RESUMO
Immunotherapy has become the fourth cancer therapy after surgery, chemotherapy, and radiotherapy. In particular, immune checkpoint inhibitors are proved to be unprecedentedly in increasing the overall survival rates of patients with refractory cancers, such as advanced melanoma, non-small cell lung cancer, and renal cell carcinoma. However, inhibitor therapies are only effective in a small proportion of patients with problems, such as side effects and high costs. Therefore, doctors urgently need reliable predictive biomarkers for checkpoint inhibitor therapies to choose the optimal therapies. Here, we review the biomarkers that can serve as potential predictors of the outcomes of immune checkpoint inhibitor treatment, including tumor-specific profiles and tumor microenvironment evaluation and other factors.
Assuntos
Humanos , Autoanticorpos , Sangue , Alergia e Imunologia , Biomarcadores Tumorais , Sangue , Alergia e Imunologia , Imunoterapia , Neoplasias , Sangue , Terapêutica , Microambiente TumoralRESUMO
Objective To observe the immune effect of PD-1 (programmed death-1) antibody in the treatment of patients with advanced cancer .Methods From October 2015 to March 2016 ,18 patients with advanced tumor were selected to receive the PD-1 antibody treatment in Eastern Hepatobiliary Surgical Hospital .Clinical efficacy ,adverse reactions and progression free survival time were monitored .The quality of life were compared before and after the treatment .Results Among 18 cases , PR 5 cases ,SD 7 cases and PD 6 cases .The KPS scores for quality of life was significantly increased (P<0 .05) after treat-ment .At the end of follow-up ,5 patients died ,2 patients were lost in follow-up ,11 patients survived .The median progression free survival was 2 .6 months (95% CI:1 .8-3 .3 months) .No serious adverse reactions and abnormal laboratory results were reported .Conclusion PD-1 antibody is a safe and effective treatment for advanced tumors .It is well tolerated and has less ad-verse reactions .The randomized control studies with larger samples are needed to further confirm our conclusions .
RESUMO
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 .
RESUMO
Objective To investigate whether decreasing affinity of CAR-T cells can increase their therapeutic outcome or not .Methods Moderate affinity La-G3HER2-CAR and high affinity Ha-G3HER2-CAR were constructed ,and electroporated to modify T cells .Western blot assay ,FCM assay and the RTCA DP cytotoxic equipment were applied to test the CAR expres-sion and cytotoxic function of CAR-T cells .Results 43000 and 58000 exogenous CD3ζfragments were expressed by both La-G3HER2-CAR-T cells and Ha-G3HER2-CAR-T cells with 58 .1% and 69 .0% transfection rate respectively .High affinity Ha-G3HER2-CAR-T cells effectively killed all target tumor cells by which HER2 was expressed at variable expression levels , while moderate affinity La-G3HER2-CAR-T cells specifically killed HER2 high-level expressing SK-OV-3 and BT474 cells ,and showed weaker cytotoxicity on HER2 moderate-level expressing MDA-MB-231 and HCC-202 cells ,and showed no cytotoxicity on HER2 low-level expressing MCF-7 and 293 cells .The underlying mechanic investigation found that La-G3HER2-CAR-T cells and Ha-G3HER2-CAR-T cells were differentially activated by co-culture with MDA-MB-231 (CD107a:8 .2% vs 71 .6% , IFN-γ:66 .3% vs 83 .4% ,TNF-α:73 .4% vs 94 .1% ) .Conclusion Moderate affinity La-G3HER2-CAR-T cells have en-hanced specific cytotoxicity toward target tumor cells compared to high affinity Ha-G3HER2-CAR-T cells ,decreasing affinity of CAR-T cell is a promising strategy to increase the therapeutic outcome of CAR-T cell based immunotherapies .
RESUMO
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.
RESUMO
Chronic hepatitis B is a worldwide infectious diseases caused by hepatitis B virus (HBV) .HBV infection is an important reason for liver cirrhosis and liver cancer in our country .Currently ,the interferon and nucleoside analogs antiviral drugs (nucleotides) is widely used in clinical practice .These drugs inhibit the replication of the virus and disease development to a certain extent ,but not fundamentally eliminate the virus .Various therapeutic vaccines have also made certain curative effect in anti HBV ,but the effect is not perfect clinically .At present ,many research results demonstrate that biological immu-notherapy can successfully eliminate HBV virus in the body , therefore it has brought a new hope for the treatment of hepatitis B .
