་ ඹ ᵝ ᘧ 㸯
Ꮫ ㄳ ồ ㄽ ᩥ ࡢ ෆ ᐜ ࡢ せ ᪨
ㄽ ᩥ ᥦ ฟ ⪅ Ặ ྡ ែ ไ ᚚ ⛉ Ꮫ 㡿 ᇦ ᾘ ჾ ෆ ⛉ ศ 㔝 ᾘ ჾ ⾑ ᾮ ෆ ⛉ Ꮫ ㅮ ᗙ Ặ ྡ ⏣ ୰ ዉ ಖ Ꮚ
㸦 ㄽ ᩥ 㢟 ┠ 㸧Vitamin A deficiency impairs host resistance to Listeria
Monocytogenes infection through excessive apoptosis of macrophages
ࣅ ࢱ ࣑ ࣥ $ Ḟ ஈ ≧ ែ ࡣ ࣐ ࢡ ࣟ ࣇ ࣮ ࢪ ࡢ 㐣 ࡞ ࣏ ࢺ ࣮ ࢩ ࢫ ࡼ ࡾ ࣜ ࢫ ࢸ ࣜ ࣭ࣔ
ࣀ ࢧ ࢺ ࢤ ࢿ ࢫ ࡢ ឤ ᰁ ᢠ ᛶ ࡀ ῶ ᙅ ࡍ ࡿ
㸦 ෆ ᐜ ࡢ せ ᪨ 㸧
࠙ せ ᪨ ࠚListeria monocytogenes ࡣ ࢢ ࣛ ࣒ 㝧 ᛶ ᱰ ⳦ ࡛ ࠊ࣐ ࢡ ࣟ ࣇ ࣮ ࢪ ࢆ ࡣ ࡌ ࡵ ࡋ ࡓ 㣗 ⣽ ⬊ ࡢ ⣽ ⬊ ㉁ ෆ ࡢ ධ ࠊ ቑ Ṫ ࢆ ㉳ ࡇ ࡍ ⣽ ⬊ ෆ ᐤ ⏕ ⳦ ࡛ ࠶ ࡿ ࠋ ở ᰁ ࡉ ࢀ ࡓ 㣗 ≀ ࡢ ᦤ
ྲྀ ࡼ ࡾ య ෆ ධ ࡋ ࠊ⭠ ⟶ ࡽ ⾑ ὶ ࡾ ᩋ ⾑ ࡸ 㧊 ⭷ ⅖ ࠊᚰ ෆ ⭷ ⅖ ࢆ ㉳ ࡇ ࡍ ࠋࡲ ࡓ ࠊ ࡋ ࡤ ࡋ ࡤ ࣐ ࢡ ࣟ ࣇ ࣮ ࢪ ࡢ ㈎ 㣗 ࣭ ẅ ⳦ ⬟ ࠸ ࡗ ࡓ ᶵ ⬟ ゎ ᯒ ⏝ ࠸ ࡽ ࢀ ࡿ ࠋ ୍ ᪉ ࠊ ࣅ ࢱ ࣑
ࣥ A ࡣ ⬡ ⁐ ᛶ ࣅ ࢱ ࣑ ࣥ ࡢ ୍ ࡘ ࡛ ࠊᵝ ࠎ ࡞ ⏕ ⌮ ά ᛶ ࢆ ᭷ ࡍ ࡿ ࠋࣅ ࢱ ࣑ ࣥ A Ḟ ஈ(Vitamin A deficiency: VAD)ࡣ ⭠ ⟶ ୖ ⓶ ࡢ ࢿ ࢡ ࣟ ࣮ ࢩ ࢫ ࡢ ቑ ຍ ࡞ ࡼ ࡾ ࠊ ࣂ ࣜ ᶵ ⬟ ࡢ ప ୗ ࢆ ㉳ ࡇ ࡍ ࠋVAD࣐ ࢘ ࢫ ࡛ L. monocytogenes ࡢ ឤ ᰁ ᢠ ᛶ ࡀ ῶ ᙅ ࡍ ࡿ ࡀ ࠊall-trans retinoic acid (ATRA)ࢆ ᢞ ࡍ ࡿ ࠊ ឤ ᰁ ᢠ ᛶ ࡀ ᅇ ࡍ ࡿ ࡇ ࢆ ᡃ ࠎ ࡣ ሗ ࿌ ࡋ ࡚ ࠸ ࡿ ࠋ ࡇ ࡢ ᶵ ᗎ ࡣ ⮬ ↛ ච ࡀ 㛵 ࡋ ࡚ ࠸ ࡿ ࠸ ࠺ ௬ ㄝ ᇶ ࡙ ࡁ ࠊL. monocytogenes ឤ ᰁ ୗ ࡢ VAD
