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.

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.

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.

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.

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.

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.

Increasing the frequency of CIK cells adoptive immunotherapy may decrease risk of death in gastric cancer patients

AIM: To analyze the correlation between cytokineinduced killer (CIK) cells adoptive immunotherapy and cancer-related death in gastric cancer patients. METHODS: One hundred and fifty-six gastric cancer patients after operation at the Third Affiliated Hospital of Soochow University were enrolled in this study. Their clinical data including demographic characteristics, operation time, tumor size, pathological type and staging, tumor metastasis, outcome of chemotherapy or CIK cells adoptive immunotherapy, survival time or time of death were collected with a standard structured questionnaire. Kaplan-Meier method was used to estimate the median survival time, and the 2- and 5- year survival rates. Hazard risk (HR) and 95% confidence interval (95% CI) of CIK cells adoptive immunotherapy for gastric cancer were calculated using the two-stage time-dependent covariates Cox model. RESULTS: The survival time of gastric cancer patients was longer after CIK cells adoptive immunotherapy than after chemotherapy (χ 2 = 10.907, P = 0.001). The median survival time of gastric cancer patients was also longer after CIK cells adoptive immunotherapy than after chemotherapy (49 mo vs 27 mo, P < 0.05). The 2- and 5-year survival rates of gastric cancer patients were significantly higher after CIK cells adoptive immunotherapy than after chemotherapy (73.5% vs 52.6%, 40.4% vs 23.9%, P < 0.05). A significant difference was observed in the survival curve for patients who received CIK cells adoptive immunotherapy (0, 1-10, 11-25, and over 25 frequencies) (χ 2 = 14.534, P = 0.002). The frequencies of CIK cells adoptive immunotherapy were significantly related with the decreasing risk of death in gastric cancer patients after adjustment for sex and age of the patients, tumor stage and relapse (HR = 0.54, 95% CI: 0.36-0.80) when the first stage Cox model was used to define the subjects who remained alive beyond 36 mo as survivors. However, no correlation was observed between the frequencies of death in CIK cells adoptive immunotherapy and the risk of gastric cancer patients (HR = 1.09, 95% CI: 0.63-0.89) when the second stage Cox model was used to define the subjects who survived for more than 36 mo as survivors. CONCLUSION: The survival time of the gastric cancer patients treated with chemotherapy combined with CIK cells adoptive immunotherapy is significantly longer than that of the patients treated with chemotherapy alone and increasing the frequency of CIK cells adoptive immunotherapy seems to benefit patients more.

Clinical study of co-treatment with DC-CIK cells for advanced solid carcinomas

Objective： The aim of this study was to observe the therapeutic effect of cytokine induced killer （CIK） cells in combination with dendritic cells （DCs） on advanced solid carcinoma patients. Methods： Isolated peripheral blood mononuclear cells （PBMCs） from 110 advanced solid tumor patients. Added granulocyte-macrophage colony-stimulating factor （GM-CSF）, tumor necrosis factor-a （TNF-a） and interleukin-4 （IL-4） to adherent cells to induce DCs, and sensitized DCs with antigens of autologous tumor cells or extrinsic tumor cell lines. Cultured suspending cells with interferon-y （IFN-y）, interleukin-2 （IL-2） and CD3 monoclonal antibody （CD3 McAb） to prepare CIK cells, then co-cultured with DCs. After analyzing the phenotype and checking tumor markers and immune function, the autologous CIK cells and DCs were transfused into the cancer patients. Results： Forty-two patients with measurable nidus, 2 achieved complete remission （CR）, 9 partial remission （PR） and 15 stable disease （SD）, while 37 patients with immeasurable nidus, 25 had efficient response. The tumor markers and immune function both improved significantly compared with those before treatment. Conclusion： DCs and CIK cells combinational treatment is safe and effective on advanced solid carcinoma and provide a new and efficacious immunity therapeutic methods for the cancer patients.

