TIL与膀胱肿瘤

1986年,Rosenberg等人从肿瘤组织中分离出浸润的淋 巴细胞,将其用IL 2扩增达一定数量后回输给荷瘤小鼠,可 使肿瘤消退,这种淋巴细胞被称为肿瘤浸润淋巴细胞(tumor infiltratinglymphocyte,TIL)。10余年来,人们对膀胱肿瘤 中的TIL进行了一系列的研究,现综述如下。 1　膀胱肿瘤中TIL的细胞成分及其含量、分布和活性 Tsujihashi等[1-3]使用消化酶和菲科尔 泛影葡胺将膀胱 肿瘤组织处理后,用间断梯度离心法分离出TIL。然后再使 用流式细胞仪对TIL中细胞成分、含量进行研究,认为TIL 主要由Leu4(T细胞)、Leu10(B细胞)、Leu7(NK…     

负载肾癌肿瘤裂解物的树突状细胞诱导产生抗原特异性细胞毒T淋巴细胞

目的　利用树突状细胞呈递肿瘤抗原的特性提高细胞毒T淋巴细胞 (CTLs)对肾癌细胞的杀伤活性。　方法　肾细胞癌患者骨髓来源的有核细胞体外经GM CSF和IL 4诱导产生树突状细胞 ,负载肿瘤裂解物后诱导自体CTLs产生。用细胞毒试验和ELISA测定CTLs杀伤活性和细胞因子的分泌。　结果　肾癌患者自体来源的DC Tuly能 (1)增加CTLs增殖 ,16d时使T细胞增殖达 4 3倍 ;(2 )上调CTLs中CD3 和CD8 T细胞群 ;(3)诱导产生的CTLs对自体肾癌细胞具有高杀伤率 ,显著高于异体肾癌和异种肿瘤细胞 (P <0 .0 5 ) ;(4)上调TNF α分泌。　结论　DCs能呈递Tuly抗原。诱导产生抗原特异性CTLs,提示DC Tuly具有为肾癌患者制作疫苗和进行特异性CTLs过继免疫治疗的临床应用前景     

转Survivin基因树突状细胞抗消化道肿瘤的免疫效应研究

目的　研究转染Survivin的树突状细胞 (DC)在体外诱导高效而特异的抗消化道肿瘤免疫效应。方法　用脂质体作为介质 ,将Survivin基因转染入DC ,用Westernblot法检测培养上清Survivin的表达 ,检测这种DC分泌细胞因子白介素 (IL 12 )、肿瘤坏死因子 (TNF) α的功能 ,以及表面分子CD1a、CD83、MHcⅡ、CD80、CD86表达的高低 ,用MTT法诱导人特异的细胞毒性T淋巴细胞 (CTLs)的能力。结果　培养上清中均可以检测到Survivin表达 ;转基因DC的上清IL 12、TNF α两种细胞因子含量为 (2 65 .2± 3 2 .7)ng/L和(4 3 7.1± 83 .5 )ng/L明显比单纯DC组高(P <0 .0 5 ) ;转基因DC表面高表达CD1a、CD83、MHCⅡ、CD80、CD86;转基因的DC提呈的T细胞对胃癌细胞、结肠癌细胞、胆管癌细胞杀伤率分别为 :65 %、77%、85 % ,而未修饰的单纯DC杀伤作用较低。结论　Survivin基因转染修饰的DC能诱导细胞毒性T淋巴细胞的特异性 ,显著地提高DC的抗原提呈功能 ,体外能诱导高效而特异的抗癌免疫效应。     

脾内转染白细胞介素-2和/或12基因增强接种肝癌大鼠T细胞功能研究

目的　探讨脾内直接注射白细胞介素IL 2、12基因对血IL 2和IL 12 ,以及T细胞活性的影响。方法　构建IL 2和或IL 12基因的逆转录病毒载体。含IL 2和或IL 12基因的包装细胞于不同时间进行脾内注射转染脾细胞。比较大鼠血IL 2和IL 12浓度、T细胞活性和毒性反应。结果　IL单基因治疗后血清IL 2或IL 12明显增高。IL 2 /IL 12联合基因组中血清IL 2和IL 12增加较单基因组显著。病理示肝癌组织中淋巴细胞浸润明显增多。IL治疗组治疗后 7d ,T细胞活性较对照组显著增高 (P <0 .0 5 )。联合基因组治疗后 7d ,T细胞功能较IL单基因组增强(P <0 .0 5 )。结论　脾内直接注射IL 2和或IL 12基因可增强T细胞活性 ,IL联合基因治疗优于IL单基因。     

