Tumor antigen-loaded dendritic cells combined with cytokine-induced killer (CIK) enhance the killing activity of human esophageal cancer cells by promoting ASK1 activation / 细胞与分子免疫学杂志

Objective To clarify the effect and mechanism of tumor antigen-loaded dendritic cells (Ag-DCs) combined with cytokine-induced killers (CIKs) on the killing of esophageal cancer tumor cells. Methods Peripheral blood DCs and CIKs were induced and cultured, and the DCs were loaded with tumor antigen to obtain Ag-DCs, and Ag-DCs were co-cultured with CIKs. The experiment was divided into CIK group, DC combined with CIK group, Ag-DC combined with CIK group. Flow cytometry was used to detect the phenotype of cells. MTT assay was employed to determine the killing activity against EC9706 cells. Annexin V-FITC/PI double staining was used to detect the apoptosis rate of cells, immunofluorescence staining to detect the expression of phosphorylated apoptotic signal-regulated kinase 1 (p-ASK1) and Western blot analysis to detect the expression of ASK1 pathway related proteins. A nude mouse model of esophageal cancer transplantation tumor was constructed and divided into control group, DC combined with CIK group and Ag-DC combined with CIK group. The corresponding immune cells were injected into the tail vein for treatment and the tumor volume was measured every 2 days. After 21 days, all nude mice were sacrificed with the tumors taken out. HE staining was used to observe the tumor pathological changes and immunohistochemical staining was performed to detect the expression of ki67 and ASK1 in the tumor tissue. Results Comparedwith the CIK group alone and the DC combined with CIK group, the ratio of CD3+ CD8+ and CD3+ CD56+ in the cells significantly increased after Ag-DCs and CIKs co-culture, along with the increased killing rate of EC9706 cells, increased apoptosis rate of EC9706 cells, and the improved activation level of ASK1. Compared with the CIK group and the DC combined with CIK group, the growth of the transplanted tumor in nude mice treated with Ag-DCs combined with CIKs was significantly inhibited, and after 21 days, it was observed that the tumor tissue mass in this group was relatively smaller, with sparsely arranged cells in the tumor tissue and a decline in the positive rate of ki67 in tumor tissue, while the positive rate of ASK1 was significantly increased. Conclusion Co-cultivation of tumor antigen-loaded DCs with CIKs can significantly increase the killing activity of esophageal cancer tumor cells. The mechanism of action may be related to the activation of the ASK1 pathway.

Recent Advances and Future Directions in Immunotherapeutics for Hepatocellular Carcinoma

Systemic target therapeutic drugs, such as sorafenib, lenvatinib, or regorafenib are the only drugs that are known to be effective against advanced hepatocellular carcinoma (HCC). However, these agents show a limited efficacy in killing residual tumors. Immunotherapy is an alternative approach to this treatment and has been used to successfully treat different cancers, including HCC. HCC is an inflammation-induced cancer and represents a very interesting target for immunotherapeutics. Immunotherapies aim to reverse the immune tolerance and suppression found in tumor microenvironments and include approaches, such as adoptive cell therapy, immune checkpoint inhibition, and cancer vaccination. Adoptive cell therapy uses autologous natural killer or cytokine-induced killer cells by cultivating them ex vivo and subsequently reinfusing them into the patient. Immune checkpoint inhibitors reactivate tumor-specific T cells by suppressing checkpoint-mediated inhibitory signaling. Cancer vaccination induces a tumor-specific immune response by activating effector T lymphocytes. A wide range of potential immunotherapy-related adverse events occur; therefore, a multidisciplinary collaborative management is required across the clinical spectrum. This review summarizes the current status of immunotherapy for HCC and provides a perspective on its future applications.

Efficacy of Autologous DC-CIK Cells Combined with Other Immune Cells in the Treatment of Hematological Malignancies -Retrospective Study / 中国实验血液学杂志

