Antitumor activities of human autologous cytokineinduced killer(CIK)cells against hepatocellular carcinoma cells in vitro and in vivo

AIM: To characterize the anticancer function of cytokine-induced killer cells (CIK) and develop an adoptive immunotherapy for the patients with primary hepatocellular carcinoma (HCC), we evaluated the proliferation rate, phenotype and the antitumor activity of human CIK cells from healthy donors and HCC patients in vitro and in vivo. METHODS: Peripheral blood mononuclear cells (PBMC) from healthy donors and patients with primary HCC were incubated in vitro and induced into CIK cells in the presence of various cytokines such as interferon-gamma (IFN-gamma), interleukin-1 (IL-1), IL-2 and monoclonal antibody (mAb) against CD3. The phenotype and characterization of CIK cells were identified by flow cytometric analysis. The cytotoxicity of CIK cells was determined by (51)Cr release assay. RESULTS: The CIK cells were shown to be a heterogeneous population with different cellular phenotypes. The percentage of CD3+/CD56+ positive cells, the dominant effector cells, in total CIK cells from healthy donors and HCC patients, significantly increased from 0.1-0.13% at day 0 to 19.0-20.5% at day 21 incubation, which suggested that the CD3+ CD56+ positive cells proliferated faster than other cell populations of CIK cells in the protocol used in this study. After 28 day in vitro incubation, the CIK cells from patients with HCC and healthy donors increased by more than 300-fold and 500-fold in proliferation cell number, respectively. CIK cells originated from HCC patients possessed a higher in vitro antitumor cytotoxic activity on autologous HCC cells than the autologous lymphokine-activated killer (LAK) cells and PBMC cells. In in vivo animal experiment, CIK cells had stronger effects on the inhibition of tumor growth in Balb/c nude mice bearing BEL-7402-producing tumor than LAK cells (mean inhibitory rate, 84.7% vs 52.8%, P【0.05) or PBMC (mean inhibitory rate, 84.7% vs 37.1%, P【0.01). CONCLUSION: Autologous CIK cells are of highly efficient cytotoxic effector cells against primary hepatocellular carcinoma cells and might serve as an alternative adoptive therapeutic strategy for HCC patients.

Ex vivo Expansion of Cytokine-Induced Natural Killer Cells from Tumor Patients for Autologous Therapy

To optimize the expansion conditions for cytokine-induced natural killer cells (CINK), including major CD3 - CD56 + NK cells, and by using the selected method to expand the CINK cells from patients with tumors for the application in the autologous immunotherapy, peripheral blood mononuclear cells (PBMC) from healthy donors were cultivated in stem cell growth medium supplemented with 5% human AB serum, and monoclonal anti-CD3 antibody, IL-2 and phytohaemagglutin (PHA) at different concentrations were added to select the optimal cultural conditions for the CINK cells. The optimized method was used to expand the CINK cells from 12 patients with various malignant tumors, the proportions of CD3 + and CD56 + cells in the expanded cells were determined by flow cytometry, and the cytotoxicity of the CINK cells from patients was detected by MTT assay in comparison with those of cytokine-induced killer (CIK) cells or anti-CDC3 antibody-activated killer (CD3AK) cells. It was found that the expansion of CINK cells could be obtained when these cells were activated with 500 ng/ml of monoclonal anti-CD3 antibody and 100?U/ml of IL-2. The highest degree of cell proliferation (51.3 folds) could be attained in the presence of PHA after 14?d cultivation of cells, and the expanded cell populations included CD3 - CD56 + NK cells (49.8%±7.2%) and CD3 + CD56 + T cells (14.7%±3.9%) could lyse 83.7% of K562 cells and 55.8% of Raji cells at a 10∶1 of effector/target ratio. Also the CINK cells of clinical scale could be expanded from tumor patients by using the optimized method with a similar expansion efficiency and cytotoxicity. These cells showed a higher cytotoxic activities against target cell lines (K562 and Raji cell lines) than those of CIK and CD3AK cells. This improved method has several advantages over other methods, such as the easy manipulation, low costs and the production of functionally active NK cells on a clinical scale from cancer patients without the need for special equipments. This might facilitate the realization of further protocol for evaluating the clinical effects of NK cells in autologous immunotherapies of tumor patients.