࣐ ࢘ ࢫ ࠾ ࡅ ࡿ ࣐ ࢡ ࣟ ࣇ ࣮ ࢪ ᶵ ⬟ ࢆ ᳨ ウ ࡋ ࡓ ࠋ
࠙ ᪉ ἲ ࠚC57BL/6J࣐ ࢘ ࢫ ࠊSevere Combined Immunodeficiency (SCID)࣐ ࢘ ࢫ ࣅ ࢱ ࣑ ࣥ A Ḟ ஈ 㣵 ࢆ ࠼ ࠊ10 㐌 㱋 ࡲ ࡛ 㣫 ⫱ ࡋ ࡓ ࣐ ࢘ ࢫ ࢆ VAD ࣐ ࢘ ࢫ ࡋ ࡓ ࠋC57BL/6J ࡢ VAS (vitamin A sufficient)ࠊVAD࣐ ࢘ ࢫ L. monocytogenesࢆ ឤ ᰁ ࡉ ࡏ ࠊ ឤ ᰁ 1
᪥ ᚋ ࡢ ⭁ ⮚ ࣭ ⫢ ⮚ ࢆ ᥇ ྲྀ ࡋ ࡓ ࠋ ⭁ ⮚ ࡣ Transferase-mediated UTP-biotin nick end labeling (TUNEL)ᰁ Ⰽ ࢆ ⾜ ࠸ ࠊ ࣏ ࢺ ࣮ ࢩ ࢫ ⣽ ⬊ ࢆ ほ ᐹ ࡋ ࡓ ࠋࡉ ࡽ ࠊ᥇ ྲྀ ࡋ ࡓ ⭁ ⮚ ࡢ CD11b㝧 ᛶ ⣽ ⬊ ࢆ ศ 㞳 ࡋ ࠊࣇ ࣟ ࣮ ࢧ ࢺ ࣓ ࢺ ࣜ ࣮ ࢆ ⏝ ࠸ ࡚ Annexin V 7-AADࢆ ᰁ Ⰽ ࡋ ࠊ ᪩ ᮇ ࣏ ࢺ ࣮ ࢩ ࢫ ⣽ ⬊ ࡢ ྜ ࢆ ィ ࡋ ࡓ ࠋ ࡲ ࡓ ࠊ ⭁ ⮚ ࣭ ⫢ ⮚ ෆ ࡢ ⮚ ჾ ෆ ⳦ ᩘ ࢆ ィ ࡋ ࡓ ࠋḟ ࠊC57BL/6J VASࠊVAD࣐ ࢘ ࢫ 4% ࢳ ࢜ ࢢ ࣜ ࢥ ࣞ ࣮ ࢺ 2ml ࢆ ⭡ ⭍ ෆ ᢞ ࡋ ࠊ 4᪥ ᚋ ⭡ ⭍ ෆ ᾐ ₶ ࡋ ࡓ PECs (peritoneal exudate cells)ࢆ ᥇ ྲྀ ࡋ ࡓ ࠋ᥇ ྲྀ ࡋ ࡓ PECs
RAR 㜼 ᐖ ࡛ ࠶ ࡿ Ro41-5253 ࢆ ᢞ ࠊL. monocytogenes ࢆ ឤ ᰁ ࡉ ࡏ ࡓ 24 㛫 ᚋ
ୖ Ύ ࢆ ᅇ ࡋ ࠊ ࣃ ࣟ ࢺ ࣮ ࢩ ࢫ 㛵 ࡋ ࡚ ࠸ ࡿ ୖ Ύ ୰ IL-1E ࢿ ࢡ ࣟ ࣮ ࢩ ࢫ 㛵 ࡋ ࡚
࠸ ࡿ ୖ Ύ ୰ TNF-Dࢆ ࡑ ࢀ ࡒ ࢀ ELISA ࡛ ィ ࠊ ᪩ ᮇ ࣏ ࢺ ࣮ ࢩ ࢫ ࡛ ⏕ ࡌ ࡿ DNA fragmentation ࢆ cell death ELISA kit ࢆ ⏝ ࠸ ࡚ ィ ࡋ ࡓ ࠋIn vitro ࡋ ࡚ ࠊ ࣐ ࢡ ࣟ ࣇ
