恶性肿瘤患者外周血CIK细胞体外增殖能力及其杀伤活性

[目的]研究恶性肿瘤患者外周血自体细胞因子诱导杀伤细胞(CIK)体外增殖情况及杀伤活性,为肿瘤自体CIK细胞治疗提供实验依据.[方法]从恶性肿瘤患者的外周血中分离出CIK细胞进行体外培养,使用细胞计数板计数不同时间点CIK细胞的数目及MTT法检测其对不同肿瘤细胞株的杀伤活性.[结果]低年龄组较高年龄组CIK细胞体外增殖及杀伤活性强( P ＜0.01),不同分期的肿瘤患者CIK细胞体外增殖能力及杀伤活性均较强,不同分期之间无显著性差异( P ＞0.05);不同性别的恶性肿瘤患者CIK细胞的体外增殖能力及杀伤活性无明显差别( P ＞0.1).恶性肿瘤患者的CIK细胞在体外对不同肿瘤细胞均具有较强的杀伤活性,对人乳腺癌细胞(MCF-7),肺癌细胞株(SC-1680),前列腺癌细胞株(PC-3),宫颈癌细胞株(Hela)的体外杀伤活性均超过50%.[结论]患者年龄是影响其外周血CIK细胞体外增殖能力及杀伤活性的重要因素,而肿瘤分期、患者性别对CIK细胞的体外增殖能力及杀伤活性无明显影响;CIK细胞具有较广的抗肿瘤细胞的特点.     

细胞因子诱导杀伤细胞活性影响因素的研究进展

细胞因子诱导杀伤(CIK)细胞是一种过继免疫细胞,因其具有来源广泛,体外易于扩增,非MHC限制性,GVHD效应弱,副作用小及对多种肿瘤具有强大的杀伤活性等优点而成为近年来过继免疫治疗的焦点.但是关于如何进一步提高CIK细胞的增殖、杀伤活性,为患者带来更多益处？这一问题一直为研究者所关注.现从CIK细胞的培养体系,CIK细胞与其他细胞间的相互作用,改变CIK特异性及其他因素等方面来讨论影响CIK细胞活性的因素.     

细胞因子诱导的杀伤细胞在临床抗肿瘤治疗中的研究进展

细胞因子诱导的杀伤细胞(cytokine-Induced killer cells,CIK)是继淋巴因子激活的杀伤细胞(lymphokine activated killer cells,LAK)、肿瘤浸润性淋巴细胞(tumor infiltrating lymphocyte,TIL)和CD3Mc Ab激活的杀伤细胞之后,具有增殖速度快、杀瘤活性高、杀瘤谱广及选择性高等优势的新一代抗肿瘤过继免疫细胞。CIK细胞对多种肿瘤细胞系均表现出强大的杀伤活性,也可提高肿瘤患者的自身免疫功能,被认为是抗肿瘤过继细胞免疫治疗的首选方案。目前以CIK细胞为主的过继免疫治疗在国内外临床广泛应用,有望联合多种抗肿瘤治疗技术成为未来肿瘤综合治疗的新趋势。     

DC—CIK细胞过继免疫治疗恶性肿瘤的护理体会

目的：总结树突状细胞一细胞因子诱导的杀伤细胞（DcCIK细胞）过继细胞免疫治疗恶性肿瘤的护理效果。方法：DC—CIK过继细胞免疫治疗护理流程化。治疗护理后无1例患者出现不良反应,63例中有51例出现食欲增加,32例睡眠和体力改善,18例疼痛患者中有13例疼痛减轻,33例化疗放疗毒副反应不同程度减轻,有效地提高了患者的生活质量。结论：用DC—CIK细胞过继免疫治疗恶性肿瘤患者能有效提高细胞免疫的治疗效果。     

细胞因子诱导的杀伤细胞在感染性疾病免疫治疗中的作用

细胞因子诱导的杀伤细胞(cytokine-induced killer cells,CIK细胞)是体外采用多种细胞因子扩增的异质性细胞群,具有增殖活性高、杀瘤活性高、非MHC限制性、安全性高等特点。此外,CIK细胞治疗可明显改善肿瘤患者免疫抑制状态,提高机体的免疫功能。CIK细胞在乙型肝炎、丙型肝炎、EB病毒相关的移植后淋巴组织增生性疾病等非肿瘤性疾病中也有应用研究的相关报道。CIK细胞过继免疫治疗有助于提高机体免疫功能,可有效抑制肝炎病毒复制、降低肝炎病毒载量、减轻肝脏炎症。本文就CIK细胞在感染相关性疾病治疗中的应用作一综述。     