OBJECTIVE@#To investigate the safety and clinical efficacy of autologous DC-CIK cells combined with other immune cells for patients with hematological malignancies and analyze patient prognosis.@*METHODS@#50 patients with hematological malignancies who received cellular immunotherapy from September 2014 to April 2016 were retrospectively studied in the First Affiliated Hospital of Xi'an Jiaotong University, 115 cases times of cellular immunotherapy were performed. According to the selected treatment, the patients were divided into the dual cell group (DC-CIK cell treatment) and the multi-cell group (DC-CIK cell combined with other immune cells); According to the treatment course, the patients were divided into the single course group (completed by ＜3 times) and the multiple course group. The changes of T lymphocyte subsets, blood routine indicators and KPS scores as well as the overall survival time before and after treatment were compared and analyzed.@*RESULTS@#[WTB1]The difference of general conditions before treatment including the number of patients, sex, age, T lymphocyte subsets, blood routine indicators, KPS scores and so on in 2 groups divided according to 2 kinds of treatment methods were not statistically significant, indicating that the 2 groups were comparable. Grouped by selected treatment, the CD4/CD8 ratio, Hb and Plt levels decreased in the dual cell group, compared with those before treatment(P＜0.05). The CD3CD4 ratio after treatment in multiple cell group decreased, compared with that before treatment (P＜0.05). The 3-year survival rate of patients in dual cell and multiple cell groups was 61.3% vs 69.8%, the overall survival time of patients in 2 groups was 32.4 months vs 39.6 months, there were no statisticall differences between 2 groups(P>0.05). Grouped by treatment course, the CD3 ratio after treatment increased, while the Hb level after treatment decreased in single course group, compared with level before treatment(P＜0.05). The CD3CD4 ratio, Plt level decreased, while the KPS scores increased after treatment in multiple course group, compared with those before treatment(P＜0.05). The 3-year survival rate in single course and multiple course groups was 52% vs 76.4%, the overall survival time was 28.7 months vs 40.9 months respectively, statistically significant with difference (P＜0.05).@*CONCLUSION@#Autologous DC-CIK cells combined with other immune cells in the treatment of hematological malignancies can change the immune function of the patients and improve the antitumor activity. The multi-course treatment can improve the quality of life, prolong the overall survival time, thus worthing clinical promotion.

Two Cases of Adjuvant Immunotherapy with Cytokine-Induced Killer Cells for Relapsed or Refractory Neuroblastoma / 임상소아혈액종양

The treatment outcomes of relapsed or refractory neuroblastoma have been unsatisfactory till date. We reported two cases of adoptive immunotherapy using cytokine-induced killer (CIK) cells against relapsed or refractory neuroblastoma. CIK cell production was attempted in three patients, out of which two patients exhibited adequate levels of CIK cell production. Two patients completed full term of CIK cell infusions (weekly for 6 weeks and then biweekly for 8 wk) without serious adverse events. The progression-free survivals for the two patients were 1.9 and 4.1 months. Their overall survivals were 16.7 and 28.7 months. Although the efficacy was unclear, CIK cell infusion combined with other treatment strategies may have prolonged overall survival in refractory neuroblastoma patients. Further studies are needed to determine the exact role of CIK cell-based immunotherapy in relapsed or refractory neuroblastoma patients.

A Case of Management for Advanced Hepatocellular Carcinoma with Extrahepatic Metastasis by Autologous Natural Killer Cells Combined with Immune Checkpoint Inhibitor

Hepatocellular carcinoma (HCC) has extremely poor prognosis. Immunotherapy has emerged as a new treatment for a number of cancers. Adoptive immunotherapy is one of the important cancer immunotherapy, which relies on the various lymphocytes including cytotoxic T lymphocytes, natural killer (NK) and cytokine induced killer cells. Also, there has been advance in techniques of NK cell activation to more effectively kill the cancer cells. Of note, recently the blocking antibodies targeting programmed cell death protein 1 (PD-1) have shown promising results in diverse cancers including HCC. We report our recent experience of a patient accompanying advanced HCC with extrahepatic metastases. Disease progression had occurred after sorafenib administration, while the patient showed local tumor control and tumor marker decrease by NK cell immunotherapy combined with PD-1 inhibitor therapy. Though, there was no definite survival advantage due to impaired liver function, which might be caused by treatment related toxicities as well as cancer progression.

