热休克肿瘤细胞抗原负载的树突状细胞对结肠癌小鼠的治疗作用

背景与目的:目前通过树突状细胞诱导机体抗肿瘤免疫已经成为肿瘤免疫治疗的一种重要途径,热休克处理肿瘤细胞可提高肿瘤细胞抗原负载的树突状细胞分化成熟,增强其体内抗肿瘤作用.本实验研究热休克肿瘤细胞抗原负载的骨髓来源的树突状细胞(dendritic cell,DC)对结肠癌小鼠的治疗作用.方法:20只小鼠随分为4组,将小鼠结肠癌细胞CT26热休克处理后超声破膜,以其细胞裂解液负载BALB/c小鼠骨髓来源的DC,观察DC诱导肿瘤特异性细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)杀伤活性;并将DC接种于荷瘤小鼠皮下,观察其对肿瘤生长的抑制作用及对荷瘤小鼠生存期的影响.结果:与冻融抗原-DC组、DC组、PBS组相比,热休克抗原-DC诱导的CTL对CT26肿瘤细胞具有显著的杀伤作用,相同效靶比(P=0.00)下其肿瘤特异性细胞毒活性强于前者{(0.99±0.19)g vs.[(1.27±0.28)g、(2.19±0.35)g、(2.14±0.27)g],P均=0.00};用其进行免疫后对小鼠肿瘤的生长具有显著的抑制作用[(128±18)mm3 vs. (313±52)mm3,P=0.04],并能显著延长荷瘤小鼠的生存时间{(57±7)d vs. [(40±3)d、(25±3)d、(24±3)d],P均=0.00}.结论:热休克肿瘤细胞抗原负载的树突状细胞对结肠癌小鼠具有显著的治疗效果.     

肿瘤抗原致敏树突状细胞治疗胶质瘤的实验研究

 目的　研究肿瘤冻融抗原致敏树突状细胞 (DC)治疗胶质瘤的疗效。方法　从大鼠骨髓中分离出单核细胞后 ,加入添加rGM CSF和rIL 4的IMDM培养基中培养后鉴定。制备大鼠C6胶质瘤冻融抗原以及DC疫苗 ,同时肿瘤冻融抗原致敏DC体外细胞毒试验 ;建立动物模型 ,体内应用DC疫苗并观察疗效。结果　单核细胞培养 8天后可获得大量的树突状细胞 ;体外细胞毒试验发现对相应的C6肿瘤细胞有明显的杀伤作用 ;动物实验发现治疗组肿瘤生长明显抑制 ,而对照组肿瘤明显生长 ,两组肿瘤体积有显著性差异P <0 .0 1。结论　建立了体外扩增大鼠骨髓DC以及制备DC疫苗的方法 ,采用冻融抗原致敏DC疫苗治疗荷瘤动物生存时间明显延长 ,肿瘤生长明显抑制 ,为研究其在脑胶质瘤临床免疫治疗打下基础     

树突细胞联合β-榄香烯对小鼠胰腺癌治疗的实验研究

背景与目的：树突细胞（Dendritic cells，DC）是目前发现的功能最强大的肿瘤抗原专职免疫递呈细胞，DC疫苗是目前最具临床应用潜能的治疗性疫苗。肿瘤细胞凋亡小体被DC的MHC—1分子提呈并诱导的特异性CTL免疫反应抗癌作用显著。本研究探讨了DC负载凋亡癌细胞抗原后诱导的免疫应答联合β-榄香烯对小鼠胰腺癌治疗作用。方法：制备C57BL／6小鼠骨髓源性DC并鉴定，负载肿瘤抗原后制备成疫苗。分别观察DC诱导的CTL和β-榄香烯对胰腺癌细胞的杀伤作用。体内观察DC疫苗联合β-榄香烯对小鼠胰腺癌的抑制作用。结果1经电镜及流式细胞鉴定可获得典型的具有抗原加工及递呈作用的DC，利用TNF-α可使成熟的树突状细胞比例增加，MHC-Ⅱ、CD83、CD86等表面分子的表达较单纯DC组显著增强。负载凋亡抗原后DC诱导的特异性CTL体外对胰腺癌细胞有显著杀伤作用，效靶比30：1时，诱导的CTL对胰腺癌细胞抑制率达93％。β-榄香烯对癌细胞增殖抑制明显，DC疫苗与β-榄香烯联合治疗可明显抑制荷瘤小鼠肿瘤的生长，使其生存期延长，联合治疗组生存期为49．4天，与单纯β-榄香烯组（24．8天）、DC疫苗（38．5天）、DC组（20．7天）及对照组（17．5天）相比有显著差异（P〈0．05）。结论：经凋亡肿瘤细胞致敏的DC回输荷瘤小鼠可激发宿主针对特异肿瘤的Th1及CTL免疫应答，以DC、介导的免疫反应联合中药的治疗方法可成为抗肿瘤的崭新且有效手段。     

