人外周血树突状细胞诱导及免疫学特性研究

[目的]从人外周血分离、纯化、扩增树突状细胞(DC),并对其形态学和免疫学特性进行初步探讨.[方法]从人外周血分离DC前体细胞(主要为CD14 细胞)用重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)和重组人白细胞介素-4(rhIL-4)联合培养,诱导扩增成熟DC.观察DC形态、分析DC表型、核型及检测DC激发同种异体淋巴细胞增殖能力.[结果]分离的DC前体经rhGM-CSF和rhIL-4共同培养1周后,可获得大量成熟DC,扩增了24.5倍,纯度达90%以上.DC高表达分化抗原CD86、CD40、HLA-DR、CD83、CDIa,能强烈激活同种异体T淋巴细胞增殖.[结论]人外周血CD14 细胞经体外诱导培养,可以生成大量功能成熟的DC,从而为进一步开展DC的基础研究和临床应用打下基础.     

肺癌患者外周血树突状细胞的生物学特性研究

目的：探讨肺癌患者外周血来源的树突状细胞（dendritic cells, DC）生物学特性。方法：从肺癌患者外周血分离获得单个核细胞，用细胞因子GMCSF (100 μg/L)，IL4 (50 μg/L)体外培养诱导。凋亡肿瘤细胞负载24 h后加入TNFα（10 μg/L）或CD40激发型单抗于培养DC中，继续诱导3～4 d。分别测定DC的表型、DC摄取抗原能力；混合淋巴细胞反应（MLR）对T细胞增殖能力的测定；细胞计数和3HTdR掺入观察DC对T细胞的激发和扩增效应，并与健康志愿者外周血来源的DC进行比较。结果： 患者外周血单个核细胞和正常人外周血单个核细胞来源的DC均高表达CD1α，CD83，CD80，CD86和HLADR等DC的相关抗原和共刺激分子；患者的未成熟DC能有效摄取FITCDextran，经TNFα或CD40激发型单抗激发诱导后，成为成熟和有功能的DC,几乎完全失去对抗原的摄取能力，与健康人外周血来源DC组相比无明显差别(P>0.05)；患者单个核细胞来源的DC在体外具有激发自体和同种异体外周血T细胞增殖的能力。结论：肺癌患者的外周血来源单个核细胞可以诱导成为具有功能的成熟DC。     

人外周血树突状细胞的体外诱导和鉴定

目的：建立从人外周血体外诱导扩增树突状细胞(DC)的方法：方法：从正常人外周血F1细胞分离单个核细胞，经三种细胞因子——重组人粒细胞一巨噬细胞集落刺激因子(rh GM—CSF)、重组人白细胞介素4(rh IL-4)和重组人肿瘤坏死因子(rh TNFα)联合诱导培养，10天后收获细胞，利用光学显微镜、透射电镜观察其外部形态和细胞超做结构，流式细胞仪检测其细胞表面抗原，自体混合淋巴细胞反应检测其刺激T细胞的增殖活性：结果：培养收获了高纯度的DC，这些细胞表面有大量的不规则突起，核也呈不规则状，核仁小，核膜异染色质聚集，含有丰富的细胞器，如核糖体、内质网、溶酶体等；细胞表面高水平表达HLA—DR、CD1α、CD80、CD83和CD86；自体混合淋巴细胞反应表明诱导所得的DC具有很强的激发同种T细胞增殖的能力。结论：人外周血单个核细胞经细胞因子诱导培养可以得到大量的成熟DC。     

肺癌患者外周血树突状细胞的体外扩增及鉴定

目的 探索体外扩增成熟树突状细胞(dendritic cell，DC)的方法，并进一步鉴定Dc的形态及表型，为开展肿瘤临床生物治疗奠定基础。方法 取肺癌患者外周血，经淋巴细胞分离液分离得到DC前体细胞，加入生长因子DCGF诱导DC生成，并加入自体肿瘤相关抗原刺激。收集细胞用透射电镜观察，并用流式细胞仪测定细胞表型。结果 应用该法扩增细胞可以得到大量成熟DC。电镜观察DC具有典型的形态。流式细胞仪细胞表型测定CD80为81．8％，HLA—DR为98．3％，CD86为69．8%，CDla为19．7％，CD14为66．9．1％，CD80和HLA-DR较未成熟DC的表达明显增加。结论 肺癌患者外周血体外培养可成功地获得成熟的DC，进一步为肿瘤临床生物治疗奠定了基础。     

