Assessment of lymphocytic function in vitro stimulated by specific tumor polypeptide combined with dendritic cells / 北京大学学报(医学版)

OBJECTIVE@#To assess the activation function of specific tumor polypeptide for dendritic cell vaccine on lymphocytes proliferation, production of cytokines and killing activity in vitro by using dendritic cells as antigen presenting vector.@*METHODS@#Peripheral blood dendritic cells (DC) and cytokine-induced killer (CIK) were isolated and cultured by adherent culture method; CCK-8 method was used to assess the proliferation function of lymphocytes and the killing function of lymphocytes to tumor cells; enzyme-linked immunospot assay method was used to evaluate the secretion function of cytokines. The experiment was divided into tumor polypeptide group (peptide with DC-CIK), DC-CIK group and CIK group.@*RESULTS@#With presence of interleukin-2 (IL-2) in the culture system, the lymphocyte proliferation of the three groups was obvious. The absorbance at 450 nm of tumor polypeptide group was significantly higher than that of CIK group at the time points day 4 and day 6 (day 4: Z=-3.79, P < 0.001; day 6: Z =-2.95, P < 0.01). The absorbance at 450 nm of group tumor polypeptide was significantly higher than that of DC-CIK group on day 4 (Z=-2.02, P < 0.05). Without IL-2 in the culture system, lymphocytes proliferated slowly in all the three groups, and there was no significant difference in 450 nm absorbance at each time point. The levels of IL-4 (Z=-2.61, P < 0.01), granulocyte-macrophage colony-stimulation factor (GM-CSF, Z=-3.85, P < 0.001), interferon- γ (IFN- γ, Z=-3.56, P < 0.001) and tumor necrosis factor-α (TNF-ɑ, Z=-3.40, P < 0.001) of tumor polypeptide group were higher than those of CIK group. There was no significant difference in the production of cytokines except IL-4 (Z=-2.15, P < 0.05) when tumor polypeptide group was compared with DC-CIK group. The production of IFN-γ (Z=-2.44, P < 0.05), TNF-ɑ (Z=-2.26, P < 0.05) and GM-CSF (Z=-3.73, P < 0.001) in DC-CIK group were higher than those of CIK group. Although there was no significant difference in killing activity between tumor polypeptide group, DC-CIK group and CIK group at hour 18 and hour 24, and the killing activity of tumor polypeptide group was higher than that of the other two groups.@*CONCLUSION@#Tumor peptide combined with dendritic cells can improve the proliferation activity of CIK cells in vitro, and increase the secretion of several cytokines.

Influence of PD-1 monoclonal antibody on anti-lung cancer effect of cytokine-induced killer cells (CIK) induced and expanded in vitro / 中国胸心血管外科临床杂志

