DC-CIK治疗大肠癌的研究进展

大肠癌是消化道常见的恶性肿瘤之一,发病率和死亡率均较高。过继免疫治疗是当今肿瘤治疗的热点,已逐步成为一些肿瘤的首选治疗方法。树突状细胞(DC)是目前已知功能最强大的抗原呈递细胞,具有呈递肿瘤抗原和抵制肿瘤细胞免疫逃逸及刺激T淋巴细胞产生免疫应答的作用。细胞因子诱导的杀伤细胞(CIK)由多种细胞因子诱导而成,具有T淋巴细胞及NK细胞抗肿瘤作用的特点。DC和CIK细胞有效结合可以同时促进DC细胞的增殖和免疫功能及加强CIK细胞的抗肿瘤作用。本文就近年来国内外应用DC-CIK治疗大肠癌的研究进展进行综述。     

脐血CIK细胞抗肿瘤研究及临床应用进展

肿瘤的生物治疗尤其是用免疫活性细胞输注的过继免疫治疗是目前研究热点之一,是继手术、放疗、化疗三大常规治疗的又一新的治疗肿瘤的方法。此疗法不仅是常规抗肿瘤治疗的补充,更是为晚期不宜手术或无法承受放疗化疗所带来的副作用的患者开辟了一个新的治疗途径,可取得常规方法无法达到的疗效,成为人类抗击肿瘤最有希望的战略措施之一。细胞因子诱导的杀伤细胞是用于肿瘤过继免疫治疗较为有效的免疫效应细胞之一,是目前所知杀伤活性最高的肿瘤杀伤细胞,具有增殖速度快、杀瘤活性高、杀瘤谱广的特点,对正常骨髓造血前体毒性小,已大量应用于临床。脐血CIK细胞增殖速度、杀瘤活性优于外周血CIK细胞,移植后移植物抗宿主病发生率更低等优点,因而受到广泛关注。本文就脐血CIK细胞的抗肿瘤研究的特点及临床应用进展作一综述。     

脐血CIK细胞抗肿瘤研究及临床应用进展

肿瘤的生物治疗尤其是用免疫活性细胞输注的过继免疫治疗是目前研究热点之一,是继手术、放疗、化疗三大常规治疗的又一新的治疗肿瘤的方法。此疗法不仅是常规抗肿瘤治疗的补充,更是为晚期不宜手术或无法承受放疗化疗所带来的副作用的患者开辟了一个新的治疗途径,可取得常规方法无法达到的疗效,成为人类抗击肿瘤最有希望的战略措施之一。细胞因子诱导的杀伤细胞是用于肿瘤过继免疫治疗较为有效的免疫效应细胞之一,是目前所知杀伤活性最高的肿瘤杀伤细胞,具有增殖速度快、杀瘤活性高、杀瘤谱广的特点,对正常骨髓造血前体毒性小,已大量应用于临床。脐血CIK细胞增殖速度、杀瘤活性优于外周血CIK细胞,移植后移植物抗宿主病发生率更低等优点,因而受到广泛关注。本文就脐血CIK细胞的抗肿瘤研究的特点及,临床应用进展作一综述。     

自体DC、CIK细胞在急性髓细胞白血病治疗中的研究进展

树突状细胞(DC)是初级免疫应答的激发者,是最有活力的抗原递呈细胞(APC),可以有效地抑制白血病细胞逃逸。未成熟DC细胞从细胞外捕获各种抗原信息,成熟DC细胞传递各种抗原信息给宿主淋巴结的T细胞,激活抗原相关的主要组织相容性复合体(MHC)限制性特异性免疫应答,另外,亦可通过影响B细胞的增殖,不同程度的活化体液免疫应答。细胞因子诱导的杀伤细胞(CIK)是一组具有细胞毒作用的异质细胞群,是较LAK细胞溶瘤活性更强的一种免疫活性细胞,对包括白血病在内的多种恶性肿瘤具有抗肿瘤效应,具有非MHC限制性的杀瘤特点。二者联合培养及应用又增强了各自的活性。DC细胞与CIK细胞对于白血病的疗效不仅在实验室得以证实,而且已经逐步应用于临床,其在清除微小残留病以及预防造血干细胞移植后复发中取得了良好的效果。随着细胞制备技术的完善和研究的进一步深入,自体DC、CIK细胞治疗急性髓细胞白血病逐渐获得众多专家的认可。中国卫生部已经把自体免疫细胞治疗技术做为第三类医疗技术应用于临床,批准号200984。本文就目前DC、CIK、DC-CIK细胞免疫疗法在急性髓细胞白血病中的应用进展加以综述。     

