B Cell Repertoire Analysis

B 细胞谱分析

• 批准号: 6823112
• 负责人: Peter E Lipsky
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ARTHRITIS AND MUSCULOSKELETAL AND SKIN DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6823112
• 关键词: B cell receptor; B lymphocyte; Sjogren's syndrome; autoantibody; autoimmune disorder; cell proliferation; gene rearrangement; human tissue; immunoglobulin genes; immunoglobulins; leukocyte activation /transformation; patient oriented research; receptor expression; single cell analysis; systemic lupus erythematosus; tissue /cell culture

The molecular basis of autoimmunity remains largely unknown. One hypothesis is that molecular abnormalities in the means by which the B cell immunoglobulin receptor is assembled or subsequently selected play a role in the tendency to develop autoantibodies. To examine this possibility, a technique has been developed to analyze the B cell receptor expressed by individual B cells without bias. Using this technique, the B cell repertoire expressed by normal individuals has been analyzed in detail. This normal data base has been employed for comparison to determine abnormalities in patients with autoimmune disease. The objective of the current study is to use single cell analysis to delineate the nature of abnormalities in the B cell immunoglobulin repertoire that might underlie autoimmune disease. The characteristics of B cells of patients with systemic lupus erythematosus (SLE) and Sjogrens's syndrome (SS) have been analyzed. The lupus repertoire shows distinct abnormalities, including evidence of enhanced mutational activity, increased receptor editing/revision and clonal expansion, consistent with intense T cell dependent stimulation. By contrast, the B cell repertoire of Sjogren's syndrome patients showed different abnormalities, including restriction of the repertoire, no enhanced mutational activity and limited receptor editing, as well as a pattern of mutations consistent with intense T cell independent stimulation. In neither disease was evidence of molecular abnormalities in receptor assembly obtained. Rather, the data are most consistent with distinct abnormalities in peripheral B cell stimulation, resulting in autoimmunity in these two autoimmune diseases. The capacity to analyze the B cell repertoire without bias has provided new insights into the nature of the abnormalities that predispose to autoimmunity. In systemic and organ-specific autoimmune diseases the B cells of interest circulate between primary and secondary lymphoid tissue and sites of inflammation. In SS, the B cells accumulating in inflamed parotid glands are polyclonal and the repertoire resembles that of the B cells in circulation. However, there is an influx or preferential homing of mutated memory B cells to the inflamed glands, which become sites of B cell proliferation and VH clonal expansion. Representative members of at least one glandular VH clone were among the circulating B cells. Circulating mutated B cells express CD27, which has become a convenient marker for identifying the memory B cell subset. In SLE patients in highly active states of disease, the number and frequency of circulating CD27 high plasma cells was greater than in SLE patients with inactive disease. The proportion of CD27+ B cells related to the duration of disease and therapeutic regiment more than the humoral levels of anti-ds DNA antibodies and circulating immune complexes, suggesting that analysis of this B cell subset may be useful to evaluate and monitor autoimmune disease activity. Ongoing Projects: 1. Homing receptor expression on T & B cell subsets 2. B cell repertoire and mutation analysis of HED-ID, xHIM, CD40def, AID & XP 3. Peripheral & tonsillar B cell subset mutation analysis 4. Absence of memory B cells in ALPS 5. B cell repertoire and mutation analysis in post-transplantation RA 6. Repertoire & mutation analysis of human splenic B cells 7. Repertoire & mutation analysis of anergic B cells in SLE 8. Role of T-regulatory suppressor cells in SLE 9. Synergy of NIK & RAF in NFkB activation 10. B cell repertoire & mutation analysis of SpA superantigen treated macaque 11. B cell repertoire & mutation analysis of ProtL & SpA superantigen treated transgenic huIg murine model 12. B cell repertoire & mutation analysis human stem cell reconstituted SCID/NOD

自身免疫的分子基础在很大程度上未知。一个假设是，在组装B细胞免疫球蛋白受体的手段中，分子异常被组装或随后选择在发展自身抗体的趋势中起作用。为了检查这种可能性，已经开发了一种技术来分析单个B细胞无偏见表达的B细胞受体。使用此技术，已详细分析了由正常个体表达的B细胞库。该正常数据库已被用于比较来确定自身免疫性疾病患者的异常。当前研究的目的是使用单细胞分析来描述可能是自身免疫性疾病的B细胞免疫球蛋白库中异常的性质。已经分析了全身性红斑狼疮（SLE）和Sjogrens综合征（SS）患者B细胞的特征。狼疮曲目显示出明显的异常，包括增强突变活性的证据，受体编辑/翻修和克隆扩张的增加，与强烈的T细胞依赖性刺激一致。相比之下，Sjogren综合征患者的B细胞库显示出不同的异常，包括限制曲目，没有增强的突变活性和受体编辑有限，以及与强烈的T细胞独立刺激一致的突变模式。在这两种疾病中，都表明获得的受体组装中分子异常。相反，数据与周围B细胞刺激的明显异常是最一致的，导致这两种自身免疫性疾病的自身免疫性。无偏见分析B细胞库的能力为自身免疫性的异常性质提供了新的见解。在全身和器官特异性的自身免疫性疾病中，感兴趣的B细胞在原发性和继发性淋巴组织和炎症部位之间循环。在SS中，在发炎的腮腺中积累的B细胞是多克隆的，并且曲目类似于循环中B细胞的B细胞。但是，突变的记忆B细胞向发炎的腺体有涌入或优先归巢，后者成为B细胞增殖和VH克隆膨胀的部位。至少一个腺体VH克隆的代表成员是循环的B细胞之一。循环突变的B细胞表达CD27，这已成为识别内存B细胞子集的方便标记。在高度活跃的疾病状态下的SLE患者中，循环CD27高浆细胞的数量和频率比患有非活性疾病的SLE患者大。与抗DS DNA抗体的体液水平和循环免疫复合物相比，与疾病和治疗团持续时间相关的CD27+ B细胞的比例更大，这表明对该B细胞子群的分析可能有助于评估和监测自身免疫性疾病活性。 正在进行的项目： 1。在T＆B细胞子集上的归巢受体表达 2。B细胞库和HED-ID，XHIM，CD40DEF，AID和XP的突变分析 3。外围和扁桃体B细胞子集突变分析 4。阿尔卑斯山中没有记忆B细胞 5。B细胞库和移植后RA中的突变分析 6。人脾脏B细胞的曲目和突变分析 7。SLE中的无性B细胞的曲目和突变分析 8。T调节抑制细胞在SLE中的作用 9。NIK＆RAF在NFKB激活中的协同作用 10。B细胞库库和Spa超抗原处理的猕猴的突变分析 11。B细胞曲目和Protl＆Spa超抗原治疗的转基因Huig Murine模型的突变分析 12。B细胞库和突变分析人类干细胞重建的SCID/NOD