LAK细胞和IL－2联合化疗治疗晚期肺癌的近期疗效观察

ＬＡＫ细胞和ＩＬ－２是目前常用的肿瘤生物制剂。自１９９２年以来，对１８例晚期肺癌，男１５例，女３例，年龄２９～６４岁，进行ＬＡＫ细胞和重组ＩＬ－２联合化疗治疗。选择胎脾、胸腺的淋巴细胞做前体细胞，体外用重组ＩＬ－２诱导制备ＬＡＫ细胞，每输３次ＬＡＫ细胞为一疗程、每次输入细胞数０．５～１．０×１０９。化疗采用环磷酰胺，长春新碱，阿霉素为主的方案。治疗结果：完全缓解（ＣＲ）５例，部分缓解（ＰＲ）７例，无效（ＮＲ）３例，病情平稳３例，有效率ＣＲ＋ＰＲ达６６％。采用本疗法后病人精神及饮食好转，能有效缓解胸痛、减轻病人痛苦。提示ＬＡＫ细胞和重组ＩＬ－２联合化疗对晚期肺癌是一种可行的有效治疗，但在临床应用中要注意毒性反应。     

肝动脉化疗栓塞结合LAK／IL—2治疗肝癌

用IL—2和胸腺因子D(TFD)联合诱导的自体LAK 4～8×10~(6～7)和同种异体LAK4～8×10~(8～9)对40例肝癌采用肝动脉化疗栓塞结合LAK/IL—2·TFD治疗.肝动脉灌注顺铂、阿霉素、丝裂霉素和部分病人用40%碘化油栓塞,LAK细胞10次为1疗程,包括静脉滴注8次～9次,肝动脉灌注1次～2次(肝动脉灌注包括LAK、IL—2和TFD),治疗期间每日肌注IL—2 500Ou～10000u和TFD10mg.疗效达到PR:52.5%、MR:30%、SD PD:17.5%.AFP下降占94.1%,半年生存率占80%.多数病人症状改善,免疫力增强,肝功能好转,生活质量提高.少数病人使用LAK细胞出现一过性寒战、发热.     

癌症患者群体LAK细胞的体外抗肿瘤的细胞毒活性

本研究应用~(125)I-UdR释放实验,测定了大量随机取样于111位癌症患者的外周血淋巴细胞以及从它们中诱导出的处于不同培养期的淋巴因子激活的杀伤细胞对K562和Raji肿瘤细胞的体外细胞毒活性.对560个有效数据的统计分析,获得以下结果:1.经过培养系统的作用,对K562细胞的细胞毒活性,从培养前的34.78±25%上升到8～13天培养期的68.04±17.3%之后基本稳定于近70%的水平,直至23～27天的培养期.此细胞毒活性的构成比模式表明,在培养之前,样品分布于很广的活性区域(10～90%);培养 8～13天和以后的培养期,约85%的样品集中到高活性区域(50～95%).2.对 Raji细胞的细胞毒活性,从培养前的8.9±8%上升到8～13天培养期的42.1±22%之后,在随后的培养期中,维持于约35%的水平.对Raji的细胞毒活性的构成比显示,在培养前,全部样品处于低活性区域(<25%);在培养过程中,部分样品(约30%)向高活性区域发生明显的扩展(50～90%),但另一部份样品(约40%)一直位于低活性区域(<35%).这样形成高低两个分布区域.这种现象的机理值得进一步探讨.     

