モノクロナ-ル抗体,遺伝子を用いた耐性癌の基礎的な診断と治療に関する研究

1991 Fiscal Year Annual Research Report

• Project/Area Number: 03151028
• Research Institution: Mie University
• Principal Investigator: 白川 茂 三重大学, 医学部, 教授 (20026850)
• Co-Investigator(Kenkyū-buntansha): 曽根 三郎 徳島大学, 医学部附属病院, 講師 (40145024) ; 玉置 憲一 東海大学, 医学部, 教授 (50055860) ; 上田 龍三 愛知県がんセンター研究所, 化学療法部, 部長 (20142169) ; 下山 正徳 国立がんセンター病院, 薬物療法部, 部長 (50110002) ; 鶴尾 隆 東京大学, 応用微生物研究所, 教授 (00012667)
• Keywords: 多剤耐性腫瘍 / P糖蛋白(Pーglycoprotein;Pーgp) / 抗Pーgp抗体MRK16 / 抗Pーgp抗体C219 / 多剤耐性遺伝子(mdr1) / mdr1mRNA発現 / ヒトマウスキメラ抗体 / magneticーaffinity cell sorring(MACS)法

Research Abstract

(1)基礎的に抗P糖蛋白(Pーgp)抗体MRK16(鶴尾)の反応性とmdr1遺伝子のmRNA発現程度を培養細胞株で検討し,中等度以上のmRNA発現細胞株でMRK16陽性であった(下山)。(2)ヒト腫瘍nude mouse移植株47株のうち多剤耐性を示す33株中21株でRTーPCR法によりmdr1遺伝子の発現を認め,このうち8株が抗Pーgp抗体C219陽性であった。なおMRK16とC219による固型腫瘍材料の免疫染色至適条件が決定された(玉置)。(3)臨床材料につきmdr1遺伝子発現を6種類のamplimerを用いてRTーPCR法で検索し,amplimerは200〜300bp前後のものが感度が高く,少量の試料で解析可能であった(向山)。(4)白血病,悪性リンパ腫などの造血器腫瘍でのPーgpの関与はin vivoの耐性細胞株で確認された。in vivoの症例検索がなされ,128例の造血器腫瘍でMRK16 F(ab')_2抗体(鶴尾)を用いた間接法で11例陽性,直接法では3例(リンパ腫再発例)の化学療法不応例のみに陽性であった(上田)。また64例の各種白血病症例で同上抗体でのFACS解析陽性例は8例で,うち5例は単球性性格を示し,経過を追跡し得た治療抵抗性8例でmdr1遺伝子発現を経過中に認めたものはPh(+)ALL,MDS各1例で臨床上検出限界に問題が残った(白川)。(5)固型癌については,MRK16の肺癌培養細胞42株の検討では全例陰性であった(上田)が,C219の免疫染色で肺癌,転移性肺腫瘍76例中26例(24%)に陽性(広橋),大腸癌ではC219で高率(68%)に陽性(玉置),また大腸癌,未分化甲状腺癌,肝癌,胎盤など(菅原),乳癌,卵巣癌など(向山)でPTーPCR法によるmdr1mRNAの有意な発現があり,化学療法抵抗性と関連があった。(6)MRK16とMRK17はADCC誘導に際し,リンパ球はMRK16においてのみeffectorとして,単球は両抗体において働くことが判明した。またヒトマウスキメラ抗体の臨床応用への可能性が示唆された(曽根,鶴尾)。(7)MRK16とmagnetic particleをcouplingしたbeadsを用いるmagneticーaffinity cell sorting(MACS)法が開発され,Pーgp発現多剤耐性細胞の分離に成功した。

Research Products

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