Objective:The aim in this study was to identify the allelic frequencies of the chemokine (SDF1-3'A) and chemokine receptor (CCR5△32, CCR5m303 and CCR2-64I) genes resistant to HIV-1 infection and/or disease progre...Objective:The aim in this study was to identify the allelic frequencies of the chemokine (SDF1-3'A) and chemokine receptor (CCR5△32, CCR5m303 and CCR2-64I) genes resistant to HIV-1 infection and/or disease progression in indigenous Chinese populations. Methods: By using QIAamp DNA Blood Mini Kit, the genomic DNA samples were purified from whole peripheral blood of healthy individuals (n=2067) from Han, Uygur, Mongolian and Tibetan ethnic groups, as well as Han patients including HIV-1 carriers (n=330), patients with other sexually transmitted diseases (STDs, n=259) and intravenous drug users (IVDUs, n=125). The allelic polymorphisms were identified by means of PCR or PCR-RFLP analyses. The sequences of randomly selected amplified PCR products were further confirmed by direct DNA sequencing. Results: The mutant frequencies were identified to be 0%-3.48% for CCR5△32, 0% for CCR5m303,19.15%-28.79% for CCR2-64 and 19.10%-28.73% for SDF1-3'A alleles, respectively, in Chinese healthy individuals from four ethnic groups. Our findings indicated the allelic frequencies vary among the different ethnic groups. Furthermore, the HIV-1 carriers, STD cases and IVDUs (all of Han ethnicity) were found to have the allelic frequencies of 0%-0.19% (CCR5A32), 0% (CCR5m303), 19.31%-20.45% (CCR2-64) and 25.61%-26.83% (SDF1-3'A) with minor variations in their frequencies between the patients and healthy Han groups. There was no CCR5-m303 mutation found in any subject in this study. Conclusion: The examined subjects of four Chinese ethnic origins showed lower frequencies of CCR5A32 and CCR5m303 alleles, but higher frequencies of mutant CCR264I and SDF1-3'A alleles compared to those identified in northern-European and American Caucasians. The significance of the different frequencies and polymorphisms of the above alleles in Chinese populations needs to be further examined in HIV-1/AIDS diseases.展开更多
Objective: To evaluate the humoral immune induction in rats of a candidate AIDS vaccine expressing the gag p24 gene from a subtype B HIV-1 isolate. Methods: The amplified p24 gene was inserted into an eukaryotic expre...Objective: To evaluate the humoral immune induction in rats of a candidate AIDS vaccine expressing the gag p24 gene from a subtype B HIV-1 isolate. Methods: The amplified p24 gene was inserted into an eukaryotic expression vector to form the supercoiled DNA vaccine. The linearized expressed DNA vaccine was preparedfrom the expression plasmid by polymerase chain reaction(PCR). The antigen gene expression in rats of the linearized and supercoiled DNA vaccines were in vitro and in vivodetected. Results: In vitro transcription and Northern hybridizationshowed that the linearized DNA vaccine could synthesizeamounts of p24 mRNA similar to the supercoiled DNA vaccine.Antibody assays of inoculated rats confirmed that thelinearized expression DNA could induce a slightly higherantibody titer than the expression plasmid, while the highestantibody titer had been induced by plasmid plus adjuvantinoculation. Conclusion: The construction of a candidate AIDS vaccinebased on the p24 gene could shed light on a potential HIVvaccine, meriting evaluation in a rhesus macaque SHIV-AIDSmodel.展开更多
Objective:To amplify antigen genes from patients with human immunodeficiency virus type 1 (HIV-1) in Guangdong Province for candidate AIDS vaccine design. Methods:Viral nucleic acid was isolated from 10 HIV-1 infected...Objective:To amplify antigen genes from patients with human immunodeficiency virus type 1 (HIV-1) in Guangdong Province for candidate AIDS vaccine design. Methods:Viral nucleic acid was isolated from 10 HIV-1 infected individuals' peripheral blood collected during 1995-2000 in Guangdong Province. The viral gag p24 gene and env gp120 gene were amplified by nested-PCR and sequenced. The homologies among HIV-1 isolates were compared with HIV-BLAST. Results: Among 10 HIV-1 isolates, nine are homologous to viruses of subtype B, and one is homologous to viruses of subtype E. Conclusion: Subtype B viruses of HIV-1 are predominantly present in Guangdong Province.展开更多