含甲胎蛋白肽段的重组腺病毒转染树突状细胞诱导细胞毒性T细胞对肝癌细胞的体外杀伤效应

目的 研究人外周血单核细胞来源的树突状细胞(DC)转染含甲胎蛋白(AFP,137-145)片段的重组腺相关病毒后所诱导的特异性T细胞对肝癌细胞株HepG2和SMMC-7721的体外杀伤作用.方法 抽取健康志愿者外周血,分离单核细胞,体外培养,使用含AFP片断的重组腺相关病毒转染未成熟DC,诱导特异性T细胞.检测体外培养的DC和细胞毒性T细胞(CTL)活性,应用四甲基偶氮唑蓝(MTT)法检测CTL对HepG2和SMMC-7721细胞的杀伤作用.结果 转染或未转染的体外培养的成熟DC高表达CD40、CD86和IL12,成熟DC诱导的CTL高表达IFNγ;修饰成熟后的DC后体外能诱导特异性CTL,该CTL在休外对肝癌细胞株HepG2和SMMC-7721均有杀伤作用.结论 重组腺相关病毒转染DC,不明显改变DC表型和刺激淋巴细胞增殖和分化功能,可诱导自体CTL增殖,含AFP(137～145)片断的腺相关病毒转染DC诱导自体CTL对肝癌细胞株HepG2和SMMC-7721细胞有明显杀伤作用,DC疫苗可以作为肝癌患者免疫治疗的有效补充.     

宫颈癌患者外周血树突状细胞抗宫颈癌免疫反应

[目的]研究自宫颈癌患者外周血分离、培养而来的树突状细胞体外对宫颈癌Hela细胞的抑制作用.[方法]应用细胞因子诱导培养从宫颈癌患者外周血分离的单核细胞,获得成熟DC.宫颈癌组织来源抗原致敏DC,激活T细胞增殖分化为细胞毒性T淋巴细胞(CTL).MTT法检测比较负载抗原DC与未负载抗原DC诱导的CTL对Hela细胞和SKOV3细胞不同的杀伤效应.[结果]宫颈癌细胞抗原致敏的DC,激活肿瘤抗原特异性CTL,对宫颈癌Hela细胞产生高效而特异的杀伤作用(56.410%),对卵巢癌SKOV3细胞仅有一定的杀伤作用(24.901%);而未致敏DC激活的CTL对以上两种肿瘤细胞只有很低的杀伤作用,分别为10.256%和20.151%.单纯肿瘤抗原刺激下的T细胞,对Hela和SKOV3细胞产生非特异性杀伤作用,其效率仅为1.865%及15.613%.[结论]宫颈癌患者外周血DC在体外能诱导高效而特异的抗宫颈癌细胞免疫反应,提示肿瘤抗原激活的DC作为疫苗在宫颈癌的免疫治疗中具有重要意义.     

肝癌细胞抗原致敏树突状细胞诱导的体外抗肿瘤作用

目的　探讨原发性肝癌患者外周血树突状细胞 (DC)经自体肝癌细胞抗原致敏后诱导的体外抗肿瘤作用。方法　对肝癌患者外周血采用密度梯度离心法分离 ,获得DC前体细胞 ,用重组人粒细胞 巨噬细胞集落刺激因子 (rhGM CSF)和重组人白细胞介素 4(rhIL 4)联合培养 ,诱导扩增DC。制备自体肝癌细胞抗原 ,体外脉冲DC ,检测DC诱导自体T细胞增殖能力及细胞毒性T细胞 (CTL )在体外对自体肝癌细胞的杀伤活性 ,并检测肿瘤抗原致敏DC分泌的IL 12水平。结果　经自体肝癌细胞抗原致敏的DC能分泌IL 12和诱导较强的自体T细胞增殖 ,且能诱导特异性CTL ,该CTL对自体肝癌细胞具有很强的杀伤活性 ,杀伤率显著高于DC、未经肝癌细胞抗原致敏的DC激活的CTL及T淋巴细胞的杀伤率 ,而对CT 2 6细胞、BEL 740 2细胞无明显的杀伤作用。结论　肝癌患者外周血DC经自体肝癌细胞抗原致敏后能诱导高效而特异的抗肝癌免疫 ,其机制可能与增强T细胞应答和诱导机体产生肿瘤特异CTL从而发挥特异性的抗肿瘤作用有关。     

