Intraperitoneal perfusion of cytokine-induced killer cells with local hyperthermia for advanced hepatocellular carcinoma

AIM:To study the effect and tolerance of intraperitoneal perfusion of cytokine-induced killer(CIK) cells in combination with local radio frequency(RF) hyperthermia in patients with advanced primary hepatocellular carcinoma(HCC).METHODS:Patients with advanced primary HCC were included in this study.CIK cells were perfused intraperitoneal twice a week,using 3.2 × 10 9 to 3.6 × 10 9 cells each session.Local RF hyperthermia was performed 2 h after intraperitoneal perfusion.Following an interval of one month,the next course of treatment was administered.Patients received treatment until disease progression.Tumor size,immune indices(CD3 +,CD4 +,CD3 + CD8 +,CD3 + CD56 +),alpha-fetoprotein(AFP) level,abdominal circumference and adverse events were recorded.Time to progression and overall survival(OS) were calculated.RESULTS:From June 2010 to July 2011,31 patients diagnosed with advanced primary HCC received intraperitoneal perfusion of CIK cells in combination with local RF hyperthermia in our study.Patients received an average of 4.2 ± 0.6 treatment courses(range,1-8 courses).Patients were followed up for 8.3 ± 0.7 mo(range,2-12 mo).Following combination treatment,CD4 +,CD3 + CD8 + and CD3 + CD56 + cells increased from 35.78% ± 3.51%,24.61% ± 4.19% and 5.94% ± 0.87% to 45.83% ± 2.48%(P = 0.016),39.67% ± 3.38%(P = 0.008) and 10.72% ± 0.67%(P = 0.001),respectively.AFP decreased from 167.67 ± 22.44 to 99.89 ± 22.05 ng/mL(P = 0.001) and abdominal circumference decreased from 97.50 ± 3.45 cm to 87.17 ± 4.40 cm(P = 0.002).The disease control rate was 67.7%.The most common adverse events were low fever and slight abdominal erubescence,which resolved without treatment.The median time to progression was 6.1 mo.The 3-,6-and 9-mo and 1-year survival rates were 93.5%,77.4%,41.9% and 17.4%,respectively.The median OS was 8.5 mo.CONCLUSION:Intraperitoneal perfusion of CIK cells in combination with local RF hyperthermia is safe,can efficiently improve immunological status,and may prolong survival in HCC patients.

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 marked T cell-dependent, synergistic anti- tumor effect of the combined therapy was observed （1968 ± 491 mm3 ys 3872 ＋ 216 mm3; P = 0,003）, Preconditioning chemotherapy with DDP augmented the infiltration of CD3＋ T lymphocytes into the tumor mass and reduced the percentage of both intratumoral and splenic Treg cells. CONCLUSION： Preconditioning with DDP markedly enhances the efficacy of adoptively transferred CIK cells, providing a potential clinical modality for the treatment of patients with colorectal cancer.

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.

Adjuvant treatment for triple-negative breast cancer: a retrospective study of immunotherapy with autologous cytokine-induced killer cells in 294 patients

Objective: To examine the efficacy and safety of a sequential combination of chemotherapy and autologous cytokine-induced killer(CIK) cell treatment in triple-negative breast cancer(TNBC) patients.Methods: A total of 294 post-surgery TNBC patients participated in the research from January 1, 2009 to January 1, 2015. After adjuvant chemotherapy, autologous CIK cells were introduced in 147 cases(CIK group), while adjuvant chemotherapy alone was used to treat the remaining 147 cases(control group). The major endpoints of the investigation were the disease-free survival(DFS) and overall survival(OS). Additionally, the side effects of the treatment were evaluated.Results: In the CIK group, the DFS and OS intervals of the patients were significantly longer than those of the control group(DFS:P = 0.047;OS: P = 0.007). The multivariate analysis demonstrated that the TNM(tumor-node-metastasis) stage and adjuvant CIK treatment were independent prognostic factors for both DFS [hazard ratio(HR)= 0.520, 95% confidence interval(CI):0.271-0.998, P = 0.049;HR = 1.449, 95% CI:1.118-1.877, P = 0.005, respectively] and OS(HR=0.414, 95% CI:0.190-0.903, P = 0.027;HR= 1.581, 95% CI:1.204-2.077, P = 0.001, respectively) in patients with TNBC. Additionally, longer DFS and OS intervals were associated with increased number of CIK treatment cycles(DFS: P = 0.020;OS: P = 0.040). The majority of the patients who benefitted from CIK cell therapy were relatively early-stage TNBC patients.Conclusion: Chemotherapy in combination with adjuvant CIK could be used to lower the relapse and metastasis rate, thus effectively extending the survival time of TNBC patients, especially those at early stages.