IL-18基因修饰增强肿瘤细胞裂解物致敏的树突状细胞的抗肿瘤效应

目的:观察经肿瘤细胞裂解物致敏的白细胞介素18(IL-18)基因修饰的树突状细胞(DC)体内诱导的抗肿瘤免疫应答反应.方法:体外培养的小鼠骨髓树突状细胞经IL-18重组腺病毒感染后(IL18-DC),再经Hepal-6肝癌细胞裂解物冲击致敏后通过皮下注射用于荷瘤小鼠的治疗.用ELISA检测细胞因子,4 h51Cr释放法检测NK细胞活性及CTL杀伤活性.结果:致敏IL18-DC组体内诱导NK细胞活性与未致敏IL18-DC组无明显差别(P＞0.05),但明显高于致敏DC组和DC组(均P＜0.01);致敏IL18-DC组体内诱导特异性CTL杀伤活性明显高于IL18-DC组、致敏DC组和DC组(均P＜0.01);致敏IL18-DC组免疫治疗作用明显优于未致敏IL18-DC组、致敏DC组和DC组(均P＜0.01).结论:肿瘤细胞裂解物致敏的IL-18基因修饰的DC疫苗进行体内免疫治疗,能诱导出显著的抗肿瘤免疫反应,为DC介导的肿瘤基因治疗开辟了新的途径.     

肝癌细胞对重组人HSP70联合肝癌组织冻融抗原修饰树突状细胞的免疫应答

目的:研究重组人热休克蛋白70(rhHSP70)联合肝癌组织冻融抗原修饰的树突状细胞(dendritic cell,DC)诱导对肝癌细胞的免疫杀伤效应.方法:外周血单个核细胞经粒-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素4 (IL-4)诱导生成DC,负载冻融抗原的同时加入rhHSP70,不同分组致敏的DC激活淋巴细胞生成肿瘤抗原特异性细胞毒性T淋巴细胞(cytotoxic T cells,CTL),四甲基偶氮唑蓝(MTT)法及3H-TdR法检测DC刺激淋巴细胞增殖能力, MTT法检测CTL对肝癌细胞的体外杀伤活性,酶联免疫吸附试验(ELISA)测定细胞因子的分泌,流式细胞术(FCM)检测DC表型变化.结果:冻融抗原致敏的DC可明显促进淋巴细胞增殖,能有效呈递肝癌冻融抗原,诱导产生抗原特异性CTL,联合rhHSP70能进一步增强CTL对肝癌细胞的杀伤作用.结论:肝癌冻融抗原联合rhHSP70修饰的DC诱导CTL对肝癌细胞能产生高效杀伤作用.     

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.     

DC肿瘤融合瘤苗抗肿瘤效应的实验研究

目的:观察DC与肿瘤细胞融合后的瘤苗体内诱导的抗肿瘤免疫应答以及对荷瘤小鼠的治疗作用.方法:应用免疫磁珠分选和贴壁培养方法收集融合细胞,应用3H-TdR掺入法、4h51Cr释放法观察T细胞增殖反应的能力和CTL活性,并观察瘤苗对荷瘤小鼠保护性免疫反应和免疫治疗作用.结果:DC肿瘤融合瘤苗具有强烈的激活T细胞增殖和抗原提呈的能力,在体外、体内诱导出更强的特异CTL细胞毒活性,使免疫小鼠产生一定的免疫保护作用,抵抗Hepa1-6肝癌细胞的再次攻击,使治疗的小鼠肿瘤的生长明显缓慢,具有更明显的治疗作用.结论:DC与肿瘤细胞融合后进行体内免疫和治疗,能诱导出显著的抗肿瘤免疫反应,为DC介导的肿瘤免疫治疗开辟了新的途径.     