膀胱癌免疫治疗相关肿瘤微环境及其在人和小鼠肿瘤中的组成特征初探

目的 应用生物信息学分析鉴定膀胱癌免疫治疗中起关键作用的肿瘤微环境（TME）组成特征,初步分析这些组分在人膀胱癌组织和BBN（N-butyl-N-（4-hydroxybutyl）-nitrosamine）诱导的自发小鼠膀胱肿瘤中表型的相似性。方法 分析晚期膀胱癌PD-L1阻断治疗IMvigor210数据集,利用DESeq2筛选与疗效相关的差异基因,通过GO和KEGG通路富集分析找出差异富集的通路;采用IOBR分析TME中的细胞浸润特征。通过TCGA数据库分析差异浸润细胞基因集与膀胱癌预后和分期的关系,在我院32例膀胱癌组织标本中进行免疫组化验证。建立BBN诱导的自发小鼠膀胱肿瘤模型,通过H&E染色和免疫组化对BBN诱导的自发小鼠膀胱肿瘤和人膀胱癌进展相似性及其TME主要组分进行比较。结果 在IMvigor210数据集中存在521个与免疫治疗疗效相关的差异基因,其中44个差异基因富集在含胶原细胞外基质上,提示成纤维细胞与免疫治疗疗效相关。TME分析发现细胞周期、组蛋白等肿瘤细胞基因集与NK细胞、CD8⁺ T细胞基因集在膀胱癌免疫治疗有效组中富集,而炎症性肿瘤相关成纤维细胞基因集在免疫治疗无效组中富集。TCGA分析显示：NK细胞和CD8⁺ T细胞基因集与膀胱癌患者总体生存时间呈正相关,CD56表达与膀胱癌病理分期呈正相关;PDGFRB表达与膀胱癌病理分期呈正相关。相比于低分期膀胱癌,PDGFRB⁺成纤维细胞在高分期膀胱癌中数量更多。在BBN小鼠模型中,BBN诱导的自发小鼠膀胱肿瘤发生率随着诱导时间逐渐升高,最终发展为肌层浸润性膀胱癌。免疫组化染色显示：小鼠和人膀胱肿瘤的TME中NK细胞和CD8⁺ T细胞以及成纤维细胞的浸润特征相似。结论 本研究结合生信分析和免疫组化评分发现：膀胱癌中NK细胞、CD8⁺ T细胞和成纤维细胞与免疫治疗疗效和预后具有相关性;这些细胞在BBN诱导的自发小鼠膀胱肿瘤与人膀胱癌组织中的浸润表型相似。这些结果可为研究膀胱癌免疫治疗的机制提供理论依据和动物模型。     

肿瘤浸润性淋巴细胞对乳腺癌患者细胞免疫功能的影响

目的 探讨肿瘤浸润性淋巴细胞（TIL）对乳腺癌患者细胞免疫功能的影响及临床意义。方法 从肿瘤组织分离培养的TIL,对25例乳腺癌患者进行治疗,检测治疗前后T淋巴细胞亚群、自然杀伤细胞（NK细胞）活性及血清可溶性白介素－2受体（sIL-2R)水平的变化。结果 经TIL治疗后患者外周血CD3、CD4、CD4／CD8和NK细胞活性均明显增高,CD8、sIL-2R水平明显降低。结论 TIL能提高乳腺癌患者的细胞免疫功能。     

消化系肿瘤浸润淋巴细胞不同输注途径对免疫功能的影响

从肿瘤组织中分离出TIL,经rIT-2培养扩增后进行自体回输,输注途径包括:(1)外周静脉;(2)局部(肿瘤供血动脉或腹腔);(3)静脉加局部、TIL输注总量为1.5×10~9以上.治疗前后抽取外周静脉血检测T细胞亚群和NK细胞活性.18例患者(胃癌9例、大肠癌5例、肝癌4例)经TIL治疗后,88.9%的患者NK活性增强,77.7%～94.4%的患者CD3 、CD4 和CD8 细胞比例升高.不同输注途径对患者的免疫功能影响亦不同.作者认为,静脉输注的TIL在体内分泌细胞因子,从而增强免疫功能.为了既增强全身免疫功能,又能使TIL较多地集中在肿瘤病灶,TIL以静脉输注加局部注射相结合为最佳途径.     

