负载自体热休克凋亡癌细胞抗原的DC治疗三阴性乳腺癌的随机对照多中心临床试验

评价自体热休克凋亡肿瘤细胞抗原负载DC对Ⅰ到Ⅳ期ER、PR、Her2三阴性乳腺癌（triple-negative breast cancer, TNBC）患者治疗的疗效和患者对该治疗的耐受性。方法：入选南京、常州、无锡三地3个医院的TNBC患者168例,按照2∶1的比例使用随机数字表将患者随机分组为DC免疫组112例、对照组56例。DC疫苗治疗每周1次,4次为1个疗程,疗程间间隔1个月,共3个疗程。疫苗治疗前、后检测患者外周血中细胞因子谱（IL-2、IL-10、IL-12、TNF-α和IFN-γ）水平和肿瘤特异性CD8+IFNγ+T细胞比例以及进行DTH试验。末次治疗后每3个月随访1次,随访2年,统计患者疾病无进展生存率（PFSR）。结果：Ⅰ~Ⅲ期TNBC患者DC免疫治疗1个疗程后,IL-2、TNF-α和IFN-γ水平即较治疗前（基线）显著升高（PIL-2=0.038 4、PTNF-α=0023 7、PIFN-γ=0.022 1）,且随疗程增加其水平不断升高;而Ⅳ期TNBC患者治疗3个疗程后细胞因子水平均无明显提高。Ⅰ~Ⅲ期TNBC患者外周血CD8+IFN-γ+T细胞比例提升速度与幅度较为缓慢,3个疗程后提升幅度才显示有统计学意义（P<0.05）。患者的DTH试验阳性率伴随疗程数的增加而提升,两者呈正相关关系（r=0.973）;早期（Ⅰ期和Ⅱ期）TNBC患者DTH平均阳性率明显高于中晚期患者（Ⅲ期和Ⅳ期）。2年随访期内Ⅰ~Ⅱ期TNBC患者病情均较稳定,生存率100%;Ⅲ~Ⅳ期TNBC患者DC治疗组PFSR明显高于对照组（71.43% vs 32.73%,P<0.05）。Ⅰ~Ⅳ期DTH阳性患者的PFSR明显高于DTH阴性患者（87.30% vs 51.02%,P<0.05）。治疗组112例TNBC患者对DC治疗耐受良好,未发现Ⅱ级以上不良反应。结论：自体热休克凋亡肿瘤细胞抗原负载DC可有效诱导早期TNBC患者产生Th1型免疫应答反应,分泌高水平Th1型抗瘤因子,3个疗程后可激发明显的肿瘤抗原特异性CTL反应,DTH试验可作为DC免疫有效性的评价指标之一。该DC免疫治疗方法可抑制晚期TNBC患者疾病进展,从而提高PFSR,患者耐受性良好。     

负载HER-2/neu多肽的树突状细胞激发特异性CTL反应

目的:探讨以HER-2/neu为靶抗原、树突状细胞(dendritic cell,DC)为抗原载体激发特异性细胞毒性T淋巴细胞(cytotoxic T lymphocytes,CTL)反应的能力及研制治疗型乳腺癌疫苗的可行性。方法:采集17例HLA-A201+HER-2/neu+乳腺癌患者外周血,分离单个核细胞与外周血淋巴细胞(peripheral blood lymphocyte,PBL),并诱导为成熟DC(mature dendriticcell,mDC);人工合成HER-2/neu多肽[E75(KIFGSLAFL)和GP2(IISAVVGIL)2条]负载mDC后体外反复致敏PBL(3次,每周1次),检测其激发HER-2/neu特异性CTL的能力与CTL的杀伤活性。同时于患者腹股沟淋巴结富集区皮内注射负载HER-2/neu多肽的DC,每周1次,共接种4次,检测接种前后患者外周血细胞因子和特异性的CTL水平变化,并进行DTH试验。结果:患者外周PBL经过负载HER-2/neu多肽DC共3轮致敏后,HER-2/neu多肽特异的CTL平均比例比对照组(未负载HER-2/neu多肽DC组)明显增高[(5.41±1.44)%vs(0.41±0.12)%,P<0.05];致敏后PBL对负载HER-2/neu多肽T2靶细胞的杀伤率明显高于对照组(未负载DC诱导的CTL)[效靶比为30∶1时,(35.5±4.7)%vs(11.2±1.4)%,P<0.05]。接种负载HER-2/neu多肽的DC后,患者体内血清中细胞因子IL-2、IL-12、IFN-γ水平较治疗前显著升高[(409.09±89.39)vs(148.79±28.32)ng/ml,(56.23±14.08)vs(24.49±56.23)ng/ml,(146.57±25.97)vs(67.77±39.35)ng/ml;均P<0.05],TNF-α和IL-10水平较治疗前变化不大(P>0.05)。患者DTH试验阳性率为47%(8/17),DTH阳性患者外周血中特异性CTL比例明显上升。结论:负载HER-2/neu多肽的DC体内、外均具有激发特异性CTL反应能力,可诱导Th1型细胞因子的分泌,未发生临床不良反应。     

