Department of Ophthalmology, Peking University Third Hospital, Beijing, China.
Beijing Key Laboratory of Restoration of Damaged Ocular Nerve, Beijing, China.
Ocul Immunol Inflamm. 2023 May;31(4):748-759. doi: 10.1080/09273948.2023.2182327. Epub 2023 Mar 3.
The ocular surface and lacrimal gland have a frontline position in mucosal immunology. However, there have been few updates to the immune cell atlas of these tissues in recent years.
To map the immune cells in murine ocular surface tissues and lacrimal gland.
Central and peripheral corneas, conjunctiva, and lacrimal gland were dissociated into single cell suspensions, followed by flow cytometry. Discrepancy of immune cells between the central and peripheral corneas was compared. In the conjunctiva and lacrimal gland, myeloid cells were clustered by tSNE and FlowSOM based on the expression of F4/80, Ly6C, Ly6G, and MHC II. ILCs, type 1 immune cells, and type 3 immune cells were analyzed.
The number of immune cells in peripheral corneas was about 16 folds of that in central corneas. B cells accounted for 8.74% of immune cells in murine peripheral corneas. In the conjunctiva and lacrimal gland, most myeloid cells tended out to be monocytes, macrophages, and classical dendritic cells (cDCs). ILC3 were 6.28% and 3.63% of ILCs in the conjunctiva and lacrimal gland, respectively. Th1, Tc1, and NK cells were predominant type 1 immune cells. γδ T17 cells and ILC3 outnumbered Th17 cells among type 3 T cells.
B cells resident in murine corneas were reported for the first time. Additionally, we proposed a strategy of clustering myeloid cells to better understand their heterogeneity in the conjunctiva and lacrimal gland based on tSNE and FlowSOM. Furthermore, we identified the ILC3 in the conjunctiva and lacrimal gland for the first time. Compositions of type 1 and type 3 immune cells were summarized. Our study provides a fundamental reference and novel insights for ocular surface immune homeostasis and diseases.
眼表面和泪腺在黏膜免疫学中具有前沿地位。然而，近年来这些组织的免疫细胞图谱几乎没有更新。
绘制小鼠眼表面组织和泪腺中的免疫细胞图谱。
将中央和周边角膜、结膜和泪腺分离成单细胞悬液，然后进行流式细胞术分析。比较中央和周边角膜之间免疫细胞的差异。在结膜和泪腺中，基于 F4/80、Ly6C、Ly6G 和 MHC II 的表达，通过 tSNE 和 FlowSOM 对髓系细胞进行聚类。分析 ILC、1 型免疫细胞和 3 型免疫细胞。
周边角膜中的免疫细胞数量约为中央角膜的 16 倍。B 细胞占小鼠周边角膜免疫细胞的 8.74%。在结膜和泪腺中，大多数髓系细胞倾向于单核细胞、巨噬细胞和经典树突状细胞（cDC）。结膜和泪腺中的 ILC3 分别占 ILC 的 6.28%和 3.63%。Th1、Tc1 和 NK 细胞是主要的 1 型免疫细胞。γδ T17 细胞和 ILC3 在 3 型 T 细胞中数量多于 Th17 细胞。
首次报道了驻留在小鼠角膜中的 B 细胞。此外，我们提出了一种基于 tSNE 和 FlowSOM 的聚类髓系细胞的策略，以更好地了解它们在结膜和泪腺中的异质性。此外，我们首次在结膜和泪腺中鉴定出 ILC3。总结了 1 型和 3 型免疫细胞的组成。我们的研究为眼表面免疫稳态和疾病提供了基本参考和新的见解。