Hepatocellular carcinoma-specific immunotherapy with synthesized α1,3-galactosyl epitope-ulsed dendritic cells and cytokine-induced killer cells

AIM： To evaluate the safety and clinical efficacy of a new immunotherapy using both α-Gal epitope-pulsed dendritic cells （DCs） and cytokine-induced killer cells. METHODS： Freshly collected hepatocellular carcinoma （HCC） tumor tissues were incubated with a mixture of neuraminidase and recombinant αl,3-galactosyltrans- ferase （αI,3GT） to synthesize α-Gal epitopes on car- bohydrate chains of the glycoproteins of tumor mem- branes. The subsequent incubation of the processed membranes in the presence of human natural anti-Gal IgG resulted in the effective phagocytosis to the tumor membrane by DCs. Eighteen patients aged 38-78 years with stage 111 primary HCC were randomly chosen for the study; 9 patients served as controls, and 9 patients were enrolled in the study group.RESULTS： The evaluation demonstrated that the pro- cedure was safe; no serious side effects or autoimmune diseases were observed. The therapy significantly pro- longed the survival of treated patients as compared with the controls （17.1 ± 2.01 mo vs 10.1 ±4.5 mo, P = 0.00121）. After treatment, all patients in the study group had positive delayed hypersensitivity and robust systemic cytotoxicity in response to tumor lysate as measured by interferon-y-expression in peripheral blood mononuclear cells using enzyme-linked immunosorbent spot assay. They also displayed increased numbers of CD8-, CD45RO- and CD56-positive cells in the peripheral blood and decreased α-fetoprotein level in the se- rum. CONCLUSION： This new tumor-specific immunotherapy is safe, effective and has a great potential for the treat- ment of tumors.

Effect of dendritic cell/cytokine-induced killer cell immunobiological cancer therapy combined with adjuvant chemotherapy in patients with triple-negative breast cancer

Objective The aim of the present study was to investigate the effect of dendritic cell(DC)/cytokine-induced killer cell(CIK) immunobiological cancer therapy in patients with triple-negative breast cancer(TNBC) who underwent adjuvant chemotherapy. Methods From January 2010 to October 2013, 120 patients with postoperative TNBC were recruited and included in the study. Patients were enrolled in one of two groups according to whether they accepted DC/CIK immunobiological cancer therapy during adjuvant chemotherapy; the patients in the DC/CIK group underwent adjuvant chemotherapy combined with DC/CIK immunobiological cancer therapy, and the control group underwent adjuvant chemotherapy alone. When six cycles of adjuvant chemotherapy and six cycles of DC/CIK immunobiological cancer therapy had been completed, differences between the two groups with regard to quality of life(Qo L), immunological indicators(CD3, CD4, CD8, and NK cell levels), disease-free survival(DFS), and side effects of chemotherapy and DC/CIK treatment were evaluated.Results In the DC/CIK group, the proportion of NK cells and CD3+ and CD4+ T-cell subgroups significantly increased, and the proportion of CD8+ cells decreased when they were compared before and after DC/CIK therapy(P < 0.05). However, there were no significant changes in the control group. By the final follow-up, DFS of the treatment group and the control group was 38.4 and 34.2 months, respectively. The Qo L improved in the patients treated with chemotherapy plus DC/CIK therapy compared with the patients treated with chemotherapy alone, and the difference between groups was significant(P < 0.05). The side effects of two groups were tolerable and not significantly different between the two groups.Conclusion The DC/CIK treatment had potential benefits for patients with TNBC compared with the control group, and was not associated with any obvious side effects. Therefore, DC/CIK therapy is a safe and effective method for the treatment of TNBC.