@#Objective    To investigate the influence of programmed cell death protein-1 (PD-1) monoclonal antibody on the anti-lung cancer effect of cytokine-induced killer cells (CIK) which were programmed in vitro. Methods    Peripheral blood mononuclear cells from 20 patients (8 males and 12 females with an average age of 56.45±5.89 years ranging from 42 to 65 years) diagnosed with advanced lung cancer from January to May 2019 at the Department of Oncology of Dalian Central Hospital were collected and induced to amplify into CIK cells in vitro. PD-1 monoclonal antibody combined with CIK cell culture group, individual cell culture group and PD-1 monoclonal antibody group were set up to detect the cell killing activity of CIK cells against lung cancer under different effective target ratio conditions, and the ratio of perforin and granzyme positive expression in PD-1 monoclonal antibody combined CIK cell culture group and individual CIK cell culture group was detected by flow cytometry. ELISA method was used to detect the interleukin-2 (IL-2), interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) cytokine secretion levels in the two groups. Results    The killing effect of CIK cells on A549 lung cancer cells increased with the increase of effective target ratio by CCK8, and PD-1 monoclonal antibody increased the killing effect of CIK cells on A549 lung cancer cells under different effective target ratio, E∶T=5∶1 (28.5%±1.9% vs. 20.3%±1.8%), 10∶1 (40.6%±2.4% vs. 31.7%±2.1%), 20∶1 (57.4%±3.5% vs. 44.7%±3.8%), 40∶1 (74.1%±8.3% vs. 60.8%±5.3%). The killing effect of PD-1 monoclonal antibody combined with CIK cells and CIK cells alone on A549 lung cancer cells was statistically different (P<0.05). The killing effect of cells in both groups on lung cancer A549 cells was stronger than that of the PD-1 monoclonal antibody group (P<0.01). The results of flow cytometry showed that PD-1 monoclonal antibody increased the positive ratio of perforin and granzyme release in CIK cells, and the positive ratios of perforin release (46.7%±3.5%% vs. 35.1%±2.2%) and granzyme release (34.6%±3.8% vs. 25.7%±3.3%) in PD-1 monoclonal antibody combination with CIK cells group and CIK cells group were statistically different (P<0.05). Similarly, the secretion levels of IL-2, TNF-α, and IFN-γ cytokines were also increased in the PD-1 monoclonal antibody combined with CIK cells group compared with the CIK group (5 409.0±168.8 pg/mL vs. 4 300.0±132.3 pg/mL, 252.7±16.7 pg/mL vs. 172.5±8.6 pg/mL, 327.2±23.5 pg/mL vs. 209.7±16.0 pg/mL, P<0.05). Conclusion    PD-1 monoclonal antibody can promote the release of tumoricidal substances in CIK cells and improve the killing effect of CIK cells on lung cancer A549 cells. It is speculated that the infusion of PD-1 monoclonal antibody before CIK cell adoption in lung cancer patients may be more beneficial to the treatment of disease. PD-1 monoclonal antibody combined with CIK cell therapy is promising as a new type of lung cancer immunotherapy.

Long-term efficacy of dendritic cell vaccines combined with cytokine-induced killer cells in the treatment of metastatic renal cell carcinoma / 中国肿瘤生物治疗杂志

@# Objective: To evaluate the long-term clinical efficacy and follow-up of dendritic cell (DC) vaccines in combination with cytokine-induced killer cell (CIK) treatment in metastatic renal cell carcinoma. Methods: From January 2011 to December 2013, 29 patients with metastatic renal cell carcinoma (pathologically confirmed as renal clear cell carcinoma) were treated by DC vaccines-CIK at the Department of Hematopoietic Stem Cell Transplantation, the Fifth Medical Center of Chinese PLA General Hospital. The 29 patients included 24 male and 5 female, with a median age of 57(32-81) years old. Mature DC vaccine was obtained by gene transfection technology and CIK cells were obtained by i n v i t r o culture; and DC vaccine-CIK was infused back to patients through lymphatic drainage area and vein by each course. Twelve patients received first line treatment, 6 patients received second line treatment after the disease progression by targeted drug therapy or cytokine therapy, and 11 patients received third-linetreatment or above. The long-term clinical efficacy and overall survival rate were evaluated. Results: The median follow-up time was 5 (1-7) years. Treatment cycle was over 2 (2-23) cycles. One case (3.4%) achieved complete remission, 9 cases (31%) achieved partial responses, 13 cases (44.8%) demonstrated stable disease over 3 months and 6 patients (20.7%) developed progressive disease. The objective response rate was 34.4%,and the disease control rate was 79.2%. Stable disease for more than one year realized in 19 cases (65.5%). The 1-, 3- and 5-year survival rates were 93.1% (27/29), 65.5% (20/29) and 51.7% (15 / 29), respectively. Neither the median progression-free survival (PFS) nor the median survival time was achieved. No adverse reactions above grade 3 were observed during treatment. Conclusion: DC vaccines-CIK therapy for the treatment of metastatic renal cell carcinoma is affirmative; it achieved good disease control and long-term survival with controllable safety, and prolonged the survival time for advanced renal cell carcinoma patients.