The effects of CIK cells on the treatment of renal cell carcinoma

Objective： To observe the effects of cytokine-induced killer （CIK） cells on the treatment of renal cell carcinoma. Methods： Twenty-eight postoperative cases with stage Ⅰ or stage Ⅱrenal cell carcinoma were admitted in our hospital from January 2002 to June 2006, all cases were pathologically confirmed, and were divided into group A （18 cases） and group B （10 cases）. Group A was administrated 3-12 periods of CIK cells treatment combined with 5-7 cycles of IL-2 and INFα-2b, together with 4 cycles of chemotherapy （5-Fu ＋ CF）. Group B was given 4 cycles of chemotherapy （5-Fu ＋ CF） and 5-7 cycles of IL-2 and INFa-2b. Results： Three cases in group A had metastatic masses in two lungs within 1 year and died within 2 years postoperatively. The other 15 cases are still alive and in good health. Six cases in group B had metastatic masses in two lungs or/and in abdominal cavity within 1 year, and 4 of them died within 2 years. All the 6 cases died within 3 years. The other 4 cases are still alive and in good health. Conclusion： CIK cells are safe and effective for the treatment of stage I or stage II renal cell carcinoma, which should be further widely used.

The clinical effects of DC-CIK cells combined with chemotherapy in the treatment of advanced NSCLC

Objective: The aim of the study was to evaluate the safety and therapeutic effects of autologous dendritic cells co-cultured with cytokine-induced killer cells (DC-CIK) combined with chemotherapy in advanced non-small cell lung cancer (NSCLC) patients. Methods: Fifty patients with advanced NSCLC (stages III to IV), who had received therapies in our Center (Department of Biotherapy, Affiliated to Cancer Hospital of Shanxi Medical University, Taiyuan, China) from August 2008 to January 2010, were treated by DC-CIK + chemotherapy as the combined treatment group; fifty advanced NSCLC patients treated with chemotherapy at the same time served as controls. The immunologic function, short-term therapeutic effects, the 1-year survival rate, the life quality, the chemotherapy side effects were compared between the two groups, the safety and therapeutic effects of DC-CIK cells therapy were observed too. Results: There was no obvious change of subsets of T cells in peripheral blood before and after therapy in DC-CIK + chemotherapy group, and IFN-γ was improved after therapy in this group (P < 0.05); in chemotherapy alone group, the ratios of CD3+CD4+, CD3+CD8+, CD3-CD56+ cells and the secretion of IL-2, TNF-α decreased significantly after therapy (P < 0.05); the ratios of CD3+CD8+, CD3+CD56+ were improved after cell culture (P < 0.05). The disease control rate (DCR) of DC-CIK + chemotherapy group was higher than that in the chemotherapy alone group (78.0% vs 56.0%, P < 0.05); the 1-year survival rates of DC-CIK + chemotherapy group and chemotherapy alone group were 50% and 44% respectively, had no significant difference. Compared with chemotherapy alone group, the occurrence of chemotherapy side effects (including bone marrow suppression, nausea and vomiting, peripheral nerve toxicity) was less in the DC-CIK + chemotherapy group (P < 0.05). The physical and appetite were better in DC-CIK + chemotherapy group after therapy. Conclusion: To compare with simple chemotherapy, DC-CIK + chemotherapy for advanced NSCLC is safe and effective, and it can improve patients' life quality and remission rate, and prolong their survival time.

Research on the biological activity and anti-tumor effect against lymphoma cells of DC-CIK cells

