Vaccination of multiple myeloma patients with idiotype-pulsed dendritic cells: immunological and clinical aspects

Multiple myeloma (MM) is characterized by a clonal proliferation of malignant plasma cells in the bone marrow secreting a monoclonal immunoglobulin (paraprotein) with specific antigenic determinants, the idiotype (Id), which can be regarded as a tumour-associated antigen (TAA). In order to analyse the impact of a dendritic cell (DC)-based vaccine, 11 patients with advanced MM were treated with CD34 stem cell-derived dendritic cells that were pulsed with Id peptides. Subsequently, the patients received three boost immunizations every other week with a combination of Id and granulocyte-macrophage colony-stimulating factor (GM-CSF) (nine patients) or with Id peptide-pulsed dendritic cells again (two patients). The treatment was well tolerated with no side-effects. The present clinical study was a proof of concept analysis of dendritic cell-based vaccines in MM. The capacity of the dendritic cells to activate idiotype-specific T cells was verified by in vitro stimulation experiments before the vaccination therapy. Immunological effects of the Id vaccination were analysed by monitoring changes in anti-idiotype antibody titres and idiotype-specific T-cell activity. After vaccination, three out of 10 analysed patients showed increased anti-idiotype antibody serum titres, indicating the induction of an idiotype-specific humoral immune response. The idiotype-specific T-cell response analysed by ELISpot was increased in four out of 10 analysed patients after vaccination, and one patient had a decreased plasma cell infiltration in the bone marrow. In conclusion, five out of 11 patients showed a biological response after vaccination. Thus, our data indicate that immunotherapy with Id-pulsed DCs in MM patients is feasible and safe. DC generated from CD34+ progenitor cells can serve as a natural adjuvant for the induction of clinically relevant humoral and cellular idiotype-specific immune responses in patients suffering from advanced MM.     

Enhanced cytotoxicity of IL-24 gene-modified dendritic cells co-cultured with cytokine-induced killer cells to hepatocellular carcinoma cells

As a novel human cytokine found recently, IL-24 could selectively kill tumor cells by multiple ways. Dendritic cells (DCs) are the major antigen-presenting cells. Recent studies have revealed that IL-24 can promote the antigen-presenting function of DCs. In this study, we evaluated the antitumor effect and mechanism of co-cultured cytokine-induced killer (CIK) cells and autologous DCs modified with IL-24 gene on hepatocellular carcinoma (HCC) cells. DCs and CIK cells were prepared routinely from human peripheral blood mononuclear cells. Recombinant adenovirus AdVGFP/IL-24 (Ad-IL24) was constructed expressing IL-24. IL-24 gene was transduced into DCs via Ad-IL24, the cells obtained were named DC-IL-24. We demonstrated that the expression rates of CD80, CD83, CD1a, HLA-DR, CD40, CXCR4 on DC-IL-24 were significantly increased compared with those of the control group. DC-IL-24 produced markedly higher levels of IL-24, IL-12 and TNF-α as compared with those of DCs. On comparison with non-transfected DCs co-cultured with CIK cells, transfected DCs co-cultured with CIK cells had a significant higher lytic activity against SMMC7721 cells, a HCC cell line.     

Clinical efficacy of immunotherapy of dendritic cell and cytokine-induced killer cell combined with chemotherapy for treatment of multiple myeloma

Objective This research was aimed to evaluate the immune mechanism and clinical effect of immunotherapy of dendritic cells(DC) and cytokine-induced killer cell(CIK) combined with chemotherapy on multiple myeloma(MM). Methods 60 patients with MM were randomly     

树突状细胞联合细胞因子诱导的杀伤细胞对胃癌细胞杀伤作用

目的探讨树突状细胞联合细胞因子诱导的杀伤细胞对胃癌细胞的杀伤作用。方法采用胃癌患者自身血液中单个核细胞(peripheral blood mononuclear cells,PBMC),经体外诱导分别扩增出DC和CIK细胞,二者共同培养后,利用MTT法检测DC细胞联合CIK细胞体外杀伤人胃癌细胞株(MNK-45、MNK-28、SG-7901)的活性。结果DC与CIK细胞共培养后得到的细胞群高表达CD3 CD56 ,平均值达到(56.74±7.63)%。通过彼此相互作用诱导出的细胞群体对胃癌细胞株MNK-45、MNK-28、SG-7901有杀伤作用,且杀伤活性随着效靶比的增加而增强。结论DC与CIK细胞共培养后有很强的增殖能力,对胃癌细胞具有杀伤活性,且其杀伤作用与胃癌细胞类型无相关性。     