Trispecific killer engager 161519 enhances natural killer cell function and provides anti-tumor activity against CD19-positive cancers

Objective:Natural killer(NK)cells have gained considerable attention due to their potential in treating"cold tumors,"and are therefore considered as one of the new strategies for curing cancer,by using worldwide development of their new possibilities and interventions with NK cell-related therapeutic products.Methods:We constructed a trispecific killer engager(TriKE)consisting of anti-CD16,IL-15,and anti-CD19.This TriKE was designed to attract CD19^(+)tumor cells to CD16^(+)NK cells,whereas IL-15 sustained the proliferation,development,and survival of NK cells.Results:Treatment with 161519 TriKE in the presence of CD19^(+)targets upregulated expression of CD69,CD107 a,TRAIL,IFN-γ,and TNF-α in NK cells,and significantly improved the proliferation and cytotoxicity of NK cells.NK cells"armed"with 161519 TriKE showed stronger cytolysis against CD19+targets compared with that of"unarmed"NK cells.A preclinical model of B-cell lymphoma in human peripheral blood mononuclear cell-reconstituted xenograft mice showed significant inhibition of tumor growth and prolonged overall survival after treatment with 161519 TriKE,when compared with that in control mice or mice treated with 1619 BiKE.Combined use of IL-2 was a more effective treatment with 1619 BiKE,when compared with that using 161519 TriKE.Conclusions:The newly generated 161519 TriKE enhanced the proliferation,activation,cytokine secretion,and cytotoxicity of NK cells in the presence of CD19+tumor cells.The 161519 TriKE aided inhibition of tumor growth and prolonged the overall survival of murine xenografts,and could be used to treat CD19-positive cancers.

Revolutionizing gastric cancer treatment:The potential of immunotherapy

Gastric cancer,a prevalent malignancy worldwide,ranks sixth in terms of frequency and third in fatality,causing over a million new cases and 769000 annual deaths.Predominant in Eastern Europe and Eastern Asia,risk factors include family medical history,dietary habits,tobacco use,Helicobacter pylori,and Epstein-Barr virus infections.Unfortunately,gastric cancer is often diagnosed at an advanced stage,leading to a grim prognosis,with a 5-year overall survival rate below 5%.Surgical intervention,particularly with D2 Lymphadenectomy,is the mainstay for early-stage cases but offers limited success.For advanced cases,the National Comprehensive Cancer Network recommends chemotherapy,radiation,and targeted therapy.Emerging immunotherapy presents promise,especially for unresectable or metastatic cases,with strategies like immune checkpoint inhibitors,tumor vaccines,adoptive immunotherapy,and nonspecific immunomodulators.In this Editorial,with regards to the article“Advances and key focus areas in gastric cancer immunotherapy:A comprehensive scientometric and clinical trial review”,we address the advances in the field of immunotherapy in gastric cancer and its future prospects.

Targeting immune checkpoints:how to use natural killer cells for fighting against solid tumors

Natural killer(NK)cells are unique innate immune cells that mediate antiviral and anti-tumor responses.Thus,they might hold great potential for cancer immunotherapy.NK cell adoptive immunotherapy in humans has shown modest efficacy.In particular,it has failed to demonstrate therapeutic efficiency in the treatment of solid tumors,possibly due in part to the immunosuppressive tumor microenvironment(TME),which reduces NK cell immunotherapy’s efficiencies.It is known that immune checkpoints play a prominent role in creating an immunosuppressive TME,leading to NK cell exhaustion and tumor immune escape.Therefore,NK cells must be reversed from their dysfunctional status and increased in their effector roles in order to improve the efficiency of cancer immunotherapy.Blockade of immune checkpoints can not only rescue NK cells from exhaustion but also augment their robust anti-tumor activity.In this review,we discussed immune checkpoint blockade strategies with a focus on chimeric antigen receptor(CAR)-NK cells to redirect NK cells to cancer cells in the treatment of solid tumors.

Natural killer cell lines in tumor immunotherapy

Natural killer(NK)cells are considered to be critical players in anticancer immunity.However,cancers are able to develop mechanisms to escape NK cell attack or to induce defective NK cells.Current NK cell-based cancer immunotherapy is aimed at overcoming NK cell paralysis through several potential approaches,including activating autologous NK cells,expanding allogeneic NK cells,usage of stable allogeneic NK cell lines and genetically modifying fresh NK cells or NK cell lines.The stable allogeneic NK cell line approach is more practical for quality-control and large-scale production.Additionally,genetically modifying NK cell lines by increasing their expression of cytokines and engineering chimeric tumor antigen receptors could improve their specificity and cytotoxicity.In this review,NK cells in tumor immunotherapy are discussed,and a list of therapeutic NK cell lines currently undergoing preclinical and clinical trials of several kinds of tumors are reviewed.