࣮ ࢪ ⣽ ⬊ ᰴ ࡛ ࠶ ࡿ RAW264.7 ࢆ ⏝ ࠸ ࠊRo41-5253 ࢆ ๓ ᢞ ࡋ ࠊL. monocytogenesࢆ ឤ ᰁ ࡉ ࡏ ࠊ ྠ ᵝ cell death detection ELISA ࢆ ⏝ ࠸ ࡚ DNA fragmentation ࢆ ィ ࡋ ࡓ ࠋ ⤫ ィ 㛵 ࡋ ࡚ ࡣ unpaired-t test ࢆ ⏝ ࠸ ࡚ ⾜ ࡗ ࡓ ࠋ
࠙ ⤖ ᯝ ࠚL. monocytogenes ᮍ ឤ ᰁ ࠊ ឤ ᰁ 1 ᪥ ᚋ ࡢ C57BL/6JࠊSCID ࣐ ࢘ ࢫ ࡢ ⭁ ⮚ ࢆ TUNEL ᰁ Ⰽ ࡛ ほ ᐹ ࡋ ࡓ ࠋ ᮍ ឤ ᰁ ࡛ ࡣ C57BL/6JࠊSCID ࣐ ࢘ ࢫ ୧ ⪅ ࡛ VAS࣭VAD ࡛ TUNEL 㝧 ᛶ ⣽ ⬊ ࡣ ㄆ ࡵ ࡽ ࢀ ࡞ ࡗ ࡓ ࠋឤ ᰁ 1᪥ ᚋ ࡛ ࡣ ࠊC57BL/6JࠊSCID ࣐ ࢘ ࢫ ୧ ⪅
࡛ VAS ࣐ ࢘ ࢫ ẚ ࡋ ࡚ VAD࣐ ࢘ ࢫ ࡛ TUNEL㝧 ᛶ ⣽ ⬊ ࡢ ቑ ຍ ࢆ ㄆ ࡵ ࡓ ࠋࡇ ࡢ ⤖ ᯝ ࡽ ࠊ L. monocytogenes ឤ ᰁ ୗ VAD࣐ ࢘ ࢫ ࡛ ࡣ T࣭B ⣽ ⬊ ௨ እ ࡢ ⣽ ⬊ ࡛ ࣏ ࢺ ࣮ ࢩ ࢫ ࡀ ቑ ຍ ࡋ ࡚ ࠸ ࡿ ࡇ ࡀ ♧ ၀ ࡉ ࢀ ࠊHE ᰁ Ⰽ ࡛ ࡣ ࠊ ࣏ ࢺ ࣮ ࢩ ࢫ ࢆ ㉳ ࡇ ࡋ ࡚ ࠸ ࡿ ⣽ ⬊ ࡣ ࣐ ࢡ ࣟ ࣇ
࣮ ࢪ ࡛ ࠶ ࡿ ⪃ ࠼ ࡽ ࢀ ࡓ ࠋ ࣇ ࣟ ࣮ ࢧ ࢺ ࣓ ࢺ ࣜ ࣮ ࡛ ࡣ ࠊVAD ࣐ ࢘ ࢫ ࠾ ࡅ ࡿ
L. monocytogenes ឤ ᰁ 2 㛫 ᚋ ࠊ6 㛫 ᚋ ࡢ CD11b 㝧 ᛶ ⣽ ⬊ ࡛ ࡣ ࠊAnnexin V 㝧 ᛶ ࠊ 7-AAD 㝜 ᛶ ࡢ ᪩ ᮇ ࣏ ࢺ ࣮ ࢩ ࢫ ࡀ ቑ ຍ ࡋ ࡚ ࠸ ࡿ ࡇ ࡀ ᫂ ࡽ ࡞ ࡗ ࡓ ࠋcell death detection ELISA ࡛ ࡣ ࠊL. monocytogenes ឤ ᰁ ୗ ࠾ ࠸ ࡚ Ro41-5253ῧ ຍ RAW264.7
