୍ ࡘ ࡛ ࠶ ࡾ ࠊ ᨺ ⨨ ࡍ ࡿ ࡜ ⫹ Ồ ᛶ ⫢ ◳ ኚ ⑕ ࠿ ࡽ ୙ ྍ ㏫ ⓗ ࡞ ⫢ ୙ ඲ ࡟ ⮳ ࡿ ࠋ ⤌ ⧊ Ꮫ ⓗ ࡟ ࡣ ⫢ እ ⫹ ⟶ ࡢ ◳ ໬ ᛶ ⫹ ⟶ ⅖ ࡜ ⥺ ⥔ ᛶ 㛢 ሰ ࡀ ᇶ ᮏ ⑓ ែ ࡛ ࠶ ࡿ ࡀ ࠊ ࡑ ࡢ ⑓ ᅉ ࡣ ᮍ ࡔ ୙ ᫂

་ ඹ ᵝ ᘧ 㸯

Ꮫ ఩ ㄳ ồ ㄽ ᩥ ࡢ ෆ ᐜ ࡢ せ ᪨

ㄽ ᩥ ᥦ ฟ ⪅ Ặ ྡ ⭘ ⒆ ไ ᚚ ⛉ Ꮫ 㡿 ᇦ ᾘ ໬ ჾ እ ⛉ Ꮫ ศ 㔝 Ặ ྡ ᓥ ⏣ ᣅ

㸦 ㄽ ᩥ 㢟 ┠ 㸧

CCL5 is induced by TLR 3 signaling in HuCCT1 human biliary epithelial cells:

possible involvement in the pathogenesis of biliary atresia

(HuCCT1 ࣄ ࢺ ⫹ ⟶ ୖ ⓶ ⣽ ⬊ ࡟ ࠾ ࠸ ࡚ CCL5 ࡣ TLR 3 ࢩ ࢢ ࢼ ࣝ ࡟ ࡼ ࡾ ㄏ ᑟ ࡉ ࢀ ࡿ 㸸

⫹ 㐨 㛢 㙐 ⑕ ࡢ Ⓨ ⑕ ᶵ ᗎ ࡬ ࡢ 㛵 ୚ ࡢ ྍ ⬟ ᛶ )

࠙ ⫼ ᬒ ࠚ

⫹ 㐨 㛢 㙐 ⑕㸦 BA; biliary atresia㸧ࡣ ࠊ᪂ ⏕ ඣ ᮇ ࡟ ࠾ ࡅ ࡿ 㞴 ἞ ᛶ ࡢ ⫢ ⫹ 㐨 ⣔ ⑌ ᝈ ࡢ

୍ ࡘ ࡛ ࠶ ࡾ ࠊ ᨺ ⨨ ࡍ ࡿ ࡜ ⫹ Ồ ᛶ ⫢ ◳ ኚ ⑕ ࠿ ࡽ ୙ ྍ ㏫ ⓗ ࡞ ⫢ ୙ ඲ ࡟ ⮳ ࡿ ࠋ ⤌ ⧊ Ꮫ ⓗ ࡟ ࡣ ⫢ እ ⫹ ⟶ ࡢ ◳ ໬ ᛶ ⫹ ⟶ ⅖ ࡜ ⥺ ⥔ ᛶ 㛢 ሰ ࡀ ᇶ ᮏ ⑓ ែ ࡛ ࠶ ࡿ ࡀ ࠊ ࡑ ࡢ ⑓ ᅉ ࡣ ᮍ ࡔ ୙ ᫂

࡛ ࠶ ࡿ ࠋ

⏕ ᚋ 㛫 ࡶ ࡞ ࠸ ࣐ ࢘ ࢫ ࡟ RNA ࢘ ࢖ ࣝ ࢫ ࡢ ୍ ࡘ ࡛ ࠶ ࡿ ࣞ ࢜ ࢘ ࢖ ࣝ ࢫ ࢆ ே Ⅽ ⓗ ࡟ ឤ ᰁ ࡉ ࡏ ࡿ ࡜ ࠊ ⫹ 㐨 㛢 㙐 ⑕ ࡟ 㢮 ఝ ࡋ ࡓ ⑓ ែ ࢆ ࿊ ࡋ ࠊ ⫹ 㐨 㛢 㙐 ⑕ ࡢ ື ≀ ࣔ ࢹ ࣝ ࡜ ࡋ ࡚ ౑ ࢃ