抗白血病树突状细胞疫苗的制备

目的探索抗白血病异体树突状细胞疫苗的制备方法.方法从健康者外周血中诱生DC,用不同方法负载K562抗原(化学融合法,冻融抗原冲击法,肿瘤与DC共培养法),比较这些DC疫苗在递呈抗原激活T细胞增殖及杀伤功能方面的差异.结果从健康者外周血中可诱生出具有正常免疫表型和抗原递呈功能的DC.不同抗原负载方法致敏的DC均能激活T细胞特异性杀伤K562细胞.冻融抗原负载的DC能有效刺激T细胞增殖,其活化的CTL对K562的杀伤毒性最强.结论异体DC可有效递呈肿瘤抗原,激活T细胞杀伤肿瘤,冻融抗原负载的DC激活T细胞对肿瘤的杀伤最强,为今后临床DC疫苗的选择提供了实验依据.     

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。     

超声抗原树突状细胞瘤苗抗肿瘤效应的体外研究

 目的　探讨结肠癌肿瘤超声抗原致敏的树突状细胞（DC）疫苗体外对结肠癌细胞的杀伤作用。方法　分离HLA-A*0201型健康供者的外周血单个核细胞（PBMC）,重组人细胞因子 rhGM-CSF、rh IL-4、rh TNF-α体外诱导DC成熟,负载SW480超声抗原后体外刺激同一供者的细胞毒性T淋巴细胞（CTL）前体,MTT法检测CTL杀伤SW480（HLA-A*0201阳性）、K562（HLA-A*0201阴性）的细胞毒实验。结果　在效靶比20∶1、10∶1、5∶1、2.5∶1作用时,CTL对SW480的杀伤率分别为51.12 %、46.67 %、52.60 %、40.00 %;对K562的杀伤率分别为29.26 %、31.98 %、39.46 %、31.30 %,证明在各种效靶比作用时,CTL对SW480的作用均强于K562（P＜0.05）。结论　肿瘤超声抗原致敏的DC瘤苗能激发具有特异杀伤功能的CTL,并且体外杀伤靶细胞具有MHC限制性。     

Compliance with guidelines and correlation with outcome in patients with advanced germ-cell tumours

PurposeTo evaluate prior compliance with guidelines in patients treated with salvage chemotherapy for advanced germ-cell tumours (GCT).Patients and methodsData concerning the initial management of patients requiring salvage chemotherapy for GCT at Institut Gustave Roussy between 2000 and 2010 were obtained and correlated with recommendations for treatment. Criteria of non-compliance were defined based on guidelines. Compliance with guidelines, predictive factors for non-compliance and the impact on outcome were analysed.ResultsAmong 82 patients treated in the salvage setting, guidelines to initial treatment were followed in only 41 cases (50%). The most common non-compliance criteria were non-adherence to the planned dose (16%), an inappropriate interval between first-line chemotherapy cycles (16%), the lack of post-chemotherapy surgery (16%) and a long interval to post-chemotherapy surgery (48%). Compliance with standard care was better in cancer centres than in other hospitals (private or public) (Odd Ratio (OR): 6.9, P = 0.001). A poor-risk status according to the International Germ Cell Cancer Collaborative Group (IGCCCG) was also predictive of compliance in univariate but not in multivariate analysis. No significant difference in outcome after salvage chemotherapy was observed. Patients relapsing after non-compliant first-line therapy tended to be more easily salvaged, which is consistent with the fact that their initial treatment was inadequate. Some of these relapses were therefore probably not due to true biologically refractory disease.ConclusionGuidelines for first-line treatment are adhered to in only half the patients requiring salvage chemotherapy. As the only predictive factor for non-compliance was the treating centre, centralisation of patients with GCT in well-trained hospitals should be recommended.     