多肽负载DC联合CIK治疗激素难治性前列腺癌的疗效

目的:研究多肽负载树突状细胞(dendritic cell,DC)联合细胞因子诱导的杀伤细胞(cytokine-induced killer cell,CIK)对激素难治性前列腺癌(hormone refractory metastatic prostate cancer,HRPC)患者免疫治疗的效果。方法:选择无锡市第四人民医院中西医结合科收治的HLA-A2+HRPC患者26例,分离外周血单个核细胞,其中贴壁细胞经GM-CSF、IL-4联合诱导培养为成熟DC,负载前列腺癌特异性抗原(prostate specific antigen,PSA)、前列腺酸性磷酸酶(prostatic acid phosphatase,PAP)、前列腺特异性膜抗原(prostate specific membrane antigen,PSMA)三个多肽,制备成DC疫苗,经患者腹股沟皮内注射;未贴壁细胞经IFN-γ、IL-2、抗CD3单抗、IL-1体外诱导培养成CIK,经静脉回输给患者。在治疗后1周进行迟发型超敏反应(delayed type hypersensitivity,DTH)检测,在患者治疗前后进行血清中细胞因子和PSA检测,治疗结束后4周进行短期疗效评价。结果:26例HRPC患者对DC联合CIK治疗的耐受良好。治疗后患者血清中IL-2、IL-12、IFN-γ水平较治疗前显著升高(上升幅度分别为65.07%、67.69%和125.38%,P<0.05或P<0.01),TNF-α和IL-10水平变化不大;DTH的阳性率为43.5%(10/23);7例患者的CD8+IFN-γ+T细胞比例较治疗前显著提高[(8.95±2.74)%vs(0.39±0.15)%,P<0.01];8/26例患者的PSA下降,降幅为13%～66%。26例患者短期疗效评价,3例PR、4例PD、19例SD,所有患者治疗中未出现明显不良反应。结论:多肽负载DC联合CIK治疗HRPC能激发患者的免疫应答、诱导Th1型细胞因子的分泌,近期疗效良好,是一种安全的治疗方法。     

转移性肾癌自体DC-CIK细胞免疫治疗疗效观察

目的:观察自体树突状细胞（dendritic cell,DC）激活的细胞因子诱导的杀伤（cytokine-induced killer,CIK）细胞治疗转移性肾癌的临床效果及安全性。方法:32例转移性肾癌患者,采集外周血单个核细胞（peripheral blood mononuclear cell,PBMC）,体外制备成DC-CIK细胞,12天后连续三天回输,输注结束后次日开始下一个疗程,共计3个疗程。观察治疗前、治疗后6个月外周血中T细胞亚群及细胞因子IL-2、IFN-γ、IL-4及IL-10的变化,并统计治疗3年后生存率以及生活质量评分（KPS）,观察不良反应。结果:治疗前后比较CD4+无显著差异,CD8+显著降低,CD3+、CD56+以及CD4+/CD8+有显著差异（P<0.05）。两组治疗后6个月 IL-2、IFN-γ与治疗前比较均升高（P<0.05）。IL-4、IL-10与治疗前比较均降低（P<0.05）。治疗前、治疗后2周患者WBC、ALT、AST以及BUN、Cr比较无显著性差别（P>0.05）。3年生存率90.63%,治疗后1年生活质量评分平均升高24.7,显著高于治疗前的11.8(P<0.05)。不良反应轻微。结论:自体DC-CIK细胞输注后可显著提升转移性肾癌（RCC）患者免疫水平,安全性良好,可作为晚期RCC患者可选择的治疗方法。     