Efficiency of Cytokine-Induced Killer Cells in Combination with Chemotherapy for Triple-Negative Breast Cancer / 한국유방암학회지

PURPOSE: The treatment of triple-negative breast cancer (TNBC) remains challenging, due to the absence of estrogen, progesterone, and human epidermal growth factor receptors. This study was designed to evaluate the efficiency and safety of cytokine-induced killer (CIK) cell immunotherapy, following regular chemotherapy, for patients with TNBC. METHODS: A total of 340 patients with postmastectomy TNBC, from January 1, 2010 to June 30, 2014, were included in this retrospective study. Seventy-seven patients received CIK cell immunotherapy, following regular chemotherapy (arm 1), and 263 patients received regular chemotherapy alone (arm 2). The primary aim was overall survival (OS) and disease-free survival (DFS), and the treatment responses and adverse events were also evaluated. RESULTS: The 5-year DFS and OS rates in arm 1 were 77.9% and 94.3%, compared with 69.8% and 85.6% in arm 2, respectively (p=0.159 and p=0.035, respectively). This clearly shows that there was no statistical difference in the 5-year DFS between the two groups. Multivariate analyses of arm 1 indicated that a Karnofsky performance score (KPS) ≥90 and stage I/IIA disease were significantly associated with a prolonged DFS period (hazard ratio [HR], 0.25; 95% confidence interval [CI], 0.09–0.74; p=0.012; and HR 0.21; 95% CI, 0.06–0.82; p=0.024, respectively), but a KPS ≥90 and stage I/IIA disease were not independent prognostic factors for OS. Toxicity was mild in patients who received the CIK therapy. CONCLUSION: The data suggested that CIK cell immunotherapy improved the efficiency of regular chemotherapy in patients with TNBC, and the side effects of CIK cell immunotherapy were mild.

Efficiency of Inducing CIK from Cryopreserved PBMNC by Using Immune Cell SR / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the efficiency of inducing CIK from peripheral blood mononuclear cells(PBMNC) by using immune cell serum replacement(immune cell SR), so as to provide a new strategy for the industrialized production of immune cells.</p><p><b>METHODS</b>The PBMNC of healthy volunteers were collected, and these cells were thawed after short-term cryopreservation and cultured to induce CIK cells. The cells viability was measured by trypan blue exclusion, the phenotypes were analyzed by flow cytometry, and the cytotoxicity was determined by Calcein-AM/PI double staining.</p><p><b>RESULTS</b>In cryopreserved PBMNC, the control group cells failed to normally proliferate. Cell proliferation ratio was low in 2% SR group in comparison with the fresh group, and the difference was significant (P<0.05), however, differences were not statistically significant between 5% SR and fresh group or between 10% AP and fresh group. CD3, CD3CD8 and CD3CD56 cell subsets were not significantly different before and after cryopreservation (P>0.05). After being cultured, CD3, CD3CD4, CD3CD8, CD3CD56 and CD3CD56 subsets and the cytotoxicity in vitro were not significantly different among all group(P>0.05).</p><p><b>CONCLUSION</b>5% SR without the protein of animal origin can be safely used as a substitute for autologous plasma in CIK induced from cryopreserved PBMNC by culture, thus providing a basis for the application of cryopreservation technique of immune cells to cell therapy.</p>

Progress in Treatment of POEMS Syndrome--Review / 中国实验血液学杂志

POEMS syndrome is a rare multiple organ involvement of the parasympathetic syndrome associated with abnormal plasma cells, mostly with high-dose chemotherapy and stem cell transplantation for the treatment. Recently, more treatment attempts to treat POEMS syndrome have been utilized so as to improve the efficacy and safety for the patients with POEMS syndrome, such as immunomodulator, alkylating agent, cytokine-induced killer cells and so on. Lenalidomide has a significant effect on relapse/refractory POEMS syndrome and patients with endocrinopathy. Cytokine-induced killer cells are also a safe and effective regimen for the treatment of POEMS syndrome. This review described the efficacy and safety of immunomodulatos, alkylators, cytokine-induced killer cells, ASCT, proteasome inhibitors and monoclonal antibodies for POEMS syndrome, and the newest clinical research and progress of POEMS syndrome ware summarized briefly.

Research Progress on Dendritic Cells Combined with Cytokine Induced Killer Cells Immunotherapy for Hematological Malignancies --Review / 中国实验血液学杂志

In recent years, with the incidence of malignant tumors increasing year by year, its treatment has been made great progress on the basis of traditional treatments such as surgery, radiotherapy and chemotherapy, it mainly focuses on rapid development of cellular immunotherapy. The efficacy of dendritic cells combined with cytokine induced killer cell (DC-CIK) immunotherapy for cancer patients is positive, it shows good antitumor activities and the abilities to reconstruct and enhance the immune system of tumor patients in clinical application, it has potential application value. In the treatment of hematologic malignancies which are chemotherapy-based and high-grade malignant, what is the effect of DC-CIK cells immunotherapy for them? In this review, we searched Chinese and abroad literatures from 2012 to 2018 and further focused on 18 clinical research articles about hematologic malignancies in order to analyze the characteristics of DC-CIK cells immunotherapy. It was found that DC-CIK cells can be an effective and promising treatment for patients with hematologic malignancies, and thus, if the process and unified plan are further standardized and improved, the efficacy will be more obvious. For this purpose, this paper reviews the biological characteristic of DC cells, CIK cells and the clinical research progress of DC-CIK cell immunotherapy for hematological malignancies.

