冻融人树突状细胞基因疫苗体外抗肿瘤免疫效应

目的：观察人外周血和人脐血来源的冻融树突状细胞（dendritic cell,DC）基因疫苗的形态、表型及体外诱导的CTL抗肿瘤活性。方法：细胞因子扩增人外周血和脐血DC,分别将EBV-LMP2、HPV16E6基因转染2种来源的冻融DC制备疫苗。动态形态学观察和流式细胞术检测疫苗表面分子表达,体外诱导并测定CTL活性。结果：人外周血和脐血冻融DC疫苗均高表达CD80、CD86和CD83,低表达CD14,高表达CD1a,体外均能诱导高效的CTL活性（P=0.001）,并与新鲜DC疫苗差异无统计学意义,P=0.138。结论：负载肿瘤相关病毒抗原基因的冻融DC疫苗保持了功能成熟DC的形态特征,且能诱导高效的特异性抗肿瘤免疫应答。     

树突状细胞体外高效诱导白血病特异性细胞毒性T淋巴细胞生成

[目的]研究体外诱导产生的细胞毒性T淋巴细胞(CTL)及其抗白血病反应.[方法]应用mGM-CSF及mIL-4细胞因子从小鼠骨髓细胞扩增出成熟树突状细胞(DC),使其负载冻融法制备的白血病细胞相关抗原(TAA),通过观察DC诱导的白血病特异性CTL的免疫表型,MTT分析其对于L7212细胞的抑制率,利用ELISA评价IL-2和IL-4水平.[结果]骨髓单个核细胞经mGM-CSF、mIL-4的联合作用7天后光镜及扫描电镜下观察到大量成熟DC生成.经负载TAA的DC活化后T细胞中CD3 、CD8 、CD25 细胞明显增多.FCM显示CD3 、CD8 、CD25 T细胞显著增多,CD8 细胞多于CD4 细胞.活化后T细胞对L7212细胞有特异性杀伤活性,在效靶比为50:1培养72 h后杀伤率达90.1%±2.7%.DC与T细胞共培养上清中IL-2的分泌水平为4.656±0.62pg/ml,明显高于普通T细胞培养组的1.436±0.11pg/ml(P=0.011),IL-4水平则无明显变化(P＞0.05).[结论] mGM-CSF及mIL-4配伍诱导生成的DCs经L7212冻融抗原负载后可在体外高效诱导白血病特异性CTL生成.     

粒细胞-巨噬细胞集落刺激因子基因修饰的树突状细胞疫苗增强体外抗肿瘤免疫效应

Objective: The aim of the study was to investigate whether dendritic cell (DC) precursors, recruited by injection of chemokine ligand 3 (CCL3), induce enhanced anti-tumor immunity after granulocyte-macrophage colony stimulating factor (GM-CSF) transfection in mice ex vivo. Methods: The 615 mice were injected with CCL3 via the tail vein. Freshly isolated B220–CD11c+ cells were cultured with cytokines. For adenoviral (Ad)-mediated gene transduction, DCs were transferred AdGM-CSF gene at different ratios of mu...     

树突状细胞疫苗的制备及其体外诱导细胞毒性T淋巴细胞对宫颈癌CaSki细胞的杀伤作用

目的:制备树突状细胞(dendritic cells,DCs)疫苗并观察其在体外诱导细胞毒性T淋巴细胞(cytotoxic T lymphocytes,CTLs)对宫颈癌CaSki细胞的杀伤效应。方法:分离培养小鼠未成熟DCs,FCM检测小鼠未成熟DCs表面标志物CD40、CD86、主要组织相容性复合体-Ⅱ(major histocompatibility complex-Ⅱ,MHC-Ⅱ)和CD11c;将已成功构建的携带人乳头状瘤病毒(human papillomavirus,HPV)16E6E7基因的重组腺病毒pAd-E6E7感染体外培养的小鼠未成熟DCs,提取CaSki细胞裂解物负载DCs,制备DC疫苗,激光共聚焦显微镜下观察pAd-E6E7感染的小鼠未成熟DCs绿色荧光蛋白表达,Western印迹法检测E6蛋白的表达;DCs疫苗诱导产生CTLs后与CaSki细胞共培养,CCK8(cell countingkit8)法检测其对CaSki细胞的杀伤效应。结果:成功分离培养了小鼠未成熟DCs,并制备获得了HPV16E6E7特异性DCs疫苗。DCs疫苗诱导产生的CTLs对CaSki细胞具有杀伤作用,pAd-E6E7感染组对CaSki的杀伤效应明显高于CaSki细胞裂解物负载组及未处理DCs组(P<0.05)。结论:以携带HPV16E6E7基因的重组腺病毒感染DCs制备基因修饰的DCs疫苗,具有诱导CTLs体外杀伤子宫颈癌CaSki细胞的作用。     