自体热休克凋亡细胞负载树突状细胞治疗大肠癌的疗效观察

目的:探讨自体热休克凋亡细胞负载的树突状细胞(dendritic cells,DC)治疗大肠癌的临床疗效。方法:采用酶消化法从手术切除的14例大肠癌新鲜组织获得单细胞悬液,热休克处理后用桦脂酸诱导其凋亡制备成细胞抗原;采集外周静脉血,分离单个核细胞,经GM-CSF与IL-4体外诱导成未成熟树突状细胞,负载细胞抗原后制备成DC肿瘤疫苗;对14例大肠癌患者进行4个疗程的DC免疫治疗,观察患者不良反应、敏感程度、生存质量、生存时间、癌胚抗原等临床指标。结果:DC治疗大肠癌未出现明显不良反应,5例患者的DTH检测阳性;14例患者的平均CEA水平由治疗前的99.5ng/ml降至治疗后的71.4ng/ml(P<0.05);治疗后血清中IL-2、IL-12及IFN-γ浓度均显著高于治疗前,具有统计学差异(P<0.05);所有患者均生存1年,其中1例患者出现了短暂的后腹膜淋巴结缩小,持续3个月,第2年由于肿瘤的进展生存率下降,由80%降至20%;患者的KPS评分上升,与治疗前比较有所改善(P<0.05)。结论:自体热休克凋亡细胞负载的DC治疗大肠癌在临床治疗中具较高安全性,可改善患者免疫功能,临床观察显示患者近期临床效果明显;为大肠癌的临床综合治疗提供了一种新方法。     

人结肠癌主动特异性免疫治疗的初步研究

目的:探讨自体肿瘤疫苗的作用机制及临床意义。方法:50例进展期结肠癌病人术后,以自体肿瘤细胞疫苗辅助主动免疫治疗。术后第4周开始免疫接种(共4次,每次间隔7~10天);接种前3天及第4次接种后1周,采集外周血。采集血清监测血清IFN-γ、IL-10水平;用PPD及自体肿瘤抗原做皮肤迟发型过敏试验(DTH),48小时后测量红斑、硬结大小(mm);对DTH反应部位皮肤活检,用免疫组化染色,了解局部免疫活性细胞浸润;临床随访。结果:自体肿瘤细胞疫苗治疗后:血清IFN-γ水平升高,由(6.01±2.30)pg/ml升至(12.98±4.65)pg/ml;而IL-10水平由(19.80±10.15)pg/ml降至(8.92±4.60)pg/ml,差异有非常显著性(P<0.05);治疗前后病人对自体肿瘤抗原的特异性DTH反应明显增强(P<0.01);DTH反应部位,CD8 T细胞、CD4 T细胞及DC细胞浸润明显增多;病人耐受性良好,无溃疡等严重副作用发生;随访结果显示:术后辅助自体肿瘤疫苗主动特异性免疫治疗,可延长结肠癌病人的无瘤生存时间,降低复发率及死亡率。结论:自体肿瘤疫苗可激发病人特异性细胞介导的免疫反应;可改善肿瘤病人的抗瘤免疫反应;自体肿瘤疫苗主动特异性免疫治疗,对于杀灭残留癌细胞、抗转移及复发有重要作用。     

自体热休克凋亡细胞负载树突状细胞治疗大肠癌的疗效观察

目的：探讨自体热休克凋亡细胞负载的树突状细胞（dendritic cells,DC）治疗大肠癌的临床疗效。方法：采用酶消化法从手术切除的14例大肠癌新鲜组织获得单细胞悬液,热休克处理后用桦脂酸诱导其凋亡制备成细胞抗原;采集外周静脉血,分离单个核细胞,经GM-CSF与IL-4体外诱导成未成熟树突状细胞,负载细胞抗原后制备成DC肿瘤疫苗;对14例大肠癌患者进行4个疗程的DC免疫治疗,观察患者不良反应、敏感程度、生存质量、生存时间、癌胚抗原等临床指标。结果：DC治疗大肠癌未出现明显不良反应,5例患者的DTH检测阳性;14例患者的平均CEA水平由治疗前的99.5ng/ml降至治疗后的71.4ng/ml（P〈0.05）;治疗后血清中IL-2、IL-12及IFN-γ浓度均显著高于治疗前,具有统计学差异（P〈0.05）;所有患者均生存1年,其中1例患者出现了短暂的后腹膜淋巴结缩小,持续3个月,第2年由于肿瘤的进展生存率下降,由80%降至20%;患者的KPS评分上升,与治疗前比较有所改善（P〈0.05）。结论：自体热休克凋亡细胞负载的DC治疗大肠癌在临床治疗中具较高安全性,可改善患者免疫功能,临床观察显示患者近期临床效果明显;为大肠癌的临床综合治疗提供了一种新方法。     