Functional crosstalk and regulation of natural killer cells in tumor microenvironment: Significance and potential therapeutic strategies

Natural killer (NK) cells eliminate a large variety of tumor cells and abnormal cells. However, NK cells in the tumor microenvironment (TME) are often functionally depleted. A few subsets of NK cells even promote tumor growth. This study reviewed the biological properties of NK cells, the dynamic phenotypic changes of NK cells in the TME, and the communication between NK cells and other immune and nonimmune cells.

Immunotherapy of Cancer—A Historical Note

We examined the possibility that the anti-estrogens, tamoxifen (TX) and toremifen (TO) interacted?with the immune system. Indeed, both TX and TO stimulated cells mediated cytotoxicity reactions by various killer cells: killer T (TK), natural killer (NK), lymphokine activated killer (LAK) cells. Both TX and TO inhibited the growth of tumors that express estrogen receptors. Thus these antiestrogens inhibited tumor growth and stimulated killer cells for cytotoxicty on such tumors. Therefore these agents were presumed to stimulate tumor immunity. We tested the P815 mouse mastcytoma with TK, LK, and TX or TO. A therapeutic effect was observed in both experiments. The SL2-5 murine lymphoma was tested with NK and TX cells or TO cells and successful immunotherapy was observed.?We digested human breast carcinomas and lung tumors with collagenase. The killer cells were separated from tumor cells on Ficoll gradients. TX and TO enhanced the cytotoxic effect of autologous killer cells on the corresponding tumor cells. This experiment indicates that the results obtained in animals are also valid for human malignant disease.

自然杀伤细胞2组成员A、程序性死亡因子配体1表达检测对PD-L1阻断免疫治疗肌层浸润性膀胱癌反应性的预测价值

目的:探究自然杀伤细胞2组成员A(NKG2A)及程序性死亡因子配体1(PD-L1)表达检测对PD-L1阻断免疫治疗肌层浸润性膀胱癌反应性的预测价值。方法:选取100例肌层浸润性膀胱癌患者作为研究对象,对其行PD-L1阻断免疫治疗后,根据治疗反应性将其分为缓解组和未缓解组;对缓解组患者随访3个月,将其分为复发组和未复发组。比较两组患者NKG2A和PD-L1在CD4^(+)和CD8^(+)上的表达水平,采用ROC曲线分析NKG2A和PD-L1预测膀胱癌患者治疗反应性的价值。结果:100例研究对象中,缓解组患者72例(72.00%),未缓解组患者28例(28.00%),两组性别、年龄比较无统计学差异(均P>0.05),但缓解组患者肿瘤直径小于未缓解组,NKG2A、PD-L1/CD4^(+)和PD-L1/CD8^(+)表达水平均低于未缓解组(均P<0.05)。膀胱癌患者NKG2A、PD-L1/CD4^(+)和PD-L1/CD8^(+)表达预测免疫治疗后缓解的AUC值分别为0.771、0.724、0.710;联合诊断的AUC为0.836。72例缓解患者中,出现复发29例(40.28%),未出现复发43例(59.72%),两组性别、年龄以及肿瘤直径比较无统计学差异(均P>0.05),但复发组NKG2A、PD-L1/CD4^(+)和PD-L1/CD8^(+)表达水平均高于未复发组(均P<0.05)。NKG2A、PD-L1/CD4^(+)和PD-L1/CD8^(+)表达预测膀胱癌患者缓解后复发的AUC值分别为0.775、0.740、0.728;联合诊断的AUC为0.874。结论:NKG2A、PD-L1/CD4^(+)和PD-L1/CD8^(+)在不同治疗反应性肌层浸润性膀胱癌患者外周血中表达水平不同,三者对于膀胱癌患者PD-L1阻断免疫治疗反应性均有一定的预测价值,且三者联合预测效能最佳。

肿瘤浸润性自然杀伤细胞在免疫治疗中的研究进展

自然杀伤(NK)细胞是一种具有强大细胞毒性和活性的先天淋巴细胞。基于NK细胞的免疫疗法在血液系统肿瘤的治疗中取得显著疗效,但在实体恶性肿瘤的治疗中疗效较低。该文就NK细胞的功能、表型及其肿瘤免疫治疗的研究进展作一综述。