Costs associated with complications are lower with capecitabine than with 5‐fluorouracil in patients with colorectal cancer

BACKGROUND:

Capecitabine, an oral alternative to 5‐fluorouracil (5‐FU) in patients with colorectal cancer (CRC), has equal clinical efficacy and a favorable safety profile; however, its use may be limited because of unit cost concerns. In this study, the authors measured the cost of chemotherapy‐related complications during treatment with capecitabine‐ and 5‐FU–based regimens.

METHODS:

Patients with CRC who received at least 1 administration of capecitabine or 5‐FU during 2004 and 2005 were identified from the Thomson MarketScan research databases. Monthly frequency and cost for 23 complications were recorded. Logistic regression was used to predict complication probability. General linear models were used to predict monthly complication cost and total monthly expenditure.

RESULTS:

In total, 4973 patients with CRC met the inclusion criteria for this analysis. Although the most frequently observed complications were the same between capecitabine and 5‐FU (nausea and vomiting, infection, anemia, neutropenia, diarrhea), each was observed with greater frequency in 5‐FU–based regimens. The mean predicted monthly complication cost was significantly higher (by 136%) with 5‐FU monotherapy than with capecitabine monotherapy (difference, $601; 95% confidence interval [95% CI], $469‐$737). In addition, the mean predicted monthly complication cost for 5‐FU+oxaliplatin was higher than the cost with capecitabine plus oxaliplatin (difference, $1165; 95% CI, $892‐$1595). When acquisition, administration, and complication costs were taken into consideration, there were no significant differences in the total cost between capecitabine regimens and 5‐FU regimens.

CONCLUSIONS:

Capecitabine compared well with 5‐FU–based therapy in patients with CRC and was associated with lower complication rates and associated costs. Cancer 2009. © 2009 American Cancer Society.     

Living with secondary breast cancer: coping with an uncertain future with unmet needs

JOHNSTON S.R.D. (2010) European Journal of Cancer Care 19 , 561–563 Living with secondary breast cancer: coping with an uncertain future with unmet needs     

Experience with Impedance Phlebography Compared with Venography

Venography is a particularly reliable method for the diagnosis of deep venous thrombosis but is not suitable as a screening test. Impedance phlebography represents another attempt to discover a simple, non-invasive and reliable method of detecting deep venous thrombosis. It does not, however, meet these criteria.     

再放疗并同步使用Docetaxel在鼻咽癌放疗后复发病人中的应用

目的:不能手术切除的鼻咽癌放疗后再复发的病人,其治疗困难,化疗疗效差,而单独再放疗只能挽救一小部分病人,本文探讨再放疗并同步使用多西紫彬醇(Docetaxel)在鼻咽癌首次放疗后复发病人中可行性及毒副反应,并评价其疗效。方法:对11例鼻咽癌足量放疗后经组织病理学证实复发、而无法行手术及腔内放疗的患者进行了同步放化疗。放疗采用三维适形放疗,外照射鼻咽部,分次量为1.8Gy,总剂量为36Gy-39.6Gy。化疗采用Docetaxel,15mg/m2,每周一次,静脉滴注。结果:10%、33%的患者分别出现Ⅲ度、Ⅳ度皮肤反应,18%、10%的病人分别出现Ⅲ度、Ⅳ度黏膜反应,18%患者出现Ⅲ度恶心呕吐,27%的患者出现Ⅲ度-Ⅳ度白细胞下降,10%患者出现Ⅲ度血小板下降。1例患者因严重的黏膜反应致使治疗延迟2周。治疗结束后,9例(82%)患者达到CR,2例(18%)达到PR,反应率为100%。结论:对于放疗后局部复发的鼻咽癌患者,采用同步放化疗,3D-CRT同时每周使用Docetaxel是可行的,其毒性反应在可以接受的范围内,短期疗效显著。     

Protothecosis successfully treated with amikacin combined with tetracyclines

Summary We report a case of protothecosis in an 18-year-old female student caused by Prototheca zopfii successfully treated with amikacin combined with tetracyclines. Zusammenfassung Es wird über eine Protothecose, verursacht durch Prototheca zopfii, bei einer 18-j?hrigen Studentin berichtet, die erfolgreich mit Amikacin in Kombination mit Tetracyclinen behandelt wurde.