臺灣博碩士論文加值系統

中文摘要
霍奇金病是一種淋巴瘤，而在病人的淋巴結會有不正常的細胞，這些細胞被命名為Reed-Sternberg細胞；CD30起初被描述當為Hodgkin/Reed-Sternberg (H-RS) 細胞的一個標識。CD30與它的同源配體CD153，分別屬於腫瘤壞死因子接受器(TNFR)以及腫瘤壞死因子(TNF)的家族成員之一，CD30被表現在霍奇金淋巴瘤的腫瘤細胞上，而藉由anti-CD3或是PHA的活化下，CD153能被誘導表現於周邊血液T細胞．在本篇研究，我們提出CD30逆訊息傳導 (reverse signaling) 在T細胞上的效應，藉由Hodgkin/Reed-Sterberg細胞株 (KMH-2) 與刺激anti-CD3抗體或是PHA的PBMC一起共同培養，我們發現T細胞的增生有被抑制情形，此種抑制現象不是憑藉於KM-H2細胞所釋放的激素或受質，因為KM-H2細胞在共同培養之前，已事先被福馬林固定。我們進一步的使用表現CD30的CHO細胞研究CD30在T細胞增生的效應。在CD30-陽性的CHO細胞存在下，PHA刺激的PBMC無法獲得有效之增生，假使刺激PHA的PBMC被培養在含CD30-Fc融合蛋白的培養基中，觀察到有相似的抑制效應。總結以上實驗，我們發現CD30之逆訊息能有抑制效應在CD153表現的T細胞。除此之外，為了描繪在CD30逆訊息反應之蛋白表現情形，使用蛋白質體學的技術來分析。並發現有一些蛋白被調控可能藉由CD30的結合反應，於這些蛋白中我們利用MALDI-TOF質譜儀鑑定出其中一個蛋白為錳-超氧化物歧化酶(Mn-SOD)。
我們推斷藉由CD30-CD30L互相影響，H-RS細胞能夠抑制活化T細胞的增生，而這也許是導致一個微觀環境有利於癌細胞的生長與存活。

英文摘要
Hodgkin''s disease is a type of malignant lymphoma, characterized by the presence of abnormal cells, named Reed-Sternberg cells, in patient’s lymph nodes. CD30 was originally described as a marker of Hodgkin/Reed-Sternberg cells. CD30 and its cognate ligand, CD153, belong to members of the TNFR and TNF superfamilies, respectively. CD30 is expressed on the surface of Hodgkin/Reed-Sternberg (H-RS) cell lines (KM-H2), while expression of CD153 can be induced on the surface of peripheral blood T cells by anti-CD3 or PHA activation. In this study, we addressed the effect of CD30 reverse signaling on T cells. By co-cultures of KM-H2 and PBMC activated by anti-CD3 or PHA, we observed T cell proliferation was inhibited. The inhibition was not dependent on cytokines or substances released from KM-H2, because KM-H2 cells were fixed with paraformaldehyde before the co-culture. We further study the effect of CD30 on T cell proliferation with CD30-expressing Chinese Hamster Ovary (CHO) cells to. In the presence of CD30-positive CHO cells, PHA-treated PBMC failed to achieve significant proliferation. Similar effects were observed if PHA-treated PBMC were cultured in the medium containing chimeric CD30-Fc fusion proteins. Taken together, we discover the inhibitory effect of CD30 reverse signaling on CD153-positive T cells. Furthermore, in order to characterize the protein expression profile in response to CD30 reverse signaling, proteomic technology was used. Several candidate spots were found to be likely regulated by CD30 engagement. One of these spots was identified as Mn-SOD by the use of MALDI-TOF MS.
We conclude that H-RS cells are able to inhibit the proliferation of activated T cells through the CD30-CD153 interaction, which may lead to a microenvironment in favor of the growth and survival of the tumor cells.

目錄
縮寫-----------------------------------------------1
中文摘要------------------------------------------.3
英文摘要-------------------------------------------4
緒論-----------------------------------------------5
第一節 前言--------------------------------------5
第二節 霍奇金病(Hodgking''s disease)------------- 5
第三節 腫瘤壞死因子群(TNF superfamily)--------- 12
第四節 腫瘤壞死因子受體群(TNFR superfamily)---- 13
第五節 逆訊息傳導(reverse signaling)------------13
第六節 CD30與CD30L------------------------------13
第一章 動機與目的---------------------------------17
第二章 實驗材料-----------------------------------18
第三章 實驗方法-----------------------------------21
第一節 細胞分離與培養---------------------------21
第二節 共同培養(co-culture)試驗-----------------24
第三節 融合蛋白的純化---------------------------27
第四節 融合蛋白 (CD30-Fc) 對細胞的影響----------35
第五節 蛋白質體學分析---------------------------35
第四章 實驗結果-----------------------------------43
第一節 細胞分離與培養---------------------------43
第二節 共同培養(co-culture)試驗-----------------43
第三節 融合蛋白的純化---------------------------45
第四節 融合蛋白 (CD30-Fc) 對細胞的影響----------46
第五節 蛋白質體學分析---------------------------46
第五章 討論---------------------------------------48
第一節CD30之功能性角色----------------------------48
第二節CD30與錳型SOD之間的關連---------------------49
第三節 未來展望-----------------------------------51
第六章 參考文獻-----------------------------------53
第七章 圖表---------------------------------------57

第六章 參考
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