胃癌肿瘤裂解物修饰的树突状细胞体外诱导抗原特异性细胞毒T淋巴细胞

目的　利用树突状细胞呈递肿瘤抗原的特性提高细胞毒T淋巴细胞 (CTLs)对胃癌细胞的杀伤活性。方法　胃细胞癌患者外周血来源的有核细胞体外经GM -CSF和IL -4诱导产生树突状细胞 ,负载肿瘤裂解物后诱导自体CTLs产生。用细胞毒试验检测CTLs杀伤活性 ,和用ELISA测定细胞因子的分泌。结果　胃癌患者自体来源的DC裂解物能诱导产生的CTLs对自体胃癌细胞具有高杀伤率 ,可达 83 % ;致敏的DC组中IL -12与TNF -α的浓度 ( 1161± 2 3 9pg/ml,10 44± 3 12 pg/ml)显著高于未致敏的DC组 ( P <0 .0 5 )。结论　DC能呈递胃癌裂解物 ,诱导产生抗原特异性CTLs。     

新型肿瘤抗原gp96在肝癌小鼠体内的抗瘤效应

目的　观察从肿瘤细胞中提取的热休克蛋白gp96在小鼠体内的特异抗瘤效应。 方法　BALB/C小鼠肝脏成功接种肿瘤后 ,分成 4组。第Ⅰ组单纯切除肿瘤 ,不做免疫治疗 ;第Ⅱ组切除肿瘤 ,用gp96进行免疫治疗 ;第Ⅲ组不切除肿瘤 ,用 gp96进行免疫治疗 ;第Ⅳ组为对照组。第2 8天检测血清中白细胞介素 (IL) 10、干扰素 (IFN ) γ水平 ;CD4、CD8和IFN γ、IL 10双阳性细胞比例。用51Cr释放法测定各组小鼠脾细胞对亲本H2 2 肝癌细胞的杀伤活性。结果　第Ⅱ组血清IFN γ为 13 .3 3ng/L ,CD8+ IFNγ+ 和CD4+ IFNγ+ 细胞比例分别为 5 .79%和 2 .91% ,较其余组升高 (P <0 .0 1) ;血清IL 10为 5 .2 8ng/L ,CD8+ IL 10 + 和CD4+ IL 10 + 细胞比例分别为 4.5 4%和0 .75 % ,低于其他各组 (P <0 .0 1)。效靶比为 10 0∶1时 ,Ⅱ组小鼠脾细胞体外杀伤亲本H 2 2癌细胞的杀伤率为 3 9.3 % ,显著高于其他各组 (P <0 .0 5 )。结论　gp96可激发特异性细胞介导的免疫反应 ,改善抗肿瘤免疫反应。     

免疫治疗预防大肠癌术后肝转移

目的 探讨肿瘤浸润淋巴细胞（TIL）、重组白细胞介素2（rIL－2）和环磷酰胺经门静脉输入治疗对进展期大癌根治术后患者免疫功能的影响及术后肝转移的预防作用。方法 将124例DukesB及C期大肠癌患者于根治术后随机分为免疫治疗组和对照组。对照组59例,术后使用氟尿嘧啶（5－Fu）、丝裂老素C（MMC）经静脉化疗;免疫治疗组65例,术后使用TIL,rIL－2和环磷酰胺经门静脉插管输入,以后的治疗与对照组相同。所有病例均随访3年。结果 免疫治疗组术后3年内大肠肝转移8例（12．3％）,而对照组为19例（33．2％）,二者差异显著（P＜0．05）。免疫治疗后,患者外周血自然杀伤组织（NK）、白细胞介素（IL－2）活性及T细胞亚群CD3,CD4/CD4/CD8都显著上升（P＜0．05）。结论 TIL,rIL－2联合环磷酰胺免疫治疗对提高进展期大肠癌术后患者抗肿瘤细胞免疫力,预防术后肝转移具有明显效果。     

