Dysregulation of anergy-related factors involved in regulatory T cells defects in Systemic Lupus Erythematosus patients: Rapamycin and Vitamin D efficacy in restoring regulatory T cells
Dysregulation of anergy-related factors involved in regulatory T cells defects in Systemic Lupus Erythematosus patients: Rapamycin and Vitamin D efficacy in restoring regulatory T cells
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DOI：
10.1111/1756-185x.12509
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发表时间：
2016-12-01
影响因子：
2.5
通讯作者：
Matache, Cristiana
中科院分区：
文献类型：
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作者：
Banica, Leontina M.;Besliu, Alina N.;Matache, Cristiana
目的：系统性红斑狼疮（SLE）患者存在T细胞活化功能障碍和无反应性。因此，本研究的目的是探讨SLE患者CD 4（+）T细胞中无能量相关因子的表达与调节性T细胞（Treg）频率的关系，并确定纠正这些缺陷的策略。方法：分析外周血单个核细胞（PBMC）中Casitas B细胞淋巴瘤B（Cbl-B）和“淋巴细胞无反应性相关基因”（GRAIL）蛋白来自SLE患者和健康供体（HD）的免疫印迹。通过实时聚合酶链反应评价SLE和HD PBMC和CD 4（+）T细胞中cbl-b、grail、生长反应因子（EGFR）2和egr 3信使RNA（mRNA）。通过流式细胞术进行CD 4（+）T细胞的表型和功能表征。体外扩增方案包括在有或无雷帕霉素（Rapa）或1，25-（OH）2D 3（维生素D：VitD）的情况下，用抗CD 3和抗CD 28单克隆抗体、人重组白细胞介素（hrIL）-2激活CD 4（+）T细胞14或21天。SLE PBMC和CD 4（+）T细胞均表达低水平的egr 2/3 mRNA。SLE患者的TCLs数量减少，抑制活性受损。确定了CD 4（+）T细胞中egr 2 mRNA水平与T细胞百分比之间的相关性。在hrIL-2和Rapa或VitD存在下，CD 4（+）T细胞的实验性活化诱导SLE T细胞的扩增。然而，在长期的，只有拉帕接触SLE CD 4（+）T细胞产生大量的Tclimate与持续suppressor activity.Conclusion：我们的研究结果表明，一个新的策略，以纠正缺陷的CD 4（+）T细胞耐受机制，可能证明有益于SLE。
Aim: Systemic Lupus Erythematosus (SLE) patients display dysfunctions in T cell activation and anergy. Therefore the aims of our study were to explore the expression of anergy-related factors in CD4(+) T cells in relationship with regulatory T cells (Tregs) frequency in SLE patients and to identify strategies to redress these defects.Method: Casitas B-cell lymphoma b (Cbl-b) and 'gene related to anergy in lymphocytes' (GRAIL) proteins were analyzed in peripheral blood mononuclear cells (PBMCs) from SLE patients and healthy donors (HD) by immunoblotting. cbl-b, grail, growth response factors (egr)2 and egr3 messenger RNAs (mRNAs) were evaluated by real-time polymerase chain reaction in SLE and HD PBMCs and CD4(+) T cells. Phenotypic and functional characterization of CD4(+) T cells was performed by flow cytometry. Tregs expansion protocol consisted in culturing CD4(+) T cells for 14 or 21days of experimental activation with anti-CD3 and anti-CD28 monoclonal antibodies, human recombinant interleukin (hrIL)-2, in the absence or presence of rapamycin (Rapa) or 1,25-(OH)2D3 (vitamin D: VitD).Results: SLE PBMCs expressed low levels of Cbl-b and GRAIL proteins. Both SLE PBMCs and CD4(+) T cells expressed low levels of egr2/3 mRNAs. SLE patients had a reduced number of Tregs with impaired suppressive activity. An association between egr2 mRNA level in CD4(+) T cells and Tregs percentage was identified. Experimental activation of CD4(+) T cells in the presence of hrIL-2 and Rapa or VitD induced the expansion of SLE Tregs. However, on long-term, only Rapa exposure of SLE CD4(+) T cells yielded high numbers of Tregs with sustained suppressive activity.Conclusion: Our results suggest a new strategy to correct defects in CD4(+) T cell tolerance mechanisms that may prove beneficial in SLE.