Treatment efficacy of DC-CIK with different loading combined with 5-FU on nude mice bearing colon cancer HT-29 cell transplanted tumor / 中国肿瘤生物治疗杂志

@# Objective: To investigate the therapeutic effect of dendritic cell-induced killer cells (DC-CIK) combined with 5-fluorouracil (5-FU) and loaded with CD133+ HT-29 cell lysate or RNA on mice bearing colon cancer HT-29 cell transplanted tumor, and to explore the underlying mechanism. Methods: Colon cancer xenograft model was established in BALB /c nude mice by using human colon cancer HT-29 cells at logarithmic growth phase; Antigen-free DC-CIK, 5-FU+DC-CIK, R+DC-CIK (loaded with total RNA of CD133+ cells), L+DC-CIK (loaded with CD133+ cell lysate), 5-FU and normal saline were respectively injected into transplanted mice, and the treatment efficacies on the growth of transplanted tumor in each group after three treatment cycles were observed, and the tumor growth curve was drawn. The nude mice were sacrificed by cervical dislocation and the tumor volume and body weight were measured. qPCR was used to detect the expression of AKT mRNA in transplanted tumor tissue, and WB was used to detect the expression of phosphorylated AKT protein. Results: After treatment, the body mass of nude mice in R+DC-CIK group, L+DC-CIK group and 5-FU+DC-CIK group increased steadily, while the body mass of nude mice in DC-CIK group and 5-FU group decreased gradually; the tumor growth speed of nude mice in R+DC-CIK group, 5-FU+DC-CIK group and L+DC-CIK group was significantly slower than that of the control group (P<0.05). Compared with 5-FU and DC-CIK alone, the combined treatment with loaded lysate/RNA had more sig nificant effect on mRNA and protein expressions of AKT(P<0.05). Conclusion: The effect of DC-CIKwithdifferentloadingoritscombinationwith5-FUisbetterthanthatofchemotherapy alone. One of the mechanisms is related to the down-regulation ofAKT level.

Inhibitory effects of CIK on lung cancer A549 cells intensified by chemotherapyinduced damaged associated molecular pattern / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of DAMP (damaged associated molecular pattern) on the inhibition of RAS-mutant A549 lung adenocarcinoma cells by CIK cells and its mechanism. Methods: Human peripheral blood mononuclear cells were isolated in vitro and CIK cells were cultured. A549 cells were treated with cisplatin (DDP) and doxorubicin (ADM) alone or in combination, and the morphology of A549 cells was observed under a microscope. The supernatant of A549 cells was co-cultured with CIK cells. Flow cytometry was used to detect the CIK cell immunophenotype after co-culture. MTT assay was used to detect the inhibition of A549 lung cancer cell proliferation induced by A549 cell supernatant. The concentration of chemotherapeutic drugs kills A549 cell supernatant CRT, ATP, HMGB1 content. Results: Low-level chemotherapeutic drugs showed more immunogenic death characteristics after killingA549 cells. The ratio of CD8+ and CD56+ in CIK cells was significantly higher than that in control CIK cells (P<0.05). The inhibition rate of CIK cells induced byA549 cells after injury onA549 lung adenocarcinoma cells was significantly higher than that of the same dose chemotherapy group [DDP group (31.34±1.51)% vs (5.97±1.74)%, ADM group (45.46±1.78)% vs (6.22±1.34)%, DDP+ ADM group (45.78±1.14)% vs (11.94±3.11)%, all P<0.05], and low-mass chemotherapeutic agents killed C549 induced by A549 cell supernatant on A549 The inhibition rate of the cells was higherthan that of the supernatant induced by the higher concentration of chemotherapeutic drugs (all P<0.05). The level of CRT,ATP, and HMGB1 in immunogenicity-related molecules in the supernatant ofA549 cells was significantly increased by low-concentration chemotherapy drugs (all P<0.05). In the low-concentration group, the supernatant-induced inhibition of the proliferation of A549 lung adenocarcinoma cells increased with the increase of CRT, ATP, and HMGB1 levels. Conclusion: The combination of lower concentration of DDP and ADM alone or in combination could more easily induce the immunogenic death of A549 cells and release higher levels of DAMP molecules, which could promote the inhibitory effect of CIK on lung cancerA549 cells.