PD-L1-driven efficient enrichment and elimination of circulating cancer cells by magnetic MoSe_(2) nanosheet

Circulating tumor cells(CTCs)are important biomarkers in the development and progression of lung cancer because they can reach other organs through the blood circulation and form distant metastases,exacerbating lung cancer progression.The presence of CTCs is also the main reason for the failure of nanomedicine-based lung cancer treatments.Therefore,magnetic MoSe_(2) nanosheets loaded with programmed death-ligand 1(PD-L1),named PD-L1-MFP NS,were employed here to precisely capture lung cancer CTCs in the blood circulation through the tumor-targeting effect of PD-L1 killing CTCs with highly effective photothermal therapy(PTT).In addition,by increasing the expression of cytomegalovirus UL16-binding protein(ULBP)ligands on tumor cells,the PD-L1-MFP NS further activated natural killer(NK)cells and triggered NK cell-induced cancer immunotherapy,thereby enhancing the overall tumor-killing effect.In summary,this material designed to capture CTCs provides a substantial advancement for personalized PTT-triggered immunotherapy and has great clinical translational potential.

基于自然杀伤(NK)细胞的肝癌免疫疗法研究进展

肝细胞肝癌(HCC)异质性高,晚期诊断和化疗耐药使其成为我国难治性肿瘤之一。自然杀伤(NK)细胞在机体的免疫监视过程中充当重要角色。但在肝癌的进展过程中,NK细胞的功能受损致肿瘤免疫逃逸。该文总结了HCC中基于NK细胞的不同免疫治疗策略,包括直接输注过继性NK细胞免疫疗法、基因工程NK细胞免疫疗法、靶向NK细胞受体免疫疗法、改变免疫抑制微环境免疫疗法、细胞因子增强NK细胞肿瘤毒性免疫疗法以及中医药增强NK细胞肿瘤毒性免疫疗法,这些基于NK细胞的免疫治疗策略已显示出巨大的治疗潜力。

自然杀伤细胞在抗肿瘤免疫中的应用

自然杀伤细胞是具有细胞毒性的淋巴样细胞,能够直接和间接发挥抗肿瘤作用。既往研究证明过继自然杀伤细胞治疗肿瘤具有一定的可行、安全及有效性,深入研究自然杀伤细胞过继疗法的抗实体瘤潜力成为现今研究热点之一。本文对当前自然杀伤细胞抗肿瘤免疫及细胞过继疗法中的研究进展作一综述。

Natural killer cell-derived extracellular vesicle significantly enhanced adoptive T cell therapy against solid tumors via versatilely immunomodulatory coordination

Insufficient tumor tropism,MHC classⅠmolecules(MHC-I)defects of tumor cells,and immunosuppressive tumor microenvironment(TME)seriously imperil the efficacy of adoptive T cell therapy on solid tumors.Here,natural killer cell-derived extracellular vesicle(Nev)is used as a versatile toolkit to synergistically improve adoptive T cell therapy for solid tumors.Specifically,Nev is modified with dibenzocyclooctynes(DBCO)linked with p H-sensitive benzoic-imine bonds;meanwhile,cytotoxic T lymphocyte(CTL)is decorated with azide groups.Then CTL is armed with Nev(CTL-Nev)through the click chemistry reaction.After systematic administration,Nev obviously promotes the tumor-targeting accumulation of CTL coming from its native tumor-tropism capability.Then,the cleavage of benzoic-imine bonds in the slightly acidic TME leads to the release of Nev,which not only directly induces tumor apoptosis but also promotes the action of CTL via multiplex pathways,such as up-regulating the MHC-I expression on tumor cells,reprogramming tumor-associated macrophages from pro-tumoral M2 phenotypes to tumoricidal M1 phenotypes.The all-around coordination of Nev with CTL results in potent tumor repression.

