The chemokine receptorCX3CR1 mediates homing of MHC class II -: Positive cells to the normal mouse corneal epithelium
The chemokine receptorCX3CR1 mediates homing of MHC class II -: Positive cells to the normal mouse corneal epithelium
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DOI：
10.1167/iovs.06-0746
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发表时间：
2007-04-01
影响因子：
4.4
通讯作者：
McMenamin, Paul G.
中科院分区：
文献类型：
--
作者：
Chinnery, Holly R.;Ruitenberg, Marc J.;McMenamin, Paul G.
目的。最近的研究表明，角膜基质和上皮中存在巨噬细胞和树突状细胞（DC）群体，尽管精确的表型和分布仍存在争议。 CX(3)CR1 是趋化因子 fractalkine 的唯一受体，由这些单核细胞衍生的细胞表达。转基因 CX(3)CR1(GFP) 小鼠，其中 CX(3)CR1 基因的一个（杂合）或两个（纯合）拷贝被增强型绿色荧光蛋白 (eGFP) 取代，用于表征小鼠角膜中的单核细胞衍生细胞，并确定该受体的表达是否影响这些细胞招募到正常细胞中。 角膜。方法。使用针对表型标记物主要组织相容性复合物 (MHC) II 类、CD169、CD68、CD11b 和 CD45 的抗白细胞抗体对整体角膜进行免疫染色，并通过落射荧光和共聚焦显微镜进行分析。对野生型、CX(3)CR1(+/GFP) 杂合子、CX(3)CR1(GFP/GFP) 纯合子和 CX(3)CR1 敲除小鼠的上皮内 MHC II+ 类细胞密度进行定量。结果。与野生型小鼠相比，CX(3)CR1缺陷型小鼠角膜上皮中MHC II+类细胞（假定的DC）数量显着减少（P < 0.009），并且存在的少数细胞不具有典型的树状形态。上皮细胞中未发现 GFP(+) MHC IF 类细胞。对杂合子和纯合子（CX(3)CR1 缺陷）小鼠的角膜进行双重免疫染色，结果显示 GFP+ 细胞在整个角膜基质中具有更多的多形性形态。其为CDIIb(+) CD169+，并且具有不同程度的CD68和MHC II类表达。这些基质细胞的免疫表型和形态强烈指示巨噬细胞表型。结论。本研究确定了 CX(3)CR1 在 MHC II 类 + 推定 DC 正常募集到角膜上皮中的作用，并建立了研究角膜疾病期间单核细胞衍生细胞和 fractalkine/CX(3)CR1 相互作用的模型。
PURPOSE. Recent investigations have revealed that populations of macrophages and dendritic cells (DCs) are present in the stroma and epithelium of the cornea, although the precise phenotype and distribution are still controversial. CX(3)CR1, the sole receptor for the chemokine fractalkine, is expressed by these monocyte-derived cells. Transgenic CX(3)CR1(GFP) mice, in which either one (heterozygous) or both (homozygous) copies of the CX(3)CR1 gene were replaced by enhanced green fluorescent protein (eGFP), were used to characterize monocyte-derived cells in the mouse cornea and to determine whether the expression of this receptor influences the recruitment of these cells into the normal cornea.METHODS. Wholemount corneas were immunostained with anti-leukocyte antibodies to the phenotypic markers major histocompatibility complex (MHC) class II, CD169, CD68, CD11b, and CD45 and analyzed by epifluorescence and confocal microscopy. The density of intraepithelial MHC class II+ cells was quantified in wild-type, CX(3)CR1(+/GFP) heterozygous, CX(3)CR1(GFP/GFP) homozygous, and CX(3)CR1-knockout mice.RESULTS. There was a significant reduction in the number of MHC class II+ cells (putative DCs) in the corneal epithelium of CX(3)CR1-deficient mice (P < 0.009) compared with wild-type mice, and the few cells that were present did not possess classic dendriform morphology. No GFP(+) MHC class IF cells were noted in the epithelium. Dual immunostaining of corneas in both heterozygous and homozygous (CX(3)CR1-deficient) mice revealed GFP+ cells with a more pleomorphic morphology throughout the entire corneal stroma. that were CDIIb(+) CD169+, and had variable degrees of expression of CD68 and MHC class II. The immunophenotype and morphology of these intrastromal cells is strongly indicative of a macrophage phenotype.CONCLUSIONS. This study has identified a role for CX(3)CR1 in the normal recruitment of MHC class II+ putative DCs into the corneal epithelium and establishes a model for investigating monocyte-derived cells and fractalkine/CX(3)CR1 interactions during corneal disease.