ࢀ ࡚ ࠸ ࡿ ࠋ ࡇ ࡢ ࣐ ࢘ ࢫ ࡢ ⫹ 㐨 㛢 㙐 ⑕ ࣔ ࢹ ࣝ ࡢ ⫹ ⟶ ୖ ⓶ ⣽ ⬊ ࡛ ࠊ ࠸ ࡃ ࡘ ࠿ ࡢ ࢣ ࣔ ࢝ ࢖

ࣥ ࡀ 㐣 ๫ ࡟ Ⓨ ⌧ ࡋ ࡚ ࠸ ࡿ ࡇ ࡜ ࡀ ሗ ࿌ ࡉ ࢀ ࡚ ࠸ ࡿ ࠋ ࡋ ࡓ ࡀ ࡗ ࡚ ࠊ RNA ࢘ ࢖ ࣝ ࢫ ࡢ ឤ ᰁ

࡜ ⫹ ⟶ ୖ ⓶ ⣽ ⬊ ࡛ ࡢ ࢣ ࣔ ࢝ ࢖ ࣥ ࡢ 㐣 ๫ ࡞ Ⓨ ⌧ ࡀ ࠊ BA ࡢ ⑓ ែ ᙧ ᡂ ࡢ ཎ ᅉ ࡟ 㛵 ୚ ࡋ ࡚ ࠸

ࡿ ࡜ ᥎   ࡉ ࢀ ࡚ ࠸ ࡿ ࠋ ࡋ ࠿ ࡋ ࠊ ࡑ ࡢ ヲ ⣽ ࡣ ᮍ ࡔ ᫂ ࡽ ࠿ ࡟ ࡉ ࢀ ࡚ ࠸ ࡞ ࠸ ࠋ

⏕ య ࡟ ౵ ධ ࡋ ࡚ ࡁ ࡓ ࢘ ࢖ ࣝ ࢫ ࡢ ᵓ ᡂ ᡂ ศ ࡣ ࠊ ✀ ࠎ ࡢ ⣽ ⬊ ࡟ ࠾ ࠸ ࡚ ࣃ ࢱ ࣮ ࣥ ㄆ ㆑ ཷ ᐜ య ࡟ ࡼ ࡗ ࡚ ㄆ ㆑ ࡉ ࢀ ࠊ ࡑ ࡢ ୗ ὶ ࡛ ⮬ ↛ ච ␿ ⣔ ࡀ ά ᛶ ໬ ࡉ ࢀ ࡿ ࠋ ࣄ ࢺ ࡢ ⫹ ⟶ ୖ ⓶ ⣽

⬊ ࡟ ࠾ ࠸ ࡚ ࡣ ࠊ RNA ࢘ ࢖ ࣝ ࢫ ࡢ ឤ ᰁ ࡜ ⮬ ↛ ච ␿ ᛂ ⟅ ࡟ 㛵 㐃 ࡋ ࡓ ࢣ ࣔ ࢝ ࢖ ࣥ ࡢ Ⓨ ⌧ ື

ែ ࢆ ㄪ ࡭ ࡓ ◊ ✲ ࡣ ࠊ ࡇ ࢀ ࡲ ࡛ Ꮡ ᅾ ࡋ ࡞ ࠸ ࠋ

ࡑ ࡇ ࡛ ࠊ ᮏ ◊ ✲ ࡛ ࡣ ࠊ ࣄ ࢺ ⫹ ⟶ ୖ ⓶ ⣽ ⬊ ⒴ ⏤ ᮶ ⣽ ⬊ ᰴ ࡛ ࠶ ࡿ HuCCT1 ࢆ ⏝ ࠸ ࡚ ࠊ BA ࡬ ࡢ 㛵 ୚ ࡀ ♧ ၀ ࡉ ࢀ ࡿ ࢣ ࣔ ࢝ ࢖ ࣥ ࡢ ࠺ ࡕ ࠊCCL5 (ู ྡ ࠊ RANTES; regulated on activation, normal T cell expressed and secreted)࡟ ὀ ┠ ࡋ ࠊ RNA ࢘ ࢖ ࣝ ࢫ ឤ ᰁ ࡜ ࡢ 㛵 㐃 ࠾ ࡼ ࡧ CCL5 ࡢ Ⓨ ⌧ ࡟ 㛵 ୚ ࡍ ࡿ ⣽ ⬊ ෆ ࢩ ࢢ ࢼ ࣝ ఏ 㐩 ࢆ ᳨ ウ ࡋ ࡓ ࠋ