CB6F1小鼠树突状细胞诱导细胞毒性T淋巴细胞的杀伤作用

[目的]探讨CB6F1小鼠脾树突状细胞(DC)的培养及其诱导针对小鼠路易斯肺癌(LLC)的细胞毒性T淋巴细胞(CTL)对肿瘤的杀伤效应.[方法]应用CB6F1小鼠脾细胞在GM-CSF、IL-4等细胞因子作用下培养出DC,反复冻融法制备LLC抗原致敏DC,与淋巴细胞及IL-2混合培养诱导出肿瘤特异性CTL,利用乳酸脱氢酶法检测CTL的杀伤活性.[结果]用GM-CSF、IL4联合培养小鼠脾细胞第4d,可见细胞形态发生改变,培养第8d,可见典型的刺突样DC,通过流式细胞术检测了DC表型高表达CD80占76.5％,CD86占60.0％,MHCⅡ占67.4％,CD11C占80.6％.[结论]应用GM-CSF、IL-4、LPS等细胞因子培养CB6F1小鼠脾细胞经过肿瘤抗原冲击,可以培养出成熟DC,并且DC可以诱导出具有杀伤活性的肿瘤特异性CTL.     

体外诱导的粘蛋白抗原特异性细胞毒性T淋巴细胞抗乳腺癌的研究

目的 制备携带粘蛋白(MUC1)抗原基因的重组腺相关病毒(AAV/MUC1),研究其感染树突状细胞(DC)所诱导的细胞毒性T淋巴细胞(CTL)特异性杀伤肿瘤细胞的活性.方法 应用分子生物学方法制备高低度的重组腺相关病毒(AAV/MUC1).AAV/MUC1体外感染外周血单核细胞,诱导分化为DC.DC与T淋巴细胞混合,刺激产生CTL.流式细胞技术检测DC和CTL的分化和功能指标,MTS方法检测CTL的杀伤活性和特异性.结果 成功制备的重组病毒(AAV/MUC1)滴度为6×1010拷贝/ml.感染单核细胞率为84.27%.所获得的CTL对MUC1阳性的乳腺癌细胞株的杀伤率为(46.32±0.07)%.其杀伤作用具有MUC1抗原特异性和MHC-I类分子限制性的特征.结论 以MUC1抗原为靶点的CTL可有效地杀伤乳腺癌细胞,为乳腺癌提供了一条新的治疗途径.     

粒细胞-巨噬细胞集落刺激因子基因修饰的树突状细胞疫苗增强体外抗肿瘤免疫效应

目的 研究粒细胞-巨噬细胞集落刺激因子(GM-CSF)基因修饰树突状细胞(DC)后形态、表型及功能的变化,以及增强DC疫苗对肿瘤细胞的体外杀伤作用.方法 小鼠尾静脉注射趋化因子配体3(CCL3),分选得到B220- CDllc+细胞,经细胞因子培养诱导分化DC.在体外用含GM-CSF基因的重组腺病毒(AdGM-CSF)转染DC,酶联免疫吸附试验(ELJSA)检测转染后GM-CSF的水平.通过细胞形态学观察、表型分析及混合淋巴细胞反应(MLR),检测GM-CSF基因修饰前后DC的变化.反复冻融法制备胃癌可溶性抗原,将其与GM-CSF基因修饰的DC共同培养,制备DC疫苗,四甲基偶氮唑蓝(MTT)法检测活化的T淋巴细胞在体外对小鼠前胃癌细胞(MFC)的杀伤作用,ELISA法检测干扰素γ(INF-γ)的分泌情况.结果 CCL3注射后,外周血中B220- CD11c+细胞明显增加,48 h达到高峰[占外周血单个核细胞的(13.88±1.10)%].AdGM-CSF转染后,培养液上清中GM-CSF浓度升高,当感染复数(MOI)为1:100时达到高峰[(130.00±12.61)pg/m1].经GM-CSF.基因修饰的DC在形态上更趋成熟,MHCⅡ类分子、CD80、CD86等细胞表型明显上调,具有更强的刺激T细胞增殖的能力.荷载胃癌抗原的DC激活的T淋巴细胞对MFC细胞具有特异性杀伤作用,并产生高水平的INF-γ[(1245.00±13.75)pg/ml].结论 GM-CSF转染DC后,能大量表达GM-CSF,DC形态及细胞表型更趋成熟,刺激T细胞增殖能力明显增强.GM-CSF基因修饰的DC在体外可诱导出针对靶肿瘤细胞的特异性杀伤作用.     

