REGULATION OF IMMUNE RESPONSES IN HUMANS AND NON-HUMAN PRIMATES

人类和非人类灵长类动物免疫反应的调节

• 批准号: 2566773
• 负责人: W STROBER
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS 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/2566773
• 关键词: B lymphocyte; CD2 molecule; CD3 molecule; T cell receptor; T lymphocyte; animal tissue; apoptosis; autoimmunity; biological signal transduction; cell cell interaction; clinical research; cytokine; cytotoxic T lymphocyte; helper T lymphocyte; human tissue; immunoregulation; interferon gamma; interleukin 2; laboratory mouse; leukocyte activation /transformation; lymphocyte proliferation; tissue /cell culture

Project 1: Autoimmune Lymphoproliferative Syndrome is a disorder characterized by massive expansion of a CD4-/CD8- T cell population and autoimmunity similar to that in lpr mice. Recently, it has been shown that both ALPS patients and lpr mice have mutations of the FAS gene and Defective apoptosis. Here, we report on immune function in ALPS patients. In initial studies we showed that the circulating level of IL-10 was greatly elevated I all ALPS patients in rough proportion to their level of CD4-/CD8- T cells. In addition, while plasma levels of TNFalpha were increased 3 - 9 fold, levels of IL-1 and IL-6 were normal. I subsequent studies, we determined cytokine production (IL-4, IL-5 IFN-gamma, Il-2) in cultured ALPS T cell subpopulations stimulated with alphaCD3/alphaCD28 or alphaCD2/alphaCD28. We showed that ALPS patients CD4+/DR+ T cells produced increased amounts of IL-4 and IL-5 (10-30 fold, n=6) and decreased amounts of IFN-gamma (2-4 fold, n=6) as compared to CD4+/DR+ T cells of controls (n=4). In parallel studies, we showed that ALPS patient peripheral B (stimulated with SAC and IL-2) and T cells (stimulated as above) produced decreased amounts of IL-10 (3-10 fold) whereas patient peripheral monocytes (stimulated with LPS) produced increased amounts of IL-10 (5 fold, n=3) as compared to control cells. Finally, we determined that patient monocytes (stimulated with LPS/IFN-gamma) produces 30-fold less Il-12 than normal cells. In conclusion, ALPS is associated with the presence of DR+ T cells which respond upon stimulation with a Th2-type cytokine profile; this, plus the increased production of IL-10 defines a cytokine mileau heading to autoimmunity. Project 2: Lamina propria, (LP) T cells are partially anergic T cells that respond poorly to a proliferative stimulus delivered via the TCR/CD3 pathway, but retain considerable ability to respond to a stimulus delivered via the CD2 co-stimulatory or accessory pathway. In the present study, we showed first that unstimulated LP T cells, as compared to unstimulated peripheral blood (PB) T cells, exhibit an increased level of apoptosis which is further increased following CD2 pathway stimulation, but not following via TCR/CD3 pathway stimulation. These results are in contrast to those obtained with PB T cells where neither stimulation via TCR/CD3 nor CD2 pathways increased the level of apoptosis above low baseline levels. We next showed that IL-2 had a sparing effect of apoptosis of unstimulated LP T cells in that IL-2 decreased and anti-IL-2 increased apoptosis of these cells; in contrast, IL-2 had no effect of apoptosis of CD2-pathway-stimulated cells. Finally, we showed that the increased apoptosis of LP T cells induces by CD2-pathway stimulation is inhibited when the Fas Antigen is blocked by a non-stimulatory anti-Fas antibody. Thus, these studies suggest that LP T cells are characterized by an increased susceptibility to Fas-mediated apoptosis most due to a "downstream" change in the Fas signaling pathway.

项目1：自身免疫性淋巴增殖性综合征是一种疾病 以CD4-/CD8-T细胞群的大规模扩张为特征 自身免疫与LPR小鼠相似。最近，它被展示出来了 阿尔卑斯患者和LPR小鼠都有Fas基因突变和 有缺陷的细胞凋亡。在这里，我们报告了阿尔卑斯山的免疫功能 病人。在最初的研究中，我们发现血液循环水平 IL-10在所有阿尔卑斯患者中显著升高，大致成比例 他们的CD4-/CD8-T细胞水平。此外，虽然血浆中的 肿瘤坏死因子α升高3~9倍，IL-1、IL-6水平正常。 在随后的研究中，我们测定了细胞因子的产生(IL-4，IL-5 干扰素-γ、IL-2)对培养的Alps T细胞亚群的影响 AlphaCD3/alphaCD28或alphaCD2/alphaCD28。我们向阿尔卑斯山患者展示了 CD4+/DR+T细胞产生更多的IL-4和IL-5(10-30倍， N=6)和减少干扰素-γ的量(2-4倍，n=6) 对照组(n=4)的CD_4~+/DR~+T细胞。在平行研究中，我们发现 阿尔茨海默病患者外周血B细胞(SAC和IL-2刺激)和T细胞 (如上刺激)产生的IL-10数量减少(3-10倍) 而患者外周血单核细胞(在内毒素刺激下)产生 IL-10含量增加(5倍，n=3)，与对照细胞相比。 最后，我们确定患者单核细胞(用 脂多糖/干扰素-γ)产生的IL-12比正常细胞少30倍。在……里面 结论阿尔卑斯综合征与DR+T细胞的存在有关 对Th2型细胞因子的刺激作出反应；这，加上 IL-10的增加定义了一个细胞因子的里程碑 自身免疫力。项目2：固有层，(LP)T细胞部分 对增殖性刺激反应差的无能T细胞 通过TCR/CD3途径，但保留相当大的反应能力 通过CD2共刺激或辅助途径传递的刺激。在……里面 本研究首先表明，未经刺激的LP T细胞，AS 与未经刺激的外周血(PB)T细胞相比， 在CD2之后进一步增加的细胞凋亡水平 途径刺激，但不通过TCR/CD3途径刺激。 这些结果与用外周血T细胞获得的结果相反 无论是通过TCR/CD3还是CD2途径的刺激都不会增加 低基线水平以上的细胞凋亡。接下来我们展示了IL-2有一个 未经刺激的LP T细胞在IL-2诱导下的抗凋亡作用 减少和抗IL-2增加了这些细胞的凋亡；相反， IL-2对CD2途径刺激的细胞无凋亡作用。 最后，我们证明了LP T细胞的凋亡增加是由 当Fas抗原被阻断时，CD2途径的刺激被抑制 一种非刺激性抗Fas抗体。因此，这些研究表明 LP T细胞的特征是易感性增加 Fas介导的细胞凋亡主要是由于Fas的“下游”变化 信号通路。