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Study on the pathological crosstalk between reproduction and immunity leading to abnormal urogenital phenotypes in autoimmune disease model mice

大谷, 祐紀 北海道大学

2021.03.25

概要

哺乳類の生殖と免疫機能は密に連関し、両者の機能を相互に制御する。卵巣内には免疫細胞が常在し、卵胞の成熟や排卵、黄体形成とその退行等、正常な雌性生殖機能の維持を担う。対して精巣では、セルトリ細胞等の精巣構成細胞と常在免疫細胞が特異的な免疫抑制環境“免疫学的特権領域”を形成し、精細胞は全身免疫から保護される。また、種々の免疫細胞は性ホルモン受容体を発現し、主にエストロゲンは免疫活性に、アンドロゲンはその抑制に寄与する。また、性染色体には多くの免疫関連遺伝子が座位する。

 このような生殖と免疫機能の相互制御機構“生殖-免疫連関”において、一方の機能変化は他方の病態を誘導する。女性では、40歳未満で生殖機能を失う早発閉経患者の約30%が全身性エリテマトーデス(SLE)や自己免疫性甲状腺炎(橋本病)等の自己免疫異常を呈する。また、自己免疫疾患の病態形成には性ホルモンが強く影響し、女性のSLE罹患率は閉経前で男性よりも8-15倍高く、性成熟前や閉経後で低下する。男性の自己免疫疾患罹患者も高率に抗精子抗体を有し、ヒトと実験動物における化学物質や感染、外傷等による精巣内免疫抑制環境の破壊は、精上皮細胞死を特徴とする造精障害を導く。一方、男性SLE患者の腎炎や皮膚炎は女性患者よりも重篤化する。

 このような生殖-免疫連関の不均衡は現代のヒトと動物の共通事象である。ワクチン接種や感染症罹患は全身免疫状態を変化させ、不妊症を代表とする生殖機能障害は、ヒトでは少子化社会、動物では種の絶滅を導き、生物多様性の維持を脅かし得る種横断的な課題となる。一方、動物福祉学的観点から、日本では半数以上の飼養犬猫が不妊化され、生産獣医療分野では人社会への安定した食糧供給のため、動物の人為的な妊娠・泌乳状態を継続的に作出する。重要なことに、性腺を摘出したイヌでは甲状腺機能低下症や血小板減少症等の免疫関連疾患罹患率が増加する。これらの証拠は、生殖-免疫連関の変化に起因する双方向性の病態形成を示唆するが、その機序は不明である。そこで本研究は、生殖および免疫機能病態モデルの表現型を詳細に解析し、当該連関における相互制御機構とその変化に起因する病態形成メカニズムの解明を目的とした。

 第一章では、自己免疫疾患モデルMRL/MpJ-Faslpr/lprマウスの雌性生殖機能を経時的に観察し、自己免疫異常と卵巣形態機能の関連について考察した。第二章では、雄の自己免疫疾患モデルBXSB/MpJ-Yaaマウスの精巣について、形態的および分子生物学的に解析を行い、自己免疫異常が精巣特異的な免疫環境に与える影響とそのメカニズムについて考察した。第三章では、精巣摘出を施したBXSB/MpJ-Yaaの表現型および病態形成機序について探索し、アンドロゲンが自己免疫疾患および関連病態形成に与える影響を考察した。

 はじめに、雌MRL/MpJ-Faslpr/lprの病態解析において、自己免疫疾患発症に伴い、卵巣への顕著な免疫細胞浸潤と、原始卵胞を含む卵胞数および黄体数の有意な減少がみられた。性周期について、MRL/MpJ-Faslpr/lprは発情期の短縮と発情休止期の延長を特徴とする周期異常を対称群よりも早期に示し、雌性生殖機能の異常が示唆された。一方、雄BXSB/MpJ-Yaaの解析において、全身自己免疫異常を呈する一方、精巣内の免疫細胞数や炎症関連分子の発現量は変化しなかった。しかし、組織学的解析において、精母細胞アポトーシス数は健常群よりも有意に増加し、その増加は精上皮ステージXIIで顕著だった。精細管内ではresidual bodyの増加とセルトリ細胞の減少がみられ、全身性自己免疫異常に伴うセルトリ細胞の機能形質変化が示唆された。以上より、マウス生殖-免疫連関において、全身性自己免疫疾患は雌性および雄性生殖機能を障害することを見出した。一方、その病態形成機序は雌雄で異なり、雌では免疫細胞等の炎症因子が卵巣構成細胞に直接的に作用するのに対し、雄では全身免疫の変化が、炎症機構を介さず精巣の免疫学的環境を変化させ、生殖機能の異常をもたらす可能性が考えられた。

 他方、精巣を摘出したBXSB/MpJ-Yaaの自己免疫疾患発症は擬手術群よりも早期化傾向であったが、病態後期の重篤度は変化しなかった。一方、腎病理解析において、精巣摘出群の病態後期における糸球体硬化病変は、擬手術群よりも有意に軽減した。重要なことに、免疫染色において、アンドロゲン受容体は糸球体構成細胞にはみられず、腎小体内では糸球体包外壁の上皮細胞(PEC)の核内に局在し、さらに総PEC数および活性化PEC数は精巣摘出群で有意に減少した。上記の結果より、アンドロゲンはBXSB/MpJ-Yaaの糸球体硬化病変を増悪させ、その病理形成にはPECが関与すると考えられた。

 結論として、本研究はマウスの生殖-免疫連関の変化が尿生殖器に病態を導くことを示した。特に病理組織学的手法により、各臓器の形態機能変化をもたらす実動細胞を特定し、生殖-免疫連関を基盤とする病態形成機構において新たな知見を見出した。一方、解析した疾患モデルマウスの表現型形成には生殖-免疫連関の不均衡に加え、遺伝的素因の関与が疑われた。獣医療においても、動物種、血統または品種特異的な遺伝的多型は疾患発症の一因となり、生殖-免疫連関異常との関連性評価は意義深い。汎動物学的観点より、動物から得る医科学的知見は動物のみならずヒト疾患の診断・治療に応用でき、獣医学領域における基礎研究の発展は動物とヒトの健全性達成に欠かせない。以上、本研究は哺乳類の生殖-免疫連関における新たな基礎知見の提供を通じ、動物とヒトの免疫異常、生殖機能障害およびそれらの病態連関の解明に寄与するものである。

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