CD3AK细胞和LAK细胞治疗晚期恶性肿瘤的临床和实验研究

将５１例晚期恶性肿瘤患者（男性２３例，女性２８例）分成两组，其中一组（３１例）以ＣＤ３ＭｃＡｂ（ＣＤ３单克隆抗体）和小剂量ＩＬ－２（５００ｕ／ｍｌ）共同诱导的ＣＤ３ＡＫ细胞治疗，另一组（２０例）输注大剂量ＩＬ－２（１０００ｕ／ｍｌ）诱导的常规ＬＡＫ细胞治疗，以探讨降低ＩＬ－２用量、提高杀伤细胞细胞毒活性的可能性。结果显示ＣＤ３ＡＫ组患者生活质量改善、症状缓解均优于ＬＡＫ组。ＣＤ３ＡＫ组ＰＲ＋ＭＲ率较ＬＡＫ组高２９．０％，Ｓ＋Ｐ率和死亡率分别较ＬＡＫ组低１２．４％和９．６％。同时比较了ＣＤ３ＡＫ细胞与ＬＡＫ细胞的体外增殖和细胞毒活性，结果表明ＣＤ３ＡＫ细胞增殖率高于ＬＡＫ细胞（Ｐ＜０．０１），靶细胞抑制率二者在０．０５水平无显著差异。提示ＣＤ３ＭｃＡｂ在刺激杀伤细胞活性，尤其在提高其增殖能力方面，具有显著的作用，ＣＤ３ＡＫ／ＩＬ－２能更有效地治疗晚期恶性肿瘤。     

粘附性淋巴因子活化的杀伤细胞抗自体白血病细胞作用的研究

建立了一套培养和处理粘附ＬＡＫ细胞（Ａ－ＬＡＫ）的系统，将１２例缓解期急性髓系白血病（ＡＭＬ）患者（ＲＰＳ）的Ａ－ＬＡＫ细胞与常规制备ＬＡＫ细胞（ＲＴ－ＬＡＫ）进行了对照研究，结果显示ＲＰＳ－Ａ－ＬＡＫ细胞于培养第１０天时其扩增指数（２０．１±１３．９）较ＲＴ－ＬＡＫ细胞的扩增指数（７．５±２．１）明显提高（Ｐ＜０．０５）。形态学研究显示Ａ－ＬＡＫ细胞主要由大颗粒淋巴细胞组成，免疫表型分析显示其主要由ＣＤ１６＋的ＮＫ细胞组成，ＲＴ－ＬＡＫ细胞主要由ＣＤ３＋的Ｔ细胞组成。其杀伤活性与ＣＤ１６＋ＮＫ细胞之间有很好的相关性（ｒ＝０．８２，Ｐ＜０．０５）。这提示ＣＤ１６＋ＮＫ细胞是Ａ－ＬＡＫ细胞的主要组成细胞，代表了杀伤功能最强的亚群。     

TIL细胞与LAK细胞体外杀伤活力的比较

应用改良ＬＤＨ释放法，对ＴＩＬ细胞与ＬＡＫ细胞的前体及培养激活的细胞进行杀伤活力的检测，结果显示无论前体细胞或培养激活的细胞，对自体瘤细胞及Ｋ５６２细胞的杀伤活性均以ＴＩＬ细胞为高，表明ＴＩＬ细胞对肿瘤的特异识别及杀伤力均较ＬＡＫ细胞为强。     

人肺腺癌细胞经化疗药物作用后对LAK细胞的敏感性

在体外实验中研究了８种化疗药物作用后的个旧人肺腺癌细胞（ＧＬＣ－８２）对ＬＡＫ细胞的敏感性。ＧＬＣ－８２细胞本身对化疗药物不敏感，经药物作用２４小时后用３Ｈ－ＴｄＲ释放法检测ＬＡＫ细胞对其杀伤率。结果表明：顺铂（ＣＤＤＰ）、卡铂（Ｃａｒｂｏ）、阿霉素（ＡＤＭ）、丝裂霉素（ＭＭＣ）及长春新硷（ＶＣＲ）处理后的ＧＬＣ－８２细胞对ＬＡＫ细胞的敏感性增强；环磷酰胺（ＣＴＸ）、足叶乙甙（ＶＰ１６）及５－氟脲嘧啶（５－Ｆｕ）则无明显影响。实验提示：对化疗不敏感的肺腺癌用ＣＤＤＰ、Ｃａｒｂｏ、ＡＤＭ、ＭＭＣ及ＶＣＲ后加用ＬＡＫ细胞可提高肺癌治疗的有效率。     