RESUMO
S-1 is the third generation of fluorouracil derivative anti-cancer agent with small side effects ,efficacy ,and drug delivery is convenient .In a large number of clinical trials and continuous clinical application ,it showed the good effect for the treatment of advanced breast cancer ,which was expected to become the first-line chemotherapy drug for breast cancer treat-ment in the future .
RESUMO
In recent years ,with a large number of chemotherapy development ,the incidence of secondary leukemia in-creased year by year ,an acute myelomonocytic leukemia after chemotherapy and radiotherapy in 1 case of ovarian cancer and breast cancer was reported .The time from first chemotherapy to acute leukemia onset was 37 months .This patient had rapid onset and short survival period .Reports in the literature suggested that alkylating agents ,topoisomerase I inhibitors ,platinum were all easy to induce secondary leukemia .Hematopoietic stem cell transplantation coud reduce mortality and prolong progres-sion free survival .
RESUMO
Objective To investigate the clinical features and treatment of liver cirrhosis with hepatitis B complicated by subcutaneous panniculitis-like T-cell lymphoma (SPTCL) associated hemophagocytic syndrome (HPS) .Methods A retrospec-tive analysis of case clinical data with liver cirrhosis complicated by SPTCL associated HPS was done in August 2014 in our hospital .Results Because of different phenotypes of T cell receptor (TCR) ,the aggression ,treatment response and prognosis of the disease were significantly different .The patients with HPS had poor treatment effect and short survival period .Conclu-sion Liver cirrhosis with hepatitis B complicated by SPTCL associated HPS is rare ,Bone marrow morphology ,pathology , immunohistochemistry and gene rearrangement detection as soon as possible are important for early diagnosis .To control HBV early and effectively is particularly important .Early diagnosis and treatment are important to prolong survival .
RESUMO
Objective To investigate the influence of gemcitabine chemotherapy on levels of regulatory Tcells (Tregs) in peripheral blood for patients with pancreatic cancer and provide evidence and reference for improving the efficacy of adoptive im-munotherapy .Methods 32 patients were enrolled in this study from January 2012 to October 2014 ,among whom 16 received gemcitabine chemotherapy combined with adoptive immunotherapy (gemcitabine group) ,the other 16 patients received adoptive immunotherapy only(control group) .The level of Tregs in peripheral blood ,side effect and overall survival were observed be-fore and after the therapy .Results The number of Tregs in peripheral blood was significantly decreased after gemcitabine chemotherapy ,and it was also lower than that of the control group .The overall survival time of the gemcitabine group was 1.3 mo longer than the control group(10 .0 mo vs 8 .7 mo) .Conclusion Therapeutic regimen of gemcitabine can remarkly de-plete Tregs in peripheral blood of patients with pancreatic cancer ,effectively regulate tumor immune tolerance ,and improve the efficacy of adoptive immunotherapy .
RESUMO
Cancer‐induced immunosuppressive cells play an important immunosuppressive role during the tumor develop‐ment process ,and the development and progression of tumors are always accompanied with abnormal accumulation of cancer‐in‐duced immunosuppressive cells .Regulatory T lymphocytes (Treg) and myeloid‐derived suppressor cells (MDSC) are major components of these inhibitory cellular networks ,and they can inhibit antitumor immune response through multiple mecha‐nisms .Recent studies have provided evidence that beyond their direct cytotoxic or cytostatic effects on cancer cells ,some con‐ventional chemotherapeutic drugs and agents used in targeted therapies can promote the elimination or inactivation of suppres‐sive Tregs or MDSCs ,resulting in enhanced anti‐tumor immunity .Hence ,chemotherapeutics ,used as a preconditioning regi‐men and combined with subsequent immunotherapy ,can promote anti‐tumor immune response .Anticancer chemoimmunothera‐py strategy will change the recognization of the role for conventional chemotherapy in anticancer treatment ,and it will be help‐ful to optimize the chemotherapy strategies more reasonably .
RESUMO
Objective To study the relationship between circulating tumor cells(CTCs) in peripheral blood of the pa‐tients with hepatocellular carcinoma (HCC) and its metastasis and prognosis .Methods 35 patients with hepatocellular carci‐noma were collected as the research objects .CTCs density were enriched through gradient centrifugation and negative‐immu‐nomagnetic methods ,then the cells were detected by chromosome fluorescence in situ hybridization combined with immunofluo‐rescence tests to identify CTCs .The clinical characters were recorded and the data were statistically analyzed .Results All the patients were detected CTC positive .The number of CTC was (4 .1 ± 2 .5) .The patients were divided into 2 groups .Group Ⅰincluded patients whose CTCs were <5 ,and others were included in group Ⅱ .The difference between number of group I and group Ⅱ had significant significance(P=0 .001);Metastasis had nothing to do with patients′sex and age(P=0 .581 ,0 .531);The number of CTCs was related to metastasis and prognosis(P=0.024 ,0.01) ,and there was significant statistic significance between group I and groupⅡ .Conclusion The number of CTCs was related to tumor metastasis .The tumor may be more prone to occur metas‐tasis and may had worse prognosis ,and the patients may had shorter life time when the number of CTCs was≥5 .