肿瘤特异性个体化多靶点DC-CIK治疗原发性肝癌的临床疗效与安全性

目的:评价肿瘤特异性个体化多靶点自体树突状细胞-细胞因子诱导的杀伤细胞(DC-CIK)治疗中晚期原发性肝癌(PLC)的临床疗效与安全性。方法:回顾性分析2019年10月至2021年9月东部战区总医院肿瘤科行DC-CIK治疗的119例中晚期PLC患者的临床资料。根据患者治疗时负载DC的抗原不同将患者分为两组,一组使用患者自体特异性多肽负载为pDC-CIK组(n=21),另一组使用肿瘤细胞裂解物负载为DC-CIK组(n=98)。分析两组患者治疗前后的临床资料,包括治疗效果和治疗前后甲胎蛋白、淋巴细胞亚群、细胞因子(IL-2、IFN-γ、TNF-α和IL-6)水平的变化、不良反应发生情况等。结果:119例PLC患者治疗后,pDC-CIK和DC-CIK组两组客观缓解率均为0%,疾病控制率分别为76.1%和72.4%(P>0.05)。治疗后两组患者CD3^(+)、CD4^(+)、CD8^(+)、CD56^(+)、CD25^(+)和CD4^(+)/CD8^(+)T淋巴细胞水平无统计学差异(均P>0.05),治疗后两组患者外周血IL-2、IFN-γ、TNF-α和IL-6的平均水平均显著高于治疗前(均P<0.001),两组患者治疗后外周血IL-2、TNF-α和IL-6的平均水平无显著差异(均P>0.05),而pDC-CIK组IFN-γ水平显著高于DC-CIK组(P<0.05)。pDC-CIK组患者平均生存时间为59.84个月,高于DC-CIK组的46.54个月,但无统计学差异(P=0.16)。治疗过程中无严重不良反应的发生。结论:PLC患者行肿瘤特异性个体化多靶点DC-CIK治疗是安全有效的,并能改善免疫功能,相较肿瘤细胞裂解物负载DC-CIK有进一步获益趋势。

Autologous cytokine-induced killer cells combined with chemotherapy in the treatment of advanced colorectal cancer: a randomized control study

Objective：To evaluate the ef icacy of autologous cytokine-induced kil er （CIK） cells transfusion combined with chemotherapy in patients suf ered from advanced colorectal cancer. Methods： Sixty untreated patients with advanced colorectal cancer were randomly divided into two groups. The 30 patients in the control group received chemotherapy with the regimen of xeloda plus oxiplatin （XELOX）. The 30 patients in the trial group were treated with chemotherapy （XELOX） in combination with autologous CIK celltransfusion. T-lymphocyte subgroups were separated and measured by flow cytometry quality of life （QOL） was determined by EORTC QLQ-C30. The short-term curative ef ect was evaluated via imaging examina-tions. The patients’ median progression free survival time was estimated by Kaplan-Meier. Results：The T-lymphocyte im-mune activity was improved in patients received autologous CIK celltransfusion than those treated with chemotherapy alone. The subgroup of CD3＋CD56＋T lymphocyte was significantly increased （4.28 ± 0.45 vs 10.14 ± 1.02, P=0.01）. Short-term ef icacy evaluation revealed that there was no significant dif erence in terms of objective response rate （ORR） between the two groups, but the disease control rate （DCR） was markedly increased （86.7%vs 56.7%, P=0.020） in the group treated by chemotherapy plus CIK cells compared to the group treated with chemotherapy alone. The progression free survival time was 8.64 months （ 95%CI 6.25-9.75 months） in control group and 10.15 months （ 95%CI 7.48-12.52 months） in trial group. Compared to patients in control group, the patients in trial group had significantly longer progression-free survival （P=0.046）. The QOL assessment suggested the QOL in trial group was obviously improved than that in the control group. Compared with the control group, patients treated with autologous CIK celltransfusion scored more in the area of physical function and general health status, while the symptomatic scores in terms of pain, fatigue, nausea and vomiting and diarrhea were significantly reduced. Conclusion：Autologous CIK celltransfusion combined with chemotherapy can ef ectively enhance the immune activity of T-lymphocytes, prevent disease progression and improve the progression-free survival and QOL in patients with advanced colorectal cancer.