In vivo anti-tumor effect of DC-CIK cells on human lymphoma cell line Raji

To research on the effect of DC-CIK cells on human lymphoma cell line Raji the immunophenotype of DCCIK cells was analyzed using flow cytometry, and its proliferation inhibition effect was detected using MTT assay. 24 nude mice [4-6 weeks old] were employed and inoculated Raji cells on right axillaries for constructing human Burkitt lymphoma model. MTT results showed that DC-CIK cells had a significant inhibitory effect on Raji cells with obvious dose- and time- dependent effect. Western Blot results confirmed that DC-CIK cells could significantly down regulate the expression of BCL-2 [P<0.05]. DC-CIK cells possesses significant anti-tumor effect on human Burkitt lymphoma bearing nude mice, and down regulation of Raji induced BCL-2 cell apoptosis may be one of the inhibitory mechanisms of DC-CIK cells

Análise do perfil fenotípico e funcional das células natural Killer e linfócitos TCD8+ no Líquen plano / Analysis of phenotypic and functional profile of Natural Killer cells and CD8 + T lymphocytes in Lichen planus

Líquen plano (LP) é uma doença mucocutânea de natureza inflamatória crônica de etiologia ainda desconhecida. Alterações na resposta imune inata, como aos padrões moleculares associados à patógenos (PAMPs) e padrões moleculares associados ao dano (DAMPs) podem levar à inflamação crônica e contribuir com a patogênese do LP. OBJETIVO: Avaliar o efeito da ativação via o DAMP S100A8 e o receptor Toll-like 4 (TLR-4) em células Natural killer (NK) e TCD8 citotóxicas e suas subpopulações de memória/efetoras em pacientes com LP. MÉTODOS: Foram selecionados 25 pacientes com LP (22 mulheres, 3 homens) com idade média de 43,46 anos ± 8,46 e um grupo controle com 25 indivíduos (22 mulheres, 3 homens) com idade média de 42 anos ± 5,5. A determinação transcricional e da expressão por imunohistoquimica dos DAMPs S100A8, HMGB-1 e de TLR-4 e RAGE foi realizada em biópsias de lesões cutâneas de indivíduos com LP, e os níveis séricos de S100A8, HMGB-1, MICA e MICB foram determinados por ELISA. As células mononucleares (CMNs) de sangue periférico foram avaliadas por citometria de fluxo quanto a frequência de TNF, IL-1beta e o marcador de desgranulação CD107a em células TCD8+ e células NK CD56+ e suas subpopulações. A avaliação da via de sinalização de TLR em células TCD8+ purificadas e ativadas com S100A8 foi analisada por PCR array e a determinação da expressão de mRNA dos componentes do inflamassoma em células TCD8+ ativadas com S100A8 por PCR em tempo real. RESULTADOS: Foi evidenciado nos indivíduos com LP elevada expressão da proteína S100A8 nas lesões cutâneas e de HMGB-1, TLR-4 e RAGE na derme, em paralelo ao aumento da expressão de mRNAs para S100A8 e S100A9 e diminuição de RAGE. Além disto, uma elevação dos níveis séricos do dímero S100A8/A9 foi detectada nos pacientes comparados aos controles, ao contrário do DAMP HMGB-1 que mostrou níveis similares em ambos os grupos. A influência do S100A8 em células TCD8+ e células NK, foi analisada em CMNs pela ativação...

