The eukaryotic vectors VR1012 expressing survivin or 33 tandem repeats of human mucin 1（MUC1）（VNTRs）,namely,VR1012-S and VR1012-VNTR（VNTR=variable number of tandem repeat）,were constructed by cloning survivin an...The eukaryotic vectors VR1012 expressing survivin or 33 tandem repeats of human mucin 1（MUC1）（VNTRs）,namely,VR1012-S and VR1012-VNTR（VNTR=variable number of tandem repeat）,were constructed by cloning survivin and VNTR genes into VR1012,respectively.The eukaryotic vector pEGFP expressing survivin and MUC1 VNTRs fusion gene pEGFP-MS was also constructed.Mouse melanoma cell line（B16） stably expressing survivin and MUC1 VNTRs（MS ＋ B16） was established by Lipofectamine-mediated transfection of pEGFP-MS into B16 cells.EGFP expression in MS ＋ B16 cells was observed using a fluorescent microscope and survivin and MUC1 VNTRs（MS） expression was confirmed by means of Western blot analysis.A syngenic graft tumor model was generated by subcutaneous injection of MS ＋ B16 cells into C57/BL6 mice and tumor size increased rapidly with time in a cell number dependent manner.After the third immunization,mice were challenged subcutaneously with 5×l0 5 MS ＋ B16 cells.Compared with that of the negative control immunized with phosphate-buffered saline（PBS）,a significant reduction of tumor growth was observed in groups immunized with survivin plasmid DNA and MUC1 VNTRs plasmid DNA.Thus,the suppression of subcutaneous tumor was antigen-specific.This model is useful for the development of tumor vaccines targeting survivin and MUCI VNTRs.展开更多
AIM: To improve the immunogenicity of receptor binding site of hepatitis B virus (HBV) on preS1 antigen using HBV core antigen as an immuno-carrier. METHODS: One to 6 tandem copies of HBV preS1 (21-47) fragment were i...AIM: To improve the immunogenicity of receptor binding site of hepatitis B virus (HBV) on preS1 antigen using HBV core antigen as an immuno-carrier. METHODS: One to 6 tandem copies of HBV preS1 (21-47) fragment were inserted into HBcAg at the sites of aa 78 and 82, and expressed in E.coli. ELISA, Western blot and animal immunization were used to analyze the antigenicity and immmunogenicity of purified particulate antigens. The ability to capture HBV by antibodies elicited by chimeric particles was detected with immuno-capture PCR. RESULTS: Recombinant antigens CI, CII, CIII carrying 1-3 copies of HBV preSl (21-47) individually could form virus-like particles (VLPs), similar to HBcAg in morphology. But recombinant antigens carrying 4-6 copies of HBV preSl (21-47) were poorly expressed in E.coli. Chimeric antigens were lacking of immunoreactivity with anti-HBc monoclonal antibodies (McAbs), but still reserved good immunoreactivity with anti-HBe McAbs. CI, CII, CIII could strongly react with anti-preS1 McAb, suggesting that preS1 (21-47) fragment was well exposed on the surface of chimeric VLPs. Three chimeric VLP antigens (CI, CII and CIII) could stimulate mice to produce high-level antibody responses, and their immunogenicity was stronger than non-particulate antigen 21-47*6, containing 6 copies of preS1 (21-47). Mouse antibodies to CI, CII and CIII were able to capture HBV virions in immuno-capture PCR assay in vitro. CONCLUSION: Chimeric particulate antigens of receptor binding site-core antigen of HBV can elicit strong antibody responses to preS1. They have a potential to be developed into prophylactic or therapeutic vaccines against HBV infection.展开更多
