• 作者： 趙祖怡; 黃偉修; 葉明陽
• 作者服務機構： 國防醫學院暨三軍總醫院內科學科血液腫瘤科; 奇美醫院內科部血液腫瘤科; 國防醫學院微生物及免疫學科
• 中文摘要： 淋巴素活化殺手細胞活性從十位癌症患者之惡性腹腔積液中生成。這十位癌症患者包括五位結腸直腸癌，三位胃癌，一位卵巢癌，及一位膽道癌。惡性腹腔積液中之淋巴球，簡稱EALs，在經由密度階差法及塑膠黏著法分離出來後在含有介白質2之培養液中連續培養30日。這些淋巴球之自然殺手細胞活性及經介白質2刺激後生成之淋巴素活化殺手細胞活性在培養之第0，7，14，及30日以鉻-51釋放試驗檢測，以K-562，HL-60，及自體癌細胞作為標的細胞。所有十位癌症患者新鮮分離之EALs均無法探測到自然殺手細胞活性。然而，淋巴素活化殺手細胞活性卻可自所有十位癌症患者之EALs生成，而其活性在培養之第7日達於頂點。當EALs存於培養液中愈久其淋巴素活化殺手細胞活性愈弱。只要介白質2繼續存在，EALs便可不斷增生。細胞表型研究顯示新鮮分離之EALs由自然殺手細胞，T輔助∕引誘細胞，T殺手／抑制細胞，及B淋巴細胞組成，分別佔有22％，18％，50％，及8％。在培養一段時閒後B淋巴細胞逐漸減少至消失，T淋巴細胞則佔絕大部份，其中T輔助∕引誘細胞之比例增加。進一步之研究顯示由EALs生成之淋巴素活化殺手細胞活性主要來自自然殺手細胞。這些結果意味存在於惡性腹腔積液中之自然殺手細胞原本處於被抑制狀態，但可為介白質2刺激後增強。當EALs被考慮用作癌症之免疫治療時，我們建議以一次大量分離EALs再經由短期培養於介白質2中取代小量分離EALs再長期培養的作法，其目的是為了要保持理想的弒癌活性。
• 英文摘要： A generation of lymphokine-activated killer (LAK) cell activities from malignant peritoneal effusionswas investigated in 10 patients with abdominal carcinomatosis. Five of the 10 patients were victims ofcolorectal cancers, three of gastric cancers, and one each of ovarian cancer and cholangiocarcinoma. Lym-phocytes, the so-called effusion associated lymphocytes (EALs), were isolated from malignant peritonealeffusions by density gradient centrifugation and the plastic adherence method. These isolated EALs weresubsequently cultured in the presence of recombinant interleukin-2(rIL-2), 3,000 I.U./ml, for 30 days.Natural killer (NK) cell activities and LAK cell activities of the freshly isolated and cultured EALs wereexamined at 0, 7, 14, and 30 days of culture by means of a standard Cr-release assay using K-562, HL-60,and autologous tumor cells as target cells. The NK cell activities of the freshly isolated EALs were notdetected in any of the 10 patients. The LAK activities, however, could be generated in all of them, andthe activities were maximal at 7 days. The longer the EALs remained in the culture, the weaker were theLAK cell activities. As far as cell growth was concerned, EALs proliferated well as long as the rIL-2 werepresent in the culture. Phenotypic analysis of the freshly isolated EALs revealed the presence of NK cells(22％，CD16＋CD56＋). T helper/inducer (18％, CD4＋), T cytotoxic/suppressor (50％, CD8＋), and Bcells (8％, CD 19＋). After being cultured with rIL-2, the B lymphocytes gradually disappeared, and the Tlymphocytes predominated with an increase in the percentage of T helper/inducer cells. Additional studiesrevealed that the major precursors of the LAK cell activities were NK cells. These results indicate that NKactivity in malignant peritoneal effusions is suppressed in cancer patients, but that it can be augmented byrIL-2. When EALs are considered for the use of adoptive immunotherapy, our data suggest that it is betterto isolate a large amount of EALs from voluminous effusions with short-term culturing than to proliferateEALs in vitro for a long time in order to obtain an ideal cytotoxicity against cancers.
• 中文關鍵字： LAK cell activity; malignant peritoneal effusions.
• 英文關鍵字： --