Lichen planus (LP) is a mucocutaneous inflammatory chronic disease of unknown etiology. Alterations in the innate immune response such as the pathogen-associated molecular pattern (PAMPs) and damage-associated molecular pattern (DAMPs) can lead to chronic inflammation and contribute to the pathogenesis of LP. OBJECTIVE: Evaluate the effect of the activation trough the DAMP S100A8 and the Toll-like receptor 4 (TLR-4) on the Natural killer cells (NK) and cytotoxic TCD8 cells and their memory / effector subsets in LP disease. METHODS: We selected 25 patients with LP (22 women, 3 men) with a mean age of 43.46 years ± 8.46 and a control group of 25 subjects (22 women, 3 men) with a mean age of 42 ± 5, 5. The transcriptional determination and protein expression by immunohistochemistry of DAMPs, S100A8 and HMGB-1 as well as TLR-4 and RAGE was performed on biopsies of skin lesions from patients with LP, and serum levels of S100A8, HMGB-1, MICA and MICB were determined by ELISA. Peripheral blood mononuclear cells (PBMCs) were assessed by flow cytometry to evaluate the frequency of TNF, IL-1beta and the degranulation marker CD107a in CD8+ T cells and CD56 + NK cells and their subsets. The evaluation of the TLR signaling pathway in purified CD8 + T cells activated with S100A8 were analyzed by PCR array and the determination of mRNA expression of inflammasome components on CD8 + T cells activated by S100A8 was measured by real time PCR. RESULTS: It was shown in the LP individuals an increased expression of the S100A8 protein in the cutaneous lesions and HMGB-1, TLR-4 and RAGE in the dermis, in parallel to increased level of mRNAs for S100A8 and S100A9 and decreased expression of RAGE. Moerover, increased serum levels of the dimer S100A8 / A9 was detected in patients compared to controls, in contrast to DAMP HMGB1 that revealed similar levels in both groups. The influence of S100A8 in CD8 + T cells and NK cells, was analyzed in PBMC activating with...

Transfection Efficiency of Ad5F11p-GFP on CIK and NK-92 Cells and Its Influence on Biological Characteristics / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study transfection efficiency of Ad5F11p-GFP and its influence on biological characteristics of CIK and NK-92 cells in order to predict the application of Ad5F11p vector in immunotherapy.</p><p><b>METHODS</b>Two kinds of immune cells, cytokine-induced killer (CIK) cells and natural-killer (NK) cell line NK-92 cells, were transfected by Ad5F11p-GFP at different multiplicity of transfection (MOI), and untransfected immune cells were used as negative control. GFP expression was determined by flow cytometry, the cell morphology was observed with microscope, the cell proliferation was analyzed by trypan blue staining, specific cytotoxicity of NK-92 cells was determined by LDH assay.</p><p><b>RESULTS</b>About 90% of transfection efficiency for NK-92 cells could be achieved at a MOI of 25, while the transfection efficiency for CIK was less than 40% at a MOI of 200. In addition, the transfection efficiency basically unchanged at the same MOI for 48 h and 96 h, and the immune cells transfected with the virus trended to form agglomeration, displaying slower proliferation, increase of IFN-γ release and enhancement of tumor killing activity.</p><p><b>CONCLUSION</b>Ad5F11p- modified NK-92 shows a good prospect for adoptive immunotherapy.</p>

Combination of Jianpi Liqi Yiliu Formula with Cytokine-induced Killer Cell Treatment for Advanced Hepatocellular Carcinoma / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To evaluate the clinical efficacy of Jianpi Liqi Yiliu Formula (JLYF) combined with cytokine-induced killer (CIK) cells for treating patients with advanced hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Between January 2011 and January 2014, 60 advanced HCC patients were enrolled in this study, who were assigned to the treatment group and the control group according to their willingness for taking JLYF, 30 cases in each group. All patients received CIK cell treatment: 1 x 10⁹-3 x 10⁹ each time, by intravenous dripping from the 1st day to the 3rd day, once per day. Besides, patients in the treatment group took JLYF decoction, while those in the control group took Chinese medical decoction by syndrome typing. All patients received treatment of at least two cycles. The time to progression (TTP) , overall survival (OS), disease control rate (DCR), performance status scale (PS), Child-Pugh scale, and adverse reactions were observed, and subgroup analyzed.</p><p><b>RESULTS</b>To May 31, 2014, all patients reached the clinical endpoint. TTP was 3.5 months (95% Cl: 3.30-4.10) in the treatment group, better than that (2.5 months, 95% CI: 2.32-2.68) of the control group (P < 0.05). DCR was 36.7% in the treatment group and 30.0% in the control group (P > 0.05). OS was 5.2 months (95% CI: 4.53-5.87) in the treatment group and 4.6 months (95% CI: 4.06-5.14) in the control group (P > 0.05). The PS scale was 1.60 ± 0.10 after treatment, lower than that (1.80 ± 0.09) before treatment in the treatment group (P < 0.05). When the PS scale was 0-2 or Child-Pugh scale was class A, TTP was longer in the treatment group than in the control group (P < 0.05). No adverse reaction occurred in the two groups during the treatment course.</p><p><b>CONCLUSIONS</b>The combination of JLYF with ClK cell treatment could prolong advanced HCC patients' TTP, improve PS scale, as compared with syndrome typed Chinese medical decoction treatment group. Besides, when the PS scale was 0-2 or Child-Pugh scale was class A, it was a better treatment program for advanced HCC patients.</p>