粒细胞-巨噬细胞集落刺激因子基因修饰的树突状细胞疫苗增强体外抗肿瘤免疫效应

Objective To investigate if granulocyte-macrophage colony stimulating factor (GM-CSF) gene-modified dendritic cells ( DC) enhance antitumor immunity in vitro. Methods Mice were injected with chemokine ligand 3 (CCL3) via the tail vein. Fresh B220-CD11c+ cells were sorted from the peripheral blood mononuclear cells (PBMCs) and cultured into DCs by cytokines. DCs were transfected with AdGM-CSF gene at different ratios of multiplicity of infection ( MOI) to determine the optimal gene transfection conditions, and the expression of GM-CSF was detected after transfection. The variation of GM-CSF gene-modifiedDCs were analyzed by morphological examination, phenotype analysis, and mixed lymphocyte reaction (MLR). DCs were loaded with gastric cancer antigen obtained by freezing and thawing method. The killing effect of DCs vaccine-stimulated T lymphocytes on gastric cancer cells was assessed by MTT assay. INF-γ production was determined with the INF-γ ELISA kit. Results B220- CD11c+ cells increased obviously after CCL3 injection. The ELISA results showed that after GM-CSF gene modification, DCs could produce high level of GM-CSF. When DCs were transfected with AdGM-CSF gene at MOI equal to 100, the GM-CSF level in culture supematants reached saturation [(130.00±12.61) pg/ml]. After GM-CSF gene-modification, DCs tend to be more maturated as detected by morphological observation and phenotype analysis. At the same time, the capacity of activating the proliferation of allogeneic T lymphocytes was enhanced greatly. T lymphocytes stimulated by DCs transfected with GM-CSF gene showed a specific killing effect on gastric carcinoma cells and produced high level of INF-γ[ ( 1245. 00±13. 75) pg/ml].Conclusion After GM-CSF gene modification, DCs can produce high level of GM-CSF, which tend to be more maturated, and the capacity of activating the proliferation of allogeneic T lymphocytes is enhanced greatly. GM-CSF gene modified DCs can induce specific CTL to target tumor cells in vitro.     

树突状细胞负载MAGE-n表位肽的体外免疫学效应研究

目的:研究树突状细胞(dendritic cell,DC)负载MAGE-n表位肽QLVFGIEVV体外诱导特异性CTL的能力及其抗肿瘤效应。方法:MAGE-n表位肽以固相多肽合成技术合成,并用HPLC进行纯化,质谱法(MS)鉴定,以流式细胞仪筛选HLA-A2+人外周血PBMC,连续贴壁法分离培养人外周血来源树突状细胞,用成熟的DC负载MAGE-n表位肽QLVFGIEVV反复刺激活化诱导抗原特异性CTL,用51Cr释放法检测CTL的杀伤活性,并用抗HLA-A2分子单抗进行杀伤抑制实验。结果:用DC负载MAGE-n表位肽QLVF-GIEVV可诱导特异性CTL反应,对MAGE-n阳性表达细胞有较强的杀伤作用。结论:DC负载抗原肽QLVF-GIEVV在体外可诱发较强的特异性免疫反应。     