Objective： To investigate the proliferation capabilities, immunophenotype changes, level of secreted cytokines and activities against lymphoma cells under the condition that cytokine-induced killer （CIK） cells co-cultured with dendritic cells （DC） in vitro. Methods： DC and CIK cells were induced from peripheral blood mononuclear cells of healthy volunteers. They were co-cultured meanwhile CIK cells were cultured alone as controls. Increased number of cells were counted by tapan-blue staining, killing activities were detected by MTT assay, immunophenotype changes were analyzed by flow cytometry, the IL-12 and INF-y levels of the cultured supernatants were detected by ELISA kits. Results： The proliferation capabilities of DC-CIK cells were significantly higher than that of CIK cells （P 〈 0.05）. Under the same condition, the ratio of double positive cells such as CD3^＋ CD8^＋, CD3^＋ CD56^＋ in CIK cells was significantly enhanced by co-cultured with DC cells （P 〈 0.05）. The level of IL-12 and INF-y secreted in supernatants was increased noticeably by co-cultured DC-CIK cells on day 3 compared to CIK cells which were cultured alone （P 〈 0.01 and P 〈 0.05）. Within the effector-target ratio range between 5：1 to 40：1, the activities against lymphoma cells of DC-CIK cells were much higher than that of CIK cells （P 〈 0.05）, and this effect was showed a positive correlation with the effector-target ratio. Conclusion： The proliferation capabilities, the level of secreted cytokines and the activities against lymphoma cells of DC-CIK cells were significantly higher than those of CIK cells. The research might provides theoretical and experimental basis for clinical immunotherapy of DC-CIK cells.

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。

Effect of IL-12 on the proliferation and cytotoxicity of CIK cells to gastric adenocarcinoma cell BGC-823

Objective: The aim of the study was to evaluate the effect of interleukin-12(IL-12) on the proliferation and cytotoxicity of cytokine-induced killer(CIK) cells in vitro. Methods: Three different combinations of cytokines, IL-2, IL-12 + IL-2, and IL-12, were used to proliferate CIK cells, adding IFN-γ, IL-1 and CD3 McAb in each one. Phenotype of the cells was analyzed by flow cytometry. The cellular proliferation and cytotoxic activity were determined by cytometry and MTT assay. Results: CIK cells generated by the three methods showed high reproductive activity, and no obviously difference in inducing CD3+CD56+ cells was found among the three groups. The group of IL-2 + IL-12 evidently enhanced both the proliferation and the cytotoxicity of the CIK cells compared with the other two groups(P < 0.05). Conclusion: IL-12 could be used to induce the CIK cells as well as IL-2. CIK cells induced by combining IL-12 with IL-2 had stronger proliferative ability and higher cytotoxicity to tumor cells in vitro, which could be used as a potential anti-tumor adoptive immunotherapy in clinic.

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。

Side Effects during Treatment of Advanced Gastric Carcinoma by Chemotherapy Combined with CIK-cell Transfusion in Elderly People

OBJECTIVE To study the side effects and therapeutic results of autologous cytokine-induced killer （CIK） cell treatment in elderly patients with advanced gastric cancer. METHODS CIK cells were induced and cultured using biotechnics in vitro, and then the cells were infused back into the patients. Sixty elderly gastric cancer patients treated by chemotherapy （FOLFOX4 protocol） were followed-up. Among them, 29 patients were treated with CIK cells during application of chemotherapy. Short-term curative effects and adverse events from the CIK transfusion and chemotherapy were observed. RESULTS Eight cases developed partial remission （PR）, 9 cases moderate remission （MR）, 7 cases stable disease （SD） and 5 cases progressive disease （PD）. Out of a total of 29 patients who received chemotherapy combined with autologous CIK therapy, the total remission rate （PR ＋ MR） was 58.6%. The total remission rate following chemotherapy alone was 45.2%, including 5 PR cases, 9 MR cases, 7 SD cases, and 10 PD cases. There was a relatively lower rate of severe chemotherapic toxicities in the CIK-cell transfusion group. Side effects of autologous CIK transfusion included chills （13 cases）, fever （9 cases）, nausea and vomiting （1 case） and general malaise （3 cases）. Side effects were treated with conventional therapy resulting in their amelioration. No patients developed shock, blood capillary leakage syndrome, or abnormalities in routine blood, urine, liver and renal function tests.CONCLUSION Adoptive immunotherapy with autologous CIK cells may decrease the clinical signs and symptoms of elderly patients who suffer from advanced gastric cancer. Adverse reactions of patients can be alleviated by conventional therapy. Autologous CIK-cell transfusion may improve endurance to chemotherapy.