Cell-based Immunotherapy for Colorectal Cancer with Cytokine-induced Killer Cells

Colorectal cancer is the third leading cancer worldwide. Although incidence and mortality of colorectal cancer are gradually decreasing in the US, patients with metastatic colorectal cancer have poor prognosis with an estimated 5-year survival rate of less than 10%. Over the past decade, advances in combination chemotherapy regimens for colorectal cancer have led to significant improvement in progression-free and overall survival. However, patients with metastatic disease gain little clinical benefit from conventional therapy, which is associated with grade 3~4 toxicity with negative effects on quality of life. In previous clinical studies, cell-based immunotherapy using dendritic cell vaccines and sentinel lymph node T cell therapy showed promising therapeutic results for metastatic colorectal cancer. In our preclinical and previous clinical studies, cytokine-induced killer (CIK) cells treatment for colorectal cancer showed favorable responses without toxicities. Here, we review current treatment options for colorectal cancer and summarize available clinical studies utilizing cell-based immunotherapy. Based on these studies, we recommend the use CIK cell therapy as a promising therapeutic strategy for patients with metastatic colorectal cancer.

Clinical Efficacy of Dendritic Cells and Cytokine-induced Killer Cells Combined with Chemotherapy for Treating Newly diagnosed Multiple Myeloma and their Effect on Function of CD4(+) CD25(+) T Cells in Peripheral Blood / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy of dendritic cells and cytokine-induced killer cells (DC-CIK) combined with chemotherapy for treating newly diagnosed patients with multiple myeloma (MM) and their effect on cellular immune functions of CD4(+) CD25(+) Treg cells in peripheral blood after adoptive immunotherapy.</p><p><b>METHODS</b>Fouty two patients with MM were randomly divided into two groups: chemotherapy group and combined therapy group; 20 patients in chemotherapy group were treated by chemotherapy only, 22 patients in combined therapy group were treated by adoptive immunotherapy (DC-CIK) combined with chemotherapy, and the clinical outcomes of patients and the levels of CD4(+) CD25(+) Treg cells in peripheral blood between 2 groups were compared.</p><p><b>RESULTS</b>After treating for 3 weeks, the quality of life, clinical index and survival of patients in combined therapy group were better than those of patients in chemotherapy group (P < 0.05); the ratios of CD4(+) CD25(+)/CD4(+) and CD4(+) CD25(+) FoxP3(+)/CD4(+) CD25(+) of patients in combined therapy group were obviously lower than those of patients in chemotherapy group (P < 0.05).</p><p><b>CONCLUSION</b>The immunotherapy of DC-CIK can strengthen the activities of CD4(+) CD25(+) Treg cells, which combined with chemotherapy can be an effective and promising effects for treatment of patients with MM.</p>