Cytokine-induced killer cell combination with TACE in the treatment of hepatocellular cancers:a meta-analysis

Objective：The aim of the study was to evaluate the ef icacy and safety of cytokine-induced kil er （CIK） cellcombined with transcatheter arterial chemoembolization （TACE） therapy in the treatment of hepatocellular carcinoma （HCC）. Methods：Randomized control ed trials （RCTs） on CIK cells combination with TACE based-therapy were identified by elec-tronic searches in the Cochrane Library, MEDLINE, Pubmed, Wanfang, VIP, CNKI and other electronic databases. We in-cluded any RCTs evaluating CIK cellcombination with TACE for the treatment of hepatocellular cancers. The quality of RCTs meeting inclusion criteria was evaluated and data on short-term and long-term curative ef ects, quality of life, liver function and immunologic function were extracted. For quantitative data, we conducted meta-analysis with ReMan 5.1 software and the GRADE System was used to rate the level of evidence and strength of recommendation. For qualitative data, data mainly adopted descriptive methods. Results：The 9 RCTs involving 870 cases meeting the inclusion criteria were included. The meta-analysis showed significant survival benefit on 0.5-year survival rate （RR=1.51, 95%CI, 1.35-1.69, P〈0.00001） in fa-vor of CIK based therapy. This ef ect was consistent at other prospective dates, including 1-year survival rate （RR=2.30, 95%CI, 1.94-2.72, P〈0.00001）, 2-year survival rate （RR=7.03, 95%CI, 3.83-12.91, P〈0.0001）. Meanwhile, the CIK-based group also demonstrated a significantly prolonged time-to-progression （TTP） （SD=1.62, 95%CI, 1.30-1.94, P〈0.0001） and overal survival （OS） （SD=20.6, 95%CI, 20.2-21.18, P〈0.0001）. Moreover, a favored response rate （RR=CR＋PR） （RR=2, 95%CI, 1.65-2.43, P〈0.00001） and the quality of life improvement rate （KPS） （RR=1.76, 95%CI, 1.26-2.45, P=0.0008） were also observed in patients receiving CIK cells and TACE therapy. Furthermore, patients in the CIK group showed lower AFP （SD=-165.23, 95%CI,-178.51--151.94, P〈0.00001）, ALT （SD=-33.14, 95%CI,-40.30--36.37, P〈0.00001）, and AST （SD=-15.57, 95%CI,-17.05--14.09, P〈0.00001） levels. In terms of T-lymphocyte subsets in peripheral blood, the analysis also showed the percentage of CD3＋, CD4＋cells and the ratio of CD4＋/CD8＋cells in the CIK group increased compared with the non-CIK group. Based on GRADE, the level of evidence was GRADE 2C, and the strength of recommen-dation was 2. Conclusion：CIK cells combined with TACE therapy demonstrated a significant superiority in improving recent and forward curative ef ects, immunity function, quality of life and liver function of HCC patients. Based on GRADE, the level of evidence was GRADE 2C, and the strength of recommendation was 2. In the light of the limitations of this study, large-scale, high-quality clinical trials should be carried out to further confirmed the above conclusion.

DC联合CIK治疗156例局部晚期或晚期胰腺癌的临床疗效

目的:评价DC联合CIK治疗156例局部晚期或晚期胰腺癌的临床疗效。方法:回顾性分析2011年11月至2023年12月在东部战区总医院肿瘤科进行自体DC联合CIK治疗的156例局部晚期或晚期胰腺癌患者的临床资料。统计患者治疗前后血清肿瘤标志物、淋巴细胞亚群、细胞因子水平的变化、不良反应发生情况以及近期疗效、远期疗效。结果:156例胰腺癌患者中有92例治疗前后均进行了影像学检查,结果显示CR 0例,PR 0例,SD 42例,PD 50例,ORR为0%,DCR为45.65%。外周血CA199水平在治疗前后无显著差异,但有19例患者治疗后下降超过20%。治疗前后患者外周血CD3^(+)、CD4^(+)、CD8^(+)、CD56^(+)、CD25^(+)淋巴细胞亚群水平和CD4+/CD8+T细胞比值无统计学差异(P>0.05),治疗后患者外周血IL-2、IFN-γ的平均水平均显著高于治疗前(P<0.05),TNF-α和IL-6无显著差异(P>0.05)。156例患者mOS为8.53个月,1年累积生存率为39%,2年累积生存率为15%,3年累积生存率为15%,没有随访到5年的生存数据。治疗过程中未发生严重不良反应。结论:DC-CIK能使局部晚期和晚期胰腺癌患者产生抗肿瘤免疫反应,取得一定的客观疗效并可能使患者生存期延长。

Autologous cytokine-induced killer cells in equal to liver protectant in a patient with metastatic rectal cancer

The cytokine-induced killer (CIK) therapy was an effective treatment for many cancers. We report a patient with postoperative rectal cancer received autologous CIK therapy combined with raltitrexed chemotherapy. After the adjuvant therapy, the serum transaminase was persistently elevated, and lung metastases was observed. Due to hepatic injury, only cytokine-induced killer therapy was administered, and it rectified transaminase. The following regimens of CIK therapy and low-dose raltitrexed could diminish the metastatic lesion, improve the quality of life and prolong the survival time. It reveals that the CIK cells may repair the hepatic injury.

