T-CELL CYTOLYTIC ACTIVITY IN SLE

这一建议的基本前提是体外异常的CTL 世代反映了SLE和SLE患者体内T细胞介导的免疫异常 对该病的发病机制和/或发展具有重要意义 系统性红斑狼疮患者的免疫功能受损状态。在SLE PBMC培养中，一代 对抗CD3单抗反应的不受限制的细胞溶解活性通常是 异常，尽管相应的增殖反应正常，有两个 主导异常的抗CD3诱导的细胞溶解反应模式 在SLE患者中存在，表明有两个主要的离散 正常抗CD3诱导的二分性的机制 增殖与异常抗CD3诱导的非限制性细胞溶解 系统性红斑狼疮的活动性。 评估异常的生物学意义和细胞机制 在SLE中CTL的产生，我们将解决两个主要问题。1)是 SLE患者固有的CTL活性的异常产生，或IS 继发于疾病的CTL活性异常？我们将研究个人 系统性红斑狼疮患者随时间的变化及抗CD3抗体的相关变化 细胞溶解随着临床参数的改变而改变。的存在 这种相关性表明与系统性红斑狼疮相关的因素 疾病引起的是细胞溶解反应受损而不是异常 CTL生成是主要的。记录受损的CTL活动 在系统性红斑狼疮中不仅仅是体内“免疫激活”的反映， 非系统性红斑狼疮免疫风湿性疾病的患者也将 纵向研究的。在平行研究中，同卵双胞胎 系统性红斑狼疮的不协调性将被评估。健康的同卵双胞胎应该展示 与SLE同卵双胞胎的CTL生成相同的缺陷，它将 提示该缺陷在系统性红斑狼疮发病前就存在，可能是 易感因素。2)异常背后的机制是什么 系统性红斑狼疮CTL活性的产生？五种广泛的机械论可能性 将被考虑。A)T细胞是否存在外在缺陷 导致CTL生成异常？单核细胞的贡献和 将评估SLE患者异常CTL生成的血清因素。B)是 SLE T细胞是否存在成熟性缺陷？表型标记物 成熟度及表型成熟度降低的影响 将监测CTL与其目标的连接情况。应将全球T 发现细胞成熟缺陷，代中出现全局性异常 细胞内第二信使的数量将被寻找。C)有没有 系统性红斑狼疮T细胞分化缺陷？细胞因子的产生模式将 决心评估SLE从Th1样细胞分化的转变 Th2样细胞分化(导致T细胞辅助者增加 T细胞杀伤活性降低)。如果这样的转变 被发现，外源细胞因子将被添加到尝试培养中 克服分化异常。D)是否有缺陷限制？ 使T细胞亚群离散？我们将监控那些数字很小的T 已知的细胞亚群对细胞溶解活性的贡献不成比例 有选择性的缺陷。E)T细胞之间的串扰是否有缺陷 子集？将对T细胞亚群进行单独和定义的测试 组合，以评估存在过度的可能性 系统性红斑狼疮的抑制性串扰。

The underlying premise of this proposal is that abnormal in vitro CTL generation mirrors abnormal in vivo T cell-mediated immunity in SLE and has important implications for pathogenesis and/or development of the immunocompromised state seen in SLE. In SLE PBMC cultures, generation of unrestricted cytolytic activity in response to anti-CD3 mAb is usually abnormal despite normal corresponding proliferative responses, with two predominant abnormal anti-CD3-induced cytolytic response patterns existing among SLE patients, suggesting two predominant discrete mechanisms underlying the dichotomy between normal anti-CD3-induced proliferation vs abnormal anti-CD3-induced unrestricted cytolytic activity in SLE. To assess the biologic importance and cellular mechanisms of abnormal generation of CTL in SLE, we will address two major questions. 1) Is abnormal generation of CTL activity inherent to the SLE patient, or is abnormal CTL activity secondary to the disease? We will study individual SLE patients over time and correlate changes in anti-CD3-induced cytolytic lytic with changes in clinical parameters. The existence of such correlations would suggest that factors associated with the SLE disease were causing the impaired cytolytic response rather than abnormal CTL generation being primary. To document that the impaired CTL activity in SLE is not simply a reflection of in vivo "immune activation", patients with non-SLE immune-based rheumatic diseases will also be longitudinally studied. In parallel studies, monozygotic twins discordant for SLE will be evaluated. Should the healthy co-twin exhibit the same defect in CTL generation as does the SLE co-twin, it would suggest that the defect antedates the onset of SLE and may be a predisposing factor. 2) What are the mechanisms underlying abnormal generation of CTL activity in SLE? Five broad mechanistic possibilities will be considered. a) Is there a defect extrinsic to the T cell resulting in abnormal CTL generation? the contribution of monocytes and serum factors to abnormal CTL generation in SLE will be assessed. b) Is there a maturational defect in SLE T cells? Phenotypic markers of maturation and the effects of decreased phenotypic maturation on attachment of CTL to their targets will be monitored. Should global T cell maturational defects be found, global abnormalities in generation of intracellular second messengers will be sought. c) Is there a differentiation defect in SLE T cells? Cytokine production patterns will be determined to assess a shift in SLE from Th1-like cell differentiation to Th2-like cell differentiation (leading to increased T cell helper activity with decreased T cell cytolytic activity). Should such a shift be found, exogenous cytokines will be added to the cultures in attempt to overcome the abnormal differentiation. d) Is there a defect limited to discrete T cell subsets? We will monitor those numerically small T cell subsets known to disproportionately contribute to cytolytic activity for selective defects. e) Is there a defect in cross-talk among T cell subsets? T cell subsets will be tested individually and in defined combinations to assess the possibility of there being excessive inhibitory cross-talk in SLE.