自体脾LAK细胞经肝动脉导管输注治疗原发性肝癌

以基因重组白细胞间介素Ⅱ(rIL-2)诱导肝癌患者自体脾LAK细胞,经肝动脉,辐注。自体LAK细胞10~9×2,rIL-2 5×10~4u×2连续低剂量经肝动脉导管输注一个疗程。治疗连续观察血清AFP变化及肝功能变化。用B超、CT观察肿块变化,结果显示:本组3例治疗后血清AFP均连续下降,B超、CT显示肿块缩小。患者主观症状好转,肝功能有所恢复。自体LAK细胞输注后仅有一过性发热。     

Augmentation of the Susceptibility of Human Leukemia to Lymphokine-Activated Killer (LAK) Cells by Exposure of the Leukemic Target Cells to Cytotoxic Drugs in Vitro and in Vivo

Experimental studies have shown that interleukin-2-induced lymphokine-activated killer (LAK) cells are able to lyse fresh noncultured leukemia cells and that human leukemia cells have a distinct susceptibility to LAK-cell-mediated cytolysis. Cytolysis is considerably lower with fresh noncultured leukemia cells than with the leukemia cell lines K562 and Daudi. For therapeutic considerations it would be desirable to achieve as much cytolysis as possible. The current study revealed that incubating leukemia cells with cytotoxic drugs in vitro significantly augments their susceptibility to the lytic effect of LAK cells and, more importantly that exposing leukemia cells to anticancer agents in vivo during induction chemotherapy also increases their sensitivity to LAK-cell-mediated cytolysis. These results support a possible benefit from combining chemotherapy with immunotherapeutic approaches in leukemia treatment.     

Granulocyte-Macrophage Colony-stimulating Factor Augments Lymphokine-activated Killer Activity from Pleural Cavity Mononuclear Cells of Lung Cancer Patients without Malignant Effusion

The role of recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) in augmentation of lymphokine-activated killer (LAK) cell induction by interleukin-2 (IL-2) from pleural cavity mononuclear cells (PCMNCs) was examined in sixteen patients with resectable primary lung cancer not associated with malignant effusion. None of the patients had received any anticancer therapy prior to this study. Incubation of PCMNCs of patients without malignant effusion with GM-CSF for 4 days in the presence of IL-2 resulted in a significant increase in LAK activity against natural killer-resistant Daudi cells. This result was obtained by using the 4 h ⁵¹Cr-release assay. PCMNCs and blood mononuclear cells (BMNCs) were harvested simultaneously from pleural cavity lavage fluid and peripheral blood in lung cancer patients. The LAK activity developed from PCMNCs and BMNCs following incubation with IL-2 for 4 days, but the LAK activity from PCMNCs was significantly lower than that from BMNCs ( P < 0.05). Incubation of PCMNCs with GM-CSF augmented the LAK activity from PCMNCs to a level as high as that from BMNCs. These results suggest that the combined use of GM-CSF with IL-2 may result in augmentation of LAK activity developed from PCMNCs of lung cancer patients without malignant effusion.     

肝癌病人化疗栓塞前后LAK细胞活性变化的研究

经肝动脉化疗栓塞（ＴＡＣＥ）与免疫疗法是当前中晚期肝癌治疗中的活跃领域，但ＴＡＣＥＥ对肝癌病人肿瘤免疫的影响尚不明确。本研究观察测定了未行治行的肝癌病人，行一次ＴＡＣＥ治疗的肝癌病人及反复行ＴＡＣＥ治疗的肝癌病人的ＬＡＫ细胞活性。结果表明：单纯行一次ＴＡＣＥ治疗的肝癌病人其ＬＡＫ细胞活性未见明显变化，而反复行ＴＡＣＥ治疗的肝癌病人其ＬＡＫ细胞活性明显增强。提示反复ＴＡＣＥ疗法对肝癌病人的肿瘤免疫有增强作用，而ＴＡＣＥ治疗中化疗药物的用量是对肿瘤免疫产生影响的重要因素。     

消炎痛对膀胱癌患者抗肿瘤免疫的影响

 目的　研究消炎痛 ( IM)对膀胱癌患者 LAK细胞增殖和抗膀胱癌细胞的细胞毒作用的影响。方法　用细胞计数法观察 LAK细胞增殖 ,以膀胱癌组织标本分离的肿瘤细胞和培养膀胱癌细胞系 BIU- 87和 EJ作为靶细胞 ,用 MTT法测定 LAK细胞对肿瘤细胞的细胞毒作用。结果　培养 48至 96小时 IM明显增强白细胞介素 ( IL- 2 )诱导的 LAK细胞增殖 ,IM处理 LAK细胞可增强其对膀胱肿瘤细胞的杀伤。结论　这些研究提示 IM对 IL- 2诱导的抗肿瘤免疫呈增强调节.     