粒细胞-巨噬细胞集落刺激因子基因修饰的树突状细胞疫苗增强体外抗肿瘤免疫效应

Objective To investigate if granulocyte-macrophage colony stimulating factor (GM-CSF) gene-modified dendritic cells ( DC) enhance antitumor immunity in vitro. Methods Mice were injected with chemokine ligand 3 (CCL3) via the tail vein. Fresh B220-CD11c+ cells were sorted from the peripheral blood mononuclear cells (PBMCs) and cultured into DCs by cytokines. DCs were transfected with AdGM-CSF gene at different ratios of multiplicity of infection ( MOI) to determine the optimal gene transfection conditions, and the expression of GM-CSF was detected after transfection. The variation of GM-CSF gene-modifiedDCs were analyzed by morphological examination, phenotype analysis, and mixed lymphocyte reaction (MLR). DCs were loaded with gastric cancer antigen obtained by freezing and thawing method. The killing effect of DCs vaccine-stimulated T lymphocytes on gastric cancer cells was assessed by MTT assay. INF-γ production was determined with the INF-γ ELISA kit. Results B220- CD11c+ cells increased obviously after CCL3 injection. The ELISA results showed that after GM-CSF gene modification, DCs could produce high level of GM-CSF. When DCs were transfected with AdGM-CSF gene at MOI equal to 100, the GM-CSF level in culture supematants reached saturation [(130.00±12.61) pg/ml]. After GM-CSF gene-modification, DCs tend to be more maturated as detected by morphological observation and phenotype analysis. At the same time, the capacity of activating the proliferation of allogeneic T lymphocytes was enhanced greatly. T lymphocytes stimulated by DCs transfected with GM-CSF gene showed a specific killing effect on gastric carcinoma cells and produced high level of INF-γ[ ( 1245. 00±13. 75) pg/ml].Conclusion After GM-CSF gene modification, DCs can produce high level of GM-CSF, which tend to be more maturated, and the capacity of activating the proliferation of allogeneic T lymphocytes is enhanced greatly. GM-CSF gene modified DCs can induce specific CTL to target tumor cells in vitro.     

癌胚抗原重组痘苗病毒转染树突状细胞诱导CEA特异性T细胞免疫

 目的　研究人癌胚抗原重组痘苗病毒 (rV CEA)转染树突状细胞 (DC)在体外诱导CEA特异性的T细胞免疫。方法　将rV CEA转染外周血单个核细胞来源的DC后用于激发自体的T细胞 ,通过对T细胞的增殖及杀伤功能的检测 ,与野生型痘苗病毒 (V 76 1)转染的DC激发的T细胞进行比较。结果　经rV CEA转染的DC激活的T细胞增殖力强 ,对CEA分泌性肿瘤细胞具特异性杀伤作用。结论　rV CEA转染的DC可以诱导CEA特异性T细胞活性。     

自体宫颈癌-树突细胞疫苗激活的CTL杀伤效应

背景与目的:树突细胞(dendriticcells,DC)是目前已知的功能最强的抗原递呈细胞(antigen-presentingcell,APC),它可以在体内、外向T淋巴细胞递呈抗原,并诱发细胞毒T淋巴细胞(cytotoxicTlymphocyte,CTL)反应。本研究旨在探讨负载自体宫颈癌抗原的DC体外激发的CTL对自体宫颈癌细胞的杀伤效应。方法:先冻融宫颈癌细胞制备抗原,然后以GM-CSF、IL-4诱导自体外周血单个核细胞(peripheralbloodmononuclearcell,PBMC)获得DC并负载抗原,刺激自体T淋巴细胞制备宫颈癌抗原特异性CTL,观察CTL对宫颈癌细胞的杀伤活性。结果:负载自体宫颈癌抗原DC诱导的特异性CTL对自体宫颈癌细胞的体外杀伤率高达79.32%～89.27%,显著高于淋巴因子激活的杀伤细胞(lymphokine-activatedkillingcells,LAK)的杀伤率(t≥2.89,P<0.05);且对宫颈癌HeLa细胞株具有一定杀伤效应(40.35%～58.09%),但低于自体癌细胞组(t≥2.97,P<0.05);特异性CTL对HepG2、MCF7、A549、MGC803细胞无明显杀伤效应。结论:自体宫颈癌-树突细胞疫苗体外诱导的CTL具有高效而特异的抗自体宫颈癌细胞免疫活性,可望成为宫颈癌生物治疗的一个有力手段。     

Identification of an HLA-A*0201 cytotoxic T lymphocyte epitope specific to the endothelial antigen Tie-2

Tie-2 stabilises pericyte-endothelial interactions during angiogenesis and is highly expressed on endothelium during several diseases, including arthritis, age-related macular degeneration and cancer. A vaccine that targets endothelium overexpressing Tie-2 may result in vessel damage and stimulate an inflammatory cascade resulting in disease regression. We have identified a region unique to Tie-2 (amino acids 1-196) that is homologous in humans and mice. Using computer algorithms, several HLA-A*0201 epitopes that are identical in mice and humans were predicted within this region; however, binding assays showed that the majority of these epitopes were of low affinity. Modification of the anchor residues of 4 epitopes enhanced HLA binding. These epitopes were incorporated by site-directed mutagenesis into a Tie-2 DNA construct. Immunisation of HLA*0201 transgenic mice with one of the modified Tie-2 constructs stimulated CTLs that recognised both wild-type and modified peptide-pulsed target cells. In contrast, no CTLs were generated in mice immunised with wild-type Tie-2 construct, demonstrating that the modified epitope was necessary in the generation of CTLs. Moreover, CTLs from mice immunised with the modified construct killed HLA-A*0201 endothelial cells overexpressing Tie-2. Our study demonstrates that it is possible to break tolerance to the endothelial antigen Tie-2, suggesting that it may be feasible to design a vaccine to activate CTLs to kill endothelial cells overexpressing Tie-2.     

