T-CELL CYTOLYTIC ACTIVITY IN SLE

这一建议的基本前提是，异常的体外CTL 产生反映了SLE中体内T细胞介导的免疫异常， 对糖尿病的发病机制和/或发展有重要意义。 SLE患者免疫功能低下。 在SLE PBMC培养物中， 对抗CD 3 mAb的不受限制的细胞溶解活性通常 尽管相应的增殖反应正常，但异常，有两个 主要异常抗CD 3诱导的细胞溶解反应模式 存在于SLE患者中，提示两种主要的离散型 正常抗CD 3诱导的 增殖与异常抗CD 3诱导的非限制性细胞溶解 活动在SLE 评估异常的生物学重要性和细胞机制， 为了在SLE中产生CTL，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 继发于疾病的异常CTL活性？ 我们将研究个人 SLE患者随着时间的推移和抗CD 3诱导的 细胞溶解，临床参数发生变化。 的存在 这种相关性表明与SLE相关的因素 疾病导致细胞溶解反应受损，而不是异常 CTL生成是主要的。 为了证明受损的CTL活性 在SLE中，并不仅仅是体内“免疫激活”的反映， 非SLE免疫性风湿性疾病患者也将 纵向研究。 在平行研究中，单卵双胞胎 将评价SLE的不一致性。 健康的双胞胎是否应该 在CTL生成中的缺陷与SLE共双胞胎相同， 提示该缺陷早于SLE的发病，可能是SLE的一个潜在的病因。 诱发因素 2)异常的机制是什么 SLE中CTL活性的产生？ 五种广泛的机械可能性 将被考虑。 a）T细胞是否存在外源性缺陷 导致CTL产生异常 单核细胞的贡献， 将评估SLE中异常CTL产生的血清因素。 B）是 SLE T细胞有成熟缺陷吗？ 表型标志物 成熟以及表型成熟降低对 将监测CTL与其靶的附着。 全球T 细胞成熟缺陷被发现，全球异常的代 第二个信使将被寻找。 C.是否有A SLE T细胞的分化缺陷？ 细胞因子产生模式将 确定评估SLE从Th 1样细胞分化的转变 Th 2样细胞分化（导致T细胞辅助细胞增加） T细胞溶解活性降低）。 这样的转变 为了发现，将外源性细胞因子添加到培养物中， 以克服异常分化。 d）是否存在缺陷限制 离散的T细胞亚群 我们将监视那些数值较小的T 已知对细胞溶解活性有不成比例贡献的细胞亚群 选择性缺陷。 e）T细胞之间的串扰是否存在缺陷 子集？ T细胞亚群将单独检测，并在定义的 联合评估是否存在过度 SLE中抑制性串扰。

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.