Non HLA-Restricted Cytotoxic Cells and Their Modulation in Acute Myelogenous Leukemia

The observation of more frequent leukemias in immune deficiencies of the non-HLA restricted cytotoxic cell system and the participation of this system in the antileukemic effect of allogeneic bone marrow transplantation in acute myelogenous leukemia (AML) indicate that these cells play a role in the immune surveillance against AML. The demonstrated susceptibility of fresh AML cells to lysis by lymphokine-activated killer cells in vitro has triggered trials attempting to modulate the non-HLA-restricted cytotoxic cells in vivo in AML during the blastic phase as well as during complete remission. The relationships between AML and the non-specific cytotoxic cells and the potential modulation of these cells for the therapy of this disease are reviewed.     

LICC细胞和LAK细胞治疗晚期恶性肿瘤的疗效观察

采用淋巴因子诱导的细胞毒细胞（ＬＩＣＣ）和ＩＬ—２／ＬＡＫ细胞治疗３２例晚期恶性肿瘤患者，ＬＩＣＣ治疗２０例，ＩＬ—２／ＬＡＫ治疗１２例，结果发现６８．８％（２２／３２）卡氏得分上升，多数患者癌性症状减轻，生活质量提高，以ＬＩＣＣ组优于ＬＡＫ组，ＣＲ＋ＰＲ＋ＭＲ率，ＬＩＣＣ为４０．０％（１２／２０），ＬＡＫ组为１６．７％（２／１２）。作者认为过继免疫作为一种癌症辅助治疗方法应予肯定；ＬＩＣＣ治疗安全、副反应小，抗瘤谱广，费用低，疗效优于ＩＬ—２／ＬＡＫ，值得扩大应用性研究。     

Successful Induction of Tumor-specific Cytotoxic T Lymphocytes from Patients with Non-small Cell Lung Cancer Using CD80-transfected Autologous Tumor Cells

Cytotoxic T lymphocytes (CTL) against human lung cancer cells are difficult to induce by a conventional method using tumor cell stimulation probably due to an insufficiency of tumor antigens (TA) or costimulatory molecules such as CD80. We, therefore, investigated the potential of CD80-transfected tumor cells as stimulators of the in vitro induction of autologous tumor-specific CTL from regional lymph node lymphocytes in patients with lung cancer. Five non-small cell lung cancer cell lines (two adenocarcinomas, 1 squamous cell carcinoma, 1 large cell carcinoma and 1 adenosquamous cell carcinoma) were established from surgical specimens and were successfully transduced with a plasmid constructed with expression vector pBj and human CD80 cDNA, using a lipofection method. CD80-transfected tumor cells (CD80-AT) significantly augmented the proliferation of autologous lymphocytes from all cases as compared with non-transfected tumor cells (AT). AT-stimulated lymphocytes from 4 out of 5 cases did not show any cytotoxicity against AT; however, lymphocytes stimulated with CD80-AT exhibited substantial cytotoxicity against parental AT in all 5 cases tested. AT-stimulated lymphocytes derived from only one out of 5 cases showed major histocompatibility complex (MHC)-class I-restricted cytokine production in response to AT, while the MHC-class I-restricted responses were found in CD80-AT-stimulated lymphocytes from 4 out of 5 cases. These results indicate that CD80 on tumor cells could be a beneficial costimulatory molecule to elicit CTL against lung cancer, and also show that TA recognized by CTL was frequently expressed on lung cancer cells.     