肺癌病人CIK细胞过继免疫治疗的护理

目的 探讨中晚期肺癌病人应用细胞因子诱导的杀伤细胞(CIK)治疗的护理规范.方法 回顾性总结2009年9月～2010年10月接受CIK细胞过继免疫治疗139例肺癌病人从护患沟通、心理护理、标本采集、CIK细胞悬液回输和输注后病情观查等环节的经验.结果 139例中晚期肺癌患者接受CIK细胞过继免疫治疗后有1例出现发热,其...     

细胞因子诱导杀伤细胞抗淋巴瘤细胞作用的研究

目的:探讨体外细胞因子诱导杀伤(CIK)细胞对淋巴瘤细胞的杀伤作用.方法:采集健康人外周血单个核细胞(PBMC),经IL-2、CD3Mc-Ab、IL-1、IFN-γ诱导,制备CIK细胞,同时以淋巴因子激活的杀伤细胞(LAK细胞)作为对照,流式细胞仪检测CIK细胞的免疫表型,LDH释放法检测其对淋巴瘤细胞株的杀伤活性.结果:CIK细胞与LAK细胞增殖在前期无明显差异,CIK细胞在培养14d后大量增殖,21d后扩增能力显著高于LAK细胞(P＜0.05);表型分析表明CIK细胞主要由CD3+和CD56+双阳性细胞构成;同一效靶比时,CIK细胞对淋巴瘤细胞株的杀伤活性明显高于LAK细胞(P＜0.05).结论:CIK细胞对淋巴瘤细胞株的杀伤作用强于LAK细胞,具有较强的抗淋巴瘤细胞活性,是肿瘤过继免疫治疗中更为有效的杀瘤效应细胞.     

卵巢癌腹水自体抗原致敏DC-CIK过继转移治疗的应用研究

目的探讨CIK细胞在不同的条件下特异性杀伤卵巢癌细胞的比较。方法自体外周血CIK细胞的体外诱导和扩增,负载卵巢癌细胞株SKOV3冻融抗原致敏外周血与腹水来源的DC诱导出的CIK细胞对SKOV3细胞杀伤率比较。结果①单一CIK细胞对SKOV3卵巢癌细胞株有明显的杀伤率有明显差异(P<0.05);②外周血DC与CIK共培养后对卵巢癌SKOV3细胞杀伤率较单一CIK细胞对SKOV3细胞杀伤率明显提高;③负载卵巢癌细胞株SKOV3冻融抗原致敏的DC诱导出的CIK细胞对SKOV3细胞杀伤率明显提高,较非致敏DC联合CIK细胞对SKOV3细胞杀伤率有明显差异(P<0.05);④负载卵巢癌细胞株SKOV3冻融抗原致敏的腹水来源的DC联合CIK细胞对SKOV3细胞杀伤率,与外周血来源的DC联合CIK细胞对SKOV3细胞杀伤率无明显差异。结论卵巢癌腹水来源的DC可以考虑作为过继免疫治疗效应细胞的一个重要来源。     

细胞因子诱导的杀伤细胞体外研究及临床应用进展

随着恶性肿瘤发病率逐渐升高,对人类生命健康的威胁日趋严重,传统的放化疗及手术治疗方式虽在治疗肿瘤方面取得了一定的进步,但对于年龄较大及失去治疗机会的中晚期肿瘤患者却束手无策,因此各种肿瘤生物治疗的新方法应运而生,以细胞因子诱导的杀伤细胞(CIK)为代表的细胞过继免疫治疗即是其中一种。CIK细胞由Schmidt-Wolf等[1]于1991年     

