생명과학회지 (Journal of Life Science)

  • 제16권6호
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  • Pages.904-910
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  • 2006
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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항종양 면역반응 유도를 위한 수지상세포의 최적 활성화 조건

The Optimal Activation State of Dendritic Cells for the Induction of Antitumor Immunity

  • 남병혁 (동아대학교 의과학연구원, 임상시험연구센터) ;
  • 조월순 (동아대학교 의과학연구원, 임상시험연구센터) ;
  • 이기원 (동아대학교 의과학연구원, 임상시험연구센터) ;
  • 오수정 (동아대학교 의과학연구원, 임상시험연구센터) ;
  • 강은영 (동아대학교 의과학연구원, 임상시험연구센터) ;
  • 최유진 (동아대학교 의과학연구원, 임상시험연구센터) ;
  • 도은주 (동아대학교 의과학연구원, 임상시험연구센터) ;
  • 홍숙희 (동아대학교 의과학연구원, 임상시험연구센터) ;
  • 임영진 (동아대학교 의과학연구원, 임상시험연구센터) ;
  • 김기욱 (동아대학교 의과대학 신경외과학교실) ;
  • 정민호 (동아대학교 의과학연구원, 임상시험연구센터)
  • Nam, Byung-Hyouk (Dong-A University Medical Science Research Center, Clinical Research Center) ;
  • Jo, Wool-Soon (Dong-A University Medical Science Research Center, Clinical Research Center) ;
  • Lee, Ki-Won (Dong-A University Medical Science Research Center, Clinical Research Center) ;
  • Oh, Su-Jung (Dong-A University Medical Science Research Center, Clinical Research Center) ;
  • Kang, Eun-Young (Dong-A University Medical Science Research Center, Clinical Research Center) ;
  • Choi, Yu-Jin (Dong-A University Medical Science Research Center, Clinical Research Center) ;
  • Do, Eun-Ju (Dong-A University Medical Science Research Center, Clinical Research Center) ;
  • Hong, Sook-Hee (Dong-A University Medical Science Research Center, Clinical Research Center) ;
  • Lim, Young-Jin (Dong-A University Medical Science Research Center, Clinical Research Center) ;
  • Kim, Ki-Uk (Department of Neurosurgery, College of Medicine Dong-A University) ;
  • Jeong, Min-Ho (Dong-A University Medical Science Research Center, Clinical Research Center)
  • 발행 : 2006.10.01
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초록

수지상세포는 종양면역에서 필수적인 강력한 CTL 반응을 개시할 수 있는 유일한 세포이다 . 특히 외인성 종양항원에 대한 CTL 반응 유도는 활성화된 수지상세포의 IL-12 분비를 통한 CD4+ helper T세포의 cross-priming을 필요로 한다. 그러나 최근에 활성화된 수지상세포는 $Th_1$ 면역반응을 유도하지만 활성화 시간이 경과함에 따라 오히려 $Th_2$반응을 유도 할 수 있다. 따라서 본 연구에서는 OVA를 종양항원 모델로 설정하여 종양특이적인 CTL 반응을 형성하기 위한 최적의 수지상세포 활성화 조건을 조사하였다. 마우스 골수세포에 서 수지상세포로의 분화는 항원제시 기능을 위한 표면분자의 발현 측면에서 볼 때 배양 6일-7일 정도가 적합하였다. 수지상세포의 IL-12 생성능은 배양 6일 이상, OVA 항원 탑재 8시간 이상의 경우에 연이은 LPS 성숙자극으로 오히려 감소하는 경향을 보였다. 즉 배양 6일의 수지상세포에 OVA 항원 탑재를 8시간 수행한 경우(8-h DC)가 in vitro에서의 IL-12생성능, ex vivo에서의 세포내 $IFN-{\gamma}$를 발현하는 CD8+ T세포의 증가 및 OVA 특이적인 세포독성효과 등에서 가장 좋은 결과를 보였다. 또한 in vivo에서 종양 치료 및 예방효과에서도 8-h DC로 면역한 경우에 가장 우수한 종양형성 억제 효과와 생존기간 연장효과를 보였다. 현재 대부분의 수지상 세포를 이용한 항종양 백신에서 항원 탑재반응을 24시간 동안 수행하고 있으나, 본 실험의 결과로 볼 때, 8시간의 in vitro 항원 탑재가 보다 효과적인 종양특이적 CTL 반응과 항종양 면역반응을 유도함을 알 수 있다. 결론적으로 본 연구를 통하여 8시간 이상의 항원접촉은 수지상세포의 기능적 활성능력을 오히려 고갈시킬 수 있음을 제시한다.

Dendritic cells (DCs) are the only antigen presenting cells (APCs) capable of initiating immune responses, which is crucial for priming the specific cytotoxic T lymphocyte (CTL) response and tumor immunity. Upon activation by DCs, CD4+ helper T cells can cross-prime CD8+ CTLs via IL-12. However, recently activated DCs were described to prime in vitro strong T helper cell type 1 $(Th_1)$ responses, whereas at later time points, they preferentially prime $Th_2$ cells. Therfore, we examined in this study the optimum kinetic state of DCs activation impacted on in vivo priming of tumor-specific CTLs by using ovalbumin (OVA) tumor antigen model. Bone-marrow-derived DCs showed an appropriate expression of surface MHC and costimulatory molecules after 6 or 7-day differentiation. The 6-day differentiated DCs pulsed with OVA antigen for 8 h (8-h DC) and followed by restimulation with LPS for 24 h maintained high interleukin (IL)-12 production potential, accompanying the decreased level in their secretion by delayed re-exposure time to LPS. Furthermore, immunization with 8-h DC induced higher intracellular $interferon(IFN)-{\gamma}+/CD8+T$ cells and elicited more powerful cytotoxicity of splenocytes to EG7 cells, a clone of EL4 cells transfected with an OVA cDNA, than immunization with 24-h DC. In the animal study for the evaluation of therapeutic or protective antitumor immunity, immunization with 8-h DC induced an effective antitumor immunity against tumor of EG7 cells and completely protected mice from tumor formation and prolonged survival, respectively. The most commonly used and clinically applied DC-based vaccine is based on in vitro antigen loading for 24 h. However, our data indicated that antigen stimulation over 8 h decreased antitumor immunity with functional exhaustion of DCs, and that the 8-h DC would be an optimum activation state impacted on in vivo priming of tumor-specific CTLs and subsequently lead to induction of strong antitumor immunity.

키워드

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