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
In order to determine the combined effects of migration and gene flow on evolution of insecticide resistance in the mosquito Culex pipiens, four samples were collected in China, among which two were collected along th...In order to determine the combined effects of migration and gene flow on evolution of insecticide resistance in the mosquito Culex pipiens, four samples were collected in China, among which two were collected along the railway from Beijing to Guangzhou. Bioassay data showed that the resistance levels of the four populations to dichlorvos were high and to parathion moderate as compared with the susceptible strain and there was no significant difference among the four populations to the same organophosphate (OP) insecticide. Starch electrophoresis was done to identify the frequency of known overproduced esterases and to analyze genetic diversity among various populations by electrophoretic polymorphism of five presumably neutral loci. The results indicated that the gene flow between populations existed and the number of effective migrants (Nm) was related to collection geography (Nm from 1.67 to 40.07). In contrast with lower genetic differentiation between two nearby populations (between GZ and ZS, ZZ and SQ) and higher genetic differentiation between two distant populations (between GZ and ZZ), there was a significant and inconsistent difference in the distribution of resistance alleles, A2-B2 when explained only with active migration. This divergent situation could be straightened out when considering passive migration (such as railway transport) which increased the spread of A2-B2 along the railway, i.e., in GZ and ZZ. The resistance alleles, A2-B2, dispersing to around areas by active migration suffered from the limitation of gene flow and the speed of invasion.展开更多
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.
文摘In order to determine the combined effects of migration and gene flow on evolution of insecticide resistance in the mosquito Culex pipiens, four samples were collected in China, among which two were collected along the railway from Beijing to Guangzhou. Bioassay data showed that the resistance levels of the four populations to dichlorvos were high and to parathion moderate as compared with the susceptible strain and there was no significant difference among the four populations to the same organophosphate (OP) insecticide. Starch electrophoresis was done to identify the frequency of known overproduced esterases and to analyze genetic diversity among various populations by electrophoretic polymorphism of five presumably neutral loci. The results indicated that the gene flow between populations existed and the number of effective migrants (Nm) was related to collection geography (Nm from 1.67 to 40.07). In contrast with lower genetic differentiation between two nearby populations (between GZ and ZS, ZZ and SQ) and higher genetic differentiation between two distant populations (between GZ and ZZ), there was a significant and inconsistent difference in the distribution of resistance alleles, A2-B2 when explained only with active migration. This divergent situation could be straightened out when considering passive migration (such as railway transport) which increased the spread of A2-B2 along the railway, i.e., in GZ and ZZ. The resistance alleles, A2-B2, dispersing to around areas by active migration suffered from the limitation of gene flow and the speed of invasion.