Autologous cytokine-induced killer cells in the treatment of multiple myeloma concomitant with lung cancer and paraneoplastic dermatoses

Cytokine-induced killer (CIK) cells have been shown to be effective in the treatment of advanced cancer and minimal residual diseases. We report a multiple myeloma (MM) patient with concomitant lung cancer and paraneoplastic dermatoses, who received cellular immunotherapy with CIK cells which were derived from peripheral blood mononuclear cells (PBMCs) after being primed with anti-cluster of differentiation 3 (CD3) monoclonal antibody, interleukin-2 (IL-2), interferon-γ (IFN-γ) and IL-1. After treatment MM and lung cancer remained stable and no progression or recurrence was observed. Paraneoplastic dermatoses were obviously improved after treatment, which was first reported. No evident side effects were observed. These findings suggested that cellular immunotherapy with CIK cells was safe and effective in this patient with MM and lung cancer, and it might be a potent therapeutic option for paraneoplastic dermatoses.     

Immunotherapy with dendritic cells and cytokine-induced killer cells for hepatocellular carcinoma: A meta-analysis

BACKGROUND Hepatocellular carcinoma(HCC) has been revealed as the second most common cause of cancer-related deaths worldwide. The introduction of cell-based immunotherapy, including dendritic cells(DCs) and cytokine-induced killer cells(CIKs), has brought HCC patients an effective benefit. However, the efficacy and necessity of cellular immunotherapy after different interventional therapy remains to be further explored.AIM To investigate the efficacy of cellular immunotherapy, involving DCs and CIKs,combined with different conventional treatments of HCC.METHODS We performed a literature search on PubMed and Web of Science up to February15, 2019. Long-term efficacy(overall survival and recurrence) and short-term adverse effects were investigated to assess the effectiveness of immunotherapy with DCs and/or CIKs. Review Manager 5.3 was used to perform the analysis.RESULTS A total of 22 studies involving 3756 patients selected by eligibility inclusion criteria were forwarded for meta-analysis. Combined with the conventional clinical treatment, immunotherapy with DCs and/or CIKs was demonstrated to significantly improve overall survival at 6 mo [risk ratio(RR) = 1.07; 95%confidence interval(CI): 1.01-1.13, P = 0.02], 1 year(RR = 1.12; 95%CI: 1.07-1.17, P 0.00001), 3 years(RR = 1.23; 95%CI: 1.15-1.31, P 0.00001) and 5 years(RR =1.26; 95%CI: 1.15-1.37, P 0.00001). Recurrence rate was significantly reduced by cellular immunotherapy at 6 mo(RR = 0.50; 95%CI: 0.36-0.69, P 0.0001) and 1 year(RR = 0.82; 95%CI: 0.75-0.89, P 0.00001). Adverse effect assessment addressed that immunotherapy with DCs and/or CIKs was accepted as a safe,feasible treatment.CONCLUSION Combination immunotherapy with DCs, CIKs and DC/CIK with various routine treatments for HCC was evidently suggested to improve patients' prognosis by increasing overall survival and reducing cancer recurrence.     

Cisplatin pretreatment enhances anti-tumor activity of cytokine-induced killer cells

AIM:To investigate whether cisplatin (DDP) enhances the anti-tumor activity of cytokine-induced killer (CIK) cells in a murine colon adenocarcinoma model.METHODS:Tumor size and weight served as indicators of therapeutic response.Immunohistochemistry was performed to observe intratumoral lymphocyte infiltration and tumor microvessel density.Changes in the percentage of regulatory T (Treg) cells within the spleens of tumor-bearing mice preconditioned with DDP were monitored using flow cytometry.RESULTS:A mark...     