肝癌患者与正常人外周血树突状细胞的诱导与功能的比较研究

目的对比研究肝癌患者和正常人外周血树突状细胞（Dc）的诱导及其功能异同,探讨能否从肝癌患者外周血诱导出功能正常、具有抗肿瘤作用的DC疫苗。方法取肝癌患者和正常人外周血分离DC前体细胞,应用rhGM—CSF、rhIL-4及肝癌细胞抗原诱导DC。观察DC生长过程的形态学变化,以流式细胞术检测DC表面分子;EUSA法检测DC培养上清中IL-12水平;3H—TdR掺入法检测肝癌细胞抗原致敏DC刺激T细胞增殖的能力;乳酸脱氢酶4小时释放法检测肝癌细胞抗原致敏DC诱导的细胞毒性T淋巴细胞（CTL）的杀瘤活性。结果从肝癌患者和正常人外周血DC前体细胞诱导的DC具有相似的生长过程和细胞形态及功能,其表面分子HLA—DR、CD1a、CD83、CD80、CD86、ICAM-1的表达水平、DC培养上清中IL-12水平、DC刺激T细胞增殖的能力以及所诱生的CTL杀瘤活性等方面比较,差异均无统计学意义（P〉0．05）。结论可从肝癌患者外周血诱导出功能正常并具有抗肿瘤作用的DC瘤苗,为以DC基础的肝癌的免疫治疗提供实验依据。     

人脐血树突状细胞抗神经胶质瘤的体外免疫效应研究

目的　研究肿瘤抗原致敏的树突状细胞 (DC)对神经胶质瘤细胞的免疫杀伤效应。方法　应用免疫磁珠分选脐血 CD34细胞 ,经 SCF FL3 GM- CSF IL- 4 TNF- α的联合诱导培养 DC,采用相差显微镜观察树突状细胞的形态 ;流式细胞仪作 DC的表面标志检测 ;MTT比色法测定同种异型的混合淋巴细胞反应能力和诱导CTL 毒性的检测。结果　 (1)脐血 CD34细胞在体外经细胞因子联合刺激后呈典型的 DC形态 ;(2 )流式检测 CD4 0、CD80和 CD86等成熟 DC特异性表面标志呈高表达 ,分别与诱导前比较 ,差异均有显著性 ;(3) MTT法测得脐血来源的 DC较外周血 DC刺激同种异体 T淋巴细胞增殖能力弱 ;经肿瘤抗原负载的 DC体外诱导出较强 CTL 毒性。结论　脐血 CD34细胞经体外扩增诱导的 DC具有典型的 CTL 毒性 ,为临床应用脐血来源 DC抗神经胶质瘤的生物治疗提供了理论依据     

负载肝癌抗原的树突状细胞瘤苗诱导的抗肿瘤作用

目的 探讨原发性肝癌患者外周血树突状细胞（DC）体外经自体肝癌细胞抗原致敏后诱导的抗肿瘤作用。方法肝癌患者外周血经梯度密度离心法分离，获得DC前体细胞，用重组人粒细胞-巨噬细胞集落刺激因子（rhGM—CSP）和重组人白细胞介素-4（rhIL-4）联合培养，诱导扩增DC。制备自体肝癌细胞抗原，体外脉冲DC，检测DC诱导自体T细胞增殖能力及细胞毒性T细胞（CTL）在体外对自体肝癌细胞的杀伤活性，并检测肿瘤抗原致敏DC分泌的IL-12水平。结果经自体肝癌细胞抗原致敏的DC能分泌IL-12和诱导较强的自体T细胞增殖，且能诱导特异性CTL，该CTL对自体肝癌细胞具有很强的杀伤活性，杀伤率明显高于DC、未经肝癌细胞抗原致敏的DC激活的CTL及T淋巴细胞的杀伤率，而对3LLLEWIS肺癌细胞、H22肝癌细胞则无明显的．杀伤作用。结论肝癌患者外周血DC经自体肝癌细胞抗原致敏后能诱导高效而特异的抗肝癌免疫，其机制可能与增强T细胞应答和诱导机体产生肿瘤特异CTL而发挥特异性的抗肿瘤作用有关。     