自体免疫免疫细胞治疗技术之CIK细胞治疗癌症

近年来:自体免疫细胞治疗技术越来越广泛地被应用到临床治疗中。自体免疫细胞治疗技术是指从患者自身外周血等中分离单个核细胞,经过专项实验室进行细胞的活化与增殖后,回输入患者体内,直接杀伤肿瘤细胞或被病毒感染的细胞,同时起到调节和增强机体免疫功能的作用。随着自体免疫细胞治疗技术的发展,应用细胞因子诱导的杀伤细胞(cytokine induced killer cell,CIK细胞)进行恶性肿瘤治疗的临床试验也陆续开展。     

胃癌的过继性免疫治疗研究进展

胃癌是目前世界上发病率及致死率较高的恶性肿瘤之一,在东亚地区尤其显著。针对胃癌的治疗手段仍是传统的手术联合化疗、放疗为主,尽管靶向药物治疗提供了新的选择,但其对晚期胃癌的疗效仍然有限。胃癌的免疫治疗作为独特的治疗手段,在近十多年发展较为活跃,特别是过继性免疫治疗手段不断有创新。过继性免疫治疗主要依赖回输具有抗肿瘤活性的细胞,目前回输的细胞由具有非特异性抗肿瘤作用向具有特异性抗肿瘤作用演变,特别是嵌合性抗原T细胞治疗的出现,为进展期胃癌患者提供了有一种潜在的选择。本文对胃癌过继性免疫治疗中采用的不同免疫活性细胞的作用机制、临床应用等进行总结,并针对其不足提出利用基因工程技术增强治疗靶向性、降低免疫逃逸的研究方向。     

细胞因子诱导杀伤细胞过继免疫治疗恶性血液肿瘤的临床应用进展

细胞因子诱导的杀伤细胞(cytokine-induced killer cells,CIK cells)是将脐血、外周血和骨髓等不同来源的单个核细胞经适当浓度的IFN-γ、IL-2和CD3单抗等联合诱导后获得的异质细胞群。CIK细胞具有增殖能力强、溶瘤谱广、对耐药细胞株也有杀伤作用,且兼有T细胞强大的杀伤活性和NK细胞(nature killer cells)非主要组织相容性复合体(non-major histocompatibility complex,non-MHC)限制性的特点,使其在肿瘤治疗中具有广阔的临床应用前景。恶性血液肿瘤患者接受CIK细胞过继免疫治疗的生存时间延长,不良反应少,但仍面临许多问题。     

树突细胞联合细胞因子诱导的杀伤细胞对急性白血病细胞的体外杀伤作用

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细胞的表型、增殖活性的变化,及抗急性白血病细胞活性.方法:正常人外周血单个核细胞诱导DC和CIK细胞,将DC与CIK共培养,以CIK细胞单独培养为对照.用台盼蓝活细胞计数计算细胞扩增倍数,MTT法测定杀伤活性,流式细胞术分析免疫表型.结果:DC-CIK细胞增殖能力明显高于CIK细胞(P＜0.05); DC、CIK细胞共培养后,CD3+ CD8+、CD3+ CD56+双阳性细胞比率较同条件下CIK细胞组显著增多(P＜0.05);在2.5∶1-20∶1的效靶比范围内,DC-CIK共培养物对AML细胞的杀伤率显著高于CIK细胞(P＜0.05),且杀伤率与效靶比呈正相关.结论:DC-CIK细胞的增殖能力、对AML细胞的杀伤活性均高于CIK细胞,为DC-CIK细胞免疫治疗提供了实验和理论依据.     

以T细胞受体为基础的免疫疗法研究进展

T细胞是机体抗肿瘤免疫的核心,以T细胞功能调控为基础的免疫检查点疗法已经在多种肿瘤的临床治疗中取得了重大突破,以基因工程化T细胞为基础的过继性免疫细胞疗法在血液瘤治疗中取得了重要进展,免疫治疗已经对肿瘤的临床治疗产生了深刻变革,成为肿瘤临床治疗策略的重要组成部分。T细胞受体（T cell receptor,TCR）赋予了T细胞识别肿瘤抗原的特异性,能够识别由主要组织相容性复合体（major histocompatibility complex,MHC）呈递的包括胞内抗原在内的广泛肿瘤抗原,具有高度的抗原敏感性,因而具有广泛的抗肿瘤应用前景。2022年第一款TCR药物的上市开启了TCR药物开发的新纪元,多项TCR药物临床研究表现出潜在的肿瘤治疗价值。本文综述了以TCR为基础的免疫治疗策略研究进展,包括T细胞受体工程化T细胞（T cell receptor-engineered T cell,TCR-T）和TCR蛋白药物,以及基于TCR信号的其他免疫细胞疗法,以期为以TCR为基础的免疫治疗策略开发提供参考。     