AIM: To investigate the association between curative effects of interferon-α and partial human leucocyte antigen (HLA)Ⅱ alleles in chronic viral hepatitis B.METHODS: Sixty patients with chronic viral hepatitis B in ...AIM: To investigate the association between curative effects of interferon-α and partial human leucocyte antigen (HLA)Ⅱ alleles in chronic viral hepatitis B.METHODS: Sixty patients with chronic viral hepatitis B in Shanghai were treated with a standard course of treatment with interferon-α for 6 mo. HLA-DRB1, -DQA1, and -DQB1 alleles were detected by polymerase chain reaction-sequence specific primer (PCR-SSP) method. RESULTS: Frequencies of HLA-DRB1*04(P＜0.025) and HLA-DQA1*0303 (P＜0.01) in non-responders were significantly higher than those in partial and complete responders. Frequencies of HLA-DQAI*0505(P＜0.025) and HLA-DQB1*0301(P＜0.005) in partial and complete responders were significantly higher than those in non-responders.CONCLUSION: Non-response to interferon-α therapy is positively correlated with HLA-DRB1*04 and HLA-DQA1*0303， and negatively correlated with HLA-DQA1*0505 and -DQB1*0301 in patient with chronic viral hepatitis B.HLA Ⅱ genes of the identification alleles provide a method for evaluating outcome of interferon-α treatment.展开更多
AIM. To investigate the influence of HLA-DRB1 alleles and HBV genotypes on inberferon-α therapy for chronic hepatitis B. METHODS： HLA-DRBI＊03, ＊07, ＊09,＊12, ＊15 alleles were determined using polymerase chain re...AIM. To investigate the influence of HLA-DRB1 alleles and HBV genotypes on inberferon-α therapy for chronic hepatitis B. METHODS： HLA-DRBI＊03, ＊07, ＊09,＊12, ＊15 alleles were determined using polymerase chain reaction/sequence specific primer （PCR/SSP） technique in 126 patients with chronic hepatitis B and 76 normal control subjects in Shandong Province, and HBV genotypes were determined by nested-PCR analysis using type-specific primers in 126 patients. RESULTS： The positivity of HLA-DRB1＊07 allele in chronic hepatitis B group was significantly higher than that in normal control group （X^2 = 6.33, P〈0.025, RR = 2.37）. Among the 126 patients, genotype B was found in 38 （30.2%）, genotype C in 69 （54.8%）, and mixed genotype （B＋C） in 19 （15.0%）, genotypes D-F were not found. Among the 46 DRB1＊07（＋） patients, 7 were responders and 39 were non-responders among them （X^2 = 6.71, P〈0.05）. The positivity of HLADRB1＊07 and prevalence of HBV genotype C were significantly higher in non-responders than in responders. CONCLUSION： High positivities of HLA-DRB1 ＊07 allele and HBV genotype C are closely associated with the lower response to interferon-α therapy for chronic hepatitis B.展开更多
OBJECTIVE The aim of this investigation was to assess the association between single nucleotide polymorphisms （SNPs） in HLA-DRB1 and the risk of developing cervical cancer. Our study focused on the second exon of th...OBJECTIVE The aim of this investigation was to assess the association between single nucleotide polymorphisms （SNPs） in HLA-DRB1 and the risk of developing cervical cancer. Our study focused on the second exon of the HLA-DRB1 alleles, which have most of the SNP sites on HLA-DRB1. METHODS We examined 30 cervical cancer patients and 66 control patients using the sequence-based typing polymerase chain reaction technique （PCR-SBT） to type 55 single nucleotide polymorphisms （SNPs） and haplotypes in the second exon of HLA-DRB1. The Chi-square test and the Bonferroni correction method were utilized for the statistical analysis of the data. An association between the alleles and cervical cancer was examined by the linkage disequilibrium test and the odds ratio （OR）. RESULTS Compared with the control group, among the 55 SNPs we studied in the second exon of HLA-DRB1, 4 showed an evident association with cervical cancer. Rs17880292 （P = 0.033, OR = 0.322） and rs1059586 （P = 0.029, OR = 2.657） had positive significance for the risk of developing cervical cancer, while rs17879702 （P = 0.016, OR = 0.222） and rs17882525 （P = 0.025, OR = 0.128） were negative. The difference in frequency of the 5582A-5592A-5667T haplotype between cervical cancer patients and the controls was significant （P = 0.043, OR = 2.735）. CONCLUSION The rs17880292 G/A genotype and the rs1059586 A/A genotype could be linked to an increased risk of cervical cancer. Rs17879702 and rs17882525 might be haplotype-tag SNPs （htSNPs） or belong to some other haplotypes, which might exert a protective effect against cervical cancer in combination. The 582A-5592A-5667T haplotype was shown to be a marker for susceptibility to cervical carcinogenesis.展开更多