卵巢癌患者外周血来源树突细胞的生物学特性

背景与目的：树突细胞（dendritic cells,DCs）是体内功能最强的抗原递呈细胞．在抗肿瘤免疫治疗中起着重要的作用。既往对DCs生物学特性的研究数据大多来自健康个体,而来自肿瘤患者的DCs的特性尚不清楚。本研究旨在探讨卵巢癌患者外周血单核细胞来源的树突细胞（monocyte—derived dendritic cells,MoDCs）的生物学特性。方法：收集8名上皮性卵巢癌患者及13名健康女性志愿者的外周静脉血标本,从中分离出单核细胞,将其置于含人白细胞介素4（interleukin4,IL4）及粒细胞．巨噬细胞集落刺激因子（granulocyte-macrophag ecolony stimulating factor,GM-CSF）的RPMI-1640完全培养基中培养,用肿瘤坏死因子-α（tumornecrosisfactor-α,TNF-α）刺激其成熟。对诱导7d后的MoDCs进行细胞形态、表型及刺激异体淋巴细胞增殖能力分析。结果：卵巢癌患者及健康妇女外周血单核细胞培养7d后,均诱导出形态学上典型的成熟MoDCs。两组MoDCs均表达丰富的HLA-ABC（MHC-Ⅰ）、HLA-DR（MHC-Ⅱ）和大量共刺激分子CD86和CD80。卵巢癌组HLA-ABC、HLA-DR、CD86及CD80的平均荧光强度（mean fluorescence intensity,MFI）与健康妇女组差异无统计学意义（P〉0．05）。混合淋巴细胞反应（mixed leukocytes reaction,MLR）结果显示,卵巢癌各组（按MoDCs与淋巴细胞的比例分组）的吸光度值均明显低于健康妇女相应各比例组（P〈0．05）。结论：卵巢癌患者的MoDCs刺激异体淋巴细胞增殖能力较健康妇女降低,提示其存在一定程度的免疫学功能异常。     

CD1c+ and CD303+ dendritic cells in peripheral blood, lymph nodes and tumor tissue of patients with non-small cell lung cancer

Dendritic cells (DCs) are the most potent antigen presenting cells, which can stimulate a cellular immune response against malignant tumor cells. Many authors have described the phenomenon of tumor infiltration by dendritic cells and emphasized an immunosuppressive tumor influence on DC function. In the present study, we examined the presence of myeloid CD1c+ (BDCA-1+) dendritic cells and lymphoid/plasmacytoid CD303+ (BDCA-2+) dendritic cells in peripheral blood, lymph nodes and cancer tissue of patients with non-small cell lung cancer (NSCLC). Fifty male patients treated surgically for NSCLC stages I-IIIa without neoadjuvant chemotherapy were included. Employing a multiparameter flow cytometry for CD1c, CD19, CD123 and CD303, we observed an accumulation of immature DCs in the tissues involved in the neoplasmatic process with the predominance of lymphoid/plasmacytoid over myeloid DCs. Moreover, in peripheral blood NSCLC patients had a significantly lower percentage of CD1c+ DCs than healthy donors. Our results suggest that NSCLC cells might hamper the maturation of DCs, thus escaping an efficient immune response.     

TRANCE- and CD40 ligand-matured dendritic cells reveal MHC class I-restricted T cells specific for autologous tumor in late-stage ovarian cancer patients.

PURPOSE: The use of mature dendritic cells (DCs) presenting tumor-associated antigens (TAAs) to trigger tumor-specific T cells in vivo or in vitro represents a promising approach for cancer immunotherapy. We hypothesized that tumor antigens, mostly unidentified, are present on ovarian tumor cells and that mature DCs could be used to generate tumor-specific responses in unprimed patients. We also sought to measure preexisting antitumor immunity in patients with advanced ovarian cancer. EXPERIMENTAL DESIGN: Autologous DCs from 10 patients with ovarian cancer were pulsed with killed autologous primary tumors as a source of TAAs. DCs were then cultured in the presence of tumor necrosis factor alpha + TRANCE (tumor necrosis factor-related activation-induced cytokine) to induce maturation. Mature TAA-pulsed DCs were used in vitro to stimulate tumor-specific peripheral blood T cells. RESULTS: TRANCE and CD40 ligand were effective at maturing DCs. T-cell lines were generated in vitro that were capable of secreting IFN-gamma in response to autologous tumor. These tumor-specific T cells were MHC class I restricted. The frequency of tumor-specific T cells in uncultured cells from malignant ascites fluid and peripheral blood was measured in the same patients. CONCLUSIONS: IFN-gamma-secreting tumor-specific T cells were demonstrated at baseline in uncultured T cells from some unvaccinated ovarian cancer patients; however, the T cells could not kill autologous tumor. These data demonstrate that mature DCs presenting tumor antigens from engulfed autologous tumors can be used to augment antitumor immunity in vitro in patients with epithelial ovarian cancer. The results support the feasibility of therapeutic vaccination of ovarian cancer patients.     