Clinical efficacy of DC-CIK immunotherapy in treatment of advanced pancreatic adenocarcinoma / 中国肿瘤生物治疗杂志

@#［ ［Abstract］ ］Objective: To analyze and compare the clinical efficacy and safety of dendritic cell cytokine-induced killer cells (DCCIK) combined with palliative therapy or chemotherapy in the treatment of advanced pancreatic carcinoma. Methods: A retrospective study was carried on 50 patients with advanced pancreatic carcinoma who were hospitalized in department of oncology of Shanxi Dayi Hospital during September 2012 to February 2016. The patients were divided into four groups according to the therapy they received (palliative treatment group, palliative+DC-CIK treatment group, chemotherapy group and chemotherapy+DC-CIK treatment group); the immunological function, quality of life and survival time of patients were analyzed; and the efficacy and safety of DC-CIK cell therapy was also evaluated. Results: The percentages of CD8+ T cells and NKT cells in DC-CIK combined therapy groups were significantly improved compared with that of pre-treatment, and the percentages of CD3+, CD8+, NK, NKT cells were increased compared with control groups (P<0.05). The quality of life of patients was significantly improved (P<0.05), while median PFS and median OS were improved but without statistical significance (P>0.05). Conclusion: Compared with palliative therapy and chemotherapy alone, combined DC-CIK immunotherapy can effectively improve the cellular immunity function and quality of life in patients with advanced pancreatic cancer. However, there was no significant extension in overall survival.

Clinical observation on Astragalus Polysaccharide Injection combined with CIK cells treating NSCLC patients with qi deficiency syndrome / 中草药

Objective: To explore the synergetic effect and observe the safety of Astragalus Polysaccharide Injection combined with cytokine-induced killer cells (CIK cells) in the treatment of advanced NSCLC patients with qi deficiency syndrome. Methods: A total of 75 advanced NSCLC patients with qi deficiency syndrome in oncology department of First Teaching Hospital of Tianjin University of TCM enrolled in the study were randomized into two groups: the control group (CIK cells group) and the combined group (Astragalus Polysaccharide Injection + CIK cells group) by the random number table method. The control group: CIK cells were transfused in vein (100 mL each time, once every Monday, Wednesday, and Friday, five times in total, the total number of cells > 1× 1010/mL). The combined group: CIK cells treatment combined with astragalus polysaccharide injection intravenous drip (250 mg Qd, 10 d). Ten days is one cycle, both groups were received the second cycle treatment in a month later. Results: The combined group's disease control rate (DCR=CR + PR + SD) was 69.4%, higher than that of control group's (36.1%), and the difference was statistically significant (P < 0.05).The KPS of combined group's effective rate was 77.8%, higher than 55.6% in the control group, the difference was statistically significant (P < 0.05). After treatment, shortness of breath, spiritlessness, hypodynamia, spontaneous perspiration and speaking reluctantly had taken a turn for the better in combined group, especially in spiritlessness, hypodynamia, spontaneous perspiration, gospeaking reluctantlyt to improve obviously, and the difference was statistical significance (P < 0.05). The haematologic toxicity in II, III, IV degree and hepatic and renal toxicity in III, IV degree did not appear in two groups. Conclusion: The Astragalus Polysaccharide Injection combined with CIK cells could control tumor lesion's progress, and improve the patient's immune function, the symptom of qi deficiency syndrome, and body functional status due to itsbetter security.

Killing effect of Huaier combined with DC-CIK on nude mice bearing colon cancer HT29 stem cells / 中国中药杂志

To compare the therapeutic effects of different treatment methods on the nude mice bearing colon cancer HT29 cells. BalB/C nude mice colon cancer stem cell models were established and randomly divided into the following four groups, with 8 nude mice in each group: blank control group, DC-CIK group, Huaier group, and Huaier combined with DC-CIK group (combined treatment group). The mice in DC-CIK group and combined treatment group received 1×10⁶ DC-CIK cells treatment by tail vein injectionafter the tumor stem cells were inoculated for 4 days,2 times a week for three weeks. The mice in Huaier group and combined treatment group received intragastric administration at the dose of 20 g/60 kg body weight, 0.2 mL/time, once a day for a total of three weeks. The mice in control group received equal volume of normal saline. Tumor size and body weight of nude mice were measured every 2 days during treatment for three weeks in each group. After the treatment, the nude mice were sacrificed to measure the tumor weight and the tumor inhibition rate was calculated. The RT-PCR method was used to detect the expression levels of the key genes in the signal pathway. After the end of the treatment, the quality of the tumor in the Huaier group, DC-CIK group and combined treatment group was significantly lower than that in the control group; the quality in combined treatment group was significantly lower than that in Huaier group and DC-CIK group.Among them, the tumor inhibition rate reached 46.77% in the combined treatment group. In respect of changes in expression levels of key genes in the signaling pathway, the mRNA expression levels of key genes PI3KR1 and Akt in PI3K/Akt pathway, key genes Wnt1 and CTTNB1 in Wnt/-catenin pathway, and key genes Notch1, Notch2, Notch3 in Notch pathway in the combined treatment group were lower than those in DC-CIK group and Huaier group. The Huaier combined with DC-CIK group showed best therapeutic effect among different treatment methods for HT29 stemcell colon tumors in nude mice, providing a new idea for clinical treatment of colon cancer.

