| Transcutaneous DNA immunization of HPV-16 E7 with hair-follicle specific promoter in mice. |
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Chan Joo Kim, Tae Woo Kim, Kyung Hee Noh, Keun Ho Lee, Hae Nam Lee, Yong Seok Lee, Tae Chul Park, Sung Eun Namkoong, Jong Sup Park |
1Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, Catholic University Medical College, Seoul, Korea. jspark@catholic.ac.kr
2Laboratory of Infection and Immunology, Graduate School of Medicine, Korea University, Ansan, Korea. |
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| Abstract |
OBJECTIVE
Previous reports have shown that transcutaneous immunization (TCI) with proteins or peptides in combination with adjuvants efficiently induces specific cellular and humoral immune responses. We compared the immune response after TCI with new construct which was derived from HPV-16 E7opt+K and pK6hf promoter instead of pCMV promoter and various adjuvant. METHODS: First, we made new construct ligated with HPV-16 E7 opt+K to Hair-follicle Specific pK6hf Promoter. Second, we applied pk6hf-E7 opt+K DNA with or without Lipofectamine 2000 and a combination of cholera toxin (CT) and CpG oligodeoxynucleotide (CpG) onto cold wax-depilated and hydrated bare skin of C57 BL/6 mice. To assess the ability of CTL(cytotoxic T-lymphocyte) activity, we performed intracellular cytokine staining with flow cytometric analysis to determine the number of E7-specific IFN-gamma- secreting CD8+ T cells generated in vaccinated mice with the DNA vaccine. RESULTS: Female C57BL/6 mice immunized by TCI methods with 30 microgram of pk6hf-E7 opt+K DNA with Lipofectamine2000 and CT efficiently generated E7-specific CD8(+) T cells compared with the group of pk6hf-E7 opt+K DNA only or DNA with Lipofectamine2000. CONCLUSION: Our results demonstrate that TCI of the linkaged-E7 DNA , E7 opt+K DNA to pk6hf, and Lipofectamine2000 and CT induced an antigen-specific CTL response. This result is of potential relevance for the development of therapeutic HPV-specific DNA vaccines with TCI and pK6hf promoter can be used safely. |
| Key Words:
DNA vaccine, HPV E7, Hair-follicle specific promoter, Transcutaneous Immunization (TCI), CTL (cytotoxic T-lymphocyte) activity |
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