RESUMO
Objective To construct an oncolytic adenovirus CNHK600-IL24, and to observe the in vivo effects of CNHK600-IL24 in treating breast cancer. Methods The IL-24 gene was cloned into adenovirus shuttle vector SG502-△CR2, and CNHK600-IL24 was obtained by cotransfection of SG502-INSIL24 and pPE3 plasmids and subsequent recombination in 293 cells. Based on the establishment of the athymic mice model of breast cancer in situ and imitated metastatic breast cancer by injection in the vena caudalis and the left artrium, we administered the virus by the tail vein. We used the optical imaging in vivo system to monitor the effects. Results The oncolytic adenovirus CNHK600-IL24 was correctly constructed and confirmed by restriction DNA sequence analysis and PCR. The titer of CNHK600-IL24 reached 1.9 ×1010pfu/ml. Establishing athymic mice model of breast cancer in situ, the volume and photon number of the tumors in the control group was significantly larger than those of the CNHK600-IL24 group( P ＜0. 05). The tumor had conspicuous necrosis after the treatment of CNHK600-IL24. There was noticeable apoptosis of the tumor cells. Immunohistochemistry showed the expression of IL-24 and the Hexon protein in the tumor cells.In athymic mice model of imitated metastatic breast cancer by infusion into the vena caudalis, most of the mice in the control group died before 38 days, the mice of the CNHK600-IL24 group survived significantly longer(P ＜0. 05 ). Using athymic mice model of imitated metastatic breast cancer by infusion in the left artrium, the optical imaging in vivo system showed obvious difference between the control group and the CNHK600-IL24 group. Conclusions The high-titer oncolytic adenovirus CNHK600-IL24 was successfully constructed and purified. The oncolytic adenovirus had obvious antitumor effect on breast cancer.
RESUMO
The regulation of a target gene expression is very important in gene therapy. However, constitutive or inappropriate expression of the genes with traditional expression system may interfere with the effect of the gene therapy, even may lead to lethal side effect. We constructed an RU486 inducible eukaryotic vector carrying DsRed protein and evaluated its regulatable effect in vitro. The single vector named PDC-RURED was constructed with molecular biological methods, which contained DsRed gene, promoter and RU486-inducible system. To minimize any potential interference, we spaced the two transcriptional elements with a 1.6 kb insulator. The vector was identified by different enzyme restrictions, sequencing analysis and PCR assay. We demonstrated the regulatable expression of this vector after transfection in HEK293 cells by fluorescence microscopy and flow cytometry. In the absence of RU486, no significant DsRed protein activation was observed, whereas in the presence of RU486 up to 40 fold activation of the DsRed protein was observed in cultured cells. The data show that the novel eukaryotic expression plasmid vector can be used to regulate the expression level of genes of interest in appropriate time under the control of RU486. This inducible expression vector provides a powerful tool for the research of gene regulation and gene therapy.
Assuntos
Humanos , Linhagem Celular , Corantes Fluorescentes , Metabolismo , Terapia Genética , Métodos , Vetores Genéticos , Genética , Rim , Biologia Celular , Proteínas Luminescentes , Genética , Mifepristona , Farmacologia , Regiões Promotoras Genéticas , Genética , TransfecçãoRESUMO
Objective To construct adenovirus vector with agiostatinK1-5 gene and to investigate the function of suppression to proliferation and migration for vascular endothelial cells.Methods With the use of gene recombination and clone technology, we constructed the adenovirus vector with the gene of angiostatin K1-5. In vitro vascular endothelial eclls proliferation assay and migration activity were performed through direct infection,MTT and transwell chemotaxis assay. Results 50% TCID indicated that the condence of resultant viruses was 1.5?109PFU/mL. It was purified by CsCL banding,final yield were generally 1.1?1010 PFU/mL plaquing-forming units. Through indirect infect assay and MTT, we found angiostatin K1-5 inhibited human vascular endothelial cells proliferation. We utilized human vascular endothelial cells to study the effect angiostatin K1-5 on cell migration,the result showed that adenoviruse vector with angiostatin K1-5 significantly inhibited HUVEC migration.Conclusion We successfully constructed adenoviruse vector with angiostatin K1-5 and demonstrated its inhibitory effect on proliferation andmigration of HUVEC.