Phenotypes of CIK cells prepared by ATG-F culture system and its killing effect against K562 cells / 中国肿瘤生物治疗杂志

@# Objective: To investigate the function of CIK (cytokine induced killer) cells cultured using ATG-F (anti-human T lymphocyte rabbit immunoglobulin-Fresenius) and IFN- γ, IL-2 system and its feasibility in clinical practice. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from healthy donors and were used to culture CIK cells by different activating antibodies; the total cell count was calculated on Day 7 and 14. The CIK cell composition, cell surface activation and proportion of inhibitory receptor molecular in ATG-F group, CD3 group and TG (Thymoglobulin) group were analyzed by Flow cytometry, and the cytotoxicity of CIK cells against K562 cells were also determined by flow cytometry at day 14 in ATG-F high-dose group, CD3 group and TG group. Results: CIK cells were successfully cultured by ATG-F, IFN-γ, IL-2 system. The proliferation rate of ATGF high-dose group was significantly higher than that in TG group (27.25±1.25 vs 16.60±1.72, P <0.01), but the proportion of CD3+ CD56+ cells showed no statistical difference compare with the CD3 group ( P >0.05). The percentage of CD3-CD56+ NK cells in ATG-F high-dose group was significantly higher than that in TG group and CD3 group [(11.19±2.60）% vs（5.66±1.00）%,（1.42± 0.51）% ， P <0.01], while the proportion of CD4+T cells was significantly lower than that in CD3, TG group [(4.35±1.47）% vs （26.88±5.01）%,（14.52±6.22）%, P <0.01]; the proportion of CD56+CD94+, CD56+CD158a+, CD56+CD158b cells was significantly higher than those in CD3 group (all P <0.01). The ATG-F high does group showed significantly higher cytotoxicity against K562 cells than that of CD3 group at the target/effect ratio of 1∶10. Conclusion: CIK cells cultured by ATG-F culture system has higher NK cell proportion than other ordinary culture system, and its activated receptor has more stronger cytotoxicity against K562 cells.

Study on the effect of IL-12 and IL-15 on the proliferation of CIK cells from of peripheral blood and the activity of killing SMMC-7721 hepatoma cell line in vitro / 国际检验医学杂志

Objective To observe the influence of IL-12,IL-15 on CIK cell in the normal culture;to observe the anti-tumor effect in the circumstance of different combination of cytokines,and to provide a new insight for preparing high effective and qualified CIK cell in vitro.Methods The optimal concentrations of IL-2,IL-12 and IL-15 were determined,respectively.After the peripheral blood from healthy blood donors was collected,monocytes were selected and co-cultured with different cytokines into different groups,as group A(IL-2 normal culture group),group B(IL-2 and IL-12 group),group C(IL-2 and IL-15 group),group D(IL-2,IL-12 and IL-15 group),and group E(cytokine control group).The monocytes in different groups were calculated by globulimeter,the activity of cells was detected by Trypan blue staining,positive ratio of CD3,CD8,CD56 on the celluar membrane was detected by flow cytometry,and the anti-tumor effect of CIK to SMMC-7721 was detected by MTSmethod,inthedayof0,5,10,15,20 after the culture.Results Statistical analysis indicated that,the proliferation multiplication of CIK cells was significantly higher in group B,group C and group D after 10,15 and 20 days of culture than those in group A(P＜0.05);and group D had higher proliferation multi-plication than that of group C(P＜0.05).The percentage of CD3 + CD8 +,CD3 + CD56+ in CIK cell membrane in group B,C,D was significantly higher than that in group A after 15 and 20 days of culture (P＜0.05).The percentage of CD3+ CD8+,CD3+ CD56+ in CIK cell membrane in group D was significantly higher than that in group B after 15 and 20 days of culture (P＜0.05).The killing rate of CIK cells for liver cancer in each group at 10,15,20 days of culture was significantly higher than that of group A when the target target ratio was 5 ∶ 1 (P＜0.05).The killing rate of CIK cells for liver cancer in group D,C at 10,15,20 days of culture was significantly higher than that of group B(P＜0.05).Conclusion IL-12and IL-15 could improve the proliferation of CIK cells,and IL-15 also has the effect of enhancing CIK cells the tumor-killing to SMMC-7721 activity.