࠙ ᪉ ἲ ࠚ

HuCCT1 ⣽ ⬊ ࢆ ᇵ 㣴 ࡋ ࠊ ࢘ ࢖ ࣝ ࢫ RNA ࢆ mimic ࡍ ࡿ ヨ ⸆ ࡛ ࠶ ࡿ ே ᕤ ⓗ ࡞ ஧ ᮏ 㙐 RNA ࡢ polyinosinic-polycytidylic acid (poly IC)࡛ ฎ ⌮ ࡋ ࠊ CCL5 ࡢ Ⓨ ⌧ ࢆ ㄪ ࡭ ࡓ ࠋ CCL5 mRNA ࠾ ࡼ ࡧ ࢱ ࣥ ࣃ ࢡ ㉁ ࡢ Ⓨ ⌧ ゎ ᯒ ࡣ ࠊᐃ 㔞 RT-PCR ἲ ࠾ ࡼ ࡧ ELISA ἲ ࢆ ⏝

࠸ ࡓ ࠋ ࡲ ࡓ ࠊ RNA ᖸ ΅ ἲ ࢆ ⏝ ࠸ ࡚ ࠊ ஧ ᮏ 㙐 RNA ࡟ ᑐ ࡍ ࡿ ࣃ ࢱ ࣮ ࣥ ㄆ ㆑ ཷ ᐜ య ࡛ ࠶

ࡿ Toll-like receptor 3 (TLR3) ࠊ ⮬ ↛ ච ␿ ᛂ ⟅ ࡢ key cytokine ࡛ ࠶ ࡿ Interferon (IFN)-ǃࠊ ㌿ ෗ ᅉ Ꮚ ࡛ ࠶ ࡿ nuclear factor- NjB (NF-NjB) ࡢ p65 ࢧ ࣈ ࣘ ࢽ ࢵ ࢺ ࠾ ࡼ ࡧ IFN regulatory factor 3 (IRF3) ࡢ ࣀ ࢵ ࢡ ࢲ ࢘ ࣥ ࢆ ⾜ ࠸ ࠊCCL5 ࡢ Ⓨ ⌧ ࡟ ࠾ ࡅ ࡿ ࡇ ࢀ

