IL-24对CIK细胞体内杀伤肿瘤细胞活性的影响

目的 研究IL-24对细胞因子诱导的杀伤细胞(CIK)体内杀伤肿瘤细胞活性的影响。方法 (1)从健康人外周血中提取单个核细胞,第一天加入IFN-γ(1 000 IU/mL),第二天加入IL-1(100 IU/mL)、CD3mAb(50 ng/mL)、IL-2(300 IU/mL)诱导CIK细胞,另一组与IL-1、CD3mAb、IL-2同时加入IL-24(1 000 IU/mL);(2)建立HepG₂肿瘤细胞株荷瘤小鼠皮下移植瘤模型,分为五组,每三天分别测量肿瘤大小。结果 IL-24诱导CIK细胞治疗组荷瘤小鼠实体瘤体积小于其它组(P＜0.05)。结论 在CIK细胞诱导过程中加入IL-24能明显增强其体内的抗肿瘤活性,对临床上把CIK细胞用于生物治疗有一定的指导意义。     

当归多糖对CIK细胞生物学特性及其杀伤活性的影响

目的:探讨当归多糖(Angelica sinensis polysaccharide,ASP)体外对人外周血来源的细胞因子诱导的杀伤(cytokine induced killer,CIK)细胞的增殖、免疫表型及其对白血病K562细胞杀伤活性的影响。方法:分离健康志愿者外周血单个核细胞(peripheral blood mononuclear cell,PBMC),第0天加入IFN-γ(1 000 IU/ml),24 h后再加入IL-2(1 000 IU/ml)、抗CD3单抗(50 ng/ml)继续培养,此后每隔3 d根据添加不同浓度的ASP分为5组:A组(不加ASP)、B~E组(分别加ASP 12.5、25、50、100μg/ml)。全自动血细胞计数仪计数各组CIK细胞的增值情况,流式细胞术检测各组CIK细胞中CD3+CD4+、CD3+CD8+、CD3+CD56+细胞的百分比,酶联免疫吸附(ELISA)法检测各组CIK细胞上清液中IFN-γ及TNF-α的分泌水平,CCK-8法检测各组CIK细胞对K562细胞的杀伤活性。结果:联合ASP诱导培养CIK细胞至第16天时,与A组相比,E组CIK细胞的增殖能力明显增加[(56.98±3.00)×106vs(43.81±2.14)×106,P<0.05)],各组细胞活率都保持在90%以上;各组CIK细胞CD3+CD4+、CD3+CD8+细胞比例无明显差异,而D组、E组CD3+CD56+细胞比例与A组相比存在差异[(26.65±3.71)%、(28.36±4.28)%vs(20.75±3.56)%,P<0.05)];E组CIK细胞在效靶比为40∶1、20∶1、10∶1时对K562细胞的杀伤活性比A组更强[(84.19±5.88)%vs(68.05±5.95)%、(76.69±6.54)%vs(64.55±9.44)%、(72.32±9.22)%vs(61.45±8.22)%,均P<0.05〗。结论:随着培养时间的延长,高浓度的ASP体外对CIK细胞的增殖及杀伤活性有一定的促进作用。     

DC-CIK对急性髓细胞白血病干细胞的杀伤作用

目的:研究树突状细胞(dendritic cell,DC)联合同源细胞因子诱导的杀伤细胞(cytokine-induced killer cell,CIK)对急性髓细胞白血病细胞株KG-1a中白血病干细胞(leukemic stem cell,LSC)的体外杀伤和诱导凋亡作用。方法:分离健康人外周血单个核细胞,贴壁细胞用GM-CSF和IL-4诱导培养DC,悬浮细胞用IL-2、IL-1、IFN-γ和CD3 mAb诱导培养CIK。将KG-1a细胞冻融物作为抗原负载DC(即Ag-DC),与CIK共培养作为实验组(Ag-DC-CIK),无抗原负载的DC与CIK共培养作为对照组(DC-CIK),单独CIK作为空白对照组,与KG-1a共育后流式细胞术检测各组细胞中CD34+CD38-CD123+白血病干细胞的比例。DC-CIK与KG-1a细胞共培养,流式细胞术检测各组细胞中KG-1a细胞与CD34+CD38-CD123+细胞的凋亡率。结果:外周血单个核细胞成功诱导DC。CIK组、DC-CIK组及Ag-DC-CIK组中CD3+CD56+细胞比例为(17.36±4.44)%、(28.22±3.66)%和(36.16±5.88)%,依次升高(P<0.05)。与对照组相比,Ag-DC-CIK组与DC-CIK组细胞中CD34+CD38-CD123+细胞比例显著降低[(8.78±0.62)%vs(3.95±0.53)%、(3.03±0.62)%,P<0.01〗。DC-CIK可诱导KG-1a细胞凋亡,凋亡率由(2.34±0.74)%上升至(12.27±1.01)%,但对其中CD34+CD38-CD123+细胞无明显的诱导凋亡作用。结论:DC联合CIK能杀伤急性髓细胞白血病干细胞,但无明显的诱导凋亡作用。     