Clinical observing of advanced non-small cell lung cancer patients treated with CIK cell combined with chemotherapy / 中国生化药物杂志

Objective To observe the efficacy of advanced non-small cell lung cancer patients treated with CIK cell combined with chemotherapy.Methods65 cases of advanced lung cancer patients admitted in our hospital from February 2013 to January 2015 were retrospectively analyzed.The patients were randomly divided into observation group (34 cases) and control group (n=31).The levels of vascular endothelial growth factor (VEGF), matrix metalloproteinase-9 (MMP-9) and carcinoembryonic antigen (CEA) in the two groups were observed before and after treatment.The patients in the control group were treated with chemotherapy on the basis of chemotherapy.Cytokeratin 19 (CYFRA21-1) levels, and clinical efficacy.Results①The patients with VEGF and MMP-9 values after treatment (298.67±36.49)ng/L, (1102.31±86.48)ng/mL, than the control group (459.19±45.28)ng/L, (1394.83±121.06)ng/mL, and the difference was significant (P<0.05).②CEA and CYFR21-1 value patients in the observation group were (14.16±1.08) ng/mL, (4.45±0.45) ng/mL, than the control group (17.86±1.84) ng/mL, (5.76±0.64) ng/mL, and the difference was significant (P<0.05).③The total observation group was significantly higher than patients 94.18% 74.19% efficiency, and the difference was statistically significant (P<0.05).ConclusionCIK cell biological therapy combined with chemotherapy for advanced lung cancer patients with a certain improvement is worthy of further research and application.

Therapeutic effect of DC-CIK cells combined with S-1 in the treatment of non-small-cell lung cancer / 国际生物医学工程杂志

Objective To observe the clinical effect of DC-CIK cells combined with S-1 in the treatment of non-small-cell lung cancer.Methods 76 patients with non-small-cell lung cancer were randomly divided into the observation group and the control group on average.The control group was treated with S-1,and the observation group was treated with DC-CIK cells combined with S-1.For the observation group,the peripheral venous blood was collected before the treatment for DC cells and CIK cells cultivation.The expression of CD4+/CD8+,CD4+ and NK cells in the peripheral blood of the two groups was detected by flow cytometry before the treatment and after one week of the treatments.Besides,the number of white blood cells and platelet count were also measured and the symptoms of non-small-cell lung cancer were observed.Results The percentage of CD4+/CD8+,CD4+ and NK cells in the peripheral blood of the observation group after the treatment was (1.65±1.03),(34.56±8.90) and (18.68±7.98),respectively,which was significantly higher than (1.32±0.70),(29.07±7.15) and (15.28±8.23) in the control group (all P＜0.05).There was no significant difference in the percentage of CD8+ cells between the observation group (25.56± 8.90) and the control group (26.64±6.77) (P＞0.05).The vomiting,leukopenia and thrombocytopenia in the observation group were less than those in the control group (all P＜0.05).The one-year survival rate was 52.63％ in the observation group which was significant higher than 42.11％ in the control group (P＜0.05).Conclusions DC-CIK cells combined with S-1 is a safe and effective treatment for non-small-cell lung cancer,which can effectively improve the clinical efficacy and prolong the survival time of patients.

