Transcutaneous DNA vaccination increase the CTL response: Preliminary results for the therapeutic HPV DNA vaccination.
- Author:
Chan Joo KIM
1
;
Eun Kyoung YIM
;
Hae Nam LEE
;
Keun Ho LEE
;
Tae Chul PARK
;
Sung Eun NAMKOONG
;
Jong Sup PARK
Author Information
1. Department of Medical Bioscience, Graduate School of Catholic University, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
DNA vaccine;
HPV;
Hair-follicle specific promoter;
Transcutaneous Immunization (TCI);
CTL
- MeSH:
Animals;
beta-Galactosidase;
Blotting, Western;
Cholera Toxin;
DNA*;
Enzyme-Linked Immunospot Assay;
Epithelium;
Immunization;
Injections, Intramuscular;
Mice;
Ovum;
RNA, Messenger;
Skin;
Vaccination*;
Vaccines, DNA
- From:Korean Journal of Gynecologic Oncology
2005;16(3):242-255
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
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-beta gal and pK6hf-beta gal 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 beta-Galactosidase activity by X-Gal staining was detected in the epithelium of the mice skin after pK6hf-beta 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 HPV- specific DNA vaccines with TCI and pK6hf promoter can be used safely.