ࡽ ࡢ ศ Ꮚ ࡢ 㛵 ୚ ࢆ ㄪ ࡭ ࡓ ࠋp65 ࡜ IRF3 ࢱ ࣥ ࣃ ࢡ ㉁ ࡢ Ⓨ ⌧ ࡣ ࠊwestern blotting ࡛ ㄪ

࡭ ࡓ ࠋ

ࡲ ࡓ ࠊ 㐣 ཤ ࡟ ᮏ Ꮫ 㝃 ᒓ ⑓ 㝔 ࡟ ࡚ ᡭ ⾡ ࢆ ⾜ ࡗ ࡓ ࣄ ࢺ ⫹ 㐨 㛢 㙐 ⑕ 2 ⑕ ౛ ࡢ ⫢ 㛛 㒊 ⤌ ⧊

ࡢ ࣍ ࣝ ࣐ ࣜ ࣥ ᅛ ᐃ ࣭ ࣃ ࣛ ࣇ ࢕ ࣥ ໟ ᇙ ษ ∦ ࢆ ⏝ ࠸ ࡚ ࠊ CCL5 ࡢ ච ␿ ⤌ ⧊ ໬ Ꮫ ࢆ ⾜ ࡗ ࡓ ࠋ

ᑠ ඣ ⫢ ⱆ ⭘ 1 ⑕ ౛ ࡢ ⫢ ษ 㝖 ᶆ ᮏ ࡢ 㠀 ⅖ ⑕ 㒊 ศ ࢆ ᑐ ↷ ⩌ ࡜ ࡋ ࡚ ⏝ ࠸ ࡓ ࠋ

࠙ ⤖ ᯝ ࠚ

ᇵ 㣴 ࡋ ࡓ HuCCT1 ⣽ ⬊ ࢆ poly IC ࡛ ฎ ⌮ ࡍ ࡿ ࡜ ࠊ CCL5 ࡢ mRNA ࠾ ࡼ ࡧ ࢱ ࣥ ࣃ ࢡ

㉁ ࡢ Ⓨ ⌧ ࡀ ஹ 㐍 ࡋ ࡓ ࠋࡲ ࡓ TLR3ࠊp65ࠊIRF3 ࡢ ࣀ ࢵ ࢡ ࢲ ࢘ ࣥ ࡟ ࡼ ࡾ ࠊpoly IC ฎ ⌮

࡟ ࡼ ࡿ CCL5 ࡢ Ⓨ ⌧ ஹ 㐍 ࡣ ῶ ᙅ ࡋ ࡓ ࡀ ࠊ IFN- ǃ ࡢ ࣀ ࢵ ࢡ ࢲ ࢘ ࣥ ࡣ CCL5 ࡢ Ⓨ ⌧ ࡟ ᙳ 㡪 ࢆ ୚ ࠼ ࡞ ࠿ ࡗ ࡓ ࠋ

CCL5 ࡢ ච ␿ ⤌ ⧊ ໬ Ꮫ ࡛ ࡣ ࠊ ࣄ ࢺ ⫹ 㐨 㛢 㙐 ⑕ ⑕ ౛ ࡢ ⫢ 㛛 㒊 ⤌ ⧊ ࡢ ⫹ ⟶ ୖ ⓶ ⣽ ⬊ ࡟ CCL5 ࡀ ᙉ ࡃ Ⓨ ⌧ ࡋ ࡚ ࠸ ࡓ ࡀ ࠊ ࣄ ࢺ ṇ ᖖ ⫢ ⤌ ⧊ ࡢ ⫹ ⟶ ୖ ⓶ ⣽ ⬊ ࡛ ࡣ Ⓨ ⌧ ࡣ ㄆ ࡵ ࡽ ࢀ

࡞ ࠿ ࡗ ࡓ ࠋ

࠙ ⪃ ᐹ ࠚ

௒ ᅇ ࠊ ᡃ ࠎ ࡣ HuCCT1 ⣽ ⬊ ࢆ poly IC ࡛ ฎ ⌮ ࡍ ࡿ ࡜ CCL5 ࡢ Ⓨ ⌧ ࡀ ㄏ ᑟ ࡉ ࢀ ࡿ ࡇ ࡜ ࢆ ぢ ฟ ࡋ ࡓ ࠋ ࡇ ࡢ CCL5 ࡢ Ⓨ ⌧ ㄏ ᑟ ࡣ ࠊ TLR3ࠊ NF- NjB p65ࠊ IRF3 ࡢ ࣀ ࢵ ࢡ ࢲ

࢘ ࣥ ࡟ ࡼ ࡾ ᢚ ไ ࡉ ࢀ ࡓ ࡇ ࡜ ࠿ ࡽ ࠊ poly IC ࡣ TLR3 ࢩ ࢢ ࢼ ࣜ ࣥ ࢢ ࢆ ά ᛶ ໬ ࡋ ࠊ ㌿ ෗ ᅉ Ꮚ ࡢ NF-NjB ࡜ IRF3 ࢆ ά ᛶ ໬ ࡋ ࡚ CCL5 ࡢ Ⓨ ⌧ ࡀ ㄏ ᑟ ࡉ ࢀ ࡓ ࡜ ⪃ ࠼ ࡽ ࢀ ࡓ ࠋ୍ ᪉ ࠊ CCL5 ࡢ Ⓨ ⌧ ࡣ IFN-E ࡟ ౫ Ꮡ ࡋ ࡞ ࠸ ࡜ ⪃ ࠼ ࡽ ࢀ ࡓ ࠋ

ࡇ ࢀ ࡽ ࡢ ⤖ ᯝ ࡣ ࠊ RNA ࢘ ࢖ ࣝ ࢫ ࡢ ឤ ᰁ ࡀ ࣄ ࢺ ࡢ ⫹ ⟶ ୖ ⓶ ⣽ ⬊ ࡟ ࠾ ࠸ ࡚ CCL5 ࡢ Ⓨ

⌧ ࢆ ஹ 㐍 ࡉ ࡏ ࡿ ࡇ ࡜ ࢆ ♧ ၀ ࡋ ࡚ ࠸ ࡿ ࠋࡲ ࡓ ࠊ NF-NjB ࡜ IRF3 ஧ ࡘ ࡢ ㌿ ෗ ᅉ Ꮚ ࡣ CCL5 ࡢ Ⓨ ⌧ ࢆ ஹ 㐍 ࡉ ࡏ ࡿ ࡇ ࡜ ࡟ ࡼ ࡾ BA ࡢ ⑓ ែ ࡟ 㛵 ୚ ࡋ ࡚ ࠸ ࡿ ྍ ⬟ ᛶ ࡀ ♧ ၀ ࡉ ࢀ ࡓ ࠋ