In vivo antitumor effect of Chinese yam extract combined with DC-CIK on nude mice bearing colon cancer HT29 stem cells / 中草药

Objective: To compare the therapeutic effects of different treatments on the nude mice bearing colon cancer HT29 cells. Methods: BalB/C nude mice colon cancer stem cell model was established and the mice were randomly divided into following four groups: blank control group, DC-CIK group, yam group, and yam combined with DC-CIK group (combined treatment group), each group of 10 nude mice. In DC-CIK group and combined treatment group, after tumor stem cells were inoculated in nude mice for 4 d, 1 × 106 DC-CIK cells were used through the tail vein injection for treatment, twice a week for three weeks. The mice in Chinese yam group and combined treatment group were ig administered with 125 mg/kg Chinese yam extract, once daily for three weeks, and the mice in control group were treated with equal volume of saline instead. Tumor size and body weight of nude mice were measured every 2 d during treatment for three weeks in each group. After the treatment, the nude mice were sacrificed to take out the tumor weight and the tumor inhibitory rate was calculated. The RT-PCR method was used to detect the expression levels of the key genes in signal pathway. Results: After treatment, the quality of tumor in yam group, DC-CIK group and combined treatment group was significantly lower than that in control group. Among them, the inhibitory rate was 51.26% in combined treatment group. In the changes of expression levels of key genes in signaling pathway, the key genes of PI3KR1 in PI3K/Akt pathway, the key gene of Wnt1 in Wnt/beta-catenin pathway, and the mRNA expression of key gene Notch1 in Notch pathway in combined treatment group were decreased compared with DC-CIK group and yam group. Compared with the DC-CIK group, the mRNA expression changes were not statistically significant. Conclusion: The effect of Chinese yam combined with DC-CIK group is the best on the treatment of HT29 cell stem cell tumor of colon cancer in nude mice. It provides a new idea for clinical treatment of colon cancer.

Effects on phenotype and function of mouse CIK cells treated with carboxylated single-walled carbon nanotubes / 中国免疫学杂志

Objective:To investigate the effect on the phenotype and function of CIK cells induced from splenic lymphocyte in mice treated with carboxylated single-walled carbon nanotubes.Methods:CIK cells were treated with carboxylated single-walled carbon nanotubes at different concentrations.Cell growth status was observed using inverted fluorescence microscopy.The killing effect was de-termined by MTS method.Immunophenotype was analyzed by flow cytometry.Results: With the increase of the effect target ratio,the killer rate of CIK was gradually enhanced and the optimal effect target ratio was 20:1.With the increase of the dose of carboxylated single-walled carbon nanotubes,the proportion of Treg cells decreased.When CIK cells were treated with 0.5 μg/ml carboxylated single-walled carbon nanotubes(CNTs),the proportion of CD3+CD4+and CD3+CD8+was significantly higher than that of the control group(P<0.05),and the killing effects of CIK anchieved best results to B16 cells,H22 cells and RM-1cells.Conclusion: The carboxylated single-walled carbon nanotubes enhanced the ability of CIK cells to kill tumor cells,significantly which provide has potential value in tumor drug development.

Research progress of cellular immunotherapy for hematological malignancies / 实用肿瘤学杂志

In recent years,considerable progress has been made in the treatment of hematologic malig-nancies.However,most patients will eventually relapse and suffer from severe side effects caused by chemotherapy or radiation.Thus,as a well tolerated,safe,effective and innovative therapy,adoptive cellular immunotherapy e-merges at this very moment.This therapy can kill tumors or control recurrence by reinfusing the in vitro amplifi-cated or treated immune effective cells into the patients.It has been actively used in the treatment of various he-matological malignancies and achieved significant effects,which brings hope for more and more patients with re-lapsed or refractory hematological diseases.