Administration route-dependent vaccine efficiency of murine dendritic cells pulsed with antigens

Dendritic cells (DCs) loaded with tumour antigens have been successfully used to induce protective tumour immunity in murine models and human trials. However, it is still unclear which DC administration route elicits a superior therapeutic effect. Herein, we investigated the vaccine efficiency of DC2.4 cells, a murine dendritic cell line, pulsed with ovalbumin (OVA) in the murine E.G7-OVA tumour model after immunization via various routes. After a single vaccination using 1 x 10(6)OVA-pulsed DC2.4 cells, tumour was completely rejected in the intradermally (i.d.; three of four mice), subcutaneously (s.c.; three of four mice), and intraperitoneally (i.p.; one of four mice) immunized groups. Double vaccinations enhanced the anti-tumour effect in all groups except the intravenous (i.v.) group, which failed to achieve complete rejection. The anti-tumour efficacy of each immunization route was correlated with the OVA-specific cytotoxic T lymphocyte (CTL) activity evaluated on day 7 post-vaccination. Furthermore, the accumulation of DC2.4 cells in the regional lymph nodes was detected only in the i.d.-and s.c.-injected groups. These results demonstrate that the administration route of antigen-loaded DCs affects the migration of DCs to lymphoid tissues and the magnitude of antigen-specific CTL response. Furthermore, the immunization route affects vaccine efficiency.     

In vitro antitumor cytotoxic T lymphocyte response induced by dendritic cells transduced with DeltaNp73alpha recombinant adenovirus

DeltaNp73alpha, the N-terminal truncated form of p73alpha is a candidate tumor antigen because of its selective expression in many human cancers and lack of expression in normal tissues. Therefore, we investigated the effects of dendritic cells infected with adenoviral DeltaNp73alpha (DNp73alpha) on breaking immune tolerance and induction of immunity against DNp73alpha-expressing (A549 lung cancer, K-562 leukemia) and non-expressing (MCF-7 breast cancer) cell lines. Immature dendritic cells generated in the presence of interleukin-4 and granulocyte/macrophage colony-stimulating factor from a human umbilical cord blood were transduced with a recombinant adenoviral (Ad) vector encoding full-length human DNp73alpha cDNA (Ad-DNp73alpha) or a control vector Ad-EGFP, using the centrifugal force method. Induction of DNp73alpha-specific CTL response was evaluated by a cytotoxic assay against the three human tumor cell lines with different DNp73alpha expression levels. The viability and activation status of transduced dendritic cells were assessed by flow cytometry. The dendrocyte/Ad-DNp73alpha-activated cytotoxic T lymphocytes showed significantly higher cytotoxicity against the cell lines A549/DNp73alpha, K-562 that expressed DNp73alpha than the DNp73alpha-null MCF-7 cells. The DCs/Ad-DNp73alpha showed higher survival rates than the DCs/Ad-EGFP or untransduced DCs, presumably due to the inhibition of cell death. These findings, with potential applications for immunotherapy, demonstrate that dendrocytes transduced with Ad-DNp73alpha can induce specific and sustained T cell responses against tumors expressing this variant p53-related gene.     

A method to generate mature dendritic cells from cryopreserved PBMC

To established methods for cryopresserving peripheral blood mononuclear cells(PBMCs)and producing DCs from cryopresserved PBMCs.Methods:Mature DCs were generated from cryopreserved PBMCs by using IL-4,GM-CSF,TNFα,IL-1β,IL-6,pgE2 and LPS.The phenotype of the resultant DCs was investigated by flow cytometry.The functions of the resultant DCs were verified by Elispot assay.Results:The resultant DCs expressed high levels of HLA ABC,HLA DR,costimulatory molecules and the DC maturation marker CD83.The mature DCs we generated from frozen PBMCs were able to prime CD8 T cells into long term IFN-γ producing peptide specific CTL.Conclusion:The DCs we developed from cryopreserved PBMC were fully mature and had the capability to stimulate immune reaction.Thus,we developed a method to generate functional mature DC from cryopreserved PBMC.     