基金Supported by the National Natural Science Foundation of China(No.30872396)the Scientific Research Foundation of Jilin Province,China(Nos.20080709,200905169)the Jilin University Basic Research Project,China(No.200903255)
文摘The eukaryotic vectors VR1012 expressing survivin or 33 tandem repeats of human mucin 1（MUC1）（VNTRs）,namely,VR1012-S and VR1012-VNTR（VNTR=variable number of tandem repeat）,were constructed by cloning survivin and VNTR genes into VR1012,respectively.The eukaryotic vector pEGFP expressing survivin and MUC1 VNTRs fusion gene pEGFP-MS was also constructed.Mouse melanoma cell line（B16） stably expressing survivin and MUC1 VNTRs（MS ＋ B16） was established by Lipofectamine-mediated transfection of pEGFP-MS into B16 cells.EGFP expression in MS ＋ B16 cells was observed using a fluorescent microscope and survivin and MUC1 VNTRs（MS） expression was confirmed by means of Western blot analysis.A syngenic graft tumor model was generated by subcutaneous injection of MS ＋ B16 cells into C57/BL6 mice and tumor size increased rapidly with time in a cell number dependent manner.After the third immunization,mice were challenged subcutaneously with 5×l0 5 MS ＋ B16 cells.Compared with that of the negative control immunized with phosphate-buffered saline（PBS）,a significant reduction of tumor growth was observed in groups immunized with survivin plasmid DNA and MUC1 VNTRs plasmid DNA.Thus,the suppression of subcutaneous tumor was antigen-specific.This model is useful for the development of tumor vaccines targeting survivin and MUCI VNTRs.
基金Supported by the Excellent Scholar Incubation Plan of Ministry of Education, China
文摘AIM: To improve the immunogenicity of receptor binding site of hepatitis B virus (HBV) on preS1 antigen using HBV core antigen as an immuno-carrier. METHODS: One to 6 tandem copies of HBV preS1 (21-47) fragment were inserted into HBcAg at the sites of aa 78 and 82, and expressed in E.coli. ELISA, Western blot and animal immunization were used to analyze the antigenicity and immmunogenicity of purified particulate antigens. The ability to capture HBV by antibodies elicited by chimeric particles was detected with immuno-capture PCR. RESULTS: Recombinant antigens CI, CII, CIII carrying 1-3 copies of HBV preSl (21-47) individually could form virus-like particles (VLPs), similar to HBcAg in morphology. But recombinant antigens carrying 4-6 copies of HBV preSl (21-47) were poorly expressed in E.coli. Chimeric antigens were lacking of immunoreactivity with anti-HBc monoclonal antibodies (McAbs), but still reserved good immunoreactivity with anti-HBe McAbs. CI, CII, CIII could strongly react with anti-preS1 McAb, suggesting that preS1 (21-47) fragment was well exposed on the surface of chimeric VLPs. Three chimeric VLP antigens (CI, CII and CIII) could stimulate mice to produce high-level antibody responses, and their immunogenicity was stronger than non-particulate antigen 21-47*6, containing 6 copies of preS1 (21-47). Mouse antibodies to CI, CII and CIII were able to capture HBV virions in immuno-capture PCR assay in vitro. CONCLUSION: Chimeric particulate antigens of receptor binding site-core antigen of HBV can elicit strong antibody responses to preS1. They have a potential to be developed into prophylactic or therapeutic vaccines against HBV infection.