负载自体肿瘤细胞裂解物的DC疫苗联合CIK治疗晚期肾癌的临床观察——附10例报告

背景与目的:肾癌的主要治疗手段是手术,但晚期肾癌术后复发率高,加上肾癌对化疗和放疗不敏感,因此,晚期肾癌预后不佳,需要寻找新的更有效的治疗方法。本研究通过负载自体肿瘤细胞裂解物的树突细胞(dendriticcells,DC)疫苗联合细胞因子诱导的杀伤细胞(cytokine-inducedkillercells,CIK)治疗10例晚期肾癌,观察近期的临床疗效,免疫学反应及副作用。方法:分离患者外周血单核细胞,体外经GM-CSF和IL-4诱导产生DC细胞,并负载自体肿瘤细胞裂解物;T淋巴细胞经IFN-γ、IL-2、CD3单抗、IL-1α体外诱导产生CIK细胞。所有患者在切除原发病灶后,接受每周一次的皮内DC疫苗注射治疗,至少8次治疗;CIK细胞过继细胞免疫治疗,每2周一次,至少接受4次治疗。临床疗效和免疫学反应分别通过影象学检查,外周血T淋巴细胞亚群改变和迟发性超敏(delayed-typehypersensitivity,DTH)反应进行评估。结果:(1)4例有可评价病灶的患者中1例部分缓解(PR),2例疾病稳定(SD),1例进展(PD);6例没有可评价病灶的患者中1例PD,1例失访,另外4例未见疾病进展。随访时间6～20个月(中位时间11个月)。(2)与治疗前比较,治疗2个月后患者CD3+、CD4+、CD4+/CD8+、CD56+明显升高(P<0.05)。(3)包括PR患者在内的6例患者DTH反应呈现阳性。(4)除一过性的发热、畏寒外没有其它不良反应出现。结论:负载自体肿瘤细胞裂解物的DC疫苗联合CIK细胞治疗晚期肾癌有一定的近期临床疗效,能诱导出特异的抗肾癌免疫反应,并且有良好的耐受性。

NK细胞肿瘤免疫治疗的研究进展

自然杀伤细胞(NK细胞)的泛特异性天然免疫识别和快速杀伤机制,使其成为抗肿瘤治疗中除T细胞外的另一把利剑。NK细胞过继转输免疫治疗在血液恶性肿瘤临床研究治疗中取得可喜的疗效; CAR-NK显示出与CAR-T同样的抗卵巢癌效果,但毒副作用较低。特别是近来TIGIT和NKG2A免疫检验点抑制剂的研究显示,NK细胞对T细胞发挥抗肿瘤活性起到不可或缺的调节和协同作用。研究进展提示,NK细胞与T细胞各具优势,达到互补的抗肿瘤效果,基于NK细胞的免疫治疗可能成为新一代免疫治疗的突破点。

纤维连接蛋白诱导CIK细胞治疗晚期癌症患者的安全性及疗效评价

目的:评价重组人纤维连接蛋白(Retro Nectin,RN)诱导的CIK细胞治疗晚期癌症患者的安全性及疗效。方法:观察85例晚期癌症患者,经RN诱导的自体或异体血CIK细胞治疗的安全性。选择晚期非小细胞肺癌患者36例,分为2组:CIK细胞治疗组20例,化疗组16例,比较两组治疗前后的免疫功能、生活质量(用汉化生活量表QLICP系统测试)、临床疗效差异;选择晚期非小细胞肺癌患者32倒,分为CIK细胞治疗组及姑息治疗组各16例,比较两组患者的生存期。结果:安全性:自体及异体CIK细胞治疗过程中主要出现兴奋感、失眠、低热,未见明显毒副反应。近期疗效:20例患者经CIK细胞治疗:PR 1例,SD 10例,PD 9例,ORR 5.00%,DCR 55.00%;16例患者经化疗:PR 5例,SD 5例,PD 6例,ORR 31.25%,DCR 62.50%,2组的DCR差异不显著(P>0.05),但化疗组的ORR明显高于CIK组(P<0.05)。在生活质量方面,化疗后病人心理模块中情绪方面明显低落(P<0.05),CIK细胞治疗的患者多有生活质量提高;CIK细胞治疗组与化疗组患者,治疗前后免疫功能无显著差异(P>0.05)。CIK细胞治疗组(11.0个月)比姑息治疗组(6.0个月)中位生存期延长,但总生存时间无统计差异(x^2=2.301,P=0.129)。结论:RN诱导的CIK细胞自体及异体治疗均简便、安全、有效,可以改善晚期肿瘤病人的生活质量,延长生存期。

