1.Activation of tolerant TCR transgenic CD8+ T cells. TCR^Gag : a tolerance model in which TCR transgenic (TG) mice .naïve mice TCR^GagxAlb:Gag : 간세포 안에서 tumor antigen 이 발현하는 transgenic mice와 TCR transgenic mice를 cross하여 tumor antigen에 specific한 T cell receptor ( TCR ) transgenic mouse 의 형태.
Env : human immunodeficiency virus Rag1 : recombination activation genes 1 CFSE : a fluorescent cell staining dye. CFSE is not cell permeable and cannot be used to stain intracellularly
Fig. A.B.C : anti-CD3에 의해 유도되는 tolerant Rag+/+ hybrid T cell의 proliferation->더 강한 stimulation에 의해 tolerize되는 TCR 의 유도를 반영하지 않음. ->이에 반해, Rag-/-상태에서 hybrid mouse T cell이 anti-CD3에서 발현되지 않음.
2. Analysis of tolerance in CD8+ T cells expressing defined dual TCRs
3.Activation of tolerant dual-TCR T cells.
4.Tolerant TCR complex selectively exhibit proximal signaling defects
Confocal microscopy(공초점 현미경) : is an optical imaging technique used to increase micrograph contrast and/or to reconstruct three-dimensional images by using a spatial pinhole to eliminate out-of-focus light or flare in specimens that are thicker than the focal plane. 가장 대표적으로 이용되는 분야 -> immunocytochemistry 와 immunohistochemistry
5.Rescue of tolerant TCR antigen responsiveness
* LCMV : (lymphocytic chorio meningitis virus) 림프구성맥락수막염바이러스 This virus is distributed throughout the world, especially in Europe and the Americas, in the common house mice species,Mus musculus and Mus domesticus. LCMV has served as a research tool for learning about the immune system, providing an important model for studies of virus-induced immune complex disease, the role of the MHC complex, persistent infections, and immunological tolerance.
6.Repeated T cell rescue after reacquisition of tolerance
summary Rag-/- TCR ^GagxAlb:Gag T cell은 anti-CD3에 반응하지 않는다. Tolerant phenotype 은 Gag 를 인식할 수 있는 dual TCR chain 의 부재에 의한 결과가 아니다. Tolerant TCR은 synapse 형성을 mobilize 하기 위해 선택적으로 불필요하다. Dual –TCR T cell 내 TCR specific tolerant 는 Gag와 관계된다. Gag 반응성의 복구가 Gag-specific TCR 발현 내 양적인 변화를 반영하지는 않는다. Tolerant dual TCR T cell의 activation과 proliferation은 LCMV-specific TCR을 통해 유도된다. Tolerance의 requisition 후, T cell rescue가 반복된다.