The relationship between DC-CIK and colorectal cancer prognoses and its influen-cing factors / 药学实践杂志

Objective To discuss dendritic cell-cytokine-induced killer (DC-CIK ) cell therapy effects and clinical out-comes in patients with colorectal cancer in order to have better clinical treatment .Methods A retrospective analysis of the data of 66 patients with colorectal cancer from the Biological Therapy Department of the Eastern Hepatobiliary Surgery Hospital was performed from January 2012 to January 2014 ,and then was followed up .Taking gender ,age ,degree of pathological differen-tiation ,TNM staging ,surgical methods ,and targeted therapy as the research basis ,by the Kaplan-Meier single factor and Cox multiple factors analysis we mainly discuss the DC-CIK cell treatment′s effect on the prognoses of patients .Results Kaplan-Meier single factor analysis results indicate :to a certain extent ,DC-CIK cell therapy can improve the prognoses of patients ;Cox multi-factor analysis results indicate whether accepting DC-CIK cell therapy is an independent factor influencing the prog-noses of patients .Conclusion DC-CIK cells therapy can improve the prognoses of patients with colorectal cancer .

Advances in immunotherapy of gastric cancer / 中国生化药物杂志

500 thousands people died from gastric cancer in China over 2015,shortage of clinical medicines and therapeutic methods is one of the reasons for the high mortality.Tumor immunotherapy is the hottest area of research over the past decade,whose success in hematological malignancies makes people raised their confidence to “cure” cancer.In recent years,gastric cancer immunotherapy research has made some achievements,while clinical outcomes still need improvement.In this paper, the progress of clinical research of common immunotherapeutic drugs and methods in several fields of gastric cancer research are reviewed to discuss the development prospects of immunotherapy for gastric cancer .

Transarterial chemoembolization combined with autologous DC-CIK cells for the treatment of hepatocellular carcinoma of BCLC C-stage：a randomized controlled study / 介入放射学杂志

Objective To investigate the efficacy of transarterial chemoembolization (TACE) combined with autologous DC-CIK cells in treating hepatocellular carcinoma(HCC) of BCLC C-stage. Methods A total of 60 cases with HCC in BCLC C-stage were randomly and equally divided into the study group (n=30) and the control group (n=30). TACE combined with autologous DC-CIK cells was employed in the patients of the study group, while only TACE was adopted in the patients of the control group. The immune function, six-month and one-year survival rates were determined, and the results were compared between the two groups. Results In the study group, the blood T lymphocyte subsets of CD3+CD8+ were significantly increased, while CD3+CD4+ were obviously decreased. When compared with the pretreatment levels, the differences were statistically significant (P<0.05). The six-month survival rate of the study group and the control group was 67.9% and 48.1% respectively (P<0.05), and the one-year survival rate of the study group and the control group was 53.6%and 29.6%respectively (P<0.05). Conclusion For the treatment of HCC in BCLC C-stage, the therapeutic effect of TACE combined with autologous DC-CIK cells is much better than that of pure TACE. Therefore, this therapy is an effective treatment for HCC in BCLC C-stage.

CIK cells and their application in the tumor treatment / 国际肿瘤学杂志

Cytokine-induced killer (CIK) cells play important roles in the treatment of solid tumors.In particular,they have broad application prospects for the patients with recurrence or metastasis.Although recently CIK treatment strategy has achieved encouraging results in preclinical studies,there are still some difficulties in the clinical applications.Some key issues remain to be resolved,such as how to ensure that the antitumor immune effector can be a lot of amplified and can stay longer in the body,how to selectively kill tumor cells,and how to ensure the security of application.

The effect of monoclonal antibody coated with anti-human CD3 and CD28 on CIK cells growth / 重庆医学

Objective To explore the enhancing effect of monoclonal antibody coated with anti-human CD3 and CD28 on acti-vation and transformation of peripheral blood mononuclear cell (PBMC)in vitro.Methods Human peripheral blood mononuclear cells were separated.Cells was cultured in vitro,and determined by flow cytometry.The solid phase with CD3 and CD28 antibody was coated and added in.The mature CIK cells were obtained after 12 days culturing.Results The CD4 + cells was lower in group C than those in group A(P <0.05).The CD8 + cells was higher in group C than that in group A and B(P <0.05).There was signifi-cant difference of T4/T8 between group C and group A and B(P <0.05 ).There was significant difference of NK cells between group B and group C(P <0.05).The CD25 + cells was lower in group C than that in group A (P <0.01).Conclusion CD3 antibody solid coated combined with CD28 antibody added to the suspension has more strong activation than both CD3 antibody and CD28 antibody solid coating on peripheral blood mononuclear cell.