The influence of autologous cytokine-induced killer cell treatment on the objective efficacy and safety of gefitinib in advanced non-small cell lung cancer

Objective The aim of the study was to observe the influence of autologous cytokine-induced killer cell （CIK） treatment on the objective efficacy and safety of gefitinib in advanced non-small ceil lung cancer （NSCLC）. Methods Sixty-six patients with NSCLC received gefitinib as second-line treatment. They were randomly divided into 2 groups, and informed consent forms were signed before grouping. Gefitinib was administrated to the control group, and autologous CIK treatment was added to the observation group. The objective treatment and adverse reactions were evaluated in both groups. Results The objective response rate （ORR） and the disease control rate （DCR） of the observation group were slightly higher than those of the control group, although no statistical differences were found between the 2 groups （P 〉 0.05）. The incidences of diarrhea, fatigue, anorexia, oral ulcers, and myelosuppression in the observation group were much lower than those in the control group （P 〈 0.05）. However, there were no statistical differences between the incidences of skin rash, and liver and kidney toxicities （P 〉 0.05）. Conclusion Autologous CIK in combination with gefitinib is effective as second-line treatment fore ad- vanced NSCLC, and can significantly reduce adverse reactions and improve the objective efficacy.

Preliminary Study of Local Immunotherapy with Autologous Cytokine-Induced Killer Cells for Glioma Patients

OBJECTIVE Cytokine-induced killer （CIK） cells are T-cells that display effective anti-tumor activity. In this stud, we investigated the anti-tumor activity of CIK cells in vitro, and conducted a preliminary investigation using autologous CIK cells to treat glioma patients through local administration. METHODS The CIK cells were derived from peripheral blood monocytes （PBMCs） of the glioma patients. The anti-tumor activity of the CIK cells against human T98-G glioma cell was tested in vitro. In addition, the autologous CIK cells were locally administrated into the tumor cavity in the malignant glioma patients through an Ommaya reservoir which was pre-inserted during tumor resection. The 4 x 108 CIK cells in a 5 ml suspension were injected once a week 2 times per cycle. Five hundreds KU of IL-2 was injected every other day. RESULTS （i） With incubation, the CIK cells showed dual staining of CD3^＋CD56^＋ with a positive rate of 3.45% on day 10 and 55.2% on day 30. In vitro anti-tumor activity （against T98-G cells） of the CIK cells reached the highest level after 18 days of incubation with different effector/target （E：T） ratios. （ii） Six patients received autologous CIK cell treatment （10 cycles）. Two patients showed no recurrence and are still alive （24 and 10 months）, while 4 cases had a recurrence 3 of which have died. The mean survival time from the first CIK cell treatment to the end of follow-up was 12.5 months. The main side-effects of the local CIK cell treatment was brain edema, which was controlled by mannitol in most of the cases. However for one patient injection of CIK cells and IL-2 had to be discontinued. CONCLUSION In vitro CIK cells are effective anti-glioma T-cells. Local therapy with CIK cells has potential anti-glioma efficacy and tolerable side-effects.

In vitro incubation of cytokine-induced killer cells from patients with and without hepatitis B virus and a cell subset analysis

Objective The aim of the study was to explore the difference between immune cell subsets during the incubation of cytokine-induced kill cells （CIKs） from patients with and without hepatitis B virus （HBV）. Methods Peripheral blood samples were extracted from 50 tumor patients, and were divided into two groups according to the presence or absence of HBV. The proliferation rate and activity of CIK cells were examined based on counts on days 1, 5, 7, 9, 11, 13, and 15 of culture. Additionally, the CD3＋, CD4＋, CD8＋, CD3＋CD8＋, C＋）3＋CD4＋, and CD3＋CD56＋ T cell populations were analyzed by flow cytometry on days 5, 7, 10, 13, and 15 of culture. Results Proliferation over a 15-day period was higher in the HBV-positive group than in the negative group （280-fold vs. 180-fold increase, respectively）, but there was no significant difference between the two groups at each time point. The frequencies of CD3＋, CD8＋ T, CD3＋CD8＋, and CD3＋CD56＋T cells increased over time, while those of CD4＋ and CD3＋CD4＋ T cells decreased over time, and these changes were greater in the positive group than in the negative group. The differences in CD8＋ T cells and CD3＋CD4＋ T cells between the two groups were significant （P 〈 0.05）. Conclusion The proliferative capacity of CIK cells was higher for patients in the HBV-positive group than those in the HBV-negative group, and immune cell subsets were more favorable in the HBV-positive group than the neaative arouD.