肝癌患者PBMC中TH1/TH2类细胞因子mRNA表达

目的探讨TH1类细胞因子IL-2、IFN-γ及TH2类细胞因子IL-4mRNA在Ⅰ期和Ⅱ期肝癌患者外周血单个核细胞(PBMC)中的表达模式,并观察IL-2、IFN-γmRNA在手术前后的表达变化。方法采用逆转录聚合酶链反应(RT-PCR)方法测定8例Ⅰ期和10例Ⅱ期原发性肝癌患者手术前后及15例健康人外周血中IL-2、IFN-γ的表达,并用凝胶图像分析系统进行图像分析。结果在18例肝癌患者中,IL-2、IFN-γmRNA的表达率分别为16.7%、5.6%,低于IL-4的表达率88.9%(P<0.05),呈现明显的TH2偏移状态,且IL-2、IFN-γ的表达强度在0.10~0.11之间,也远低于IL-4的0.21~0.22;在8例Ⅰ期肝癌患者中,术前有25.0%(2/8)表达IL-2mRNA,12.5%(1/8)表达IFN-γ,高于对照组的13.3%(2/15)、5.6%(1/15);手术治疗后两者的表达率分别是50.0%、37.5%,高于术前组;术前组及正常对照组两种细胞因子mRNA表达强度(0.10~0.12)明显低于术后组的0.20~0.22(P<0.05)。另外,10例Ⅱ期患者手术前后术均无IFN-γm...     

过继免疫联合化疗对晚期非小细胞肺癌的疗效及免疫功能的影响

目的 探讨过继免疫联合化疗治疗非小细胞肺癌的疗效和对免疫功能的影响.方法 选择NSCLC患者112例按治疗方式分为观察组和对照组.观察组42患者采用紫杉醇加奈达铂(TP方案)化疗,21 d为1个治疗周期,第1个周期治疗结束后10 d开始回输DC-CIK细胞,1次/d,输注4次为1个疗程,每次回输前给予苯海拉明20 mg肌注防止过敏反应的发生,然后再进行2个周期的化疗.对照组70例患者仅给予TP方案化疗3个周期.结果 观察组患者DCR为66.7％,对照组患者DCR为52.9％,差异有统计学意义,P＜0.05.观察组患者治疗后外周血CD3+、D8+、D56+细胞比值的升高有统计学意义,P＜0.05,CD4+/CD8+的比值降低,与治疗前相比差异显著,P＜0.05.对照组患者治疗后外周血CD3+、D8+、D56+细胞比值的降低有统计学意义,P＜0.05,CD4 +/CD8+的比值升高,与治疗前相比差异显著,P ＜0.05.观察组患者治疗后外周血IL-2、IL-12、IFN-γ和TNF-oα水平升高显著,P＜0.05.对照组患者治疗后外周血IL-12、IFN-γ和TNF-α降低显著,P＜0.05.结论 过继免疫治疗联合化疗治疗NSCLC,可提高患者的疾病控制率,改善患者的免疫功能.     

宫颈癌患者血清Th1、Th2细胞因子表达水平及意义

目的 探讨宫颈癌患者血清Th1、Th2细胞因子表达水平及意义.方法 选取宫颈癌患者94例(观察组),同时选取健康女性90例作为对照组,检测两组患者干扰素-γ(IFN-γ)、白细胞介素-2(IL-2)、IL-4和IL-6.结果 观察组IFN-γ、IL-2、IL-4和IL-6水平分别为(52.13±9.55)pg/ml、(38.70±8.96)pg/ml、(27.61±6.22)pg/ml和(32.16±7.81)pg/ml,均高于对照组(P﹤0.05);Ⅲ~Ⅳ期患者IFN-γ、IL-2、IL-4和IL-6水平高于Ⅰ期和Ⅱ期患者(P﹤0.05),Ⅱ期患者IFN-γ、IL-2、IL-4和IL-6水平高于Ⅰ期患者(P﹤0.05);不同分化程度患者IFN-γ、IL-2、IL-4和IL-6水平比较差异无统计学意义(P﹥0.05),有无淋巴结转移患者IFN-γ、IL-2、IL-4和IL-6水平比较差异无统计学意义(P﹥0.05).结论 宫颈癌患者Th1/Th2细胞因子动态平衡紊乱,可能在肿瘤的发生发展中有一定作用.     