ࣄ ࢺ ࡢ ⤌ ⧊ ᶆ ᮏ ࢆ ⏝ ࠸ ࡓ ච ␿ ⤌ ⧊ ໬ Ꮫ ࡛ ࡣ ࠊ ⫹ 㐨 㛢 㙐 ⑕ ⑕ ౛ ࡢ ⫹ ⟶ ୖ ⓶ ⣽ ⬊ ࡟ CCL5 ࡀ ᙉ ࡃ Ⓨ ⌧ ࡋ ࡚ ࠸ ࡓ ࡀ ࠊ ṇ ᖖ ⤌ ⧊ ࡢ ⫹ ⟶ ୖ ⓶ ⣽ ⬊ ࡟ ࡣ Ⓨ ⌧ ࡋ ࡚ ࠸ ࡞ ࠿ ࡗ ࡓ ࠋ

௨ ୖ ࡢ ⤖ ᯝ ࠿ ࡽ ࠊࣄ ࢺ ࡟ ࠾ ࠸ ࡚ ࠊ஧ ᮏ 㙐 RNA ࢘ ࢖ ࣝ ࢫ ࡢ ឤ ᰁ ࡀ ⫹ 㐨 ୖ ⓶ ⣽ ⬊ ࡟ ࠾ ࡅ ࡿ TLR3 ୗ ὶ ࡢ ⮬ ↛ ච ␿ ᛂ ⟅ ࢆ ά ᛶ ໬ ࡋ ࠊ ࡑ ࢀ ࡟ ࡼ ࡾ 㐣 ๫ ࡟ Ⓨ ⌧ ࡋ ࡓ CCL5 ࡞ ࡝ ࡢ ࢣ ࣔ ࢝ ࢖ ࣥ ࡀ BA ࡢ ⑓ ែ ᙧ ᡂ ࡟ 㛵 ୚ ࡍ ࡿ ྍ ⬟ ᛶ ࡀ ⪃ ࠼ ࡽ ࢀ ࡓ ࠋBA ࡟ ᑐ ࡍ ࡿ ἞ ⒪ ࡣ

⌧ ᅾ ࡛ ࡣ እ ⛉ ⓗ ἞ ⒪ ࡋ ࠿ ࡞ ࡃ ࠊ BA ࡢ Ⓨ ⑕ ᶵ ᗎ ࡟ ╔ ┠ ࡋ ࡓ ἞ ⒪ ἲ ࡢ 㛤 Ⓨ ࡀ ᮃ ࡲ ࢀ ࡿ ࠋ CCL5 ࡞ ࡝ ࡢ ⅖ ⑕ ᛶ ࢣ ࣔ ࢝ ࢖ ࣥ ࡢ ⣽ ⬊ ෆ ࢩ ࢢ ࢼ ࣝ ఏ 㐩 ⤒ ㊰ ࡢ ゎ ᫂ ࡣ ௒ ᚋ BA ࡢ ἞ ⒪ ἲ 㛤 Ⓨ ࡟ ࡘ ࡞ ࡀ ࡿ ྍ ⬟ ᛶ ࡀ ࠶ ࡿ ࠋ

ͤ 1 எ ࡢ ሙ ྜ 㸪ۑ ۑ 㡿 ᇦ ۑ ۑ ᩍ ⫱ ◊ ✲ ศ 㔝 ࡟ ࠿ ࠼ ࡚ 㸪ᡤ ᒓ ࡢ ۑ ۑ ㅮ ᗙ ࢆ グ ධ ࡍ ࡿ ࡇ ࡜ ࠋ

ͤ 2 ㄽ ᩥ 㢟 ┠ ࡀ ⱥ ᩥ ࡢ ሙ ྜ ࡣ 㸦 㸧 ෆ ࡟ ࿴ ヂ ࢆ ௜ グ ࡍ ࡿ ࡇ ࡜ ࠋ