyper-eosinophilia in granular acute B-cell lymphoblastic leukemia with myeloid antigen expression (2012)
  • Author(s): Kobayashi D, Kogawa K, Imai K, Tanaka T, Sada A, Nonoyama S
  • Journal Title: Pediatr Int Volume: (in press)
  • Peer Reviewed
• [Journal Article] Clinical and genetic characteristics of XIAP deficiency in Japan (2012)
  • Author(s): Yang X, Kanegane H, Nishida N, Imamura T, Hamamoto K, Miyashita R, Imai K, Nonoyama S, Sanayama K, Yamaide A, Kato F, Nagai K, Ishii E, Zelm M, Latour S, Zhao X, Miyawaki T
  • Journal Title: J Clin Immunol Volume: (in press)
  • Peer Reviewed
• [Journal Article] Functional STAT3 deficiency compromises IL-12-dependent generation of human T-follicular helper cells (2012)
  • Author(s): Ma CS, Avery DT, Chan A, Batten M, Bustamante J, Boisson-Dupuis S, Arkwright PD, Minegishi Y, Nonoyama S, French MA, Choo S, Peake J, Wong M, Cook MC, Fulcher DA, Casanova JL, Deenick EK, Tangye SG
  • Journal Title: Blood Volume: (in press)
  • Peer Reviewed
• [Journal Article] Missense Mutations of the BMPR1B (ALK6) Gene in Childhood Idiopathic Pulmonary Arterial Hypertension (2012)
  • Author(s): Chida A, Shintani M, Nakayama T, Furutani Y, Hayama E, Inai K, Saji T, Nonoyama S, Nakanishi T
  • Journal Title: Circ J Volume: (in press)
  • Peer Reviewed
• [Journal Article] Clinical profile and genetic basis of Wiskott-Aldrich syndrome at Chandigarh, north India (2012)
  • Author(s): Suri D, Singh S, Rawat A, Gupta A, Kamae C, Honma K, Nakagawa N, Imai K, Nonoyama S, Oshima K, Mitsuiki N, Ohara O, Bilhou-Nabera C, Proust A, Ahluwalia J, Dogra S, Saikia B, Walker Minz R, Sehgal S
  • Journal Title: Asian Pac J Allergy Immunol Volume: 30 Pages: 1-8
  • Peer Reviewed
• [Journal Article] Factors Associated with Steroid Phobia in Caregivers of Children with Atopic Dermatitis (2012)
  • Author(s): Kojima R, Fujiwara T, Matsuda A, Narita M, Matsubara O, Nonoyama S, Ohya Y, Saito H, Matsumoto K
  • Journal Title: Pediatr Dermatol Volume: (in press)
  • Peer Reviewed
• [Journal Article] Endocrine complications in primary immunodeficiency diseases in Japan (2012)
  • Author(s): Nozaki T, Takada H, Ishimura M, Ihara K, Imai K, Morio T, Kobayashi M, Nonoyama S, Hara T
  • Journal Title: Clin Endocrinol Volume: (in press)
  • Peer Reviewed
• [Journal Article] Survey of Japanese infants younger than 3 months who were treated with oseltamivir for influenza : Safety of oseltamivir treatment (2012)
  • Author(s): Morioka I, Nonoyama S, Tanaka-Taya K, Ihara T, Sugaya N, Ueta I, Kumagai T, Okada K, Hosoya M, Okabe N, Morishima T
  • Journal Title: Scand J Infect Dis Volume: (in press)
  • Peer Reviewed
• [Journal Article] Multiple reversions of an IL2RG mutation restore T cell function in an X-linked severe combined immunodeficiency patient (2012)
  • Author(s): Kawai T, Saito M, Nishikomori R, Yasumi T, Izawa K, Murakami T, Okamoto S, Mori Y, Nakagawa N, Imai K, Nonoyama S, Wada T, Yachie A, Ohmori K, Nakahata T, Heike T
  • Journal Title: J Clin Immunol Volume: (in press)
  • Peer Reviewed
• [Journal Article] Outcomes of Childhood Pulmonary Arterial Hypertension in BMPR2 and ALK1 Mutation Carriers (2012)
  • Author(s): Chida A, Shintani M, Yagi H, Fujiwara M, Kojima Y, Sato H, Imamura S, Yokozawa M, Onodera N, Horigome H, Kobayashi T, Hatai Y, Nakayama T, Fukushima H, Nishiyama M, Doi S, Ono Y, Yasukouchi S, Ichida F, Fujimoto K, Ohtsuki S, Teshima H, Kawano T, Nomura Y, Gu H, Ishiwata T, Furutani Y, Inai K, Saji T, Matsuoka R, Nonoyama S, Nakanishi T
  • Journal Title: Am J Cardiol Volume: (in press)
  • Peer Reviewed