RESUMO
<p><b>OBJECTIVE</b>To study the injection of NKG5SV gene to inhibit growth and metastasis of hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>NKG5SV gene was inserted into retroviral vector pLXSN by normal methods. LacZ gene was used as control. LCI-D20 tumor together with saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was subcutaneously inoculated to the nude mice. Tumor formation rate and tumor size were noted 35 days after inoculation. LCI-D20 tumor was inoculated subcutaneously. Saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was intratumorally injected respectively 10 days after inoculation. Tumor growth was observed 35 days after inoculation. Liver cancer was resected 22 days after intrahepatic inoculation. Saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was respectively injected at incisal margin or intraspleen. Mice were killed 35 days after inoculation to observe tumor recurrence at incisal margin, intrahepatic metastasis and extrahepatic metastasis.</p><p><b>RESULTS</b>Tumor formation rate and tumor diameter(cm) were 1.76 +/- 0.11, 1.51 +/- 0.34, 0.33 +/- 0.04 in the control group, LacZ group, NKG5SV group respectively when tumor and different cDNA were inoculated together. Tumor diameter(cm) and weight(g) were 0.87 +/- 0.08, 0.83 +/- 0.05, 0.26 +/- 0.04; 0.43 +/- 0.06, 0.38 +/- 0.04, 0.08 +/- 0.06 in the control group, LacZ group, NKG5SV group respectively when different cDNA were injected into the LCI-D20 tumor. Sites with extrahepatic metastasis nidi, incisal margin recurrence tumor size(cm), intrahepatic metastasis nidi, metastasis involved hepatic lobes in the control group, LacZ group, NKG5SV group were 4.25 +/- 1.48, 4.25 +/- 1.04, 0.63 +/- 0.51; 1.51 +/- 0.27, 1.35 +/- 0.17, 0.81 +/- 0.17; 2.50 +/- 1.41, 2.38 +/- 1.06, 1.25 +/- 0.71; 2.13 +/- 0.99, 2.00 +/- 0.75, 1.38 +/- 0.74 respectively when NK cells were injected at incise margin. They were 4.38 +/- 1.85, 4.25 +/- 1.48, 1.00 +/- 0.75; 1.13 +/- 0.23, 0.97 +/- 0.29, 0.76 +/- 0.16; 2.50 +/- 1.41, 2.05 +/- 1.12, 0; 2.13 +/- 0.83, 1.75 +/- 0.88, 0 respectively when NK cell were injected intrasplenicly.</p><p><b>CONCLUSIONS</b>NKG5SV gene can inhibit HCC growth and postoperative metastasis and recurrence.</p>
Assuntos
Animais , Humanos , Masculino , Camundongos , Antígenos de Diferenciação de Linfócitos T , Divisão Celular , Terapia Genética , Métodos , Vetores Genéticos , Genética , Injeções , Neoplasias Hepáticas Experimentais , Genética , Patologia , Terapêutica , Camundongos Nus , Metástase Neoplásica , Receptores Imunológicos , Genética , Fisiologia , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
<p><b>OBJECTIVE</b>To develop a new kind of vector system called gene-viral vector, which combines the advantages of gene and virus therapies.</p><p><b>METHODS</b>Using recombinant technology, an anti-tumor gene was inserted into the genome of replicative virus specific for tumor cells. The cell killing effect, reporter gene expression of the green fluorescence protein, anti-tumor gene expression of mouse interleukin-12 (mIL-12) and replication of virus were observed by the methods of cell pathology, fluorescence microscopy, ELISA and electron microscopy, respectively.</p><p><b>RESULTS</b>A new kind of gene-viral vector system of adenovirus, in which the E1b-55 kD gene was deleted but the E1a gene was preserved, was constructed. The vector system, like the replicative virus ONYX-015, replicated and proliferated in tumor cells but not in normal ones. Our vector had an advantage over ONYX-015 in that it carried different kinds of anti-tumor genes to enhance its therapeutic effect. The reporter gene expression of the green fluorescence protein in tumor cells was much better than the adenovirus vector employed in conventional gene the rapy, and the expression in our vector system was as low as or even less than that in the conventional adenovirus gene therapy system. Similar results were observed in experiments with this vector system carrying the anti-tumor gene mIL-12. Replication and proliferation of the virus carrying the mIL-12 gene in tumor cells were confirmed by electron microscopy.</p><p><b>CONCLUSIONS</b>Gene-viral vectors are new vectors with an anti-tumor gene inserted into the genome of replicative virus specific for tumor cells. Because of the specific replication and proliferation of the virus in tumor cells, expression of the anti-tumor gene is increased hundreds to thousands of times. This approach takes full advantages of gene therapy and virus therapy to enhance the effect on the tumor. It overcomes the disadvantages of conventional gene therapy, such as low transfer rate, low gene expression, lack of target tropism, and low anti-tumor activity. We believe that this is a promising means for future tumor treatment.</p>