白细胞介素(IL)-2与IL-4对小鼠膀胱癌肿瘤浸润性淋巴细胞增殖及细胞毒性的协同调节作用

目的 探讨白细胞介素（ＩＬ）－２与ＩＬ－４对膀胱癌肿瘤浸润性淋巴细胞（ＴＩＬ）体外增殖及细胞毒性免疫调控的协同作用。方法 分离膀胱癌ＴＩＬ,置于含ＩＬ－２和（或）ＩＬ－２的完全培养基因中培养４周,定期计数ＴＩＬ增殖数量。四甲基偶氮唑蓝（ＭＴＴ）比色法检测ＴＩＬ细胞毒性。结果 对比单纯ＩＬ－２的培养条件,ＩＬ－２联合ＩＬ－４后４周时ＴＩＬ扩增数量是前者的１．６５倍（Ｐ〈０．０５）。在交靶比为１０：１时,ＴＩＬ对自体膀胱癌细胞（ＢＴＴ７３９）表现出高水平的杀伤活性（Ｐ〈０．０５）。联合培养的ＴＩＬ抗ＢＴＴ３９或小鼠淋巴瘤瘤株（ＹＡＣ－１）的活性与在单纯ＩＬ－２培养的条件下相比无显著改变（Ｐ均〉０．０５）。结论 ＩＬ－４对ＩＬ－２活化的膀胱癌ＴＩＬ增殖具有较强的正向调节效应,而对ＴＩＬ细胞毒性未见明显影响。     

红色诺卡氏菌细胞壁骨架对小鼠膀胱癌的治疗作用

目的　探讨红色诺卡氏菌细胞壁骨架（ＮＣＷＳ）对膀胱肿瘤的治疗效果及其机理。方法　观察皮下注射ＮＣＷＳ对小鼠同系移植膀胱癌的抗癌效应和ＮＣＷＳ对细胞免疫因子的影响。结果　给予ＮＣＷＳ的小鼠与肿瘤对照组比较肿瘤消退率为４１ ．６７ ％ ,抑瘤率为９６ ．２ ％ ;ＮＣＷＳ组ＩＬ１ 、ＩＬ２、ＩＬ６ 及ＮＫ 细胞活性和部分Ｔ细胞亚群则明显高于肿瘤对照组,但未能达到正常对照组的水平。结论ＮＣＷＳ对膀胱癌具有明显的抗癌效应,其机理为通过提高机体的ＩＬ１ 、ＩＬ２ 、ＩＬ６ 水平和ＮＫ 的活性及部分Ｔ 细胞亚群,提高机体的抗肿瘤免疫能力。     

BCG灌注对外周血免疫细胞的调节及其功能的影响

应用单克隆抗体免疫组化，细胞培养及放免分析方法对３４例膀胱癌患者ＢＣＧ灌注前后外周血免疫细胞的构成，Ｔ细胞转化水平、刺激指数、ＮＫＣ活性，ＩＬ－２诱生与活性进行了比较研究。结果表明，ＢＣＧ灌注能有效的提高外周血免疫细胞构成与活性功能。在Ｔ刺激指数、转化水平及ＮＫＣ活性调节中Ｔ４细胞数量及Ｔ４／Ｔ８比值及ＩＬ－２活性水平起重要作用。实验结果显示，ＢＣＧ灌注的抗肿瘤活性可能是细胞介导的局部免疫与宿主免疫系统功能增强的共同结果。     

Murine IL-2 secreting recombinant Bacillus Calmette-Guerin augments macrophage-mediated cytotoxicity against murine bladder cancer MBT-2

