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インフルエンザウイルス肺炎における活性イオウ分子種の役割の検討

滝田 克也 東北大学

2021.09.24

概要

背景と目的
インフルエンザウイルスは、エンベロープを持つ一本鎖 RNA ウイルスであり全世界で季節性の流行を起こしている。年間 3-5 億人の罹患者がおり 25-50 万人が死亡している。インフルエンザウイルス肺炎は、特に重要なインフルエンザウイルス感染合併症の一つであり、一部の患者は急性呼吸窮迫症候群(acute respiratory distress syndrome : ARDS)を発症し、死亡率は 40~46 %に上るためその治療方法の確立が重要である。インフルエンザウイルス感染時には活性酸素種(Reactive Oxygen Species : ROS)の産生が増加することが知られており、ROSはさらなる炎症シグナルの伝達や、抗ウイルス作用に関与する。しかしながら感染に伴って過剰に産生された ROS により肺内の酸化還元の均衡が崩れることで、むしろ宿主の細胞や肺組織を傷害し肺炎病態を悪化させることが報告されていたが、抗酸化物質を用いた治療はヒトにおいて有効性を示すことができていない。活性イオウ分子種(reactive sulfur species: RSS)は近年注目を集めている新規内因性抗酸化分子の総称であり、ミトコンドリアに局在するシステイン tRNA 合成酵素(cysteinyl-tRNA synthetase 2: CARS2)により産生される。本学ではCOPD 患者の気道被覆液や肺組織では健常者と比べてRSS 含有量が有意に低下していることを世界に先駆けて 2017 年に報告している。しかし CARS2 や RSS がインフルエンザウイルス感染症病態において果たしている役割は明らかになっていない。以上の背景より、本研究はインフルエンザウイルス感染症における酸化還元不均衡に着目し、RSS および CARS2 の役割を明らかにすることを目的とした。

方法と結果
Cars2+/-マウスと野生型マウスにインフルエンザウイルスを経気管的に投与した肺炎モデルにおいて致死率を比較したところ、Cars2+/-マウスでは致死率の増悪を認めた。さらにマウス肺の肺組織の障害の程度や肺胞洗浄液(BALF)中の細胞分画や炎症性メディイエーターを解析したところ、Cars2+/-マウスは野生型マウスより肺傷害の程度、炎症細胞数、IL-6 などの炎症性メディエーターの産生が有意に増加した。マウス肺の RSS を質量分析法で網羅的に解析したところインフルエンザウイルス感染時にはグルタチオン(Glutathione-SH : GSH)だけではなく、グルタチオンパースルフィド(glutathione persulfide: GSSH)、システインパースルフィド(cysteine persulfide: CysSSH)などの RSS が低下していた。RSS供与体であるグルタチオントライサルファイド(glutathione trisulfide : GSSSG)を前投与することでインフルエンザウイルス感染に伴う致死率の低下、肺傷害の進行や炎症性メディエーターの産生の増加が抑制された。不死化ヒト気道上皮細胞株 BEAS-2B 細胞に toll like receptor 3 のリガンドである poly(I:C) (Polyinosinic-polycytidylic acid sodium salt)およびインフルエンザウイルスを投与し、細胞内の ROS と RSS の産生量および CARS2 の発現量を検討した。poly(I:C)およびインフルエンザウイルスにより ROS の産生が増加したが、一方で CARS2発現量および RSS 含有量が減少した。GSSSG で前処理しておくと、poly(I:C)やインフルエンザウイルスによるRSS の減少が回復し、ROS の産生が抑制された。

結論
本研究の結果より、インフルエンザウイルス感染時に CARS2-RSS の経路が減弱することで生じる酸化還元の不均衡が肺炎や肺傷害の発症に関与する可能性が示唆された。外因性の RSS 補充がインフルエンザウイルス感染に伴う肺炎の予後を改善することが示唆された。CARS2-RSS 経路を標的にした治療戦略がインフルエンザウイルスによる肺傷害の新規治療法につながる可能性がある。

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