Comparison of Several Optimization Schemes for the Induction and Expansion of Antibody-Mediated High Efficiency CIK (AMHE-CIK) In Vitro / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To compare several schemes of inducing and expanding the antibody-mediated high efficiency CIK (AMHE-CIK) in vitro, so as to find out a method that can acquire a large number of cells capable to kill the tumor cells in a short time.</p><p><b>METHODS</b>Peripheral blood mononuclear cells (PBMNC) from healthy volunteers was isolated and activated with CD3 antibody, then were cultured with the addition of different cytokines (IL-2, IL-4, G-CSF, GM-CSF, IFN-γ, TNF-α) for 14 days in vitro. The morphological changes of cells were observed by light microscopy. Based on the immunophenotypes of cells in each groups analyzed by flow cytometry, the cytokines capable to induce the dendritic cells and killer cells were screened out, respectively. According to different combination of cytokines, the cells were divided 4 groups: control, IL-2, group 1 (componant A included IL-2, IL-4, and GM-CSF. Componant B included IL-2, G-CSF, IFN-γ, and TNF-α), and group 2 (componant A included IL-2, IL-4, and GM-CSF. Componant B included IL-2, IL-4, G-CSF, IFN-γ, and TNF-α). The proliferation and differentiation of CD3(+) CD8(+) and CD3(+) CD56(+) cells were measured by flow cytometry after culture in vitro for 7 days.</p><p><b>RESULTS</b>After inducing and expanding in vitro for 7 days, the cell proliferation rate of control group, IL-2 group, group 1 and group 2 were 1.57 ± 0.01, 4.17 ± 0.16, 5 ± 0.47, 7.17 ± 0.24-folds, respectively. The differences between IL-2 group, group 1, group 2 and control group were statistically significant (P < 0.05). The immunophenotype analysis showed that the proportion of CD3(+) CD8(+) induced by each protocol was 13.96 ± 0.23%, 26.33 ± 0.55%, 36.83 ± 0.34% and 35.88 ± 0.16%, respectively. The proportion of CD3(+) CD8(+) in group 1 and 2 was higher than that in IL-2 group (P < 0.05), but the difference between them was not significant (P < 0.05). The proportions of CD3(+) CD56(+) induced by each protocol were 11.03 ± 0.28%, 29.31 ± 0.60%, 39.96 ± 0.38% and 29.33 ± 0.54%, respectively, the proportion of group 1 was higher than that of IL-2 group and group 2 (P < 0.05), but the difference between IL-2 group and group 2 was not significant (P < 0.05).</p><p><b>CONCLUSION</b>The group 1 protocol obtained from this study can promote the proliferation of DC-CIK and also increase the proportion of the tumor killing cells (CD3(+) CD8(+) and CD3(+) CD56(+)).</p>

Effect and Safety of CIK Cell Infusion on Peripheral Blood Immune Cell Level in the Elderly Patients with Multiple Myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of CIK cells on the level of peripheral blood immune cells in the elderly patients with multiple myeloma and its safety.</p><p><b>METHODS</b>A total of 60 patients with multiple myeloma from April 2004 to April 2015 in our hospital were enrolled in the study. According to the treatment plan, the patients were randomly divided into control and observation group. The patients in control group was given VAD chemotherapy, the patients in observation group was treated with CIK cells on basis of the control group protocol. ELISA was used to detect the serum levels of IL-17, IL-6 and transforming growth factor (TGF); the hemoglobin, erythrocyte sedimentation rate (ESR) and serum creatinine were assayed also. The incidence of adverse reaction in patients was assayed; the therapeutic efficacy of observation and control groups was judged after treatment curses.</p><p><b>RESULTS</b>The serum levels of IL-17, IL-6 and TGF-β between two groups before treatment were not significantly different (P > 0.05), but after treatment, thier levels in two groups decreased, moreover the levels of the observation group was significantly lower than that in control group (P < 0.05). Before treatment, there was no significant difference in the levels of CD3(+) CD4(+), CD3(+) CD8(+) and CD3(+) CD4(+)/CD3(+) CD8(+) between the two groups (P > 0.05); after treatment, these levels all decreased, moreover the levels of the observation group significantlly lower than that in control group (P < 0.05). The incidence of nausea and vomiting, heart palpitations, chest tightness, increase of myocardial enzyme, amino transferase and creatinine all were not significantly different between two groups (P > 0.05). The curative efficiency of the observation group was significantly higher than that of the control group (P < 0.05).</p><p><b>CONCLUSION</b>CIK cell therapy has better curative effect in the elderly patients with multiple myeloma. The level of peripheral blood immune cells can be significantly increased by decreasing the level of immunosuppressive factor.</p>