Cytokine-induced killer cells/dendritic cells and cytokine-induced killer cells immunotherapy for the treatment of esophageal cancer: A meta-analysis

Objectives:This meta-analysis was designed to systematically evaluate whether autologous cytokine-induced killer cells (CIK) or dendritic cells and cytokine-induced killer cells (DC-CIK) immunotherapy combined with chemotherapy can improve the therapeutic effect and safety of chemotherapy in esophageal cancer (EC).Materials and methods:Randomized controlled trials (RCTs) were electronically searched databases including CNKI, WanFang, WeiPu, CBMDisc, PubMed, Web of Science, EMbase, the Cochrane Library, and Clinical Trials. The databases were searched for articles published until June 2019. Two researchers independently screened the literature, extracted data, and evaluated the quality of the included literature. Meta-analysis was performed using RevMan5.3.Results:Seventeen studies (1416 participants) were included. The differences between CIK/DC-CIK combination chemotherapy and chemotherapy alone were significant. The results displayed that the number of CD3⁺, CD4⁺, CD4⁺/CD8⁺, and NK cells was significantly increased after 1 to 2 weeks of treatment with CIK/DC-CIK cells in the treatment group (all P < .05). In addition, the results shown that 1-year overall survival was significantly prolonged (P < .0001) and quality of life was improved (P = .001) in EC chemotherapy combined with immunotherapy groups compared with conventional treatment. Furthermore, cytokine expression levels of interleukin 2 (IL-2), tumor necrosis factor α (TNF-α), and interleukin 12 (IL-12) were significantly increased (P = .0003) as well as the levels of immunoglobulins were elevated (P < .00001). Serum levels of tumor marker molecules, carcinoembryonic antigen (CEA), carbohydrate antigen (CA)-199, and CA-125 were lower in treatment groups than that of control groups (P < .00001). No fatal adverse reactions were noted (P = .04).Conclusions:It is safe and effective for patients to use chemotherapy combined with CIK/DC-CIK immunotherapy. Immunotherapy can simultaneously improve the antitumor immune response. Specifically, DC-CIK cells can increase T lymphocyte subsets, CIK cells, NK cells, and immunoglobulins in peripheral blood to enhance antitumor immunity. Therefore, combination therapy enhances the immune function and improves the therapeutic efficacy of patients with EC.     

DC与CIK或LAK联合对结肠癌细胞株SW480杀伤作用的研究

[目的]研究树突状细胞(DC)联合细胞因子诱导或未诱导的杀伤细胞(CIK)或淋巴因子激活的杀伤细胞(LAK)对结肠癌细胞株SW480的杀伤活性.提供DC联合CIK或LAK治疗结肠癌的实验依据.[方法]取人外周血分离出单个核细胞(PBMNC),诱导生成DC、CIK、LAK细胞;流式细胞仪检测DC经SW480肿瘤抗原冲击后的表型变化;以CIK+DC细胞、CIK细胞、LAK+DC细胞及LAK细胞作为效应细胞,SW480为靶细胞,以15∶1、30∶1、45∶1为效靶比,LDH释放法测定细胞杀伤试验活性;ELISA检测杀伤试验中干扰素γ(IFN-γ)、白细胞介素2(IL-2)、IL-12、IL-17的分泌水平.[结果]流式细胞仪检测DC经SW480肿瘤抗原冲击后,其表面分子HLA-DR、CD40、CD80和CD86表达分别平均为90.23％、73.68％、85.96％、57.55％,与未经肿瘤抗原冲击DC比较,DC成熟的表面标志分子表达明显增加(P＜0.01).相同效靶比下,CIK+DC细胞组对SW480的杀伤作用最强,明显高于其他细胞组(P＜0.01);CIK+ DC细胞组在效靶比为45∶1时,杀伤活性最强(P＜0.01);单独CIK细胞组的杀伤活性明显高于LAK+DC细胞组(P＜0.01);LAK+ DC细胞组的杀伤活性明显高于单独LAK细胞组(P＜0.01).效靶比为45∶1时,各杀伤试验细胞组上清液中IFN-γ、IL-2、IL-12、IL-17的分泌量,CIK+DC细胞组的IFN-γ、IL-12的分泌量显著高于其他细胞组(P＜0.05);LAK+DC、单独LAK细胞组IL-2的分泌量明显高于CIK+DC、单独CIK细胞组(P＜0.05);单独CIK细胞组IFN-γ的分泌量明显高于LAK+DC、单独LAK细胞组(P＜0.05).[结论]CIK+DC细胞组对SW480的杀伤活性明显强于单独CIK、LAK+ DC组、单独LAK细胞组.其机制可能是,SW480抗原致敏的DC分泌IFN-γ、IL-12等刺激、诱导CIK细胞的活化和增殖,明显增强CIK细胞杀伤SW480的活性.     