• [Journal Article] Delayed onset adenosine deaminase deficiency associated with acute disseminated encephalomyelitis (2012)
  • Author(s): Nakaoka H, Kanegane H, Taneichi H, Miya K, Yang X, Nomura K, Takezaki S, Yamada M, Ohara O, Kamae C, Imai K, Nonoyama S, Wada T, Yachie A, Hershfield MS, Ariga T, Miyawaki T
  • Journal Title: Int J Hematol Volume: (in press)
  • Peer Reviewed
• [Journal Article] Severe primary antibody deficiency due to a novel mutation of micro heavy chain (2012)
  • Author(s): Mohammadzadeh I, Yeganeh M, Aghamohammadi A, Parvaneh N, Behniafard N, Abolhassani H, Tabassomi F, Hemmat M, Kanegane H, Miyawaki T, Ohara O, Rezaei N
  • Journal Title: J Investig Allergol Clin Immunol Volume: 22 Pages: 78-79
  • Peer Reviewed
• [Journal Article] Quantification of κ-deleting recombination excision circles in Guthrie cards for the identification of early B-cell maturation defects (2011)
  • Author(s): Nakagawa N, Imai K, Kanegane H, Sato H, Yamada M, Kondoh K, Okada S, Kobayashi M, Agematsu K, Takada H, Mitsuiki N, Oshima K, Ohara O, Suri D, Rawat A, Singh S, Pan-Hammarstrom Q, Hammarstrom L, Reichenbach J, Seger R, Ariga T, Hara T, Miyawaki T, Nonoyama S
  • Journal Title: J Allergy Clin Immunol Volume: 128 Pages: 223-225
  • DOI: doi: 10.1016/j.jaci.2011.01.052.
• [Journal Article] Analysis of mutations and recombination activity in RAG-deficient patients (2011)
  • Author(s): Asai E, Wada T, Sakakibara Y, Toga A, Toma T, Shimizu T, Nampoothiri S, Imai K, Nonoyama S, Morio T, Muramatsu H, Kamachi Y, Ohara O, Yachie A
  • Journal Title: Clin Immunol Volume: 138 Pages: 172-7
  • DOI: doi: 10.1016/j.clim.2010.11.005.
• [Journal Article] Rapid diagnosis of FHL3 by flow cytometric detection of intraplatelet Munc13-4 protein (2011)
  • Author(s): Murata Y, Yasumi T, Shirakawa R, Izawa K, Sakai H, Abe J, Tanaka N, Kawai T, Oshima K, Saito M, Nishikomori R, Ohara O, Ishii E, Nakahata T, Horiuchi H, Heike T
  • Journal Title: Blood Volume: 118(5) Pages: 1225-30
  • DOI: doi: 10.1182/blood-2011-01-329540.
• [Journal Article] ADA-SCID with 'WAZA-ARI' mutations that synergistically abolished ADA protein stability (2011)
  • Author(s): Okura Y, Yamada M, Kobayashi I, Santisteban I, Arredondo-Santisteban G, Kato Z, Iguchi A, Yoshida M, Ohara O, Nakagawa N, Imai K, Hershfield MS, Ariga T
  • Journal Title: Br J Haematol Volume: 153(5) Pages: 675-6
  • DOI: doi: 10.1111/j.1365-2141.2011.08640.x.
• [Journal Article] Early and rapid detection of X-linked lymphoproliferative syndrome with SH2D1A mutations by flow cytometry (2011)
  • Author(s): Zhao M, Kanegane H, Kobayashi C, Nakazawa Y, Ishii E, Kasai M, Terui K, Gocho Y, Imai K, Kiyasu J, Nonoyama S, Miyawaki T
  • Journal Title: Cytometry B Clin Cytom Volume: 80 Pages: 8-13
  • Peer Reviewed
• [Journal Article] Contribution of Sarcoplasmic Reticulum Ca2+ Release and Ca2+ Transporters on Sarcolemmal Channels to Ca2+ Transient in Fetal Mouse Heart (2011)
  • Author(s): Takizawa M, Ishiwata T, Kawamura Y, Kanai T, Kurokawa T, Nishiyama M, Ishida H, Asano Y, Nonoyama S
  • Journal Title: Pediatr Res Volume: 69 Pages: 306-311
  • Peer Reviewed
• [Journal Article] Defective IL-10 signaling in hyper-IgE syndrome results in impaired generation of tolerogenic dendritic cells and induced regulatory T cells (2011)
  • Author(s): Saito M, Nagasawa M, Takada H, Hara T, Tsuchiya S, Agematsu K, Yamada M, Kawamura N, Ariga T, Tsuge I, Nonoyama S, Karasuyama H, Minegishi Y
  • Journal Title: J Exp Med Volume: 208 Pages: 235-249
  • Peer Reviewed
• [Journal Article] Severe human herpesvirus 6 associated encephalopathy in three children : Analysis of cytokine profiles and the carnitine palmitoyltransferase 2 gene (2011)