恶性胸腔积液来源树突状细胞对自体肿瘤 浸润性淋巴细胞的作用

 目的探讨恶性胸腔积液来源DCs（dentritic cells,DCs）对自体肿瘤浸润性淋巴细胞（tumor infiltration lymphocytes, TILs）增殖及杀伤肿瘤细胞能力的影响。方法用体外培养方法从16例肺癌患 者恶性胸腔积液来源分离单个核细胞,再用密度梯度离心辅以免疫磁珠分选细胞,用白细胞介素4（IL-4 ）、肿瘤坏死因子-α（TNF-α）、粒-单核细胞刺激因子（GM-CSF）等诱导出DCs,用流式细胞仪和电子 显微镜鉴定分离DCs;用IL-2、植物血凝素诱导同一患者胸腔积液单个核细胞中的TILs,用HEA-125磁珠分 选纯化肿瘤细胞,3H-TdR渗入法检测DCs对TILs增殖能力;MTT法检测TILs对肿瘤细胞杀伤活性。结果从肺 癌患者恶性胸腔积液单个核细胞中可诱导成熟DCs,电镜和光镜观察结果显示,这类DCs具有典型的DC形态 ,高表达HLA-ABC、HLA-DR、CD86,较高表达CD80、CD54,也表达CD83、CD1a。DCs可明显促进TILs增殖能 力（增加约1.7倍）。以TILs本身为对照,DCs激活的TILs对肿瘤细胞杀伤活性明显增加［从（31.80± 14.05）%提高到（51.89±13.27）%,P<0.05］。结论肺癌患者恶性胸腔积液单个核细胞中可诱导成熟DCs ,这种DCs可刺激自体TILs扩增,增加TILs杀伤肿瘤细胞的活性。     

A new strategy using autologous dendritic cells and lymphokine-activated killer cells for cancer immunotherapy: efficient maturation of DCs by co-culture with LAK cells in vitro

Among a variety of antigen presenting cells (APCs), accumulating results support that the mature dendritic cell (DC) has the potential to induce efficient cytotoxic T lymphocyte (CTL) responses in the context of peptide-based immunotherapy. DCs have been known to assume the mature form by signaling through the CD40-CD40 ligand (CD40L) interaction, which may be provided by activated CD4+ T cells expressing abundant CD40L molecules on their surfaces. Here, we report that DCs generated from peripheral blood monocytes obtained from patients with advanced cancer exhibit a mature phenotype after co-culturing with autologous lymphokine-activated killer (LAK) cells generated by the stimulation of peripheral blood mononuclear cells with anti-CD3 monoclonal antibody (mAb) and interleukin (IL)-2. Part of this process appeared to be dependent on the expression of CD40L on the surface of LAK cells, although it was also suggested that some other humoral factors produced by LAK cells may be involved in this effect as well. DCs derived from the donors, of which LAK cells demonstrated a higher Th1/Th2 ratio upon activation determined by the intracellular detection of interferon-gamma and IL-4, showed more efficient maturation upon co-culture with LAK cells than DCs from donors with a low Th1/Th2 ratio. Importantly, these matured DCs induced a two-times stronger antigen-presenting capacity measured by an allo-reactive mixed lymphocytes reaction assay as compared to immature DCs. These results imply the use of the combination of DCs and LAK cells for immunotherapy against cancer.     