Immunization against murine multiple myeloma with fusions of dendritic and plasmacytoma cells is potentiated by interleukin 12

Fusions of cancer cells and dendritic cells (DCs) are effective in the treatment of animal tumor models and patients with metastatic renal carcinoma. In this study, we have fused DCs with mouse 4TOO plasmacytoma cells. The results demonstrate that vaccination of mice with the fusion cells (FC/4TOO) is associated with induction of antitumor humoral and cytotoxic T lymphocyte (CTL) responses. Immunization with FC/4TOO cells protected mice against tumor challenge. In addition, treatment of established multiple myeloma with FC/4TOO cells was associated with prolongation of survival but not with eradication of disease. As interleukin (IL)-12 potentiates the induction of immune responses, recombinant mouse IL-12 was administered with the FC/4TOO vaccine. Treatment of mice with FC/4TOO and IL-12 was associated with increased CTL activity and T-cell proliferation responses. Treatment with FC/4TOO and IL-12 also resulted in eradication of established disease. These findings demonstrate that immunization with FC/4TOO fusion cells and IL-12 potentiates antitumor immunity and the treatment of murine multiple myeloma.     

Enhancement of the cytotoxic activity of cytokine-induced killer cells transfected with IL3PE38KDEL gene against acute myeloid leukemia cells

Cytokine-induced killer (CIK) cells, one of the feasible and effective methods of adoptive immunotherapy, have shown anti-leukemia activity in vivo and in vitro. But the strategy exhibits limited cytotoxic activity in clinical studies. In this study, CIK cells were transfected with an interleukin-3/Pseudomonas exotoxin gene (IL3PE38KDEL). RT-PCR and ELISA were used to verify the expression of IL3PE38KDEL in the transfected CIK cells. These cells released 1,186.7?±?149.6 pg IL3PE38KDEL/10⁴ cells over 48 h into the medium and the culture supernatant selectively killed IL3 receptor(IL3R)-positive HL60 cells, but not IL3R-negative K562 cells. Moreover, IL3PE38KDEL transfection did not influence phenotypes and cytokine production of CIK cells. Co-cultured with leukemia cells, IL3PE38KDEL transfected CIK cells showed enhanced cytotoxicity against IL3R-positive HL60 cells at all effector-to-target (E:T) ratios, but exerted a basal anti-leukemia activity against IL3R-negative K562 cells. Our findings demonstrate that IL3PE38KDEL gene transfection may be a novel strategy for improving anti-leukemia activity of CIK cells.     

IL-24对细胞因子诱导的杀伤细胞的作用

目的:寻求增强CIK细胞的细胞毒活性的有效方法.方法:从健康人外周血中提取出单个核细胞,第1天加入IFN-γ,第2天加入IL-1,CD3 mAb,IL-2诱导CIK细胞,另一组与IL-1.CD3 mAb,IL-2同时加入IL-24.杀伤分为4组:未加IL-24培养组、单独IL-24杀伤组、加IL-24培养组、未加IL-24的CIK细胞培养组在杀伤时加入IL-24.细胞计数法测定细胞的增殖、MTT法测定细胞杀伤活性和流式细胞术分析细胞表型.扫描电镜和透射电镜观察CIK细胞对肿瘤细胞的杀伤和肿瘤细胞的改变.结果:未加IL-24培养组CIK细胞增殖高于加IL-24培养组,两者比较有明显差异(126.34±2.14 vs 108.87±1.29,P<0.05).加IL-24培养组细胞各个效靶比杀伤活性均达到90%以上,明显高于其他各组(效靶比为10:1时95.58%±2.21% vs 27.31%±2.69%,8.74%±2.41%,38.65%±21.30%,P<0.05;效靶比为20:1时91.97%±4.21% vs 34.27%±0.85%,11.54%±2.78%,48.32%±11.72%,P<0.05;效靶比为40:1时91.84%±9.28% vs 50.67%±1.30%,23.73%±11.07%,52.89%±12.26%,P<0.05).不同时间加IL-24培养组各个细胞表型与未加IL-24培养组相比没有差别.透射电镜下观察加IL-24培养组凋亡和坏死肿瘤细胞比未加IL-24培养组明显增多.结论:CIK细胞诱导过程中加入IL-24能明显增强其杀伤活性.     