人源白细胞介素21对外周血及脐血来源CIK细胞产生和抗肿瘤活性的作用研究

目的:研究新近发现的免疫调节因子——白细胞介素21(IL-21)对外周血及脐血来源的CIK细胞产生及抗肿瘤活性的体外作用。方法:采集分离正常人的外周血及脐血单个核细胞,加用细胞因子诱导培养CIK细胞,在有无人源IL-21(200ng/μl)培养条件下,检测CIK细胞表达及杀伤K562细胞和急性白血病患者肿瘤细胞活性的变化;检测培养上清IFN-γ的浓度及杀伤活性以及RT-PCR法检测培养细胞的IFN-γRNA表达的不同。结果:在人源IL-21的作用下,培养14天时,CIK细胞的产生由17.5%升至26.5%(外周血来源);33.8%升至55.9%(脐血来源)。②CIK细胞对K562细胞的杀伤作用由24.0%升至52.2%(外周血来源);35.1%升至79.7%(脐血来源);脐血来源CIK细胞对13例急性白血病患者肿瘤细胞杀伤作用由27.4%升至58.3%。③外周血来源培养上清IFN-γ的浓度上升了近一倍,对K562的杀伤作用增加了近一倍;脐血来源培养上清IFN-γ的浓度上升了三倍多,对K562的杀伤作用增加了近二倍;④外周血和脐血来源培养细胞的IFN-γRNA表达均明显增高。结论:人源IL-21可增加外周血及脐血来源CIK细胞产生及增强其抗肿瘤活性,通过增加IFN-γ的表达产生为其作用机制之一,提示IL-21在增强肿瘤免疫治疗中具有潜在的临床应用前景。

DCIK细胞联合化疗治疗晚期消化道肿瘤的临床疗效

目的:评价树突状细胞调节的细胞因子诱导的杀伤细胞(DC activated and cytokins induced killer,DCIK)联合化疗治疗晚期消化道肿瘤的疗效。方法:实验采用武警总医院2005年6月至2007年8月应用DCIK联合全身化疗治疗晚期消化道肿瘤的患者23例作为联合治疗组。同期进行单纯化疗的20例晚期消化道肿瘤患者作为对照组。联合治疗组患者均于化疗前采集外周血单个核细胞(PBMC)。将PBMC体外培养产生DCIK细胞。联合治疗组患者首先行2周期全身化疗,化疗结束后回输质量合格的DCIK细胞。单纯化疗组仅进行2周期全身化疗。观察两组近期疗效、临床受益反应改善程度、肿瘤标志物、免疫指标及1年、3年生存率。结果:两组近期疗效无显著性差异(P>0.05)。联合治疗组治疗后KPS评分升高(P<0.05),单纯化疗组治疗后KPS评分较治疗前无改善(P>0.05)。联合治疗组患者治疗后外周血CD3+CD8+、CD3+CD56+细胞的比例大幅升高(P<0.01);而单纯化疗组患者无明显变化(P>0.05)。联合治疗组1年生存率78.3%;单纯化疗组1年生存率80%(P>0.05)。联合治疗组3年生存率52.2%;单纯化疗组3年生存率30%(P<0.01)。结论:DCIK联合化疗治疗晚期消化道肿瘤具有更好的疗效,其临床受益反应有较大的提高,免疫功能有所改善,3年生存率提高。

CH-296和IFN-γ共同培养的CIK细胞治疗晚期恶性实体瘤临床观察

目的:CIK细胞具有异质性的抗肿瘤作用,其中IFN-γ在增强CD3+CD56+细胞作用中起到关键作用。在TCR刺激信号存在的情况下,CH-296被发现能够刺激T细胞的增殖。本研究探讨CH-296和IFN-γ是否有相互促进CIK细胞表达的作用。方法:CIK细胞在CH-296包被培养板或不包括的IFN-γ中培养14 d。将CIK细胞增殖、代谢、表型、细胞毒性同传统的CIK细胞相比较。20例(其中18例为Ⅳ期)患者接受了3个周期的RN-CIKs治疗。临床反应通过KPS评分和CT检查进行评价。结果:CH-296通过减缓代谢和提高增殖以时间依赖的方式促进CIK细胞的表达。CH-296和IFN-γ共同培养带有T细胞表型的CIK细胞,有更强的细胞毒性和细胞因子分泌功能。临床试验中未发生不良事件。16例患者临床症状有所缓解。总体临床反应率(PR+SD)为65%(13/20),中位OS(16.95±6.10)个月。结论:CH-296能够促进IFN-γ提高CIK细胞的抗肿瘤作用,使CIK细胞有更强的增殖能力,细胞毒作用并减缓代谢。