树突状细胞的靶向抗肿瘤治疗进

树突状细胞(dendritic cell,DC)是目前已知体内最强的抗原提呈细胞(ARC),其在肿瘤免疫中具有重要的作用.DC的靶向抗肿瘤治疗成为当今肿瘤免疫治疗的研究热点.本文就DC的抗肿瘤机制、DC基因修饰策略及DC疫苗的临床治疗进展进行综述.     

Combined Therapy with Cytokine-Induced Killer Cells and Oncolytic Adenovirus Expressing IL-12 Induce Enhanced Antitumor Activity in Liver Tumor Model

Both adoptive immunotherapy and gene therapy hold a great promise for treatment of malignancies. However, these strategies exhibit limited anti-tumor activity, when they are used alone. In this study, we explore whether combination of cytokine-induced killer (CIK) adoptive immunotherapy with oncolytic adenovirus-mediated transfer of human interleukin-12 (hIL-12) gene induce the enhanced antitumor potency. Our results showed that oncolytic adenovirus carrying hIL-12 (AdCN205-IL12) could produce high levels of hIL-12 in liver cancer cells, as compared with replication-defective adenovirus expressing hIL-12 (Ad-IL12). AdCN205-IL12 could specifically induce cytotoxocity to liver cancer cells. Combination of CIK cells with AdCN205-IL12 could induce higher antitumor activity to liver cancer cells in vitro than that induced by either CIK or AdCN205-IL12 alone, or combination of CIK and control vector AdCN205-GFP. Furthermore, treatment of the established liver tumors with the combined therapy of CIK cells and AdCN205-IL12 resulted in tumor regression and long-term survival. High level expression of hIL-12 in tumor tissues could increase traffic of CIK cells to tumor tissues and enhance their antitumor activities. Our study provides a novel strategy for the therapy of cancer by the combination of CIK adoptive immunotherapy with oncolytic adenovirus-mediated transfer of immune stimulatory molecule hIL-12.     

IL-12 enhances efficacy and shortens enrichment time in cytokine-induced killer cell immunotherapy

Cytokine-induced killer (CIK) cells are T cell derived ex vivo expanded cells with both NK and T cell properties. They exhibit potent anti-tumor efficacy against various malignancies in preclinical models and have proven safe and effective in clinical studies. We combined CIK cell adoptive immunotherapy with IL-12 cytokine immunotherapy in an immunocompetent preclinical breast cancer model. Combining CIK cells with IL-12 increased anti-tumor efficacy in vivo compared to either therapy alone. Combination led to full tumor remission and long-term protection in 75% of animals. IL-12 treatment sharply increased the anti-tumor efficacy of short-term cultured CIK cells that exhibited no therapeutic effect alone. Bioluminescence imaging based in vitro cytotoxicity and in vivo homing assays revealed that short-term cultured CIK cells exhibit full cytotoxicity in vitro, but display different tumor homing properties than fully expanded CIK cells in vivo. Our data suggest that short-term cultured CIK cells can be “educated” in vivo, producing fully expanded CIK cells upon IL-12 administration with anti-tumor efficacy in a mouse model. Our findings demonstrate the potential to improve current CIK cell-based immunotherapy by increasing efficacy and shortening ex vivo expansion time. This holds promise for a highly efficacious cancer therapy utilizing synergistic effects of cytokine and cellular immunotherapy.     