基金This project was supported by grant from National Natural Sciences Foundation of the PR China(39770683)
文摘Objective:The aim in this study was to identify the allelic frequencies of the chemokine (SDF1-3'A) and chemokine receptor (CCR5△32, CCR5m303 and CCR2-64I) genes resistant to HIV-1 infection and/or disease progression in indigenous Chinese populations. Methods: By using QIAamp DNA Blood Mini Kit, the genomic DNA samples were purified from whole peripheral blood of healthy individuals (n=2067) from Han, Uygur, Mongolian and Tibetan ethnic groups, as well as Han patients including HIV-1 carriers (n=330), patients with other sexually transmitted diseases (STDs, n=259) and intravenous drug users (IVDUs, n=125). The allelic polymorphisms were identified by means of PCR or PCR-RFLP analyses. The sequences of randomly selected amplified PCR products were further confirmed by direct DNA sequencing. Results: The mutant frequencies were identified to be 0%-3.48% for CCR5△32, 0% for CCR5m303,19.15%-28.79% for CCR2-64 and 19.10%-28.73% for SDF1-3'A alleles, respectively, in Chinese healthy individuals from four ethnic groups. Our findings indicated the allelic frequencies vary among the different ethnic groups. Furthermore, the HIV-1 carriers, STD cases and IVDUs (all of Han ethnicity) were found to have the allelic frequencies of 0%-0.19% (CCR5A32), 0% (CCR5m303), 19.31%-20.45% (CCR2-64) and 25.61%-26.83% (SDF1-3'A) with minor variations in their frequencies between the patients and healthy Han groups. There was no CCR5-m303 mutation found in any subject in this study. Conclusion: The examined subjects of four Chinese ethnic origins showed lower frequencies of CCR5A32 and CCR5m303 alleles, but higher frequencies of mutant CCR264I and SDF1-3'A alleles compared to those identified in northern-European and American Caucasians. The significance of the different frequencies and polymorphisms of the above alleles in Chinese populations needs to be further examined in HIV-1/AIDS diseases.
基金Financially supported by Natural Science Foundation of China(No 39870725)Natural Science Foundation of Guangdong(No 980642)Research Foundation of Guangdong Education Buresu(No.20032).
文摘Objective: To evaluate the humoral immune induction in rats of a candidate AIDS vaccine expressing the gag p24 gene from a subtype B HIV-1 isolate. Methods: The amplified p24 gene was inserted into an eukaryotic expression vector to form the supercoiled DNA vaccine. The linearized expressed DNA vaccine was preparedfrom the expression plasmid by polymerase chain reaction(PCR). The antigen gene expression in rats of the linearized and supercoiled DNA vaccines were in vitro and in vivodetected. Results: In vitro transcription and Northern hybridizationshowed that the linearized DNA vaccine could synthesizeamounts of p24 mRNA similar to the supercoiled DNA vaccine.Antibody assays of inoculated rats confirmed that thelinearized expression DNA could induce a slightly higherantibody titer than the expression plasmid, while the highestantibody titer had been induced by plasmid plus adjuvantinoculation. Conclusion: The construction of a candidate AIDS vaccinebased on the p24 gene could shed light on a potential HIVvaccine, meriting evaluation in a rhesus macaque SHIV-AIDSmodel.
基金Financially supported by Natural Science Foundation of China(No 39870725)Natural Science Foundation of Guangdong(No 980642)Research Foundation of Guangdong Education Beaureu(No.20032).
文摘Objective:To amplify antigen genes from patients with human immunodeficiency virus type 1 (HIV-1) in Guangdong Province for candidate AIDS vaccine design. Methods:Viral nucleic acid was isolated from 10 HIV-1 infected individuals' peripheral blood collected during 1995-2000 in Guangdong Province. The viral gag p24 gene and env gp120 gene were amplified by nested-PCR and sequenced. The homologies among HIV-1 isolates were compared with HIV-BLAST. Results: Among 10 HIV-1 isolates, nine are homologous to viruses of subtype B, and one is homologous to viruses of subtype E. Conclusion: Subtype B viruses of HIV-1 are predominantly present in Guangdong Province.
