±¹¼ÒÀû µµÆ÷¹ýÀ» ÀÌ¿ëÇÑ Hair-follicle specific promoter¸¦ °¡Áø HPV-16 E7 DNA vaccineÀÇ È¿¿ë¼º
Transcutaneous DNA immunization of HPV-16 E7 with hair-follicle specific promoter in mice
±èÂùÁÖ ( Kim Chan-Joo ) - °¡Å縯´ëÇб³ Àǰú´ëÇÐ »êºÎÀΰúÇб³½Ç
±èÅ¿ì ( Kim Tae-Woo ) - °í·Á´ëÇб³ Àǰú´ëÇÐ °¨¿°¸é¿ªÇб³½Ç
³ë°æÈñ ( Noh Kyung-Hee ) - °í·Á´ëÇб³ Àǰú´ëÇÐ °¨¿°¸é¿ªÇб³½Ç
À̱ÙÈ£ ( Lee Keun-Ho ) - °¡Å縯´ëÇб³ Àǰú´ëÇÐ »êºÎÀΰúÇб³½Ç
ÀÌÇØ³² ( Lee Hae-Nam ) - °¡Å縯´ëÇб³ Àǰú´ëÇÐ »êºÎÀΰúÇб³½Ç
À̿뼮 ( Lee Yong-Suk ) - °¡Å縯´ëÇб³ Àǰú´ëÇÐ »êºÎÀΰúÇб³½Ç
¹ÚÅÂö ( Park Tae-Chul ) - °¡Å縯´ëÇб³ Àǰú´ëÇÐ »êºÎÀΰúÇб³½Ç
³²±Ã¼ºÀº ( Namkoong Sung-Eun ) - °¡Å縯´ëÇб³ Àǰú´ëÇÐ »êºÎÀΰúÇб³½Ç
¹ÚÁ¾¼· ( Park Jong-Sup ) - °¡Å縯´ëÇб³ Àǰú´ëÇÐ »êºÎÀΰúÇб³½Ç
Abstract
¸ñÀû: °æÇÇÀû ¸é¿ª¹ý (TCI; Transcutaneous Immunization)À̶õ, ÇǺΠǥ¸é¿¡ Á÷Á¢ ¹é½ÅÀ» µµÆ÷ÇÏ´Â ¹æ¹ýÀ¸·Î, ´Ü¹éÁúÀ̳ª, peptide¹é½ÅÀÇ °æ¿ì adjuvant¿Í ÇÔ²² Åõ¿©ÇÏ¿© ¼¼Æ÷ ¸é¿ª°ú ¾×»ó ¸é¿ª¹ÝÀÀÀ» È¿°úÀûÀ¸·Î À¯µµÇÔÀÌ º¸°íµÇ¾î ÀÖ´Ù. °æÇÇÀû ¸é¿ªÀ» ÅëÇÑ È¿°úÀûÀÎ HPV E7 DNA vaccine °³¹ßÀ» À§ÇÏ¿©, HPV-16 E7 opt+K ºÎºÐ°ú Hair-follicle ƯÀ̼º pK6hf-promoter ¸¦ À¶ÇÕÇÑ DNA°¡ ´Ù¾çÇÑ ¸é¿ª Áõ°Á¦¿Í ÇÔ²² Åõ¿©µÇ¾î È¿°úÀûÀÎ ¸é¿ª¹ÝÀÀÀ» À¯µµÇÒ ¼ö ÀÖ´ÂÁö ¾Ë¾Æº¸°íÀÚ ÇÑ´Ù.
¿¬±¸ ¹æ¹ý: ¸ÕÀú HPV-16 E7 opt+K ºÎºÐ°ú Hair-follicle ƯÀ̼º pK6hf-promoter¸¦ À¶ÇÕÇÏ¿´´Ù. ´ÙÀ½´Ü°è·Î C57/BL6 ÁãÀÇ ÇǺο¡ pK6hf-E7 +K DNA¿Í ¸é¿ª Áõ°Á¦·Î cholera toxin (CT), CpG oligodeoxy nucleotide (ODN)À» »ç¿ëÇÏ¿© °æÇÇÀû ¸é¿ª À» ½ÃÇàÇÑ ÈÄ ¼¼Æ÷µ¶¼º T Àӯı¸ (CTL; Cytotoxic T lymphocyte) ¹ÝÀÀÀ» E7-specific IFN-gamma- secreting CD8+ T cellsÀÇ ¼ö¸¦ intracellular cytokine staining ÈÄ flow cytometric analysisÇÏ¿´´Ù.
°á°ú: °æÇÇÀû ¸é¿ª¹ýÀ¸·Î pK6hf-E7 opt+K DNA ¸¸ Åõ¿©ÇÑ °æ¿ì¿Í, DNA¿Í LipofectAMINE 2000À» Åõ¿©ÇÑ ±ºº¸´Ù pK6hf-E7 opt+K DNA, Lipofectamine2000, CTÀ» Åõ¿©ÇÑ ±ºÀÌ ´õ ³ôÀº E7-specific CD8+ T cellsÀÇ ¼ö¸¦ º¸¿´´Ù.
°á·Ð: À§ÀÇ ½ÇÇè¿¡¼ E7 opt+K DNA¸¦ pK6hf¿Í ¿¬°á½ÃŲ DNA¿Í CTÀ» adjuvant·Î ÀÌ¿ëÇÑ °æÇÇÀû ¸é¿ª¹ýÀÌ Ç׿ø ƯÀ̼º CTLÀ¯µµ¸¦ ÇÒ ¼ö ÀÖÀ½À» ¹àÇô³»¾ú´Ù. ¶ÇÇÑ ÀÌ·¯ÇÑ °á°ú´Â pK6hf promoter¸¦ °¡Áø HPV-16 E7 DNA ¹é½ÅÀ» °æÇÇÀû ¸é¿ª ¹ý¿¡¼ÀÇ °¡´ÉÇÑ ¹æ¹ýÀÓÀÏ Á¦½ÃÇϰí ÀÖ´Ù.
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.
Ű¿öµå
DNA ¹é½Å;ÀÎÀ¯µÎÁ¾ ¹ÙÀÌ·¯½º;¸ð³¶ ƯÀ̼º promoter;°æÇÇÀû ¸é¿ª (Transcutaneous Immunization; TCI);¼¼Æ÷µ¶¼º¸é¿ª
DNA vaccine;HPV E7;Hair-follicle specific promoter;Transcutaneous Immunization;TCI;CTL(cytotoxic T-Lymphocyte) activity
KMID :
0358420070500030512
DOI :
¿ø¹® ¹× ¸µÅ©¾Æ¿ô Á¤º¸
µîÀçÀú³Î Á¤º¸