AIM: To perform sequencing analysis in patients with very early-onset inflammatory bowel disease (VEO-IBD) to determine the genetic basis for VEO-IBD in Chinese pediatric patients. METHODS: A total of 13 Chinese pedia...AIM: To perform sequencing analysis in patients with very early-onset inflammatory bowel disease (VEO-IBD) to determine the genetic basis for VEO-IBD in Chinese pediatric patients. METHODS: A total of 13 Chinese pediatric patients with VEO-IBD were diagnosed from May 2012 and August 2014. The relevant clinical characteristics of these patients were analyzed. Then DNA in the peripheral blood from patients was extracted. Next generation sequencing (NGS) based on an Illumina-Miseq platform was used to analyze the exons in the coding regions of 10 candidate genes: IL-10, IL-10RA, IL-10RB, NOD2, FUT2, IL23R, GPR35, GPR65, TNFSF15, and ADAM30. The Sanger sequencing was used to verify the variations detected in NGS. RESULTS: Out of the 13 pediatric patients, ten were diagnosed with Crohn's disease, and three diagnosed with ulcerative colitis. Mutations in IL-10RA and IL-10RB were detected in five patients. There were four patients who had single nucleotide polymorphisms associated with IBD. Two patients had IL-10RA and FUT2 polymorphisms, and two patients had IL-10RB and FUT2 polymorphisms. Gene variations were not found in the rest four patients. Children with mutations had lower percentile body weight ( 1.0% vs 27.5%, P = 0.002) and hemoglobin ( 87.4 g/L vs 108.5 g/L, P = 0.040) when compared with children without mutations. Although the age of onset was earlier, height was shorter, and the response to treatment was poorer in the mutation group, there was no significant difference in these factors between groups. CONCLUSION: IL-10RA and IL-10RB mutations are common in Chinese children with VEO-IBD. Patients with mutations have an earlier disease onset, lower body weight and hemoglobin, and poorer展开更多
Objective To investigate whether adenovirus mediated IL 2 gene modification can improve antitumor immunity of the tumor antigen pulsed dendritic cells(DC) in vivo. Methods DC were generated from bone marrow, then, pul...Objective To investigate whether adenovirus mediated IL 2 gene modification can improve antitumor immunity of the tumor antigen pulsed dendritic cells(DC) in vivo. Methods DC were generated from bone marrow, then, pulsed with FBL 3 erythroleukemia cell lysates and transfected with IL 2 gene simultaneously. IL 2 secretion and T cell stimulating activity of the DC were detected. The number and cell subsets of draining lymph nodes, cytotoxic lymphocyte(CTL) activity ,and the increase of protective effect for the mice s.c vaccinated with DC were observed. Results IL 2 gene modified, antigen pulsed DC could secrete high level of IL 2 in vitro, stimulate proliferation of syngeneic T cells markedly. DC vaccine could increase the number of draining lymph node and induce CTL activity, which directed to FBL 3, more significantly. After the DC vaccination, the survival time of the mice challenged with wild type FBL 3 cells were prolonged. Conclusion Antitumor immune response could be induced more potently by vaccination with IL 2 gene modified, antigen pulsed DC.展开更多
Objective To investigate the growth inhibition of interleukin 2 receptor (IL 2R) gene transduced peripheral blood lymphocytes (PBLs) on human ovarian cancer cells Methods Interleukin 2 (IL 2) and IL 2R genes ...Objective To investigate the growth inhibition of interleukin 2 receptor (IL 2R) gene transduced peripheral blood lymphocytes (PBLs) on human ovarian cancer cells Methods Interleukin 2 (IL 2) and IL 2R genes were transfected into human ovarian cancer cell line 3AO and PBLs, respectively, using the same Fugene vector Twenty four hours later transfected and nontransfected PBLs were cocultured with transfected and nontransfected 3AO for 48 hours Cytotoxity of PBLs on 3AO was detected by the MTT assay Results The morphology of IL 2 transduced 3AO and IL 2R transduced PBLs remained unchanged 3AO cells could be transfected with the IL 2 gene and expressed IL 2 mRNA, and PBLs could be transfected with the IL 2R gene and expressed IL 2R mRNA IL 2 transduced 3AO cells enhanced their response to the cytotoxity of PBLs Furthermore, growth inhibition of PBLs to 3AO cells increased significantly when the IL 2R was transfected into PBLs and when the IL 2 gene was transfected into 3AO cells and the two were combined Conclusions IL 2R gene transduced PBLs are able to enhance their cytotoxity on IL 2 gene transduced ovarian cancer cells This method may be a new way to investigate IL 2 gene therapy for ovarian cancer展开更多