AFP与HBsAg混合多肽负载树突状细胞治疗HBV阳性肝癌的研究

目的：以AFP和HBsAg为双靶标,以树突状细胞（DCs）为抗原载体与佐剂,探讨其激发特异性（CTL）反应的能力及其在HBV阳性肝癌（HCC）患者应用的可行性。方法：采集20例HLA—A201＋HBV＋AFP＋HCC患者外周血,分离单核细胞（Mo）和外周淋巴细胞（PBLs）,将Mo诱导为成熟DCs（mDC）;人工合成AFP和HBsAg多肽,负载mDC后体外致敏,检测其体外杀伤活性;同时于患者皮内注射负载AFP和HBsAg混合多肽的DCs,监测接种前后患者外周血中细胞因子和特异性CTL水平变化,并进行DTH试验,治疗后跟踪检测患者AFP水平和乙肝两对半水平的变化。结果l患者外周PBLs经DCs致敏后,对负载AFP和HBsAg多肽的T2靶细胞杀伤率明显增高,P〈0．05。接种负载AFP和HBsAg多肽的DCs后,血清中IL-2、IL-12、IFN-γ水平较治疗前显著升高,P〈0．05,TNF-a和IL-10水平较治疗前均差异不大,P〉0．05;DTH试验阳性率为41．7％（5／12）,患者外周血中特异性CTLs比例明显上升的有4例（4／12）。治疗后有4例患者AFP水平下降,9例HBeAg阳性的患者中有3例HBeAg下降,2例患者出现血清学应答。结论：负载AFP和HBsAg多肽的Dcs体内、体外均具有激发特异性cTLs反应能力,可诱导Thl型细胞因子的分泌,有望成为一种HCC治疗的新方法。     

自体树突状细胞疫苗治疗雌、孕激素受体双阴性乳腺癌的疗效观察

目的 评价自体肿瘤冻融抗原致敏的自体树突状细胞（ADC）疫苗免疫治疗,对处于Ⅱ、ⅢA期雌、孕激素受体双阴性表达乳腺癌患者的疾病进展及生存情况的影响。方法利用自体肿瘤冻融抗原致敏的CD14+前体细胞产生树突状细胞,在细胞因子的作用下开始成熟,制备ADC疫苗。选取符合入组标准的63例患者,根据治疗方案分为实验组和对照组,实验组共接受4次皮下ADC回输。测定实验组疫苗接种前后IFN-γ+／CD8+T细胞含量及两组针对抗肿瘤裂解物发生的Ⅳ型变态反应情况,随访两组的疾病进展情况。结果 疫苗接种后提高了IFN-γ+／CD8+T细胞数量,与疫苗接种前比较差异有统计学意义[（23.4±4.1）％ vs.（14.3±2.0）％, P＜0.05];实验组18例（58.1％）为Ⅳ型变态反应阳性,而对照组均为阴性,差异有统计学意义（P＜0.05）。实验组的3年无进展生存率高于对照组,差异有统计学意义（76.9％ vs. 31.0％, P＜0.05）。实验组未发生严重不良反应。结论ADC疫苗可能通过触发乳腺癌患者的免疫应答机制来治疗乳腺癌,具有较好的效果,并延长无进展生存期。     

Skin tests predict survival after autologous tumor cell vaccination in metastatic melanoma: Experience in 81 patients

Background:Currently there is no standard adjuvant treatmentfollowing surgical resection of metastatic melanoma. We investigated whethersurgery followed by autologous tumor cell-BCG vaccination was beneficial formalignant melanoma patients. In this study we focus on the prognostic valueof DTH response following vaccination therapy. Patients and methods:Eighty-one patients with AJCC stage III andIV melanoma were selected. Whenever feasible, radical metastasectomy wasperformed. ASI was initiated by the administration of three weeklyintra-cutaneous vaccinations with 10⁷ irradiated autologous tumorcells, starting four weeks after surgery. Depending on the size of DTHresponse to the first three injections, subsequent vaccinations were planned.The first two vaccines also contained 10⁷ BCG organisms as animmune stimulatory adjuvant. Results:Induration as well as erythema correlated strongly withsurvival (P< 0.0001 and P= 0.0004). After radicalmetastasectomy in stage III melanoma patients a five-year survival of48% was observed. In stage IV disease, a five-year survival of34% was seen, after radical surgery had been performed. Whenmacroscopic disease was present at start of vaccination treatment, no clinicalresponses occurred. Apart from transient skin ulceration at the site ofBCG-containing vaccinations, no serious side effects were observed. Conclusions:This study shows that large-scale preparation ofautologous melanoma cell vaccines is feasible, while vaccination results inDTH responses that correlate significantly with survival. ASI seemed to bebeneficial in stage III and stage IV melanoma when given in the adjuvantsetting, while causing only very mild side effects.     

