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

描述（由申请人提供）： 自然杀伤（NK）细胞包括一个显着比例的关节浸润细胞，但它们在类风湿关节炎（RA）的发病机制中的作用仍有待阐明。已发现产生高水平干扰素-n（IFND）并共激活巨噬细胞的独特的CD 56 bnight NK亚群优先浸润患病关节。相比之下，CD 56 dim NK亚群具有较差的炎症活性，但细胞溶解功能增强，在炎症关节中代表性不足。然而，这两个NK亚群是否代表成熟的连续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不同的生物功能，影响临床疾病的结果。我们和其他人已经表明VDJ重组，即B细胞和T细胞中抗原受体重排的过程，也发生在人和鼠NK细胞中。基于小鼠模型中NK细胞重组的分析，我们假设炎性NK细胞代表个体发生学上不同的谱系。使用不可磨灭的DNA重排作为克隆标记，我们建议确定RA患者CD 56 bright和CD 56 dim NK细胞之间的发育关系（目的1）。然后，我们利用在VDJ重组后表达绿色荧光蛋白（GFP）的转基因小鼠模型，以检查重组”（GFP+）与重组”（GFP”）NK细胞的发育动力学和活化潜力（目的2和3）。表征重组 * 炎性NK细胞的发育动力学和功能活性可以为RA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.