CIK过继免疫治疗恶性肿瘤病人的护理

细胞因子诱导的杀伤细胞(cytokine induced killers，CIK)是将人外周血单个核细胞在体外用多种细胞因子共同培养一段时间后获得的一群异质细胞。由于该细胞同时表达CD3和CD56两种膜蛋白分子，故又称为天然杀伤细胞(NK细胞)样T淋巴细胞，兼具有T淋巴细胞强大的抗瘤活性和NK细胞的非主要组织相容性复合物(MHC)限制性杀瘤优点。我院从2001年11月-2004年11月应用CIK过继免疫治疗25例，取得了良好的疗效。现将治疗中的有关护理介绍如下。     

Cytokine induced killer cells as adoptive immunotherapy strategy to augment graft versus tumor after hematopoietic cell transplantation

Donor lymphocyte infusion (DLI) is used to increase the graft versus tumor (GVT) effect after allogeneic hematopoietic cell transplant (HCT). The limited spectrum of activity and high risk of graft versus host disease (GVHD) remain major limitations of this approach. The finding of new cell populations for adoptive immunotherapy, with the ability to separate GVT from GVHD, would be useful. Here we review the main basic, preclinical and clinical research on cytokine-induced killer (CIK) cells, highlighting the aspects of their antitumor and alloreactive potentials that might favourably affect the balance between GVT and GVHD. CIK cells are ex vivo-expanded T lymphocytes sharing NK markers and endowed with a potent MHC-unrestricted antitumor activity against haematological and solid malignancies. Studies in preclinical animal models have demonstrated their low GVHD potential when infused across MHC-barriers, and recent clinical studies seem to confirm these findings in patients with hematological malignances relapsing after HCT. If consolidated with larger clinical trials, adoptive immunotherapy with CIK cells might represent an effective alternative to classic DLI, helping HCT to succesfully meet current challenges like the extension across major HLA-barriers and application to solid tumors.     

自体CIK细胞联合白介素-2治疗老年恶性血液病的疗效与护理

目的探讨CIK细胞联合白介素-2治疗老年恶性血液病患者的疗效及护理经验。方法回顾性总结16例老年恶性血液病患者实施CIK细胞联合白介素-2治疗前后的护理方法及疗效。结果12例达完全缓解,2例部分缓解;2例患者治疗期间曾出现一过性乏力,发热。治疗后一般情况改善主要表现为乏力减轻、精神状态改善、食欲增强、关节疼痛减轻。结论CIK细胞联合白介素-2治疗老年恶性血液病效果好,安全性高。治疗期间做好高质量的护理是保证治疗效果的关键。     

树突状细胞疫苗联合细胞因子诱导的杀伤细胞治疗鼻腔及鼻窦恶性黑色素瘤的近期疗效观察

目的探讨负载自体肿瘤抗原的树突状细胞(DC)疫苗联合细胞因子诱导的杀伤细胞(C IK)治疗鼻腔、鼻窦恶性黑色素瘤的疗效。方法对15例鼻腔鼻窦恶性黑色素瘤患者在常规手术后2周,通过体外培养扩增DC及C IK细胞,并静脉回输,进行免疫治疗,观察患者临床症状改善及生活质量改善状况及近期疗效,利用流式细胞仪检测回输前后患者T细胞亚群及NK细胞变化。结果15例接受自体DC及C IK细胞治疗患者中,治疗前后患者临床症状有明显改善,提高率达86%,自体回输免疫治疗后,CD3+、CD4+T细胞和NK细胞(CD16+、CD56+)及CD4+/CD8+比例均显著提高(P0.01)。结论负载自体肿瘤抗原的DC疫苗联合C IK治疗鼻腔、鼻窦恶性黑色素瘤可以提高患者免疫功能,改善临床症状,提高生存质量。     

Cytokine-induced killer (CIK) cells as feasible and effective adoptive immunotherapy for the treatment of solid tumors