肝癌患者外周血树突状细胞的体外扩增、鉴定及内吞能力观察

目的　从肝癌患者外周血培养扩增树突状细胞，观察其形态、表型、内吞及递呈抗原能力，为树突状细胞在肝癌生物治疗中的应用奠定基础。方法　从肝癌患者外周血中分离单个核细胞，在 Ｇ Ｍ Ｃ Ｓ Ｆ、 Ｉ Ｌ４ 的诱导下培养扩增树突状细胞。光学显微镜下观察其体外培养过程中的形态特征及变化，电镜观察其超微结构， Ｆ Ａ Ｃ Ｓ观察其表型特征， Ｍ Ｌ Ｒ检测其抗原递呈能力， Ｈ Ｒ Ｐ内吞实验检测其群体内吞能力。结果　肝癌患者外周血树突状细胞具典型的形态及表型特征，较巨噬细胞更强的激发同种混合淋巴细胞反应的能力。其群体的内吞能力约在培养第 ５ 天达高峰，之后有明显下降。结论　从肝癌患者外周血中可获取较大量的、高纯度的、具较强抗原递呈能力的树突状细胞，在其具有活跃的内吞能力时，以合适的肿瘤抗原致敏可望成为肝癌生物治疗的崭新手段。     

Presentation of tumor antigens by phagocytic dendritic cell clusters generated from human CD34+ hematopoietic progenitor cells: induction of autologous cytotoxic T lymphocytes against leukemic cells in acute myelogenous leukemia patients

The use of antigen-presenting dendritic cells (DCs) is currently proposed for tumor immunotherapy through generation of CTLs to tumor antigens in cancer patients. In this study, DCs were differentiated using granulocyte-macrophage colony-stimulating factor and tumor necrosis factor-alpha from CD34+ hematopoietic progenitor cells that had been mobilized into the peripheral blood. To use the phagocytic activity of DCs for processing and presentation of tumor antigens, we established DC clusters containing immature DCs by preserving proliferating cell clusters without mechanical disruption. After an 11-day culture, the developed clusters contained not only typical mature DCs but also immature DCs that showed active phagocytosis of latex particles, suggesting that the clusters consisted of DCs of different maturational stages. These heterogeneous clusters could present an exogenous protein antigen, keyhold limpet hemocyanin, to both CD4+ and CD8+ T lymphocytes. Furthermore, in three acute myelogeneous leukemia patients, clusters pulsed with autologous irradiated leukemic cells could also induce antileukemic CTLs. The mechanical disruption of clusters abrogated the induction of CTLs to leukemic cells as well as to hemocyanin. This observation gives an important information for the use of heterogeneous DC clusters derived from autologous peripheral blood CD34+ cells in the case of immunotherapy for leukemia.     

肺癌肿瘤全溶物脉冲自体树突状细胞体外诱生T细胞抗肿瘤反应的研究

目的　探讨非小细胞肺癌 (NSCLC)患者全瘤溶解物 (WTL)冲击的自体树突状细胞 (DCs)瘤苗体外诱生T细胞介导的抗肿瘤反应。方法　在含重组人粒细胞 巨噬细胞 集落刺激因子 (rhGM CSF)和重组人白细胞介素 4 (rhIL 4 )的培养条件下 ,以 10例患者的贴壁外周血单核细胞 (PBMCs)衍生不成熟的树突状细胞 (imDCs) ,分别添加肿瘤坏死因子 (TNFα)、自体WTL或WTL TNFα诱导成熟 ,即分别为DCs/TNF、DCs/WTL和 DCs/WTL ,用流式细胞仪和混合淋巴细胞反应 (MLR) ,检测细胞表面分子的表达变化及其刺激异基因或同基因的T细胞增殖能力。将 DCs/WTL和自体T细胞共培养1～ 2周 ,以诱导细胞毒性T淋巴细胞 (CTL) ,用计数γ 干扰素 (IFN γ)释放的酶联免疫斑点 (ELISPOT)试验和乳酸脱氢酶 (LDH)释放的细胞毒试验 ,分别检测该CTL中识别自体肿瘤细胞释放特异性IFN γ的T细胞数和溶瘤活性。结果　以 30 μg/ml蛋白的WTL体外冲击自体 10 6imDCs,可导致上调DC表型 ,包括CD1a、CD83和CD86以及HLA DR的分子表达 ,与常规TNFα诱导成熟的DCs表型相似。虽然两者均能显著刺激异基因T细胞的增殖 ,但其刺激自体T淋巴细胞的增殖能力 ,DCs/WTL却显著高于DCs/TNF(P <0 .0 5 )。将体外与 DCs/WTL共培养的自体T细胞作为反应或效应细胞 ,再次