IL-21 增强CIK细胞对食管癌EC9706 细胞的杀伤作用及其可能的机制

目的：探讨细胞因子IL-21 对CIK 细胞体外杀伤食管癌EC9706 细胞活性的影响,并初步分析其可能的分子机制。方法：体外无菌分离人外周血单个核细胞,分别进行常规培养和加IL-21 培养CIK细胞。流式细胞术检测2 种培养CIK细胞的免疫表型,LDH释放法检测2 种培养CIK 细胞杀伤食管癌EC9706 的活性,ELISA 法检测2 种CIK 细胞培养上清液IFN-γ 浓度。结果：常规培养和加IL-21 培养的2 种CIK细胞增殖数量无明显差异。加IL-21 培养的CIK细胞CD3+CD56+细胞比例、CD3+细胞内颗粒酶B 和穿孔素表达率均显著高于常规培养CIK 细胞[(26.95±3.53)% vs (16.18±1.04)%,(33.29±2.30)% vs (23.58±2.28)%和(59.70±1.91)% vs (45.96±2.67)%,均P<0.05）。效靶比20∶1 和30∶1 时,加IL-21 培养的CIK细胞杀伤EC9706 细胞的活性均显著高于常规培养的CIK 细胞[20∶1 时:(43.66±1.99)% vs (34.59±1.75)%;30∶1 时：57.52±2.15)% vs (42.23±1.87)%,均P<0.05]。加IL-21培养CIK细胞的上清液FN-γ 的浓度明显高于常规培养CIK细胞的上清液[(142.7±13.4) vs (42.6±3.3) ng/L,P<0.05]。结论：IL-21增加CD3+CD56+细胞比例和颗粒酶B及穿孔素的表达,提高CIK细胞分泌IFN-γ,从而增强CIK细胞对EC9706 细胞的杀伤活性。     

抗原致敏DC联合CIK在原发性肝癌治疗中的应用

目的:探讨负载自身肝癌裂解物的树突状细胞(DC)与细胞因子诱导杀伤细胞(CIK)共培养对CIK体外杀伤活性的影响,并观察抗原致敏DC(Ag-DC)联合CIK治疗原发性肝癌后患者的免疫状态、临床疗效及毒副反应.方法:选择24例原发性肝癌患者,分离外周血单个核细胞,其中贴壁细胞经GM-CSF和IL-4诱导产生DC,并负载自体肿瘤裂解物;悬浮细胞经IFN-γ、IL-2、抗CD3单抗、IL-1α体外诱导产生CIK.将Ag-Dc与CIK共培养,观察CIK在体外对肝癌细胞株SMMC-7721的杀伤活性;24例患者接受Ag-DC+CIK的过继免疫治疗,观察疗效.结果:1)Ag-DC与CIK共培养后,CIK体外肿瘤杀伤活性明显提高;2)Ag-Dc联合C1K治疗原发性肝癌可明显改善患者细胞免疫功能,提高临床疗效;3)除一过性发热、畏寒外未见其它不良反应.结论:负载自体肿瘤细胞裂解物的DC疫苗联合CIK可作为原发性肝癌常规治疗的有效辅助手段.     

