Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan.
Sanford Burnham Prebys Medical Discovery Institute, La Jolla, California, USA.
J Biol Chem. 2021 Jan-Jun;296:100419. doi: 10.1016/j.jbc.2021.100419. Epub 2021 Feb 15.
Thymus organogenesis and T cell development are coordinated by various soluble and cell-bound molecules. Heparan sulfate (HS) proteoglycans can interact with and immobilize many soluble mediators, creating fields or gradients of secreted ligands. While the role of HS in the development of many organs has been studied extensively, little is known about its function in the thymus. Here, we examined the distribution of HS in the thymus and the effect of its absence on thymus organogenesis and T cell development. We found that HS was expressed most abundantly on the thymic fibroblasts and at lower levels on endothelial, epithelial, and hematopoietic cells. To study the function of HS in the thymus, we eliminated most of HS in this organ by genetically disrupting the glycosyltransferase Ext1 that is essential for its synthesis. The absence of HS greatly reduced the size of the thymus in fetal thymic organ cultures and in vivo, in mice, and decreased the production of T cells. However, no specific blocks in T cell development were observed. Wild-type thymic fibroblasts were able to physically bind the homeostatic chemokines CCL19, CCL21, and CXCL12 ex vivo. However, this binding was abolished upon HS degradation, disrupting the CCL19/CCL21 chemokine gradients and causing impaired migration of dendritic cells in thymic slices. Thus, our results show that HS plays an essential role in the development and growth of the thymus and in regulating interstitial cell migration.
胸腺器官发生和 T 细胞发育受多种可溶性和细胞结合分子的协调。硫酸乙酰肝素(HS)蛋白聚糖可以与许多可溶性介质相互作用并使其固定,从而形成分泌配体的场或梯度。虽然 HS 在许多器官发育中的作用已被广泛研究,但对其在胸腺中的功能知之甚少。在这里,我们研究了 HS 在胸腺中的分布以及其缺失对胸腺器官发生和 T 细胞发育的影响。我们发现 HS 在胸腺成纤维细胞上表达最为丰富,在内皮细胞、上皮细胞和造血细胞上表达水平较低。为了研究 HS 在胸腺中的功能,我们通过遗传破坏对其合成至关重要的糖基转移酶 Ext1 来消除该器官中的大部分 HS。HS 的缺失大大减少了胎胸腺器官培养物中和体内(在小鼠中)胸腺的大小,并减少了 T 细胞的产生。然而,未观察到 T 细胞发育的特定阻滞。野生型胸腺成纤维细胞能够在体外物理结合稳态趋化因子 CCL19、CCL21 和 CXCL12。然而,这种结合在 HS 降解后被废除,破坏了 CCL19/CCL21 趋化因子梯度,并导致胸腺切片中树突状细胞迁移受损。因此,我们的结果表明 HS 在胸腺的发育和生长以及调节间质细胞迁移中发挥着重要作用。