Enhanced antitumor effects of DC-activated CIKs to chemotherapy treatment in a single cohort of advanced non-small-cell lung cancer patients

Cytokine-induced killer (CIK) cells show cytolytic activity against tumor. The purpose of this study was to evaluate the antitumor effect of dendritic cell (DC)-activated CIK cells in vitro and their clinical efficacy of DC-activated CIK cells in combination with chemotherapy (abbreviated below as chemotherapy plus DC + CIK) in patients with advanced non-small-cell lung cancer (NSCLC). A paired study was performed between 61 patients treated with chemotherapy alone (group 1) and 61 patients treated with chemotherapy plus DC + CIK cells (group 2). In group 2, 36 patients with adenocarcinoma and 18 patients with squamous cell carcinoma were analyzed for the survival rate. Compared to unstimulated CIK cells, DC-activated CIK cells significantly enhanced antitumor activity, increased the ratio of CD3⁺CD56⁺ cells, promoted cell proliferation and lessened cell apoptosis. In the paired study, the 1- and 2-year overall survival rates in group 2 were 57.2 and 27.0 %, which were significantly higher than that of group 1 (37.3 and 10.1 %) (P < 0.05). There was no significant difference in the survival rate between the adenocarcinoma and squamous carcinoma patients in group 2. The present study suggests that DC-activated CIK cell has enhanced antitumor effects and chemotherapy plus DC + CIK cells improved the clinical outcomes of chemotherapy for advanced NSCLC patients.     

Ten-year update of the international registry on cytokine-induced killer cells in cancer immunotherapy

Cytokine-induced killer (CIK) cells represent an exceptional T-cell population uniting a T cell and natural killer cell-like phenotype in their terminally differentiated CD3⁺CD56⁺ subset, which features non-MHC-restricted tumor-killing activity. CIK cells have provided encouraging results in initial clinical studies and revealed synergistic antitumor effects when combined with standard therapeutic procedures. We established the international registry on CIK cells (IRCC) to collect and evaluate clinical trials for the treatment of cancer patients in 2010. Moreover, our registry set new standards on the reporting of results from clinical trials using CIK cells. In the present update, a total of 106 clinical trials including 10,225 patients were enrolled in IRCC, of which 4,889 patients in over 30 distinct tumor entities were treated with CIK cells alone or in combination with conventional or novel therapies. Significantly improved median progression-free survival and overall survival were shown in 27 trials, and 9 trials reported a significantly increased 5-year survival rate. Mild adverse effects and graft-versus-host diseases were also observed in the studies. Recently, more efforts have been put into the improvement of antitumoral efficacy by CIK cells including the administration of immune checkpoint inhibitors and modification with chimeric antigen receptorc. The minimal toxicity and multiple improvements on their tumor-killing activity both make CIK cells a favorable therapeutic tool in the clinical practice of cancer immunotherapy.     

Combining Antiangiogenic Therapy with Adoptive Cell Immunotherapy Exerts Better Antitumor Effects in Non-Small Cell Lung Cancer Models

Introduction

Cytokine-induced killer cells (CIK cells) are a heterogeneous subset of ex-vivo expanded T lymphocytes which are characterized with a MHC-unrestricted tumor-killing activity and a mixed T-NK phenotype. Adoptive CIK cells transfer, one of the adoptive immunotherapy represents a promising nontoxic anticancer therapy. However, in clinical studies, the therapeutic activity of adoptive CIK cells transfer is not as efficient as anticipated. Possible explanations are that abnormal tumor vasculature and hypoxic tumor microenvironment could impede the infiltration and efficacy of lymphocytes. We hypothesized that antiangiogenesis therapy could improve the antitumor activity of CIK cells by normalizing tumor vasculature and modulating hypoxic tumor microenvironment.

Methods

We combined recombinant human endostatin (rh-endostatin) and CIK cells in the treatment of lung carcinoma murine models. Intravital microscopy, dynamic contrast enhanced magnetic resonance imaging, immunohistochemistry, and flow cytometry were used to investigate the tumor vasculature and hypoxic microenvironment as well as the infiltration of immune cells.

Results

Our results indicated that rh-endostatin synergized with adoptive CIK cells transfer to inhibit the growth of lung carcinoma. We found that rh-endostatin normalized tumor vasculature and reduced hypoxic area in the tumor microenvironment. Hypoxia significantly inhibited the proliferation, cytotoxicity and migration of CIK cells in vitro and impeded the homing of CIK cells into tumor parenchyma ex vivo. Furthermore, we found that treatment with rh-endostatin significantly increased the homing of CIK cells and decreased the accumulation of suppressive immune cells in the tumor tissue. In addition, combination therapy produced higher level of tumor-infiltration lymphocytes compared with other treatments.

Conclusions

Our results demonstrate that rh-endostatin improves the therapeutic effect of adoptive CIK cells therapy against lung carcinomas and unmask the mechanisms of the synergistic antitumor efficacy, providing a new rationale for combining antiangiogenesis therapy with immunotherapy in the treatment of lung cancer.     