来那度胺增强细胞因子诱导的杀伤细胞杀伤效应的研究

 【摘要】　目的 　探讨来那度胺调节细胞因子诱导的杀伤（CIK）细胞对淋巴瘤细胞的杀伤效应。方法　取健康志愿者外周血单个核细胞,经过干扰素-γ（INF-γ）、白细胞介素-2（IL-2）、抗CD3单抗诱导CIK细胞增殖,并对培养不同时间CIK细胞用不同浓度来那度胺进行处理。流式细胞术检测CD＋3CD＋56细胞比例;酶联免疫吸附（ELISA）法检测粒-巨噬细胞集落刺激因子（GM-CSF）、INF-γ及肿瘤坏死因子α（TNF-α）的分泌水平;用Cell Counting Kit-8（CCK-8）试剂盒检测来那度胺作用后的CIK细胞及未处理的CIK细胞对3种淋巴瘤细胞株的体外杀伤效应。 结果　CIK细胞在多种细胞因子诱导下,CD＋3CD＋56细胞比例明显增加,0.2～5.0 μmol／L的来那度胺对其扩增分化没有影响（P＞0.05）。来那度胺作用下CIK细胞分泌GM-CSF、INF-γ、TNF-α水平有显著提高（P＜0.05）。在1.0 μmol／L来那度胺作用下CIK细胞对淋巴瘤细胞的杀伤效力明显高于未经来那度胺处理的CIK细胞[（42.53±2.19）%比（15.5±3.82）%;（44.78±4.86）% 比（29.94±6.33）%;（54.71±5.31）%比( 37.43±9.75）%]。结论　在来那度胺作用下,CIK细胞分泌功能和对3种淋巴瘤细胞的杀伤效应均有显著增强。     

重组人纤维连接蛋白诱导的CIK细胞的生物学特性和对肺癌细胞株杀伤活性的体外研究

背景与目的CIK细胞是过继免疫治疗的重要手段之一,简化体外培养过程从而提高其增殖率和杀瘤活性仍是目前研究的一个热点课题。本研究观察重组人纤维连接蛋白(recombinant human fibronectin,RN)诱导CIK细胞的生物学特性,建立一种高效、简便的体外CIK细胞扩增方法。方法抽取10名健康人外周静脉血各50mL,用淋巴细胞分离液分离单个核细胞,分别采用RN诱导法和传统方法培养CIK细胞,记录细胞增殖数;用流式细胞术测定免疫细胞表型和分泌IFN-γ、IL-4、穿孔素和颗粒酶B细胞的百分比;用MTT法测定CIK细胞对4种人肺癌细胞株的体外杀伤率。结果RN诱导的CIK细胞扩增倍数为传统方法的2.0倍-3.5倍,具有统计学差异(P<0.05);RN诱导组和传统方法组CD3+CD16+CD56+细胞绝对数分别增加了3778倍和2069倍;RN诱导组细胞中CD3+CD8+细胞比例明显高于传统方法组(P<0.05);但CD3+CD4+细胞比例无统计学差异(P>0.05);对4种肺癌细胞株的体外杀伤活性无统计学差异(P>0.05)。RN诱导的CIK较诱导前:分泌IFN-γ的细胞比例明显增加;分泌IL-4的细胞...     

DC与CIK共培养后对白血病细胞杀伤活性的研究

目的研究细胞因子诱导的杀伤细胞(CIK)与同源树突状细胞(DC)共培养后CIK细胞的增殖活性、表型的变化,及其对K562、HL-60白血病细胞细胞毒作用的影响。方法采集健康供者的外周血单个核细胞(MNC),置于37℃,5%CO2培养箱培养2 h,收集非贴壁细胞用于诱导培养CIK细胞,贴壁细胞诱导分化出成熟DC,将成熟DC和CIK细胞按1 5的比例混合培养3天,用MTT法检测DC-CIK共培养细胞杀伤K562和HL-60白血病细胞株的活性。结果DC-CIK共培养后增殖速度明显快于单纯CIK细胞组(P<0.05);培养第14天,CIK中CD3+CD8+、CD3+CD56+双阳性细胞的比率分别为58.6%±7.3和26.5%±6.2,DC-CIK的CD3+CD8+、CD3+CD56+的比率分别为72.5%±4.2和38.4%±6.1,表达差异显著(P<0.05);在2.5∶1~20∶1的效靶比范围内,DC-CIK对K562、HL-60细胞的杀伤活性较单纯CIK细胞组的杀伤活性要高(P<0.05)。结论DC与CIK共培养细胞是增殖活性和细胞毒活性均高于CIK细胞的免疫活性细胞。     