Dacarbazine (DTIC) versus vaccination with autologous peptide-pulsed dendritic cells (DC) in first-line treatment of patients with metastatic melanoma: a randomized phase III trial of the DC study group of the DeCOG.

BACKGROUND: This randomized phase III trial was designed to demonstrate the superiority of autologous peptide-loaded dendritic cell (DC) vaccination over standard dacarbazine (DTIC) chemotherapy in stage IV melanoma patients. PATIENTS AND METHODS: DTIC 850 mg/m2 intravenously was applied in 4-week intervals. DC vaccines loaded with MHC class I and II-restricted peptides were applied subcutaneously at 2-week intervals for the first five vaccinations and every 4 weeks thereafter. The primary study end point was objective response (OR); secondary end points were toxicity, overall (OS) and progression-free survival (PFS). RESULTS: At the time of the first interim analysis 55 patients had been enrolled into the DTIC and 53 into the DC-arm (ITT). OR was low (DTIC: 5.5%, DC: 3.8%), but not significantly different in the two arms. The Data Safety & Monitoring Board recommended closure of the study. Unscheduled subset analyses revealed that patients with normal serum LDH and/or stage M1a/b survived longer in both arms than those with elevated serum LDH and/or stage M1c. Only in the DC-arm did those patients with (i) an initial unimpaired general health status (Karnofsky = 100) or (ii) an HLA-A2+/HLA-B44- haplotype survive significantly longer than patients with a Karnofsky index <100 (P = 0.007 versus P = 0.057 in the DTIC-arm) or other HLA haplotypes (P = 0.04 versus P = 0.57 in DTIC-treated patients). CONCLUSIONS: DC vaccination could not be demonstrated to be more effective than DTIC chemotherapy in stage IV melanoma patients. The observed association of overall performance status and HLA haplotype with overall survival for patients treated by DC vaccination should be tested in future trials employing DC vaccines.     

Delayed-type hypersensitivity response is a predictor of peripheral blood T-cell immunity after HER-2/neu peptide immunization.

Many groups that immunize cancer patients with cancer vaccines use the generation of a delayed-type hypersensitivity (DTH) response as the primary measure of the ability to immunize a patient to a tumor cell or specific tumor antigen. This study examines whether the development of a tumor antigen-specific DTH response, measured after vaccination with peptide-based vaccines, correlates to in vitro assessment of peripheral blood antigen-specific T-cell responses. The HER-2/neu protein was used as a model tumor antigen. Thirty-two patients who completed a course of immunization with HER-2/neu peptide-based vaccines were analyzed. HER-2/neu peptide-specific DTH responses (n = 93) and peripheral blood T-cell responses (n = 93) were measured 30 days after the final immunization. Size of DTH induration was correlated with HER-2/neu-specific T-cell proliferative responses assessed from peripheral blood lymphocytes isolated concurrently with peptide skin test placement. HER-2/neu peptide-specific DTH responses > or =10 mm2 correlated significantly to a measurable peptide-specific peripheral blood T-cell response defined as stimulation index >2.0 (P = 0.0006). However, antigen-specific DTH responses with magnitudes between 5 and 9 mm2 were not significantly associated with the development of systemic immunity. DTH responses between 5 and 9 mm2 carried an odds ratio of 1.3 (P = 0.61) in predicting a measurable systemic tumor antigen response. The findings presented here demonstrate that tumor antigen-specific DTH responses > or =10 mm2 correlate with measurable in vitro antigen-specific lymphocytic proliferation and are, in this model system, a reflection of systemic immunization.     

Immunization of cancer patients with a HER-2/neu, HLA-A2 peptide, p369-377, results in short-lived peptide-specific immunity.