PURPOSE: This study was to establish a more effective anti-cancer immunomodulating agent by constructing recombinant (r) Bacillus Calmette-Guérin (BCG) secreting alpha-antigen (alpha-Ag) fused murine (m) interleukin (IL)-2, and to study its biological activity on cell-mediated cytotoxicity against murine bladder cancer cell, MBT-2, in vitro. MATERIALS AND METHODS: pSO246 plasmid vector ligated with mIL-2 gene was introduced into BCG by electroporation. Thioglycollate-elicited murine peritoneal exudate cells (PEC) were stimulated in vitro with parental BCG or rBCG and their cytotoxic activity and the cytokine production was studied. Cytokines were assayed by an enzyme-linked immunosorbent assay (ELISA) and L929 bioassay. Cytotoxicity was measured by 51Cr releasing assay. RESULTS: rBCG (alpha-Ag-IL-2) secreted functional IL-2 and augmented more efficient cytotoxicity to MBT-2 and cytokines such as IL-12, tumor necrosis factor and interferon (IFN)-gamma in PEC than parental BCG did. rBCG (alpha-Ag) had the same activity as BCG. Anti-IL-2 antibody reduced rBCG (alpha-Ag-IL-2)-mediated cytotoxicity and IFN-gamma production. Exogenous IL-2 also enhanced BCG-mediated cytotoxicity, but 100 times more IL-2 was required to express the same activity as rBCG (alpha-Ag-IL-2). Anti-IL-12 neutralizing antibody and the depletion of T cells and NK cells reduced IFN-gamma production by PEC stimulated with rBCG (alpha-Ag-IL-2), suggesting that T cells, NK cells and IL-12 participate in the enhancement of IFN-gamma production. CONCLUSIONS: rBCG secreting IL-2 showed significant antitumor activity and cytokine production and this will be a promising agent for bladder cancer patient to reduce both clinical dose and side effects of BCG for immunotherapy.     

Regulatory effects of interleukin-4 on tumor-infiltrating lymphocytes derived from human renal cell carcinoma.

We have studied the effects of interleukin-4 (IL-4) on the expansion, proliferation, phenotype, and antitumor activity of tumor-infiltrating lymphocytes (TIL) derived from human renal cell carcinoma. Cultures were obtained from three primary renal tumors and one group of tumor-invaded, regional lymph nodes. IL-4 induced a significant increase in lymphocyte expansion and proliferation, but the response was dependent on the concurrent dose of IL-2 in culture. Increased growth activity was only observed in those cultures receiving low doses (20 U/ml) of IL-2 (average increase of fold expansion of 6.5, P < 0.01) with no changes in growth activity in the high dose (1000 U/ml) cultures. The combination of low dose IL-2 and IL-4 (200 U/ml) promoted lymphocyte growth significantly better than high dose IL-2 alone, the current standard growth regimen for in vitro expansion of TIL. TIL grown in the presence of IL-4 significantly reduced the level of non-specific, non-major histocompatibility complex-restricted antitumor activity (P < 0.01 for allogeneic renal, nonrenal, and NK-sensitive K562 cells), while exhibiting no effect on the level of autologous killing. This is in contrast to previous findings of significant enhancement of autologous antitumor activity using IL-4 on tumor-specific, melanoma-derived TIL. We also evaluated the effects of irradiated autologous tumor stimulation (TIL:tumor ratio, 10:1) on TIL cultures. Addition of autologous tumor cells increased expansion and proliferation of all cultures regardless of concurrent lymphokines present in the culture media (average increase fold expansion of 2.21, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Adoptive immunotherapy and metastasized cancer of the kidney. Regulator effects of interleukin-4 on tumor infiltrating lymphocytes (TIL)]

Adoptive immunotherapy is a new therapeutic approach of the treatment of advanced renal cell cancer. Experimental studies have shown that the cells with the highest cytolytic activity are tumor infiltrating lymphocytes (TIL). The effects of interleukin-4 (IL-4) on the expansion, proliferation, phenotype and antitumor activity of TIL were studied. Cultures were obtained from three primary renal tumors and one group of tumor invaded, regional lymph nodes. IL-4 induced a significant increase in lymphocytes expansion and proliferation, but the response was dependent of the concurrent dose of IL-2 in culture. TIL grown in the presence of IL-4 significantly reduced the level of non specific, non MHC restricted antitumor activity while exhibiting no effect on the level of autologous killing. The effects of irradiated autologous tumor stimulation on TIL cultures were also evaluated. Addition of autologous tumor increased expansion and proliferation of all cultures and significantly enhanced levels of autologous killing. IL-4 and autologous tumor stimulation are effective growth factors when used in combination with a lose dose IL-2 regimen and may be of significant benefit in the expansion of TIL for clinical trials.     