Clinical Analysis of Autologous Cytokine-induced Killer Cells Combined with IL-2 for Treating of Elderly Patients with B-cell Malignant Lymphoma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the values of autologous cytokine-induced killer cells combined with rhIL-2 for therapy of elderly patients with B-cell malignant lymphoma.</p><p><b>METHODS</b>Eighty-five elderly patients with B-cell malignant lymphoma were treated by cytokine induced killer cells combine with rhIL-2 (CIK+IL-2 group), 85 elderly patients with B-cell malignant lymphoma treated without cytokine induced killer cells combined with rhIL-2 were used as the control group. The patients in CIK+IL-2 group and control group were divided into 4 subgroups accerding to lymphoma types: group A: diffuse large B cell lymphoma (DLBCL), group B: mucosa-associated lymphoid tissue type (MALT), group C: lymphoplas macytic lymphoma (LPL) and group D: hodgkin's lymphma (HL). The clinical effects, T-lymphocyte, β2 microglobulin level, quality of life and long-term survival were observed.</p><p><b>RESULTS</b>The levels of CD3+, CD3+/CD8+, CD3+/CD56+ after treatment in the 4 subgroups of CIK+IL-2 group were higher than levels before treatment and the control group (P<0.05); the levels of β2 microglobulin after treatment for the 4 groups were lower than before treatment and the control group (P<0.05); with 1 case death, 16 cases were turned from CRu and PR to CR; the CR rate was not significantly different among the 4 subgroups (P>0.05); the scores of physical performance, role function, cognitive function, emotional functioning, and social function after treatment in the 4 subgroups were higher than the those before treatment (P<0.05); the survival time of patients in the CIK+IL-2 group lasted for 8-76 months, their median survival time was (22.36±5.38) months; the survival of the control group lasted for 7-55 months, their median survival time was (16.15±3.62) months. The survival time of the CIK+IL-2 group was longer than that of the control group (P<0.05).</p><p><b>CONCLUSION</b>The treatment of aged patients with B-cell malignant lymphoma by autologous cytokine-induced killer cells combined with rhIL-2 can effectively improve the T-lymphocyte subsets, β2 microglobulin level and quality of life, and can prolong survival time of patients.</p>

Biomarcadores de inflamación subclínica en pacientes infanto-juveniles con diabetes tipo 1 / Subclinical inflammation biomarkers in an infant-juvenile patients with type 1 diabetes

La diabetes tipo 1 (DT1) se asocia a un riesgo incrementado de complicaciones vasculares. Las citoquinas proinflamatorias IL-6, MCP-1 y TNF-α han sido implicadas en el desarrollo de estas complicaciones. El objetivo de este trabajo fue determinar los niveles plasmáticos de IL-6, MCP-1, TNF-α, PCRus y fibrinógeno (Fg) en pacientes infanto-juveniles con DT1 y su asociación con el grado de control glucémico y tiempo de evolución de la enfermedad. Se estudiaron 45 pacientes con DT1 (24 M/21 F), edad 11,2}1,8 años, con tiempo de evolución de la enfermedad de 3,1}3,0 años, sin complicaciones vasculares, que se compararon con 20 sujetos sanos. Se determinaron los niveles plasmáticos de IL-6, MCP-1 y TNF-α, Fg, PCRus, recuento de leucocitos, glucemia en ayunas y HbA1c. Se descartó la presencia de retinopatía y nefropatía. Los datos fueron analizados con el programa SPSS 15 para Windows. Los niños diabéticos presentaron niveles mayores de IL-6 (1,10}0,74 vs 0,68}0,19 pg/ml; p=0,005), MCP-1 (130}49 vs 95}18 pg/ml; p=0,02), PCRus (1,02}1,07 vs 0,43}026 mg/l; p=0,007), Fg (299}59 vs 246}18 mg/dl, p=0,0001), respecto de los controles. No se observaron diferencias significativas de TNF-α entre ambos grupos. Al agrupar a los diabéticos según el grado de control glucémico (HbA1c <8% y ≥8%) y el tiempo de evolución de la enfermedad (≤3 y >3 años), no se encontraron diferencias significativas en las moléculas estudiadas. En los diabéticos la HbA1c se correlaciono con IL-6, MCP-1 y PCRus. Estos resultados reflejan un estado proinflamatorio en la población diabética estudiada.

Two Cases of Early Recurred Hepatocellular Carcinoma after Surgical Resection Which Showed Different Outcomes

Hepatocellular carcinoma (HCC) has poor prognosis, even after curative resection. Early recurrence after curative treatment is a major cause of the poor prognosis. Pathologic factors such as vessel invasion, satellite nodule, size of tumor and pathologic grade are prognostic factors predicting early recurrence and poor prognosis. We share our experience of two cases which both showed early recurrence after curative hepatic resection, but eventually demonstrated different prognosis. Since the most common cause of death after potentially curative treatment is tumor recurrence, suppression of tumor recurrence might be linked to survival gain. Currently, there is no adjuvant therapy for HCC endorsed by international guidelines. However, recent studies have shown that antiviral treatment for hepatitis B virus-related HCC and immunotherapy using autologous cytokine-induced killer cell reduced HCC recurrence. Further study is needed to select patients who will benefit from adjuvant treatments.