Ideally, vaccines should be designed to elicit long-lived immunity. The goal of this study was to determine whether HER-2/neu peptide-specific CD8+ T-cell immunity could be elicited using an immunodominant HER-2/neu-derived HLA-A2 peptide alone in the absence of exogenous help. Granulocyte macrophage colony-stimulating factor (GM-CSF) was used as adjuvant. Six HLA-A2 patients with HER-2/neu-overexpressing cancers received 6 monthly vaccinations with a vaccine preparation consisting of 500 microg of HER-2/neu peptide, p369-377, admixed with 100 microg of GM-CSF. The patients had either stage III or IV breast or ovarian cancer. Immune responses to the p369-377 were examined using an IFN-gamma enzyme-linked immunosorbent spot assay. Before vaccination, the median precursor frequency (range), defined as precursors per 10(6) peripheral blood mononuclear cell, to p369-377 was 0 (no range). After vaccination, the median precursor frequency to p369-377 in four evaluable patients was 0 (0-116). Overall, HER-2/neu peptide-specific precursors developed to p369-377 in two of four evaluable subjects. The responses were short-lived and not detectable at 5 months after the final vaccination. Immunocompetence was evident, because patients had detectable enzyme-linked immunosorbent spot responses to tetanus toxoid and influenza. These results demonstrate that HER-2/neu MHC class I epitopes can induce HER-2/neu peptide-specific IFN-gamma-producing CD8+ T cells. However, the magnitude of the responses were low, as well as short-lived, suggesting that CD4+ T-cell help is required for lasting immunity to this epitope.     

DC - CIK 治疗 43 例晚期非小细胞肺癌的近期疗效观察简

目的：探讨树突状细胞（dendritic cell,DC）共培养细胞因子诱导的杀伤细胞（cytokine-induced killer,CIK）治疗晚期非小细胞肺癌（non-small cell lung cancer,NSCLC）患者的近期疗效。方法：将43例确诊的晚期非小细胞肺癌患者给予DC-CIK治疗,观察治疗前后外周血中 T细胞亚群及细胞因子的变化,卡氏评分（Karnofsky,KPS）和临床疗效,并观察其不良反应。结果：患者治疗后CD3＋、CD4＋、CD56＋和CD4＋/CD8＋的比例较治疗前增加（P＜0.05）,差异有统计学意义。免疫治疗增加了细胞因子IL-2、IL-12、IFN-γ和TNF-α的水平（P＜0.05）。治疗后疾病控制率为53.49%,KPS评分总提高率为83.72%。结论：DC-CIK治疗能提高患者的免疫功能及生活质量,有望成为非小细胞肺癌有效的过继免疫治疗方法。     

Intranodal administration of peptide-pulsed mature dendritic cell vaccines results in superior CD8+ T-cell function in melanoma patients.

PURPOSE: We evaluated the feasibility, safety, and immunogenicity of mature, peptide-pulsed dendritic cell (DC) vaccines administered by different routes. PATIENTS AND METHODS: We performed a randomized, phase I, dose-escalation study in 27 patients with metastatic melanoma receiving four autologous peptide-pulsed DC vaccinations. Patients were randomly assigned to an intravenous (IV), intranodal (IN), or intradermal (ID) route of administration (ROA). For each route, primary end points were dose-limiting toxicity, maximum-tolerated dose, and T-cell sensitization. Sensitization was evaluated through tetramer staining, in vitro peptide recognition assays, and delayed-type hypersensitivity (DTH) responses. RESULTS: Twenty-two (81.5%) of 27 patients completed all four vaccinations. Vaccinations were well tolerated; a few patients exhibited grade 1 to 2 toxicities including rash, fever, and injection site reaction. All routes of administration induced comparable increases in tetramer-staining CD8+ T cells (five of seven IV, four of seven IN, and four of six ID patients). However, the IN route induced significantly higher rates for de novo development of CD8+ T cells that respond by cytokine secretion to peptide-pulsed targets (six [85.7%] of seven IN patients v two [33%] of six ID patients v none [0%] of six IV patients; P =.005) and de novo DTH (seven [87.5%] of eight IN patients v two [33.3%] of six ID patients v one [14.3%] of seven IV patients; P =.01) compared with other routes. CONCLUSION: Administration of this peptide-pulsed mature DC vaccine by IN, IV, or ID routes is feasible and safe. IN administration seems to result in superior T-cell sensitization as measured by de novo target-cell recognition and DTH priming, indicating that IN may be the preferred ROA for mature DC vaccines.