Regulation of Cell survival Following T Cell Recognition

T 细胞识别后细胞存活的调节

• 批准号: 6906582
• 负责人: ALFRED LM BOTHWELL
• 金额: $ 36.79万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6906582
• 关键词: BCL2 gene /protein; CD antigens; SCID mouse; T lymphocyte; anergy; antigen antibody reaction; antigen presenting cell; apoptosis; cell adhesion molecules; cell cell interaction; clinical research; cysteine endopeptidases; cytoprotection; cytotoxic T lymphocyte; histocompatibility; human subject; leukocyte adhesion molecules; phlebotomy; protein structure function; tissue /cell culture; transfection; transplantation immunology; vascular endothelium; xenotransplantation

DESCRIPTION (provided by applicant): These investigators have previously shown effective strategies to achieve cytoprotection of human umbilical vein endothelial cells (HUVEC) from human cytotoxic T lymphocytes (huCTL) by genetic transduction of the anti-apoptotic caspase-resistant Bcl-2 gene. By contrast, porcine aortic endothelial cells (PAEC) transduced with Bcl-2 are resistant to some inducers of apoptosis but sensitive to xenogeneic huCTL. Comparison of the sensitivity of these transduced lines suggests that the sensitivity of porcine EC represents a gain of function when compared to human EC. The differences in sensitivity will be characterized with regard to factors regulating the Fas mediated signaling pathway and the extent of caspase activation will be characterized biochemically. Based on this characterization additional genes will be examined to further achieve cytoprotection of PAEC in vitro. The activity of both CD4 and CD8 T cells can be inhibited by cells known as regulatory T cells, which may play important roles in regulating development of autoimmunity and transplantation. The prototypic CD4+CD25+ regulatory T cells will be isolated and characterized functionally and biochemically. The mechanisms these regulatory cells utilize to inhibit allo- and xeno- interactions with CD4 and CD8 T cells will be compared. These cell types offer an additional strategy to reduce graft rejection. Finally, in vivo models using SCID/beige mice will be utilized to evaluate cytoprotective strategies and the function of regulatory T cells. A protocol has been established that very effectively shows cytoprotection of human microvessels by Bcl-2 transduction. We will utilize this model to study cytoprotective strategies using porcine microvessels.

描述（由申请人提供）：这些研究者先前已经显示出有效的策略，以通过人类细胞毒性T淋巴细胞（HUCTL）的遗传转导抗胃caspase caspase-Caspase-Caspase-Caspase-CASPase-CASPase-BCL-2 Gene来实现人类细胞毒性T淋巴细胞（HUCTL）的人体脐静脉内皮细胞（HUVEC）。相比之下，用Bcl-2转导的猪主动脉内皮细胞（PAEC）对某些凋亡的诱导剂具有抗性，但对异构性Huctl敏感。这些转导线的灵敏度的比较表明，与人类EC相比，猪EC的灵敏度代表功能的增长。关于调节FAS介导的信号通路的因素，灵敏度的差异将被表征，并且胱天蛋白酶激活的程度将在生化上表征。基于此特征，将检查其他基因，以进一步实现PAEC的体外细胞保护。 CD4和CD8 T细胞的活性都可以被称为调节T细胞的细胞抑制，这可能在调节自身免疫性和移植的发展中起重要作用。原型CD4+ CD25+调节T细胞将在功能和生化上分离并表征。这些调节细胞用于抑制与CD4和CD8 T细胞的同种和异种相互作用的机制。这些细胞类型提供了减少移植排斥的附加策略。最后，使用SCID/米色小鼠的体内模型将用于评估细胞保护策略和调节性T细胞的功能。已经建立了一项方案，该方案非常有效地显示了通过Bcl-2转导对人微血管的细胞保护作用。我们将利用该模型使用猪微血管研究细胞保护策略。