基金Supported by the Foundation of Shanghai Municipal Health Bureau,No.01444
文摘AIM: To investigate the association between curative effects of interferon-α and partial human leucocyte antigen (HLA)Ⅱ alleles in chronic viral hepatitis B.METHODS: Sixty patients with chronic viral hepatitis B in Shanghai were treated with a standard course of treatment with interferon-α for 6 mo. HLA-DRB1, -DQA1, and -DQB1 alleles were detected by polymerase chain reaction-sequence specific primer (PCR-SSP) method. RESULTS: Frequencies of HLA-DRB1*04(P＜0.025) and HLA-DQA1*0303 (P＜0.01) in non-responders were significantly higher than those in partial and complete responders. Frequencies of HLA-DQAI*0505(P＜0.025) and HLA-DQB1*0301(P＜0.005) in partial and complete responders were significantly higher than those in non-responders.CONCLUSION: Non-response to interferon-α therapy is positively correlated with HLA-DRB1*04 and HLA-DQA1*0303， and negatively correlated with HLA-DQA1*0505 and -DQB1*0301 in patient with chronic viral hepatitis B.HLA Ⅱ genes of the identification alleles provide a method for evaluating outcome of interferon-α treatment.
文摘AIM. To investigate the influence of HLA-DRB1 alleles and HBV genotypes on inberferon-α therapy for chronic hepatitis B. METHODS： HLA-DRBI＊03, ＊07, ＊09,＊12, ＊15 alleles were determined using polymerase chain reaction/sequence specific primer （PCR/SSP） technique in 126 patients with chronic hepatitis B and 76 normal control subjects in Shandong Province, and HBV genotypes were determined by nested-PCR analysis using type-specific primers in 126 patients. RESULTS： The positivity of HLA-DRB1＊07 allele in chronic hepatitis B group was significantly higher than that in normal control group （X^2 = 6.33, P〈0.025, RR = 2.37）. Among the 126 patients, genotype B was found in 38 （30.2%）, genotype C in 69 （54.8%）, and mixed genotype （B＋C） in 19 （15.0%）, genotypes D-F were not found. Among the 46 DRB1＊07（＋） patients, 7 were responders and 39 were non-responders among them （X^2 = 6.71, P〈0.05）. The positivity of HLADRB1＊07 and prevalence of HBV genotype C were significantly higher in non-responders than in responders. CONCLUSION： High positivities of HLA-DRB1 ＊07 allele and HBV genotype C are closely associated with the lower response to interferon-α therapy for chronic hepatitis B.
文摘OBJECTIVE The aim of this investigation was to assess the association between single nucleotide polymorphisms （SNPs） in HLA-DRB1 and the risk of developing cervical cancer. Our study focused on the second exon of the HLA-DRB1 alleles, which have most of the SNP sites on HLA-DRB1. METHODS We examined 30 cervical cancer patients and 66 control patients using the sequence-based typing polymerase chain reaction technique （PCR-SBT） to type 55 single nucleotide polymorphisms （SNPs） and haplotypes in the second exon of HLA-DRB1. The Chi-square test and the Bonferroni correction method were utilized for the statistical analysis of the data. An association between the alleles and cervical cancer was examined by the linkage disequilibrium test and the odds ratio （OR）. RESULTS Compared with the control group, among the 55 SNPs we studied in the second exon of HLA-DRB1, 4 showed an evident association with cervical cancer. Rs17880292 （P = 0.033, OR = 0.322） and rs1059586 （P = 0.029, OR = 2.657） had positive significance for the risk of developing cervical cancer, while rs17879702 （P = 0.016, OR = 0.222） and rs17882525 （P = 0.025, OR = 0.128） were negative. The difference in frequency of the 5582A-5592A-5667T haplotype between cervical cancer patients and the controls was significant （P = 0.043, OR = 2.735）. CONCLUSION The rs17880292 G/A genotype and the rs1059586 A/A genotype could be linked to an increased risk of cervical cancer. Rs17879702 and rs17882525 might be haplotype-tag SNPs （htSNPs） or belong to some other haplotypes, which might exert a protective effect against cervical cancer in combination. The 582A-5592A-5667T haplotype was shown to be a marker for susceptibility to cervical carcinogenesis.