基金Supported by National Nature Science Foundation of China,No.81400588
文摘AIM: To perform sequencing analysis in patients with very early-onset inflammatory bowel disease (VEO-IBD) to determine the genetic basis for VEO-IBD in Chinese pediatric patients. METHODS: A total of 13 Chinese pediatric patients with VEO-IBD were diagnosed from May 2012 and August 2014. The relevant clinical characteristics of these patients were analyzed. Then DNA in the peripheral blood from patients was extracted. Next generation sequencing (NGS) based on an Illumina-Miseq platform was used to analyze the exons in the coding regions of 10 candidate genes: IL-10, IL-10RA, IL-10RB, NOD2, FUT2, IL23R, GPR35, GPR65, TNFSF15, and ADAM30. The Sanger sequencing was used to verify the variations detected in NGS. RESULTS: Out of the 13 pediatric patients, ten were diagnosed with Crohn's disease, and three diagnosed with ulcerative colitis. Mutations in IL-10RA and IL-10RB were detected in five patients. There were four patients who had single nucleotide polymorphisms associated with IBD. Two patients had IL-10RA and FUT2 polymorphisms, and two patients had IL-10RB and FUT2 polymorphisms. Gene variations were not found in the rest four patients. Children with mutations had lower percentile body weight ( 1.0% vs 27.5%, P = 0.002) and hemoglobin ( 87.4 g/L vs 108.5 g/L, P = 0.040) when compared with children without mutations. Although the age of onset was earlier, height was shorter, and the response to treatment was poorer in the mutation group, there was no significant difference in these factors between groups. CONCLUSION: IL-10RA and IL-10RB mutations are common in Chinese children with VEO-IBD. Patients with mutations have an earlier disease onset, lower body weight and hemoglobin, and poorer
文摘Objective To investigate whether adenovirus mediated IL 2 gene modification can improve antitumor immunity of the tumor antigen pulsed dendritic cells(DC) in vivo. Methods DC were generated from bone marrow, then, pulsed with FBL 3 erythroleukemia cell lysates and transfected with IL 2 gene simultaneously. IL 2 secretion and T cell stimulating activity of the DC were detected. The number and cell subsets of draining lymph nodes, cytotoxic lymphocyte(CTL) activity ,and the increase of protective effect for the mice s.c vaccinated with DC were observed. Results IL 2 gene modified, antigen pulsed DC could secrete high level of IL 2 in vitro, stimulate proliferation of syngeneic T cells markedly. DC vaccine could increase the number of draining lymph node and induce CTL activity, which directed to FBL 3, more significantly. After the DC vaccination, the survival time of the mice challenged with wild type FBL 3 cells were prolonged. Conclusion Antitumor immune response could be induced more potently by vaccination with IL 2 gene modified, antigen pulsed DC.
基金ThisworkwassupportedbyagrantfromtheNaturalScienceFoundationofZhejiangProvince (No 3 97461)
文摘Objective To investigate the growth inhibition of interleukin 2 receptor (IL 2R) gene transduced peripheral blood lymphocytes (PBLs) on human ovarian cancer cells Methods Interleukin 2 (IL 2) and IL 2R genes were transfected into human ovarian cancer cell line 3AO and PBLs, respectively, using the same Fugene vector Twenty four hours later transfected and nontransfected PBLs were cocultured with transfected and nontransfected 3AO for 48 hours Cytotoxity of PBLs on 3AO was detected by the MTT assay Results The morphology of IL 2 transduced 3AO and IL 2R transduced PBLs remained unchanged 3AO cells could be transfected with the IL 2 gene and expressed IL 2 mRNA, and PBLs could be transfected with the IL 2R gene and expressed IL 2R mRNA IL 2 transduced 3AO cells enhanced their response to the cytotoxity of PBLs Furthermore, growth inhibition of PBLs to 3AO cells increased significantly when the IL 2R was transfected into PBLs and when the IL 2 gene was transfected into 3AO cells and the two were combined Conclusions IL 2R gene transduced PBLs are able to enhance their cytotoxity on IL 2 gene transduced ovarian cancer cells This method may be a new way to investigate IL 2 gene therapy for ovarian cancer
