OBJECTIVETo construct the Escherichia coli (E. coli) prokaryotic expression system pET9aHPV11L2E7, purify the fusion protein L2E7 and study the immunnogenicity of the protein.

METHODSThe HPV11 L2, E7 coding region was amplified from condyloma acuminata tissue specimen by PCR. The recombinant plazmid pET9aHPV11L2E7 was established and sequenced. Fusion protein L2E7 (553 amino acids) was expressed in host strain BL21 (DE3plus) by IPTG inducing and identified by using SDS-PAGE and Western blotting. Then L2E7 protein purified with CM column was inoculated to Balb/c mice and its cell-mediated and humoral immunnogenicity was assessed by IFN-gamma enzyme-linked immunospot (ELISPOT) and enzyme-linked immunosorbent assay (ELISA).

RESULTSThe E. coli prokaryotic expression system pET9aHPV11L2E7 was established and the purified fusion protein L2E7 was obtained successfully. The mice in vivo experiment indicated that the purified protein L2E7 could induce HPV11E7 specific cell-mediated immune responses and high level HPV L2E7 antibody was detected in serum.

CONCLUSIONThe purified fusion protein L2E7 could induce specific cell-mediated and humoral immune responses. It can be used as a candidate of genital wart immune therapeutic vaccine.

Animals ; Antibodies, Viral ; blood ; Capsid Proteins ; genetics ; immunology ; isolation & purification ; Escherichia coli ; genetics ; metabolism ; Female ; Gene Expression ; Human papillomavirus 11 ; genetics ; immunology ; Humans ; Mice ; Mice, Inbred BALB C ; Oncogene Proteins, Viral ; genetics ; immunology ; isolation & purification ; Papillomavirus Infections ; immunology ; virology ; Papillomavirus Vaccines ; genetics ; immunology ; isolation & purification ; Recombinant Fusion Proteins ; genetics ; immunology ; isolation & purification

OBJECTIVETo construct chimerical DNA vaccine plasmid of human papillomavirus type 11 (HPV11) L1-E7, and to evaluate its immunogenicity.

METHODSMolecular cloning techniques were used to construct recombinant plasmid pcDNA3 L1-E7 as a DNA vaccine. BALB/c mice were vaccinated with DNA recombinants through muscle injection.IL-2 and gamma-INF secreted by immunized spleens lymphocyte and HPV 11 L1 or E7 specific antibodies were assayed by ELISA method. Spleens lymphocyte proliferation was measured by MTT assay.

RESULTSThe chimerical DNA plasmid of pcDNA3 L1-E7 was constructed correctly. Specific anti-HPV11 E7 and L1 antibodies, specific lymphocyte proliferation and secretions of IL-2 and gamma-INF were detected in vaccinated mice.

CONCLUSIONSpecific immune response, including cellular immunity and humoral immunity, could been detected in mice vaccinated with chimerical DNA vaccine of pcDNA3 L1-E7.

Animals ; Antibodies, Viral ; blood ; immunology ; Base Sequence ; Female ; Genetic Engineering ; Human papillomavirus 11 ; genetics ; immunology ; Humans ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Papillomavirus Infections ; blood ; immunology ; virology ; Papillomavirus Vaccines ; administration & dosage ; genetics ; immunology ; Random Allocation ; Vaccines, DNA ; administration & dosage ; genetics ; immunology

Little is known about the prevalence and seroprevalence of low-risk human papillomavirus (HPV) and the risk factors for HPV infection in Korean women. We determined the prevalence of low-risk HPV among 902 women aged 20-59 yr and the seroprevalence of low-risk HPV subtypes 6 and 11 among 1,094 women aged 9-59 yr in the general population. Genital low-risk HPV DNA was assessed by liquid hybridization and polymerase chain reaction. Antibody titers against HPV 6 and 11 were measured by a multiplexed competitive luminex technique. The prevalence of genital low-risk HPV was 4.9%. It reached its highest peak of 10.3% at 20-29 yr of age and a second peak of 3.2% at 50-59 yr of age. The seroprevalence of HPV 6 or 11 was 9.4%. It reached its highest peak of 12.7% at 25-29 yr of age and a second peak of 12.3% at 50-59 yr of age. In multivariable analysis, the number of lifetime sexual partners and past history of sexually transmitted diseases were associated with the seroprevalence but not prevalence of HPV. It is suggested that younger women should receive prophylactic HPV vaccination before they become sexually active and exposed to HPV in their 20s. This study provides baseline data for developing HPV vaccination programs and monitoring vaccine efficacy in Korea.
Adolescent ; Adult ; Antibodies/blood/immunology ; Child ; DNA, Viral/analysis ; Female ; Human papillomavirus 11/genetics/immunology ; Human papillomavirus 6/genetics/immunology ; Humans ; Middle Aged ; Odds Ratio ; Papillomavirus Infections/diagnosis/*epidemiology ; Prevalence ; Republic of Korea/epidemiology ; Risk Factors ; Seroepidemiologic Studies ; Sexual Behavior ; Young Adult