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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 黃思誠(Su-Cheng Huang),江伯倫,許博欽(Bor-Ching Sheu) | |
| dc.contributor.author | Wen-Chun Chang | en |
| dc.contributor.author | 張文君 | zh_TW |
| dc.date.accessioned | 2021-06-13T17:26:03Z | - |
| dc.date.available | 2007-01-28 | |
| dc.date.copyright | 2005-01-28 | |
| dc.date.issued | 2005 | |
| dc.date.submitted | 2005-01-20 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/39324 | - |
| dc.description.abstract | 背景與目的
子宮内膜癌在美國是女性生殖系統最常見的侵襲癌，自1950年以來，雖然子宮内膜癌的死亡率降低超過 60%，其發生率卻明顯增加，而且年齡層有逐漸降低的趨勢。我們知道腫瘤細胞在癌化過程中，一方面需要大量繁殖增生，另一方面則需對抗來自宿主體內免疫系統的清除作用，才能達到其侵入深部組織與蔓延的目的。因此雖然在腫瘤組織發現有大量的腫瘤內浸潤淋巴球 (tumor infiltrating lymphocytes，TILs)，許多免疫逃避性機轉仍被提出，說明這些腫瘤內浸潤淋巴球對抗腫瘤的毒殺能力微弱，自然殺手細胞 (natural killer cells，NK cells) 活性低，且對於腫瘤刺激的增生反應能力小。有人發現自然殺手細胞抑制受體 (inhibitory natural killer cell receptors，iNKRs)，存在於自然殺手細胞及 T 細胞，可抑制自然殺手細胞的活性與 CD8+ T 細胞的毒殺能力。我們的研究目的在於找出子宮内膜癌細胞逃避宿主的免疫監控與自然殺手細胞抑制受體之相關性，進一步希望對於未來發展免疫調節治療有所幫助。 材料與方法 我們利用機械式研磨萃取法 (mechanical dispersal technique) 來獲得子宮内膜癌之腫瘤內浸潤淋巴球，進一步利用細胞免疫螢光染色及流體細胞儀分析子宮内膜癌腫瘤中浸潤淋巴球表面自然殺手細胞抑制受體的分布。 結果 我們在子宮内膜癌組織分離出的單核細胞球相對於正常子宮内膜組織中有較多的現象 (8450/5655)，且子宮内膜癌病患分離出的腫瘤內浸潤 CD3+ T 淋巴球比正常子宮内膜病患分離出的正常子宮内膜浸潤 CD3+ T 淋巴球有較高表現比例 (子宮内膜癌90.05%； 正常子宮内膜79.60，P = 0.086)，表示子宮内膜癌組織可能比正常子宮内膜組織含有更多的浸潤淋巴球，因此 CD3+ T 淋巴球對於子宮内膜癌之抗癌免疫反應可能扮演重要角色。在進一步的研究當中，發現從子宮内膜癌組織分離出來的浸潤 CD3+ CD8+ T 淋巴球顯著比週邊血液 CD3+ CD8+ T 淋巴球含有較高表現比例的CD94 及 NKG2A，流體細胞儀分析顯示8.40%的子宮内膜癌浸潤淋巴球表現 CD94 (子宮内膜癌，8.40% [4.95-13.73]；週邊血液，3.80% [1.30-5.00]，P = 0.013)，而15.90%的子宮内膜癌浸潤淋巴球表現 NKG2A (子宮内膜癌，15.90% [8.25-21.15]，週邊血液，2.10% [1.15-5.30]，P < 0.001)；CD94 及 NKG2A 的表現比例在子宮内膜癌浸潤淋巴球顯著高於控制組的週邊血液單核細胞球及正常子宮内膜組織，這些現象暗示著腫瘤可能分泌某種物質，以改變自然殺手細胞抑制受體在 T 細胞上的表現，而高度表現於子宮内膜癌組織浸潤淋巴球的自然殺手細胞抑制受體，可能會抑制淋巴球的毒殺反應，並且使其失去對腫瘤的局部免疫控制。除此之外，我們也觀察到 CD158b 及 NKB1 高度表現於子宮内膜癌病患的週邊血液 CD3+ CD8+ T 淋巴球 (CD158b：子宮内膜癌組織，3.80% [1.08-7.28]；週邊血液，10.70% [4.95-14.98]，P = 0.001；NKB1：子宮内膜癌組織，0.40% [0.00-0.80]；週邊血液，2.20% [0.43-3.88]，P = 0.045)，而這些現象可能會使全身免疫系統對於腫瘤的控制降低。 我們第二部分的研究，主要為利用流體細胞儀 PhiPhiLux 毒殺試驗分析高度表現於 CD3+ CD8+ T 淋巴球上的 CD94/NKG2A 對於其毒殺能力的影響。結果顯示CD94/ NKG2A 在 CD3+ CD8+ T 淋巴球上的高度表現明顯會降低其毒殺能力，而阻斷了 CD94 或 NKG2A 與 HLA class I 分子的作用，則可增加其毒殺能力，NKG2A的阻斷對於 CD3+ CD8+ T 淋巴球的毒殺能力的增加明顯高於 CD94 的阻斷，且愈高的 E/T ratio 差別愈明顯。所以高度表現於 CD3+ CD8+ T 淋巴球上的 CD94 或 NKG2A 可能與其無法有效毒殺子宮内膜癌細胞有關。 結論 由於人類子宮内膜癌腫瘤細胞可產生大量之細胞激素 (cytokines)，而免疫細胞表面自然殺手細胞抑制受體可能受細胞激素之影響，進一步抑制了免疫細胞對於子宮内膜癌的抗癌反應。因此，子宮内膜癌細胞本身可能提供必要的訊息，使對抗腫瘤的專一毒殺性淋巴球表現出自然殺手細胞抑制受體，使子宮内膜癌逃過免疫監控。我們期待在不久的將來，可釐清高度表現於子宮内膜癌組織浸潤淋巴球的自然殺手細胞抑制受體在活體內 (in vivo) 上的功能，與其對於子宮内膜癌細胞免疫監控的影響，更進一步幫助我們對於未來抗癌藥物的發展。 | zh_TW |