CD 40激发肿瘤抗原特异性DCs对CIK细胞生物学活性的影响

目的:研究CD40激发凋亡肿瘤细胞致敏的树突状细胞(dendritic cells,DCs)对细胞因子诱导杀伤(cytokine-inducedkiller,CIK)细胞的细胞表型、增殖活性及细胞毒活性的影响。方法:常规方法从健康人外周血单个核细胞中诱导DCs和CIK细胞,采用凋亡肿瘤细胞负载DCs,并用或不用激发型CD40单克隆抗体(CD40mAb)激活DCs成熟;成熟DCs与同源的CIK细胞共育5d,分别获得DC40Ag-CIK细胞及DCAg-CIK细胞;观察细胞增殖活性;流式细胞仪检测细胞表型;酶联免疫吸附实验(enzyme-linked immunosorbent assays,ELISA)法检测细胞培养上清液中IFN-γ的含量;[3H]-TdR掺入法测定细胞杀伤活性。结果:凋亡肿瘤细胞负载激发可使DCs上调表达CD1a、CD80、CD86、CD83、HLR-DR,联合CD40mAb激发可进一步促进DCs成熟;培养至第14天时,DC40Ag-CIK细胞、DCAg-CIK细胞、CIK细胞分别扩增(18.2±1.7)倍、(15.0±1.2)倍、(9.3±1.8)倍;DC40Ag-CIK细胞中的CD3 CD56 比例较DCAg-CIK细胞和CIK细胞明显上调(P<0.05);DC40Ag-CIK细胞、DCAg-CIK细胞对A549杀伤活性强于CIK细胞(P<0.05),并且DC40Ag-CIK细胞强于DCAg-CIK细胞(P<0.05);CIK细胞、DCAg-CIK细胞、DC40Ag-CIK细胞培养上清液中IFN-γ的含量递增,分别为(1494.7±246.3)pg/mL、(2706.3±197.0)pg/mL、(3676.3±335.0)pg/mL。结论:与单独凋亡肿瘤细胞负载的DCs相比,CD40激发的凋亡肿瘤细胞负载的DCs可进一步提高CIK细胞的增殖活性及细胞毒活性。     

长期冷冻保存的外周血干细胞采集物形成CIK的能力及其生物学活性的研究

目的:观察是否能从长期深低温冷冻保存的外周血干细胞采集物(PBSC)大量诱导出具有细胞毒活性的CIK细胞。方法:10例液氮中保存3~6年的PBSC标本作为实验组,7例新鲜的PBSC标本作为对照组。PBSC标本去除红细胞后用IFN-γ、CD3单抗、IL-2联合诱导CIK细胞。14天后收获细胞,检测活力、扩增倍数。流式细胞学检测CIK细胞免疫表型,MTT法检测其细胞毒活性。结果:实验组和对照组培养后CIK细胞分别扩增了41·8倍和22·8倍,活化后CD8 T细胞比例升高(P<0·05),但两组的NKT细胞(CD3 CD16/56 T细胞)纯度活化后并没有升高。两组CIK细胞对K562细胞株的杀伤作用无显著性差异。结论:从长期冷冻保存的外周血干细胞采集物中可以诱导出具有细胞毒活性的CIK细胞。     

New paradigms for therapy for osteosarcoma

Osteosarcoma is a primary bone malignancy generally affecting the young, with 60% of cases occurring before the age of 25 years and the peak incidence at 15 years. Survival has improved over the past several decades, with nonmetastatic disease having an approximately 70% chance of long-term survival. Unfortunately, patients with metastatic disease at diagnosis or those who have recurrent disease have a dismal prognosis, with approximately 20% surviving long term. In this review article we describe several new therapies in development for osteosarcoma. These include immune-based therapies, strategies to inhibit tumor growth, radiotherapy, and the introduction of new chemotherapies and targets.     

