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KMID : 1137020050160030242
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Journal of Gynecologic Oncology
2005 Volume.16 No. 3 p.242 ~ p.255
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Transcutaneous DNA vaccination increase the CTL response: Preliminary results for the therapeutic HPV DNA vaccination
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김찬주/Kim CJ
임은경/이해남/이근호/박태철/남궁성은/박종섭/Yim EK/Lee HN/Lee KH/Park TC/Namkoong SE/Park JS
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Abstract
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Objective : Transcutaneous immunization (TCI) is a novel vaccination based on the application onto bare skin. We compared the immune response after TCI with the model DNA (OVALBUMIN) to HPV E7 and various adjuvant with intramuscular injection.
We investigated the efficacy of immunization with new construct driven by K6hf promoter and compared with CMV promoter.
Methods : First, we make new construct ligated with OVA to Hair-follicle Specific pK6hf Promoter and evaluated the expression. Mouse skin was transfected with pCMV-OVA, pK6hf-OVA, pCMV- βgal and pK6hf- βgal and expression was determined by RT-PCR and X-Gal staining. OVA protein expression was analyzed by Western blot. Second, we immunized C57/BL6 mice with pCMV-OVA or pK6hf-OVA DNA and cholera toxin (CT) and/or CpG. CTL was measured by ELISPOT assay of the splenocytes from the mmunized mice with the DNA vaccine.
Results : The β-Galactosidase activity by X-Gal staining was detected in the epithelium of the mice skin after pK6hf-β gal application. The mRNA and protein expression from pK6hf-OVA were evident following transcutaneous methods. Those were weaker than pCMV-OVA. TCI with pCMV-OVA and LipofectAMINE 2000 trigered an speicific CTL and Th2 response. CpG was the adjuvant for CTL after pCMV-OVA. CT and CpG did increase the CTL after pK6hf-OVA.
Conclusion : Our data demonstrate that TCI of DNA is possible methods of CTL. CpG and CT were useful in the adjuvant for CTL. The pK6hf-OVA can induce specific CTL. This result is of potential relevance for the development of therapeutic HPVspecific DNA vaccines with TCI and pK6hf promoter can be used safely.
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