Assuntos
Humanos , Adenoviridae , Genética , Proteínas E1A de Adenovirus , Genética , Proteínas E1B de Adenovirus , Genética , Terapia Genética , Métodos , Vetores Genéticos , Genética , Interleucina-12 , Genética , Neoplasias , Terapêutica , Recombinação Genética , Células Tumorais Cultivadas , Replicação ViralRESUMO
Objective To construct a new replicating adenovirus vector CNHK600-p53 carrying anti-tumor gene p53 and investigate its effect on hepatocarcinoma cell line PLC/PRF5. Methods The methylthiazolyl tetrazolium assay (MTT) was used to observe the killing effect of Fluorouracil, Mitomycin and Epirubicin alone or in combination with adenovirus CNHK600-p53 on PLC/PRF5. Results When 5-Fu at concentration of 400?g/ml, the inhibition rate was (65?4. 2) % ; with MMC at 1?g/ml, the rate was (41?1.9)%; and it was (65?1.8)% when EPI at concentration of 10?g/ml. With PLC/PRF5 infected by adenovirus CNHK600-p53 ( MOI = 0. 625) , the inhibition rate was significantly increased to (89?5. 3)%,(60?2.3)% and (75?1.5)% respectively; When MOI was 0.3125, 0.625, 1.25, the inhibition rate in CNHK600-p53 group and Ad-p53 group was (27?2. 5)% , (30?3. 7)% , (61?4. 3)% and(4?2.7)%, (5?3.5)%, (16?4.5)% respectively. Conclusion For hepatocarcinoma cell line PLC/PRF5 the effect of CNHK600-p53 is stronger than Ad-p53.
RESUMO
Cytotoxic T lymphowcytes (CTL) play a major role in host anti-tumor immune responses. A major obstacle to the application of adoptive immunotherapy in the treatment of human maligancy is the inability to generate enough activated CTLs since the cytotoxic T cell undergoes activation induced apoptosis during destroying tumor cells. It is important to study how to limit activaton induced apoptosis of T cell so as to maximize the number of CTL and to enhance the tumor cytotoxicity. We have used CD3-induced Jurkat cell line as an activated T cell apoptosis model and introduced the anti-sense ICE cDNA into Jurkat T cells with retroviral vector. The effect on apoptosis of Jurkat cell induced by anti-CD3 or anti-Fas monoclonal antibody was evaluated after transfection with antisense human ICE. We found that the level of ICE expression in Jurkat cell transduced by the vector decreased and apoptosis was minimized in antisense ICE-transfected Jurkat cell after anti-CD3 or anti-Fas treatment. These results suggest that antisense blocking of ICE expression can partially inhibit Jurkat cell apoptosis induced by anti-CD3 or anti-Fas. Thus, applying antisense blocking of ICE to gene therapy may block the apoptosis of activated T cells, furthennore, enhance the antitumor effect.
RESUMO
We used retroviral vector pLXSN to construct recombinant retroviral vectors with the human apoptosis gene, interleukin-l? converting enzyme (ICE). The vectors were introduced into packaging cell line PA317 by electroporation method. The G418 resistant colonies were selected, and the supematants of the colonies were used to infect the human hepatocellular carcinoma cell line SMMC7721. G418 resistant colonies of SMMC7721 were named SMMC7721-MCE and SMMC7721-neo. The results of RT-PCR analysis showed that exogenous hICE gene had successfully integrated into the genome of SMMC7721-hICE cells. The proliferation rate and tumorigenicity of cells in nude mice were examined. Our data showed that the growth rate and the tumorigenicity of SMMC7721-hICE cells in nude mice were considerablely decreased comparing with parent SMMC7721 and SMMC7721-neo. These results suggested that the retroviral vector expressing hICE gene was successfully constructed and could suppress the growth ability and tumorigenicity of human hepatocellular carcinoma cells, which provided a basis for further investigation of hICE gene therapy.