Anti-myeloma activity of endogenous and adoptively transferred activated natural killer cells in experimental multiple myeloma model

OBJECTIVE: Despite advances in autologous stem cell transplantation and chemotherapy, multiple myeloma (MM) remains an incurable disease. Due to the role of natural killer (NK) cells in host resistance against several tumors, it is of interest to explore the anti-MM activity of NK cells. For this reason, we aimed to determine if NK cells provide anti-MM activity following interleukin-2 (IL-2) administration, and if ex vivo activated and intravenously administered NK cells prolong survival in MM-bearing C57BL/KaLwRij mice. METHODS: The anti-MM effect of IL-2 was tested by intraperitoneal injection into the 5T33MM-inoculated mice. Subsequently, in vivo effector cell depletions were performed by administration of anti-NK1.1 or anti-CD8 monoclonal antibodies. Finally, magnetically separated and activated NK cells from splenocytes of C57BL/KaLwRij mice were adoptively transferred to tumor-bearing mice in conjunction with IL-2 treatment. RESULTS: IL-2 administration into MM-bearing mice significantly prolonged their survival. This effect was diminished by in vivo depletion of NK cells. Adoptive transfer of activated NK cells showed a significant in vivo anti-MM effect that was dependent on cell dose. Biodistribution of the marked adoptively transferred NK cells correlated with MM cells' homing sites. CONCLUSION: These data suggest that activated NK cells have a promising potential in adoptive immunotherapy for MM.     

Fusion of dendritic cells with multiple myeloma cells results in maturation and enhanced antigen presentation

Dendritic cells (DCs) are potent antigen-presenting cells that are uniquely capable of inducing primary immune responses. Although tumour cells may directly inhibit DC maturation, exposure to tumour products may also result in their activation. Fusions of cancer cells and DCs are being explored as cancer vaccines. The effect of tumour cell fusion on DC maturation and their functional characteristics has not been defined. In the present study, immature and mature DC generated from human CD34+ and peripheral blood precursors were fused to multiple myeloma cells in the presence of polyethylene glycol. Fusion of both immature and mature DCs with tumour cells resulted in an activated phenotype. In this regard, fusion cells expressed interleukin-12, a cytokine essential for the induction of T-helper cell type 1 immunity. In contrast to immature DCs, fusion cells also strongly expressed CC-chemokine receptor R7, which is responsible for DC migration to draining lymph nodes. Fusions generated with both immature and mature DCs also potently stimulated T-cell expression of gamma-interferon and cytotoxic T lymphocyte killing of tumour targets. These findings demonstrate that tumour cell fusion induces DC maturation and the development of an activated phenotype necessary for their effectiveness as cancer vaccines.     

Natural cytotoxicity to autologous antigen-pulsed dendritic cells in multiple myeloma

To analyse autologous lymphocyte cytolytic activities of potential importance for cell-based immunotherapy in multiple myeloma (MM), in vitro differentiated dendritic cells (DCs) loaded with patient-specific monoclonal immunoglobulin (mIg) were used as autologous target cellsin cytotoxicity assays. Effector populations consisted of purified natural killer (NK) cells (CD56+, CD3-) and T cells (CD3+). The MM patients' NK cells cultured in the presence of interleukin 2 (IL-2) showed pronounced cytotoxic activity towards autologous mature DCs. Autologous MM DC targets displayed similar susceptibility to NK cell lysis, compared with allogeneic control DC targets, despite high surface expression of self major histocompatibility complex (MHC) antigens. However, some degree of classic MHC class I-mediated negative regulation was implicated in the NK-DC interactions, as indicated by class I blocking experiments. NK-mediated lysis was also discerned towards primary autologous MM cells. The results indicated that the major effector mechanism was mediated through the perforin-granzyme exocytosis pathway. In conclusion, NK cells from MM patients displayed significant and consistent cytotoxicity towards autologous mature DCs, suggesting that innate immunity could be implicated in MM and may influence the outcome of the administration of tumour antigen-pulsed DCs in treatment trials.