To construct a DNA vaccine as a prophylactic model to prevent condyloma acuminatum and detect its immu-nogenicity in mice. Methods The major capsid protein (L1) gene of human papillomavirus (HPV) 6b was inserted into ...To construct a DNA vaccine as a prophylactic model to prevent condyloma acuminatum and detect its immu-nogenicity in mice. Methods The major capsid protein (L1) gene of human papillomavirus (HPV) 6b was inserted into an eukaryotic ex-pression plasmid (pcDNA3.1). The recombinant plasmid was transfected into COS-7 cells. Western blot were performed to detect whether L1 protein can be expressed in eukaryotic cells. Eighteen female BALB/c mice were tested for immunoge-nicity study. Results The recombinant plasmid (pcDNA3.1-HPV6bL1) was verified as HPV6b L1 gene by sequencing. Western blot showed specific strip. Anti-L1 protein antibodies could be detected in the mice’s sera inoculated with pcDNA3.1-HPV6bL1. Similarly, IL-4, IL-2, and IFN-γ were increased in the same mice. Conclusion HPV6b L1 recombinant plasmid was constructed successfully which had immunogenicity for BALB/c mice. It provided experimental evidence for the research of DNA vaccine of condyloma acuminata..-展开更多
A human papillomavirus genome DNA of 7.9 kb from a Chinese woman with genital condyloma acuminata was cloned in Bam HI site of pAT153. According to the results obtained from Southern blotting, restriction mapping as w...A human papillomavirus genome DNA of 7.9 kb from a Chinese woman with genital condyloma acuminata was cloned in Bam HI site of pAT153. According to the results obtained from Southern blotting, restriction mapping as well as partial DNA sequencing, the isolated genome (HPV6BV) had obvious variance and was referred to as a new variant of HPV6 found in China the first time. HPV6BV L1 gene was successfully expressed in E. coli as a fusion protein with pUR288. The β-galactosidase/L1 fusion protein reacted with both β-galactosidase antiserum and HPV antibody using Western blot technique. The E. coli-produced fusion protein, possessing HPV antigenicity, may provide a reagent for clinical diagnosis and epidemiological survey.展开更多
目的高危型人乳头瘤病毒(high-risk human papillomavirus,HPV)感染是宫颈癌的主要致病原因,现已发现HPV感染可诱发人类的许多肿瘤和疣。文中旨在探讨南京地区肛管及肛门区尖锐湿疣(condyloma acuminata,CA)患者中HPV基因类型的感染情...目的高危型人乳头瘤病毒(high-risk human papillomavirus,HPV)感染是宫颈癌的主要致病原因,现已发现HPV感染可诱发人类的许多肿瘤和疣。文中旨在探讨南京地区肛管及肛门区尖锐湿疣(condyloma acuminata,CA)患者中HPV基因类型的感染情况及其临床意义。方法从86份肛管及肛门区CA石蜡组织标本中提取23种HPVDNA,采用基因扩增芯片技术对其组织进行23种HPV基因类型的检测,并对其患者进行临床病理资料分析。结果 86份肛管及肛门区CA患者组织标本中检出HPV阳性66例,感染率为76.74%(66/86),其中单一型的阳性检出率为56.98%(49/86);单一型的感染中HPV11型为26例,其阳性检出率为30.23%(26/86),是最主要的感染类型;其次HPV6型为23例,阳性检出率为26.74%(23/86);混合型HPV感染17例,阳性检出率为19.77%(17/86);其中HPV6+11型8例,占混合型感染的47.06%(8/17),是混合型感染的主要类型;其次是5例4种类型的混合感染,占混合型感染的29.41%(5/17)。结论 HPV11型、6型、6+11型、4种类型的混合感染是肛管及肛门区CA的主要致病类型,基因扩增芯片检测技术是一种比较适合临床进行HPV分型检测的、敏感性高和特异性好的诊断方法,尤其适合开展某种病变中HPV感染的分子流行病学的研究。展开更多
文摘To construct a DNA vaccine as a prophylactic model to prevent condyloma acuminatum and detect its immu-nogenicity in mice. Methods The major capsid protein (L1) gene of human papillomavirus (HPV) 6b was inserted into an eukaryotic ex-pression plasmid (pcDNA3.1). The recombinant plasmid was transfected into COS-7 cells. Western blot were performed to detect whether L1 protein can be expressed in eukaryotic cells. Eighteen female BALB/c mice were tested for immunoge-nicity study. Results The recombinant plasmid (pcDNA3.1-HPV6bL1) was verified as HPV6b L1 gene by sequencing. Western blot showed specific strip. Anti-L1 protein antibodies could be detected in the mice’s sera inoculated with pcDNA3.1-HPV6bL1. Similarly, IL-4, IL-2, and IFN-γ were increased in the same mice. Conclusion HPV6b L1 recombinant plasmid was constructed successfully which had immunogenicity for BALB/c mice. It provided experimental evidence for the research of DNA vaccine of condyloma acuminata..-
文摘A human papillomavirus genome DNA of 7.9 kb from a Chinese woman with genital condyloma acuminata was cloned in Bam HI site of pAT153. According to the results obtained from Southern blotting, restriction mapping as well as partial DNA sequencing, the isolated genome (HPV6BV) had obvious variance and was referred to as a new variant of HPV6 found in China the first time. HPV6BV L1 gene was successfully expressed in E. coli as a fusion protein with pUR288. The β-galactosidase/L1 fusion protein reacted with both β-galactosidase antiserum and HPV antibody using Western blot technique. The E. coli-produced fusion protein, possessing HPV antigenicity, may provide a reagent for clinical diagnosis and epidemiological survey.