A framework for rehabilitation for cancer survivors

This paper introduces a theoretical framework that recognises the rehabilitation needs of people who have cancer and offers a multi‐tiered model to meet these needs. Various models for providing survivorship care have been previously proposed, giving rise to multiple possible delivery systems. Existing cancer rehabilitation frameworks recognise different phases of illness, goals of care and the need for services at all stages of illness. The ‘Stained Glass Cancer Rehabilitation Framework’ incorporates survivor needs and rehabilitation modalities, arranged in a practical hierarchy and builds on earlier models. A broad view of rehabilitation services considers complexity, temporal and geographic factors. Recognition that needs emerge over time demands a routine long‐term approach to screening for physical, functional and psychosocial rehabilitation needs by medical and other health professionals. New methods of care delivery and coordination from specialist to primary care settings are needed, long after treatment is completed. Service delivery infrastructure supported by funding reform and training of rehabilitation professionals in delivering appropriate interventions for cancer survivors is essential, together with more research into cancer rehabilitation interventions, functional outcomes and their delivery.     

Follow-up for women after treatment for cervical cancer

Question

What is the most appropriate follow-up strategy for patients with cervical cancer who are clinically disease-free after receiving primary treatment?

Perspectives

For women with cervical cancer who have been treated with curative intent, follow-up includes identification of complications related to treatment and intervention in the event of recurrent disease. Most women who recur with cervical cancer are not curable; however, early identification of recurrence can alter disease management or treatment-planning options, and for those with a central pelvic recurrence and no evidence of distant disease, there is a potential for cure with additional therapy. Follow-up protocols in this population are variable, using a number of tests at a variety of intervals with questionable outcomes.

Outcomes

Outcomes of interest included recurrence, survival, and quality of life.

Methodology

The Gynecology Cancer Disease Site Group (dsg) conducted a systematic review of the literature and a narrative review of emerging clinical issues to inform the most appropriate follow-up strategy for patients with cervical cancer. The evidence was insufficient to specify a clinically useful recommended follow-up schedule, and therefore, the expert consensus opinion of the Gynecology Cancer dsg was used to develop recommendations on patient surveillance. The resulting recommendations were reviewed and approved by the Gynecology Cancer dsg and by the Program in Evidence-Based Care Report Approval Panel. An external review by Ontario practitioners completed the final phase of the review process. Feedback from all parties was incorporated to create the final practice guideline.

Results

The systematic review of the literature identified seventeen retrospective studies. The Gynecology Cancer dsg used a consensus process to develop recommendations based on the available evidence from the systematic review, the narrative review, and the collective clinical experience and judgment of the dsg members.

Practice Guideline

The recommendations in this practice guideline are based on the expert consensus opinion of the Gynecology Cancer dsg, informed by evidence from retrospective studies. These are some general features of an appropriate follow-up strategy:

1. At a minimum, follow-up visits with a complete physical examination, including a pelvic–rectal exam and a patient history, should be conducted by a physician experienced in the surveillance of cancer patients.
2. There is little evidence to suggest that vaginal vault cytology adds significantly to the clinical exam in detecting early disease recurrence.
3. Routine use of various other radiologic or biologic follow-up investigations in asymptomatic patients is not advocated, because the role of those investigations has yet to be evaluated in a definitive manner.
4. A reasonable follow-up schedule involves follow-up visits every 3–4 months in the first 2 years and every 6–12 months in years 3–5. Patients should return to annual population-based general physical and pelvic examinations after 5 years of recurrence-free follow-up.

     

Technique for external beam treatment for mesothelioma

A combined photon-electron beam treatment for diffuse pleural mesothelioma is discussed in this paper. The technique consists of parallel opposed 10 MV X rays prescribed to 4250 cGy using customized blocks to shield the lung. The pleura is then boosted with electrons to a dose of 3600 cGy. The combination yields a TDF of 74 ret to the pleura. As discussed in an earlier paper, this treatment method when combined with subtotal pleurectomy and I-125 implantation leads to improved survivals with minimal complications. The details of this 3-dimensional radiation treatment method were not described in detail. To improve target coverage and local control, the technique has been modified. CT is now used along with simulation plane films to define the entire pleural surface. The target volume has also been extended from the dome to the base of this diaphragm. These changes have led to improved pleural dose distributions; by blocking the liver or stomach, and boosting the crus of the diaphragm with electrons, there is little added morbidity. As is demonstrated by dose volume histograms, we have been able to deliver 4250 cGy +/- 10% to most of the pleura with 1/3 of the lung parenchyma receiving less than 2100 cGy.     

Industry gains incentives for drugs for children

Under the FDA Safety and Innovation Act, manufacturers who get a drug for a rare pediatric disease approved and on the market earn a voucher requiring the FDA to review a second drug within 6 months of submission of an application for its approval.