DC-CIK不同输注方式治疗原发性肝癌的临床疗效和预后

目的:探讨不同途径输注DC-CIK对中晚期原发性肝癌(PLC)的治疗效果和预后价值。方法:回顾性分析2018年10月至2021年9月间东部战区总医院肿瘤科DC-CIK治疗的69例中晚期PLC患者的临床资料,根据DC-CIK治疗时采用的输注方式不同将患者分为肝动脉灌注(HAI)组(n=29)和静脉输注(Ⅳ)组(n=40),比较两组患者的临床疗效、外周血T淋巴细胞亚群(CD3^(+)、CD4^(+)、CD8^(+)T和CD4^(+)/CD8^(+)T细胞比值),以及细胞因子(IL-2、IL-6、IFN-γ和TNF-α)、甲胎蛋白(AFP)的变化、总生存期(OS)和不良反应发生情况。结果:69例PLC患者经DC-CIK治疗后,HAI组患者的客观缓解率(ORR)为0%,疾病控制率(DCR)为75.8%;Ⅳ组患者的ORR为0%,DCR为72.5%(P>0.05)。两组患者治疗前后T淋巴细胞亚群指标变化差异无统计学意义(均P>0.05),两组患者治疗后外周血IL-2、IL-6、IFN-γ和TNF-α的平均水平均显著高于治疗前(均P<0.01),两组间比较无显著差异(P>0.05);HAI组患者平均OS为48.17个月,Ⅳ组OS为39.65个月,两组间比较无显著差异(P>0.05)。治疗过程中无严重不良反应发生。结论:自体DC-CIK HAI治疗PLC安全有效,较Ⅳ治疗有提升临床获益的趋势,值得临床借鉴。

肿瘤抗原负载树突状细胞联合CIK通过促进ASK1活化增强对人食管癌细胞的杀伤活性

目的探究肿瘤抗原负载的树突状细胞(Ag-DC)联合细胞因子诱导的杀伤细胞(CIK)对食管癌肿瘤细胞杀伤的影响及作用机制。方法诱导培养外周血DC和CIK,用Ag负载DC获得Ag-DC,将Ag-DC与CIK共培养。实验分为CIK组、DC联合CIK组、Ag-DC联合CIK组。流式细胞术检测细胞表型,噻唑蓝(MTT)法测定细胞对EC9706食管癌细胞的杀伤活性,异硫氰酸荧光素标记的膜联素V/碘化丙啶(annexin V-FITC/PI)双染法检测细胞凋亡率,免疫荧光染色检测磷酸化的细胞凋亡信号调节激酶1(p-ASK1)表达,Western blot法测定ASK1途径相关蛋白水平。构建食管癌移植瘤裸鼠模型,分为对照组、DC联合CIK组和Ag-DC联合CIK组。通过尾静脉注射对应免疫细胞进行治疗,每隔2 d测量一次肿瘤体积。21 d后处死所有裸鼠,取出瘤体,HE染色观察肿瘤组织病变情况,免疫组织化学染色法检测抗原KI-67(ki67)及ASK1表达。结果与单独CIK组和DC联合CIK组相比,Ag-DC与CIK共培养后,细胞中CD3+CD8+与CD3+CD56+的比例明显升高,对EC9706细胞的杀伤率明显增加,增加EC9706细胞凋亡,并促进ASK1的活化水平;与单独CIK组和DC联合CIK组比较,Ag-DC联合CIK处理的裸鼠移植瘤生长受到明显的抑制,21 d后观察到该组肿瘤组织块较小,肿瘤组织内细胞排列较为稀疏,肿瘤组织内ki67阳性率明显减少,而ASK1阳性率则明显增高。结论Ag-DC与CIK共培养后,能够明显提高对食管癌肿瘤细胞的杀伤活性,该作用机制可能与ASK1途径的激活相关。

Harnessing cytokine-induced killer cells to accelerate diabetic wound healing:an approach to regulating post-traumatic inflammation

