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Summary in Japanese
研究の背景
国際社会ではこれまでに、COVID-19 をはじめとした様々な新興・再興感染症が出現
し、公衆衛生上の脅威となっている。約 3/4 以上の新興・再興感染症は人獣共通感染症で
あり、中でもウイルス感染症の多くは野生動物からの伝播を経て出現したと考えられる。
そのため、野生動物の保有するウイルスの調査はワンヘルスの観点から重要である。近
年、メタゲノム解析技術の普及に伴い、多くの検体から様々な新規ウイルスゲノムが検出
されている。しかし、ほとんどの報告はゲノム解析に留まっており、ウイルス分離および
ウイルス学的な性状解析の試みは十分ではない。特に、プロテアーゼ依存性ウイルスは、
培養細胞での増殖時に無血清培地下でトリプシンを作用させる必要があるため、検体の接
種による細胞傷害が生じやすく、ウイルス分離が困難である。これまで、宿主 II 型膜貫通
型セリンプロテアーゼ（TTSP）発現細胞において、トリプシン非存在下においてもプロテ
アーゼ依存性ウイルスの感染増殖が可能であることが明らかになっているが、プロテアー
ゼ発現細胞を野生動物からの網羅的なウイルス分離に用いた例は少ない。そこで本研究で
は、プロテアーゼ発現細胞を用いて野生動物由来検体からウイルス分離を実施し、分離し
たウイルスのウイルス学的および疫学的性状解析を実施することを目的とした。
第一章：新型コロナウイルス（SARS-CoV-2）スパイクタンパク質を活性化す
る TTSP の検討
まず、代表的なプロテアーゼ依存性ウイルスである SARS-CoV-2 の感染を促進する
TTSP を検索した。SARS-CoV-2 は、TMPRSS2 などの TTSP を利用して、ウイルスのスパ
イクタンパク質を切断・活性化し細胞侵入効率を上昇させるが、SARS-CoV-2 の感染にお
ける TMPRSS2 以外の TTSP の役割については解明されていない。本章では、ヒト ACE2
発現 HEK293T 細胞および Vero E6 細胞を用いて 12 種類の TTSP をスクリーニングし、
TMPRSS11D および TMPRSS13 が SARS-CoV-2 の感染増殖を促進することを明らかにし
た。また、SARS-CoV-1 と SARS-CoV-2 は、ウイルス侵入過程において同様の TTSP を利
用することが明らかになった。本研究は、宿主の TTSP が SARS-CoV-2 の感染に影響を与
えることを示し、細胞や組織の指向性、病原性に影響することを示唆している。
105
第二章：潜在的レゼルボアである齧歯類動物マストミスからの脳心筋炎ウイルス
（EMCV）の分離および性状解析
EMCV は様々な哺乳類動物に感染し、脳炎、心筋炎、生殖障害、糖尿病などを引き起こ
す。特に、養豚場における繁殖障害や、動物園や野生動物保護区等での希少動物の突然死
は経済・環境の観点から重要な問題である。動物との接触によってヒトへも稀に感染し、
発熱性疾患を引き起す人獣共通感染症である。本章では、ザンビアで採取した齧歯類動物
（マストミス、Mastomys natalensis）から EMCV ZM12/14 株を分離し、性状解析を実施し
た。系統解析の結果，ZM12/14 株は P1 および P3 領域で EMCV-1 と同じ系統に分類された
が、P2 領域では既知の EMCV 株と異なる系統に分類され、特徴的な進化系統を有するこ
とが示唆された。さらに、ザンビア各地で捕獲した各種齧歯類動物（n=179）の組織およ
び血清を用いて、EMCV の RT-PCR スクリーニングと中和抗体測定を実施した。その結
果、M. natalensis のみから EMCV ゲノム（19/179 = 10.6%）および中和抗体が検出され
（33/179 = 18.4%）、RT-PCR 陽性、陰性検体どちらにおいても、高い中和抗体価（≧
320）が確認された。他種の齧歯類動物ではゲノムおよび中和抗体は検出されなかった。
また、ZM12/14 株を腹腔内接種した BALB/c マウスは無症候性の持続感染を引き起こし
た。感染マウスでは感染 2 週間後において、高い中和抗体価と脳および脾臓における高い
ウイルスゲノム量が認められ、上記の疫学調査と一致する結果が得られた。本研究では、
ザンビアで初めて EMCV を検出し、M. natalensis が自然感染宿主としてレゼルボアの役割
を担うことを示唆する結果を得た。
第三章：コウモリおよび齧歯類動物由来ロタウイルス A（RVA）の分離と性
状解析
RVA は、ヒトや様々な動物に下痢性疾患を引き起こす。近年、ヒト RVA との遺伝子再
集合を示唆するコウモリおよび齧歯類動物由来 RVA が複数報告されている。しかし、
様々な非典型的遺伝子型が含まれるコウモリおよび齧歯類動物由来 RVA のウイルス学的
性状はほとんど解明されていない。そこで、ヒト TMPRSS2/TMPRSS11D 共発現 MA104 細
胞とザンビアで採取した野生動物検体を用いて RVA のウイルス分離先行型スクリーニン
グを実施し、コウモリ（Rousettus aegyptiacus）およびマストミス（M. natalensis）から
RVA を単離した。全ゲノム配列解析の結果、コウモリ由来 RVA 16-06 株の遺伝子型は、
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