࡛ ࣏ ࢺ ࣮ ࢩ ࢫ ࡀ ቑ ຍ ࡋ ࡚ ࠸ ࡓ ࠋࡲ ࡓ ࠊVAS࣐ ࢘ ࢫ ࡢ PECs ࡛ ࡣ ࠊRo41-5253 ῧ ຍ ⩌ ࡛ L. monocytogenes ឤ ᰁ ୗ ࡛ ࡢ ࣏ ࢺ ࣮ ࢩ ࢫ ࡀ ቑ ຍ ࡋ ࡚ ࠸ ࡓ ࠋ ຍ ࠼ ࡚ ࠊVAD ࣐ ࢘ ࢫ ࡢ PECs ࡣ VAS ࣐ ࢘ ࢫ ࡢ PECs ẚ ㍑ ࡋ ࡚ ࠊL. monocytogenes ឤ ᰁ ୗ ࡛ ࡢ ࣏ ࢺ ࣮ ࢩ ࢫ ࡀ ቑ ຍ ࡋ ࡚ ࠸ ࡿ ࡇ ࡀ ♧ ࡉ ࢀ ࡓ ࠋL. monocytogenes ឤ ᰁ ୗ RAW264.7 ࡢ ୖ Ύ ୰ IL-1E࣭
TNF-Dࡣ Ro41-5253 ῧ ຍ ࡛ ᭷ ព ⏘ ⏕ ࡀ ప ୗ ࡋ ࡚ ࠾ ࡾ ࠊࣃ ࣟ ࢺ ࣮ ࢩ ࢫ ࠊࢿ ࢡ ࣟ ࣮ ࢩ ࢫ ࡢ 㛵 ࡣ ྰ ᐃ ⓗ ⪃ ࠼ ࡽ ࢀ ࡓ ࠋ ᭱ ᚋ ࠊVASࠊVAD ≧ ែ ࡢ C57BL/6J ࣐ ࢘ ࢫ ࠊSCID࣐
࢘ ࢫ ࡽ ᥇ ྲྀ ࡋ ࡓ ⭁ ⮚࣭⫢ ⮚ ࡢ ⮚ ჾ ෆ ⳦ ᩘ ࡣ ࠊឤ ᰁ 1 ᪥ ᚋ ࠊ2᪥ ᚋ ࠸ ࡎ ࢀ ࠾ ࠸ ࡚ ࡶ VAD
࣐ ࢘ ࢫ ࡛ ᭷ ព ቑ ຍ ࡋ ࡚ ࠸ ࡓ ࠋ
࠙ ⪃ ᐹ ࠚTUNELᰁ Ⰽ ࡛ VAD࣐ ࢘ ࢫ ⭁ ⣽ ⬊ ࡢ ࣏ ࢺ ࣮ ࢩ ࢫ ࡀ ቑ ຍ ࡋ ࡚ ࠾ ࡾ ࠊࡑ ࡢ ⣽ ⬊ ࡣ HE ᰁ Ⰽ ࡚ ࣐ ࢡ ࣟ ࣇ ࣮ ࢪ ࡛ ࠶ ࡿ ⪃ ࠼ ࡽ ࢀ ࡓ ࠋ ࡉ ࡽ ࣇ ࣟ ࣮ ࢧ ࢺ ࣓ ࢺ ࣜ ࣮ ࡛ ࡣ VAD࣐ ࢘ ࢫ ࡢ CD11b㝧 ᛶ ⣽ ⬊ ࠊRo41-5253 ῧ ຍ ࡢ RAW264.7 ࡸ PECs ࡛ ࣏ ࢺ ࣮ ࢩ ࢫ ቑ ຍ ࡀ ほ ᐹ ࡉ ࢀ ࡓ ࠋ ࡇ ࡢ ࡇ ࡽ ࠊ ࣐ ࢡ ࣟ ࣇ ࣮ ࢪ ࡢ 㐣 ࡞ ࣏ ࢺ ࣮ ࢩ ࢫ ࡼ ࡾ ࠊVAD
࡛ ࡣ L. monocytogenes ࡢ ឤ ᰁ ᢠ ᛶ ࡀ ῶ ᙅ ࡍ ࡿ ࡇ ࡀ ⪃ ࠼ ࡽ ࢀ ࡓ ࠋ
࠙ ⤖ ㄒ ࠚ ࡇ ࡢ ⤖ ᯝ ࡽ ࠊ ࣅ ࢱ ࣑ ࣥ $ Ḟ ஈ ≧ ែ ࡣ ࣐ ࢡ ࣟ ࣇ ࣮ ࢪ ࡢ 㐣 ࡞ ࣏ ࢺ ࣮ ࢩ ࢫ
ࡼ ࡾ L. monocytogenes ࡢ ឤ ᰁ ᢠ ᛶ ࡀ ῶ ᙅ ࡍ ࡿ ⪃ ࠼ ࡽ ࢀ ࡿ ࠋ ࣅ ࢱ ࣑ ࣥ $ ࡣ ࣐ ࢡ ࣟ
ࣇ ࣮ ࢪ ᶵ ⬟ ࢆ ㄪ ⠇ ࡍ ࡿ ࡇ ࡛ ᜏ ᖖ ᛶ ࡢ ⥔ ᣢ ᐤ ࡋ ࠊᐟ ච ࡗ ࡚ ࣅ ࢱ ࣑ ࣥ $ ࡢ ᦤ ྲྀ ࡀ 㔜 せ ࡛ ࠶ ࡿ ࡇ ࡀ ᫂ ࡽ ࡞ ࡗ ࡓ ࠋ