Study on adoptive immunotherapy with tumor-infiltrating lymphocytes (TIL) for renal cell carcinoma--amplification of IL-2-elicited TIL proliferation by OKT3-monoclonal antibody

Human autologous peripheral blood lymphocytes (PBL) and lymphocytes infiltrating renal cell carcinoma (TIL) were cultured with medium containing 1000 IU/ml of human interleukin 2 (IL-2). A high cytotoxic activity against fresh autologous as well as cultured allogenic tumor cells was developed. By culturing these lymphocytes with OKT3 monoclonal antibody during the initial 2 days of long-term culture, in terms of T cell activation signal, IL-2-driven lymphocyte proliferation was remarkably accelerated with maintenance of appreciable level of cytotoxic activity. The same culture method also induced an increase in OKT3 and IL-2 receptor positive lymphocyte population in LAK cells and TIL. This method may enable us to gain more autologous TIL in vitro for adoptive immunotherapy of renal cell carcinoma than the usual culture method with IL-2 alone. Five patients with metastatic renal cell carcinoma were treated with adoptive immunotherapy with TIL, LAK and IL-2. One patient with pulmonary metastasis has had a minor response which has lasted for 3 months so far. We have not experienced any serious side effects during the treatment.     

BCG激活膀胱肿瘤浸润淋巴细胞的抗肿瘤作用研究

为了探讨ＢＣＧ的抗肿瘤机理，从１５例手术切除的膀胱移行细胞癌新鲜组织标本中制备肿瘤浸润淋巴细胞（ＴＩＬ），分别在含ＢＣＧ或ＩＬ－２的全培养基中培养扩增，测定不同培养时间的抗瘤活性。结果：用活ＢＣＧ培养的ＴＩＬ第１２天对自体肿瘤细胞的杀伤活性达高峰，杀伤率为４８．３％；用ＩＬ－２培养的ＴＩＬ第１４天达杀伤高峰，杀伤率为４３．８％，用死ＢＣＧ培养的ＴＩＬ，其扩增结果和抗瘤活性均明显低于活ＢＣＧ及ＩＬ－２。提示ＢＣＧ对ＴＩＬ的直接激活作用可能是其抗肿瘤机理之一。     

N-CWS对小鼠膀胱癌抑癌效应的影响--白细胞介素2水平及自然杀伤细胞活性观察

目的:探讨红色诺卡氏菌细胞壁骨架（Nocardiarubracellwallskeleton,N-CWS）对膀胱肿瘤的治疗效果及免疫学机制。方法：膀胱癌荷瘤鼠注射N-CWS,观察N-CWS的抑瘤效果及N-CWS对白细胞介素2（IL-2）水平和自然杀伤（NK）细胞活性的影响。结果：注射N-CWS的膀胱癌荷瘤鼠的抑瘤率为96.2％,其IL-2水平及NK细胞活性明显高于肿瘤对照组。结论：N-CWS对膀胱癌具有明显的抑制作用,其机制可能是通过提高机体的IL-2水平和NK细胞活性促进机体的抗肿瘤免疫而实现的。     

The results of immunotherapy as an adjunct to the surgical treatment of primary lung cancers

A total of 263 primary non-small cell lung cancer patients resected in our Institute from March, 1978 to October, 1984 were utilized in order to evaluate the efficacy of transfer factor (TF) immunotherapy as an adjunct to surgical treatment and TF was significantly effective to cases with stage I diseases, but not to stages II, III and IV diseases, indicating that TF could only suppressed micrometastasis existing at the time of surgery. In order to improve the further results of immunotherapy as an adjunct to surgical treatment, we analyzed cytotoxic activity against autologous lung cancer and K562 leukemia cells in tumor bearing hosts. Furthermore, we studied the effect of interleukin 2 (IL2) activated lymphocyte dialysate on cytotoxic activity against lung cancer cells. When 3 different sources of lymphocytes including peripheral blood lymphocytes (PBL) regional lymphnode cells (LNC) and tumor infiltrating lymphocytes (TIL) were incubated with IL2 for 8 days at 37 degrees C in 5% CO2 atmosphere, relatively high cytotoxic activity was demonstrated with 2 major different patterns in PBL, LNC and TIL including one systemic predominant and the other local predominant patterns, suggesting that IL2 might be a local or systemic possible immunotherapeutic reagent. Finally, we stimulated lymphocytes from household contact family members with IL2 and MMC treated lung cancer cells. These in vitro modulated T-lymphocytes demonstrated considerable cytotoxic activity against the target cells which were used as in vitro sensitization. The dialysate of these in vitro stimulated cells showed specific activity on cytotoxicity against lung cancer cells and might be a possible reagent in stead of TF for clinical trial.