人类LAK细胞体外实验性扩增及生物学活性分析

由癌症患者外周血中分离出少量ＬＡＫ前体细胞。在ＬＡＫ细胞培养体系中加入重组人类白细胞介素２，植物血球凝集素及辅助细胞，明显提高了ＬＡＫ细胞的体外增殖能力并延长了ＬＡＫ细胞的存活时间。表型分析发现，ＬＡＫ－Ｌ细胞的ＩＬ－２受体（ＩＬ－２－Ｒ－）表达水平明显高于ＬＡＫ－Ｓ细胞，ＬＡＫ－Ｌ细胞的体外大量增殖具有以ＯｋＴ＾＋ ８表型为主的单亚群增殖特性。体外杀伤实验及裸小鼠体内成瘤抑制实验结果表明，用此法     

Role of Interleukin-2 Activated MHC-Nonrestricted Lymphocytes in Antileukemia Activity and Therapy

Although the application of biological therapy for solid tumors with cytokines and adoptively transferred activated lymphocytes has received substantial attention, this approach has not been actively explored in treatment of hematopoietic neoplasms. This review will address the possibilities of interleukin-2 (IL-2) and IL-2 activated natural killer (NK) cells and T cells in antileukemia reactivity and therapy. The new approaches to optimal activation and generation of oncolytic cells, selective propagation of lymphocyte subsets, and the role of adhesion molecules in antileukemia cytotoxicity will also be addressed. We trust that this article will be conducive to the development of new directions in leukemia research and treatment.     

硒酸酯多糖增强荷瘤小鼠免疫功能和自身肿瘤杀伤活性的作用

本文研究了硒酸酯多糖(KSC)对荷S180肉瘤小鼠NK细胞活性、LAK细胞活性、IL-2分泌能力和自身肿瘤杀伤活性(ATK)的影响及抑癌效应.结果表明,KSC(40mg/kg·d×9d,ig)能增强荷瘤鼠NK细胞和LAK细胞活性,促进脾细胞产生IL-2,增强ATK活性;加强环磷酰胺(Cy,20mg/kg·d×9d,ip)的抑瘤作用,并能拮抗Cy对免疫活性细胞的抑制效应.体外用rIL-2激活荷瘤鼠脾淋巴细胞可诱生或增强ATK活性.本研究结果提示,KSC上调肿瘤宿主NK细胞和LAK细胞活性及IL-2的分泌水平,增强ATK活性,可作为生物反应调节剂(BRM)应用于肿瘤生物治疗.     

健康人和骨肉瘤病人来源的LAK细胞体外抗瘤作用

采用５１Ｃｒ释放试验对健康人和骨肉瘤病人外周血单个核细胞（ＰＢＭ）在重组白细胞介素－２（ｒＩＬ－２）条件下，ＬＡＫ细胞的诱导形成及对４种传代瘤细胞的体外杀伤活性进行探讨。实验结果表明：２种来源ＬＡＫ细胞对Ｋ５６２（人慢性髓样红白血病细胞系）、ＳＭＭＣ７７２１（人肝癌细胞系）、ＬＡＸ（人肺腺癌细胞系）的杀伤活性均在５５％以上（按效靶比例５０：１），而对ＯＳ细胞（人骨肉瘤细胞系）的活性普遍低下，杀伤率低于３５％。结果证实：１）健康人和骨肉瘤病人的ＰＢＭ均能在ｒＩＬ－２条件下诱导形成具有广谱抗瘤活性的ＬＡＫ细胞群，二者的杀伤格局和效力相近。２）骨肉瘤细胞本身对ＬＡＫ细胞的杀伤作用存在强烈抗性。     

LAK细胞及rIL—2腹腔灌注对胃癌术后预防腹膜转移的研究

作者选用3组Ⅲ期胃癌病例，即根治术后用LAK细胞／rIL－2腹腔灌注的LAK组，用5－Fu腹腔灌注的5-Fu组，未用药的对照组。经随访观察发现LAK组出现腹膜种植转移者2年只有7％（2／27），明显少于另外两组。因此文章指出：①LAK细胞／rIL－2腹腔灌注能有效的预防胃癌根治术后的腹腔种植转移，临床推广使用俾有助于生存率的提高；②鉴于LAK细胞的杀伤特性，阐明其真正价值是用于预防癌根治后的转移；③只有LAK细胞数量超过癌细胞数时才能有效的发挥其作用。