  • Author(s): Matsumoto H, Hatanaka D, Ogura Y, Chida A, Nakamura Y, Nonoyama S
  • Journal Title: Pediatr Infect Dis J Volume: 30 Pages: 999-1001
  • Peer Reviewed
• [Journal Article] Nationwide Survey of Patients with Primary Immunodeficiency Diseases in Japan (2011)
  • Author(s): Ishimura M, Takada H, Doi T, Imai K, Sasahara Y, Kanegane H, Nishikomori R, Morio T, Heike T, Kobayashi M, Ariga T, Tsuchiya S, Nonoyama S, Miyawaki T, Hara T
  • Journal Title: J Clin Immunol Volume: 31 Pages: 968-976
  • Peer Reviewed
• [Journal Article] Primary Immunodeficiency Diseases : an update on the Classification from the International Union of Immunological Societies Expert Committee for Primary Immunodeficiency (2011)
  • Author(s): Al-Herz W, Bousfiha A, Casanova JL, Chapel H, Conley ME, Cunningham-Rundles C, Etzioni A, Fischer A, Franco JL, Geha RS, Hammarstrom L, Nonoyama S, Notarangelo LD, Ochs HD, Puck JM, Roifman C, Seger R, Tang MLK
  • Journal Title: Front Immun Volume: 2 Pages: 54
  • Peer Reviewed
• [Journal Article] Quantification of human herpesvirus-6 (HHV-6) DNA in a cord blood transplant recipient with chromosomal integration of HHV-6 (2011)
  • Author(s): Kobayashi D, Kogawa K, Imai K, Tanaka T, Hiroi S, Satoh H, Tanaka-Taya K, Nonoyama S
  • Journal Title: Transpl Infect Dis Volume: 13 Pages: 650-653
  • Peer Reviewed
• [Journal Article] Ulinastatin, a Urinary Trypsin Inhibitor, for the Initial Treatment of Patients with Kawasaki (2011)
  • Author(s): Kanai T, Ishiwata T, Kobayashi T, Sato H, Takizawa M, Kawamura Y, Tsujimoto H, Nakatani K, Ishibashi N, Nishiyama M, Hatai Y, Asano Y, Kobayashi T, Takeshita S, Nonoyama S
  • Journal Title: Circulation Volume: 124 Pages: 2822-2828
  • Peer Reviewed
• [Journal Article] Cardiopulmonary arrest caused by craniometaphyseal dysplasia (2011)
  • Author(s): Chida A, Yanagawa Y, Matsumoto H, Nonoyama S
  • Journal Title: Indian J Pediatr Volume: 78 Pages: 1010-1012
  • Peer Reviewed
• [Journal Article] Clinical and host genetic characteristics of Mendelian susceptibility to mycobacterial diseases in Japan (2011)
  • Author(s): Hoshina T, Takada H, Sasaki-Mihara Y, Kusuhara K, Oshima K, Okada S, Kobayashi M, Ohara O, Hara T
  • Journal Title: J Clin Immunol Volume: 31 Pages: 309-314
  • Peer Reviewed
• [Journal Article] A novel serum-free monolayer culture for orderly hematopoietic differentiation of human pluripotent cells via mesodermal progenitors (2011)
  • Author(s): Niwa A, Heike T, Umeda K, Oshima K, Kato I, Sakai H, Suemori H, Nakahata T, Saito MK.Jul
  • Journal Title: PLoS One Volume: 6 Pages: e22261
  • Peer Reviewed
• [Journal Article] High incidence of NLRP3 somatic mosaicism in patients with chronic infantile neurologic, cutaneous, articular syndrome : results of an International Multicenter Collaborative Study (2011)
  • Author(s): Tanaka N, Izawa K, Saito MK, Sakuma M, Oshima K, Ohara O, Nishikomori R, Morimoto T, Kambe N, Goldbach-Mansky R, Aksentijevich I, de Saint Basile G, Neven B, van Gijn M, Frenkel J, At?stegui JI, Yag?e J, Merino R, Iba?ez M, Pontillo A, Takada H, Imagawa T, Kawai T, Yasumi T, Nakahata T, Heike T
  • Journal Title: Arthritis Rheum Volume: 63 Pages: 3625-3632
  • DOI: doi:10.1002/art.30512
  • Peer Reviewed
• [Journal Article] Novel mutations of MVK gene in Japanese family members affected with hyperimmunoglobulinemia D and periodic fever syndrome (2011)
  • Author(s): Mizuno T, Sakai H, Nishikomori R, Oshima K, Ohara O, Hata I, Shigematsu Y, Ishige T, Tamura K, Arakawa H
  • Journal Title: Rheumatol Interest Volume: (in press)
  • Peer Reviewed
• [Remarks]
  • URL: http://pidj.rcai.riken.jp/