| dc.description.abstract | Background: To investigate the expression of inhibitory natural killer receptors (iNKRs) within the human tumor milieu, we directly examined the in vivo expressions of various iNKRs on tumor-infiltrating lymphocytes (TILs) derived from human endometrial carcinoma (EC).
Material and Methods: Totally 22 patients with Stage Ia–IIIa EC were enrolled. TILs were isolated from tissue specimens by means of a mechanical dispersal technique. The subpopulations of immunocytes were quantified and expressions of NKRs on CD8 | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T17:26:03Z (GMT). No. of bitstreams: 1 ntu-94-P91421014-1.pdf: 2905706 bytes, checksum: a2126293d9fdab9ab58f4cb0021e40ba (MD5) Previous issue date: 2005 | en |
| dc.description.tableofcontents | 目錄
一、中文摘要•••••••••••••••••••••••••••••••••••••••1 二、緒論 研究背景•••••••••••••••••••••••••••••••••••••••••••3 子宮内膜癌的發生率••••••••••••••••••••••••••••••••••3 免疫監督與腫瘤內浸潤淋巴球••••••••••••••••••••••••••••3 免疫逃避性••••••••••••••••••••••••••••••••••••••••••4 自然殺手細胞抑制受體•••••••••••••••••••••••••••••••••4 細胞激素與自然殺手細胞抑制受體•••••••••••••••••••••••••6 研究目的••••••••••••••••••••••••••••••••••••••••••••7 研究的假說••••••••••••••••••••••••••••••••••••••••••7 三、研究方法與材料 病例收集••••••••••••••••••••••••••••••••••••••••••••8 機械式研磨萃取•••••••••••••••••••••••••••••••••••••••8 流體細胞儀分析••••••••••••••••••••••••••••••••••••••8 表現比例的定義••••••••••••••••••••••••••••••••••••••9 細胞激素(rhIL-15)誘發CD94/NKG2A分析••••••••••••••••••••9 流體細胞儀細胞毒殺分析•••••••••••••••••••••••••••••••9 統計分析•••••••••••••••••••••••••••••••••••••9 四、結果 第一部分 子宮内膜癌組織及正常子宮内膜組織的重量及分離出的細胞•••••11 子宮内膜癌病患的週邊血液單核細胞球及腫瘤內浸潤淋巴球之比較-- CD3+ 腫瘤內浸潤淋巴球含有較高表現比例的 CD8+ T 淋巴球••11 子宮内膜癌病患的週邊血液單核細胞球分離出來的 CD3+ CD8+ T 淋巴球含有較高表現比例的CD158b及NKB1•••••••••••••••••••11 子宮内膜癌病患的腫瘤內浸潤淋巴球分離出來的 CD3+ CD8+ T 淋巴球含有較高比例的CD94/NKG2A•••••••••••••••••••••••••••12 第二部分 活體外 (in vitro) rhIL-15 刺激 CD3+ CD8+ T 淋巴球表現 CD94/NKG2A•••••••••••••••••••••••••••••••••••••••••12 rhIFN-γ可明顯誘導出 HLA class I 分子高度表現於 K562 標的細胞上•••••••••••••••••••••••••••••••••••••••••••••••12 CD94/NKG2A在 CD8+ T 淋巴球上的高度表現明顯會降低其毒殺能力••••••••••••••••••••••••••••••••••••••••••••••••12 五、討論••••••••••••••••••••••••••••••••••••••••••14 六、展望 子宮内膜癌組織內細胞激素之表現••••••••••••••••••••••17 腫瘤細胞與自體免疫細胞的混合培養分析••••••••••••••••••17 子宮内膜癌組織腫瘤內浸潤CD3+ CD8+ T 淋巴球之毒殺分析•••••18 七、論文英文簡述 (summary) Abstract••••••••••••••••••••••••••••••••••••••••••19 Introduction•••••••••••••••••••••••••••••••••••••••20 