Dendritic Cells Decreased the Concomitant Expanded Tregs and Tregs Related IL-35 in Cytokine-Induced Killer Cells and Increased Their Cytotoxicity against Leukemia Cells

Regulatory T cells (Tregs) are potent immunosuppressive cells and essential for inducing immune tolerance. Recent studies have reported that Tregs and Tregs related cytokines can inhibit the antitumor activity of cytokine-induced killer (CIK) cells, but dendritic cells co-cultured CIK (DC-CIK) cells can be used for induction of a specific immune response by blocking of Tregs and TGF-β, IL-10. As a novel identified cytokine, IL-35 is specially produced by Tregs and plays an essential role in immune regulation. However, it remains unknown whether IL-35 roles in tumor immunotherapy mediated by CIK and DC-CIK cells. In this study, we cultured CIK and DC-CIK cells from the same healthy adult samples, and investigated their phenotype, proliferation, cytotoxic activity against leukemia cell lines K562 and NB4 by FCM and CCK-8, measured IL-35, TGF-β and IL-10 protein by ELISA, detected Foxp3, IL-35 and IL-35 receptor mRNA by Real-time PCR, respectively. We found Tregs and IL-35 concomitantly expanded by a time-dependent way during the generation of CIK cells, but DC significantly down-regulated the expression of them and simultaneously up-regulated the proliferation ability as well as cytotoxic activity of CIK cells against leukemia cell lines. Therefore, our data suggested that DC decreased concomitant expanded Tregs and Tregs related IL-35 in CIK cells and might contribute to improve their cytotoxicity against leukemia cells in vitro.     

Cytokines for the induction of antitumor effectors: The paradigm of Cytokine-Induced Killer (CIK) cells

Cytokine-Induced killer (CIK) cells are raising growing interest in cellular antitumor therapy, as they can be easily expanded with a straightforward and inexpensive protocol, and are safe requiring only GMP-grade cytokines to obtain very high amounts of cytotoxic cells. CIK cells do not need antigen-specific stimuli to be activated and proliferate, as they recognize and destroy tumor cells in an HLA-independent fashion through the engagement of NKG2D. In several preclinical studies and clinical trials, CIK cells showed a reduced alloreactivity compared to conventional T cells, even when challenged across HLA-barriers; only in a few patients, a mild GVHD occurred after treatment with allogeneic CIK cells. Additionally, their antitumor activity can be redirected and further improved with chimeric antigen receptors, clinical-grade monoclonal antibodies or immune checkpoint inhibitors. The evidence obtained from a growing body of literature support CIK cells as a very promising cell population for adoptive immunotherapy. In this review, all these aspects will be addressed with a particular emphasis on the role of the cytokines involved in CIK cell generation, expansion and functionalization.     

The apoptotic and proliferative fate of cytokine-induced killer cells after redirection to tumor cells with bispecific Ab

BACKGROUND: Cytokine-induced killer (CIK) cells are ex vivo expanded T cells with co-expression of CD3 and CD56 and NK activity. They have recently been evaluated in a phase I/II clinical trial against malignant lymphoma. Bispecific Ab (bsAb) redirect CIK cells to tumor targets, thus enhancing their cytotoxicity. While bsAb may improve T-cell mediated anti-tumor activity, little is known about the fate of effector cells upon redirection to tumor targets using a bsAb. METHODS: Using ex vivo-activated CIK cells, Her2/neu expressing breast and ovarian cell lines and a F(ab')2 Her2/neu x CD3 bsAb, we investigated the anti-tumor activity and the proliferative and apoptotic outcome of CIK cells. RESULTS: When redirected to tumor targets with bsAb, there was a significant increase in anti-tumor activity as well as an increase in both CIK cell proliferation and apoptosis. The addition of agonistic Ab against CD28 did not significantly increase proliferation or apoptosis of CIK cells redirected to CD80- and CD86- tumor targets. To attempt to reduce T-cell apoptosis, we incubated CIK cells in the presence of the pan-caspase inhibitor z-VAD-fmk, which led to a partial reduction in T-cell apoptosis without increasing cellular cytotoxicity. DISCUSSION: bsAb are effective in redirecting activated T cells to tumor targets and such redirection leads to both T-cell proliferation and apoptosis that are not altered by co-stimulation through CD28. Effector cell apoptosis can be reduced by using a caspase inhibitor but this does not increase CIK cell cytotoxicity.