Impaired immunohomeostasis in diabetic wounds prolongs infiammation and cytokine dysfunction,thus,delaying or pre-venting wound-surface healing.Extensive clinical studies have been conducted on cytokine-induced killer(CIK)cells recently,as they can be easily proliferated using a straightforward,inex-pensive protocol.Therefore,the function of CIK cells in regulat-ing inflammatory environments has been drawing attention for clinical management.Throughout the current investigation,we discovered the regenerative capacity of these cells in the chal-lenging environment of wounds that heal poorly due to diabe-tes.We demonstrated that the intravenous injection of CIK cells can re-establish a proregenerative inflammatory microenviron-ment.promote larizationand.ultimatel accelerateskin healing in diabetic mice.The results indicated that CIK cell treatment affects macrophage polarization and restores the function regenerative cells under hyperglycemic conditions.This novel cellular therapy offers a promising intervention for clinical applic tions through specific inflammatory regulation functions.

肿瘤特异性个体化多靶点DC-CIK治疗晚期非小细胞肺癌的临床疗效与安全性

目的:评价肿瘤特异性个体化多靶点树突状细胞-细胞因子诱导的杀伤细胞(DC-CIK)治疗晚期非小细胞肺癌(NSCLC)患者的临床疗效和安全性。方法:回顾性分析2019年10月1日至2022年10月31日东部战区总医院生物治疗科行肿瘤特异性个体化多靶点DC-CIK治疗晚期NSCLC患者的临床资料。统计NSCLC患者的临床疗效和不良反应,分析治疗前后血清中肿瘤标志物的变化,FCM检测患者治疗前后的淋巴细胞亚群和各种细胞因子的表达情况,用质谱仪检测治疗前后靶点的变化。结果:共入组52例晚期NSCLC患者,其中女性21例、男性31例;年龄32~71岁,平均年龄(50.97±10.72)岁,中位年龄47.5岁。经DC-CIK治疗后,CR 0例,PR 0例,SD 27例,PD 25例。与治疗前比较,DC-CIK治疗后:(1)CEA和CYFRA21-1水平无显著改变,CA125水平显著低于治疗前(P<0.01);(2)治疗后患者淋巴细胞亚群无显著变化;(3)治疗后患者外周血IL-2、IL-4、IFN-γ和TNF-α水平显著升高(均P<0.01),IL-6、IL-10及IL-17水平无明显变化(;4)治疗后靶点数下降明显。DC-CIK治疗过程中无严重不良反应发生。结论:晚期NSCLC患者行肿瘤特异性个体化多靶点自体DC-CIK治疗是安全的,能使患者产生抗肿瘤免疫反应并得到一定的临床获益。

CIK细胞的体外增殖及杀瘤活性的实验研究

通过第１天在外周血淋巴细胞中加入γ－ＩＦＮ，第２天再加入ＩＬ－２、ＣＤ３单抗和ＩＬ－１，获得了已被定义为多种细胞因子诱导的杀伤细胞（Ｃｙｔｏｋｉｎｅｉｎｄｕｃｅｄｋｉｌｅｒｃｅｌｓ）即ＣＩＫ细胞。通过这种方法，使外周血中微量的ＣＤ３＋ＣＤ５６＋细胞得到大量扩增。这种ＣＤ３＋ＣＤ５６＋细胞被证明是Ｔ细胞并具有ＮＫ细胞表面标志ＣＤ５６抗原。ＣＩＫ能溶解多种肿瘤细胞，表现为非ＭＨＣ限制性杀伤，杀伤活性远高于ＬＡＫ细胞。

DC-CIK细胞的生物学活性及体外抗白血病作用的研究(英文)

本研究探讨树突状细胞(DC)对细胞因子诱导的杀伤细胞(CIK)增殖能力、免疫表型、分泌细胞因子水平及抗白血病细胞的作用。健康人外周血单个核细胞诱导DC和CIK,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,ELISA双抗体夹心法测定干扰素-γ(IFN-γ)、白介素-12(IL-12)的水平。结果表明: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+细胞比率较相同条件下CIK细胞组显著增多(p<0.05);共培养3天,DC-CIK细胞上清液中IL-12、IFN-γ水平均比CIK细胞单独培养的水平高(p<0.01,p<0.05);在5∶1-40∶1的效靶比范围内,DC-CIK细胞对白血病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增加CIK细胞的增殖能力和抗白血病活性,有可能作为一种临床有效的抗白血病免疫治疗手段。