Cytokines Required for Induction of Histocompatibility Leukocyte Antigen-class I-restricted and Tumor-specific Cytotoxic T Lymphocytes by a SART1-derived Peptide

Although there have been several reports on peptides of human tumor-rejection antigens capable of inducing histocompatibility leukocyte antigen (HLA)-class I-restricted and tumor-specific cytotoxic T lymphocytes (CTLs), it is not yet clear which cytokines are required for CTL induction. This study has investigated the cytokine combinations required for optimal induction of CTLs by SARTI^690–698 peptide, which is capable of inducing HLA-A24-restricted and tumor-specific CTLs in peripheral blood mononuclear cells (PBMCs). Pretreatment of PBMCs as a source of antigen-presenting cells (APCs) with interferon (IFN)-γ, or to some extent with IFN-α, but not with any of the other cytokines tested, augmented the peptide-induced CTL activity in HLA-A24 heterozygotes, but not in HLA-A24 homozygotes. This IFN-γ -mediated augmentation was inhibited by either interleukin (IL)-4 or IL-10. IL-2 alone in culture, along with weekly stimulation by peptide-pulsed APCs, was sufficient for the differentiation and proliferation of CTLs for the initial several weeks of culture. This IL-2-mediated activation of CTLs was inhibited by the addition of IFN-γ, IL-4, or IL-10 to the IL-2 culture. For further expansion of the CTLs, dendritic cells (DCs) induced from PBMCs with IL-4 and granulocyte macrophage colony-stimulating factor (GM-CSF) were required as APCs. These results indicate that IFN-γ and IL-2 are important in the activation of APCs and CTLs, respectively, while GM-CSF and IL-4 are needed for the induction of DCs, which in turn are required for further expansion of mature CTLs. These results are important in allowing for a better understanding of the cellular and molecular basis of tumor-specific immunity, and also for the development of peptide-based specific immunotherapy.     

肺癌gp96-多肽复合物/DC疫苗体外诱导CTL反应的实验研究

目的 研究热休克蛋白gp96－多肽复合物负载树突状细胞（dendritic cell,DC）后，能否诱导出gp96－多肽复合物特异性的细胞毒性T细胞（cytotoxic T lymphocyte,CTL）反应。方法 从一例肺癌患者肿瘤组织中提取gp96-多肽复合物和自体肿瘤细胞溶解物，分别负载从该患者骨髓血中培养的DC。以不同形式的抗原／DC疫苗分别刺由患者外周血中分离的淋巴细胞。采用ELISA法检测淋巴细胞所释放的IFN－γ量作为CTL反应的指标；以Cr^51释放实验分析致敏后的淋巴细胞对不同靶细胞的裂解和杀伤作用。结果 所有肿瘤抗原致敏淋巴细胞后均可以诱导产生CTL反应，其中以gp96－多肽复合物／DC疫苗诱导释放的IFN－γ量最高。肿瘤抗原致敏淋巴细胞后对原代培养的肿瘤细胞的杀伤作用高于PG细胞和K562细胞。结论 自体肿瘤组织中提取的gp96－多肽复合物能诱导出肿瘤特异性CTL反应，而负载DC后能激发起更强的CTL。     

IL-15对树突状细胞激活的T淋巴细胞免疫反应的作用观察

  目的  观察IL-15和IL-2对树突状细胞疫苗(dendritic cells, DC)激活的淋巴细胞免疫反应的作用。  方法  取健康小鼠的脾淋巴细胞, 在体外采用肿瘤抗原负载的DC疫苗与脾淋巴细胞混合后, 分别加入IL-15或IL-2培养7d。采用流式细胞术检测淋巴细胞免疫表型的变化; 使用酶联免疫斑点实验(ELISPOT)检测分泌干扰素(interferon, IFN)-γ的细胞数量; 利用51Cr释放法检测淋巴细胞对肿瘤细胞的杀伤活性。  结果  IL-15联合DC疫苗增强了T细胞反应, 包括淋巴细胞免疫表型和功能的改变。  结论  与IL-2相比, IL-15能更好的增强DC疫苗诱发的免疫反应, 为IL-15与DC疫苗的联合应用提供了一定的依据。