MHC class II expression and potential antigen-presenting cells in the retina during experimental autoimmune uveitis.
MHC class II expression and potential antigen-presenting cells in the retina during experimental autoimmune uveitis.
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DOI：
10.1186/s12974-017-0915-5
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发表时间：
2017-07-18
影响因子：
9.3
通讯作者：
Willermain F
中科院分区：
文献类型：
--
作者：
Lipski DA;Dewispelaere R;Foucart V;Caspers LE;Defrance M;Bruyns C;Willermain F
在实验性自身免疫性葡萄膜炎（EAU）的发展过程中，哪些细胞类型负责视网膜中的自身抗原呈递存在争议。在本研究中，我们旨在鉴定和表征EAU期间易表达主要组织相容性复合体（MHC） II类的视网膜驻留和浸润细胞。通过过继性转移irbp1 -20免疫动物的自身反应性淋巴细胞，在C57BL/6小鼠中诱导EAU。用免疫染色法观察MHCⅱ类蛋白的表达。流式细胞术（FC）分析，视网膜解剖和酶消化成单细胞悬浮液。通过FC对3个MHC II+视网膜细胞群进行分类，并对其RNA进行RNA- seq处理。免疫染色显示EAU强烈诱导MHC II类表达，特别是在炎症血管水平的视网膜内，在严重炎症的眼睛中延伸到视网膜外层和视网膜下间隙。大多数MHC II+类细胞表达造血标志物IBA1。FC定量分析表明MHC II类诱导与疾病严重程度显著相关，并与共刺激分子表达上调相关。特别是，大多数MHC iii类细胞在EAU期间表达共刺激分子。进一步的表型分析鉴定出MHC II类+视网膜细胞群：低MHC II类表达的CD45 - CD11b -非造血细胞和高MHC II类表达的CD45+CD11b+造血细胞，可进一步分为Ly6C+和Ly6C -细胞，可能与浸润性巨噬细胞和驻地小胶质细胞相对应。对这三个分类的群体进行转录组分析，发现样本聚类明显，分别在Ly6C+和Ly6C−细胞中巨噬细胞标记物和小胶质细胞标记物富集。功能注释分析显示，两种造血细胞群在MHC ii类相关抗原呈递和T细胞活化方面都比非造血细胞更有能力。我们的研究结果强调了造血细胞在局部抗原呈递中的潜力，无论其Ly6C表达如何。我们的工作进一步提供了在EAU期间表达MHC ii类视网膜细胞的首次转录组学研究，并提供了一系列可能涉及视网膜自身免疫发病机制的新候选基因。本文的在线版本（doi:10.1186/s12974-017-0915-5）包含补充材料，可供授权用户使用。
Controversy exists regarding which cell types are responsible for autoantigen presentation in the retina during experimental autoimmune uveitis (EAU) development. In this study, we aimed to identify and characterize the retinal resident and infiltrating cells susceptible to express major histocompatibility complex (MHC) class II during EAU. EAU was induced in C57BL/6 mice by adoptive transfer of autoreactive lymphocytes from IRBP1-20-immunized animals. MHC class II expression was studied by immunostainings on eye cryosections. For flow cytometry (FC) analysis, retinas were dissected and enzymatically digested into single-cell suspensions. Three MHC class II+ retinal cell populations were sorted by FC, and their RNA processed for RNA-Seq. Immunostainings demonstrate strong induction of MHC class II expression in EAU, especially in the inner retina at the level of inflamed vessels, extending to the outer retinal layers and the subretinal space in severely inflamed eyes. Most MHC class II+ cells express the hematopoietic marker IBA1. FC quantitative analyses demonstrate that MHC class II induction significantly correlates with disease severity and is associated with upregulation of co-stimulatory molecule expression. In particular, most MHC class IIhi cells express co-stimulatory molecules during EAU. Further phenotyping identified three MHC class II+ retinal cell populations: CD45−CD11b− non-hematopoietic cells with low MHC class II expression and CD45+CD11b+ hematopoietic cells with higher MHC class II expression, which can be further separated into Ly6C+ and Ly6C− cells, possibly corresponding to infiltrating macrophages and resident microglia. Transcriptome analysis of the three sorted populations leads to a clear sample clustering with some enrichment in macrophage markers and microglial cell markers in Ly6C+ and Ly6C− cells, respectively. Functional annotation analysis reveals that both hematopoietic cell populations are more competent in MHC class II-associated antigen presentation and in T cell activation than non-hematopoietic cells. Our results highlight the potential of cells of hematopoietic origin in local antigen presentation, whatever their Ly6C expression. Our work further provides a first transcriptomic study of MHC class II-expressing retinal cells during EAU and delivers a series of new candidate genes possibly implicated in the pathogenesis of retinal autoimmunity. The online version of this article (doi:10.1186/s12974-017-0915-5) contains supplementary material, which is available to authorized users.
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影响因子：
3.7
作者：
Gonzalez-Pena, Dianelys;Nixon, Scott E.;Rodriguez-Zas, Sandra L.
通讯作者：
Rodriguez-Zas, Sandra L.
影响因子：
4.1
作者：
DICK, AD;FORD, AL;SEDGWICK, JD
通讯作者：
SEDGWICK, JD
影响因子：
3.3
作者：
Lewis, Nuruddeen D.;Hill, Jonathan D.;Modis, Louise K.
通讯作者：
Modis, Louise K.
影响因子：
4.3
作者：
FUJIKAWA, LS;CHAN, CC;NUSSENBLATT, RB
通讯作者：
NUSSENBLATT, RB
影响因子：
3.9
作者：
LIGHTMAN, S
通讯作者：
LIGHTMAN, S