INTRODUCTION: Cytokine-induced killer (CIK) cells are heterogeneous ex vivo-expanded T lymphocytes with mixed T-NK phenotype and endowed with a wide MHC-unrestricted antitumor activity. CIK cells can be expanded from peripheral blood mononuclear cells (PBMC) cultured with the timed addition of IFN-γ, Ab anti-CD3 and IL2. A consistent subset of mature CIK cells presents a CD3(+)CD56(+) phenotype. The CD3(+)CD56(+) cellular subset is the main responsible for the tumor-killing activity, mostly mediated by the interaction of NKG2D receptor with MHC-unrestricted ligands (MIC A/B; ULBPs) on tumor cells. AREAS COVERED: In the present work, we described the biologic characteristics of CIK cells, focusing on those aspects that may favor their clinical translation. We reviewed preclinical data and analyzed reports from clinical trials. A specific paragraph is dedicated to future research perspectives in the field. EXPERT OPINION: CIK cells represent a realistic new option in the field of cancer immunotherapy. Crucial issues, favoring their clinical translation, are the easy availability of large amounts of expanded CIK cells and their MHC-unrestricted tumor killing, potentially effective against many tumor types. Intriguing future perspectives and open challenges are the investigation of synergisms with other immunotherapy approaches, targeted therapies or even conventional chemotherapy.     

免疫功能检测对评估细胞因子诱导的自体杀伤细胞治疗老年血液肿瘤疗效的研究

本研究探讨细胞因子诱导的自体杀伤(cytokine induced killer,CIK)细胞输注对造血系统恶性肿瘤老年患者免疫功能的影响及其与疗效的关系。采集20例造血系统恶性肿瘤老年患者外周血单个核细胞,在体外经重组干扰素-γ(IFN-γ)、白介素-2(IL-2)、抗CD3单克隆抗体诱导成CIK细胞,回输细胞数为(2-3)×109个,回输后第1-10天应用IL-2 100万单位/天,皮下注射。28天为1个疗程,共完成了136个周期的自体CIK细胞输注。观察治疗前后细胞免疫功能变化,结合肿瘤相关生物学指标、缓解情况、生活质量及生存期的变化进行疗效分析。结果表明,14例接受8个疗程的CIK细胞输注,6例接受4个疗程的输注,回输后所有患者未出现不良反应。CIK细胞治疗后外周血CD3+、CD3+CD8+、CD3+CD56+细胞比例明显升高(p0.05),β2微球蛋白、LDH水平显著下降(p0.05)。所有患者症状改善、生活质量显著提高(p0.01)。11例达完全缓解,7例达部分缓解,2例疾病稳定。结论:自体CIK细胞输注可提高造血系统恶性肿瘤老年患者的细胞免疫功能且安全有效。     

Evaluation of an automated closed fluid management device for processing expanded cytokine-induced killer cells to use in immunotherapy programs for cancer

BACKGROUND: Cytokine-induced killer (CIK) cells are a heterogeneous population of immune cells derived from peripheral blood lymphocytes with a high proliferative potential ex vivo. This study shows a rapid and reproducible protocol for adoptive immunotherapy with CIK cells in patients with hematologic malignancies. For this purpose a new automatic cell processing device (CytoMate, Baxter Oncology) was tested to improve extensive manipulations of these cells. STUDY DESIGN AND METHODS: Twenty CIK expansions obtained from healthy donors and patients with hematologic malignancies were washed and refilled with fresh medium during culture with the CytoMate. Recovery, viability, and cytotoxic activity were evaluated. Six cryopreserved CIK procedures were thawed and processed for washing out dimethyl sulfoxide automatically. Recovery of cells, viability, and early apoptosis were measured immediately after washing, and cytotoxic activity against target cell lines K562 and Daudi was tested after short culture. RESULTS: Prewash volume of CIK cultures was 3600 mL (range, 1970-6000 mL). After automatic wash, the total CIK cell recovery was 85.3 percent (range, 78.5%-97.5%), and living cells were greater than 95 percent. After thawing, the median recoveries of total nucleated cells and natural killer T (NKT) cells were, respectively, 80.7 percent (range, 65%-95.5%) and 90.5 percent (range, 70.5%-98.5%). Thawed cells preserved their cytotoxic activity after cryopreservation (approx. 50% lysis at effector:target ratio of 40:1). CONCLUSION: The automatic wash with the CytoMate showed a good recovery of viable CIK cells during expansion and allowed an efficient manipulation of thawed cells. The use of this simple and efficient washing technique is suitable for clinical-grade processing in cellular therapy protocols.     