Material and Methods••••••••••••••••••••••••••••••••22 Results•••••••••••••••••••••••••••••••••••••••••••27 Discussion••••••••••••••••••••••••••••••••••••••••31 八、參考文獻•••••••••••••••••••••••••••••••••••••••34 九、圖表••••••••••••••••••••••••••••••••••••••••••43 圖表目錄 圖一、自然殺手細胞抑制受體影響毒殺性 T 淋巴球毒殺力之機轉••••••••••••••••••••••••••••••••••••••••••••••••43 圖二、自然殺手細胞抑制受體影響毒殺性淋巴球對被病毒感染或腫瘤細胞之毒殺力的機轉••••••••••••••••••••••••••••44 圖三、流體細胞儀分析下相互比較由子宮内膜癌組織 (i-p) 或週邊血液單核細胞球 (a-h) 分離出之 CD3+ CD4+ 或 CD3+ CD8+ T 淋巴球上自然殺手細胞抑制受體 (CD158a, CD158b, 及NKB1) 的表現•45 圖四、流體細胞儀分析下相互比較CD94 及 NKG2A 於子宮内膜癌組織 (g-l) 與週邊血液單核細胞球 (a-f) 分離出之 CD3+ CD4+ 或 CD3+ CD8+ T 淋巴球上的表現•••••••••••••••••••••••••••46 圖五、活體外 (in vitro) rhIL-15 刺激 CD3+ CD8+ T 淋巴球表現 CD94••••••••••••••••••••••••••••v•••••••••••••47 圖六、活體外 (in vitro) rhIL-15 刺激 CD3+ CD8+ T 淋巴球表現 NKG2A••••••••••••••••••••••••••••••••••••••••••••••48 圖七、將 K562 細胞以 rhIFN-γ (200 ng/ml) 刺激活化2天，誘導出 HLA class I 分子的表現••••••••••••••••••••••••••••••••••••••••••••••••49 圖八、流體細胞儀分析在不同比例的效應細胞與標的細胞 (E/T ratio = 1:1，3:1，9:1) 下，PhiPhiLux 毒殺分析試驗的結果••••••••••••••••••••••••••••••••••••••••••••••50 圖九、比較在不同比例的效應細胞與標的細胞下5組毒殺分析試驗的結果•••••••••••••••••••••••••••••••••••••••••••••••51 表一、子宮内膜癌組織及正常子宮内膜組織的平均重量及分離出的細胞••••••••••••••••••••••••••••••••••••••••••••••••52 表二、子宮内膜癌病患與正常子宮内膜病患分離出的週邊血液單核細胞球及腫瘤或正常子宮内膜組織內浸潤淋巴球之比較•••••••53 表三、流體細胞儀分析比較子宮内膜癌病患與正常子宮内膜病患分離出的週邊血液單核細胞球及腫瘤或正常子宮内膜組織內浸潤淋巴球，其自然殺手細胞抑制受體 (CD158a, CD158b, NKB1, CD94, NKG2A) 於 CD8+ T 淋巴球之表現•••••••••••••••••••••••••54 | |
| dc.language.iso | zh-TW | |
| dc.subject | 腫瘤內浸潤淋巴球 | zh_TW |
| dc.subject | 自然殺手細胞抑制受體 | zh_TW |
| dc.subject | 膜癌 | zh_TW |
| dc.subject | 子宮内 | zh_TW |
| dc.subject | endometrial cancer | en |
| dc.subject | inhibitory natural killer receptors | en |
| dc.subject | tumor-infiltrating lymphocytes | en |
| dc.title | 子宮內膜癌腫瘤內免疫細胞及抑制受體分析 | zh_TW |
| dc.title | Analysis of Inhibitory Natural Killer Receptors on Tumor Infiltrating Lymphocytes Derived from Human Endometrial Cancer | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 93-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | #VALUE! | |
| dc.subject.keyword | 膜癌,自然殺手細胞抑制受體,腫瘤內浸潤淋巴球,子宮内, | zh_TW |
| dc.subject.keyword | tumor-infiltrating lymphocytes,endometrial cancer,inhibitory natural killer receptors, | en |
| dc.relation.page | 54 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2005-01-21 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 臨床醫學研究所 | zh_TW |
| 顯示於系所單位： | 臨床醫學研究所 | |
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