Clonal analysis of joint-infiltrating natural Killer cells in chronic arthritis

描述(由申请人提供)： 自然杀伤(NK)细胞是关节浸润性细胞的重要组成部分，但其在类风湿关节炎(RA)发病机制中的作用尚不清楚。一种独特的CD56bnIGNK亚群产生高水平的干扰素-n(IFND)并共同激活巨噬细胞，已被发现优先渗透到受影响的关节。相比之下，CD56dim NK亚群的炎症活性较低，但细胞溶解功能增强，在炎症关节中的表达不足。然而，这两个NK亚群是否代表了成熟的顺序阶段或不同的发育谱系仍有待研究。这是一个重要的问题，因为不同的个体发育可能反映出不同的生物学功能，影响临床疾病的结果。我们和其他人已经证明，VDJ重组，即B细胞和T细胞中抗原受体重排的过程，也发生在人和小鼠的NK细胞中。基于对小鼠模型中NK细胞重组的分析，我们假设炎性NK细胞代表了个体遗传学上不同的谱系。我们建议用不可磨灭的DNA重排作为克隆标记，以确定类风湿关节炎(RA)患者CD56bright和CD56dim NK细胞的发育关系(目标1)。然后，我们利用VDJ重组后表达绿色荧光蛋白(GFP)的转基因小鼠模型，以检测重组(GFP+)与重组(GFP)NK细胞(AIMS 2和3)的发育动力学和激活潜力。表征重组炎性NK细胞的发育动力学和功能活性可以为RA的替代疗法的发展提供见解。

DESCRIPTION (provided by applicant): Natural killer (NK) cells comprise a significant proportion of joint-infiltrating cells but their role in the pathogenesis of rheumatoid arthritis (RA) remains to be elucidated. A distinct CD56bnght NK subset that produces high levels of interferon-n (IFND) and co-activates macrophages has been found to preferentially infiltrate afflicted joints. By contrast, a CD56dim NK subset that has poor inflammatory activity but heightened cytolytic function is underrepresented in inflamed joints. However, it remains to be investigated whether these two NK subsets represent sequential stages of maturation or different developmental lineages. This is an important issue because different ontogeny may reflect distinct biological functions that influence clinical disease outcomes. We, and others have shown that VDJ recombination, the process of antigen receptor rearrangement in B cells and T cells, also occurs in human and murine NK cells. Based on analysis of recombination in NK cells in a mouse model, we hypothesize that inflammatory NK cells represent an ontogenetically distinct lineage. Using indelible DNA rearrangements as a clonal marker, we propose to determine the developmental relationship between CD56bright and CD56dim NK cells in patients with RA (Aim 1). We then take advantage of a transgenic mouse model that expresses green fluorescent protein (GFP) consequent to VDJ recombination, in order to examine the developmental kinetics and activation potential of recombination"" (GFP+) versus recombination" (GFP") NK cells (Aims 2 & 3). Characterizing the developmental dynamics and functional activities of recombination* inflammatory NK cells could provide insights into the development of alternative therapies for RA.