DC-CIK as a widely applicable cancer immunotherapy

ABSTRACT

Introduction: Immunotherapy is now a standard treatment for many malignancies. Although immune checkpoint inhibition has demonstrated substantial efficacy by enhancing T cell activation and function in the tumor microenvironment, adoptive transfer of T and NK cell products promises to provide activated cells capable of immediate and direct tumor destruction. A widely applicable, non-MHC dependent, cellular therapy, consisting of in vitro generated dendritic cells (DC) combined with cytokine-induced killer cells (CIK), is highly efficient to produce from individual patients and has demonstrated safety and efficacy alone or with chemotherapy.

Areas covered: We summarize the clinical data from studies of DC-CIK and discuss future research directions.

Expert opinion: Patients with a wide variety of tumor types who have received DC-CIK therapy may experience clinical responses. This versatile therapy synergizes with other anti-cancer therapies including chemotherapy and immunotherapy.     

In Vivo Proof of Concept of Adoptive Immunotherapy for Hepatocellular Carcinoma Using Allogeneic Suicide Gene-modified Killer Cells

Cell therapy based on alloreactivity has completed clinical proof of concept against hematological malignancies. However, the efficacy of alloreactivity as a therapeutic approach to treat solid tumors is unknown. Using cell culture and animal models, we aimed to investigate the efficacy and safety of allogeneic suicide gene-modified killer cells as a cell-based therapy for hepatocellular carcinoma (HCC), for which treatment options are limited. Allogeneic killer cells from healthy donors were isolated, expanded, and phenotypically characterized. Antitumor cytotoxic activity and safety were studied using a panel of human or murine HCC cell lines engrafted in immunodeficient or immunocompetent mouse models. Human allogeneic suicide gene-modified killer cells (aSGMKCs) exhibit a high, rapid, interleukin-2–dependent, and non–major histocompatibility complex class I-restricted in vitro cytotoxicity toward human hepatoma cells, mainly mediated by natural killer (NK) and NK-like T cells. In vivo evaluation of this cell therapy product demonstrates a marked, rapid, and sustained regression of HCC. Preferential liver homing of effector cells contributed to its marked efficacy. Calcineurin inhibitors allowed preventing rejection of allogeneic lymphocytes by the host immune system without impairing their antitumor activity. Our results demonstrate proof of concept for aSGMKCs as immunotherapy for HCC and open perspectives for the clinical development of this approach.     

Cytokine-induced killer (CIK) cells as feasible and effective adoptive immunotherapy for the treatment of solid tumors

Introduction: Cytokine-induced killer (CIK) cells are heterogeneous ex vivo-expanded T lymphocytes with mixed T-NK phenotype and endowed with a wide MHC-unrestricted antitumor activity. CIK cells can be expanded from peripheral blood mononuclear cells (PBMC) cultured with the timed addition of IFN-γ, Ab anti-CD3 and IL2. A consistent subset of mature CIK cells presents a CD3⁺CD56⁺ phenotype. The CD3⁺CD56⁺ cellular subset is the main responsible for the tumor-killing activity, mostly mediated by the interaction of NKG2D receptor with MHC-unrestricted ligands (MIC A/B; ULBPs) on tumor cells.

Areas covered: In the present work, we described the biologic characteristics of CIK cells, focusing on those aspects that may favor their clinical translation. We reviewed preclinical data and analyzed reports from clinical trials. A specific paragraph is dedicated to future research perspectives in the field.

Expert opinion: CIK cells represent a realistic new option in the field of cancer immunotherapy. Crucial issues, favoring their clinical translation, are the easy availability of large amounts of expanded CIK cells and their MHC-unrestricted tumor killing, potentially effective against many tumor types. Intriguing future perspectives and open challenges are the investigation of synergisms with other immunotherapy approaches, targeted therapies or even conventional chemotherapy.     

New adoptive immunotherapy strategies for solid tumours with CIK cells

INTRODUCTION: Despite development and introduction of new and innovative drugs, a large number of malignant diseases are associated with unfavourable prognosis. In recent years, considerable progress in cancer treatment has been obtained by the application of cytokine-induced killer (CIK) cells. AREAS COVERED: This review provides an overview and summary of recent advances in adoptive immunotherapy strategies in cancer treatment using CIK cells. A selective literature search has been performed. EXPERT OPINION: The application of CIK cells as adoptive immunotherapy plays an important role in cancer treatment. Combining CIK cells with other conventional and established therapy options represents an innovative approach and will hopefully provide new insight for the future.