In vitro stimulation of mouse spleen cells by COL17A1

Tertiary lymphoid tissues (TLTs) are formed in systemic organs manifesting chronic inflammation. Herein, we found that the renal pelvis (RP) could form urinary tract-associated lymphoid tissues (UTALTs) in a TLT-formation manner in humans and mice with chronic kidney disease (CKD), regardless of infectious pyelonephritis. Furthermore, collagen type XVII alpha 1 chain (COL17A1), localized ectopically to the transitional epithelium (TE) covering the UTALT, where it participated in TE development via immunological stimulation of UTALT-forming cells. The spleens were isolated from C57BL/6N at 6-month old, minced, washed, resuspended, and cultured. These cultured cells were stimulated with or without recombinant human COL17A1 for 3 hours. And then, RNA was collected and gene expression was analyzed.

三级淋巴组织（Tertiary lymphoid tissues, TLTs）是在伴发慢性炎症的全身器官中形成的。本研究发现，无论是否合并感染性肾盂肾炎，肾盂（renal pelvis, RP）在慢性肾病（chronic kidney disease, CKD）患者与小鼠体内均可通过三级淋巴组织形成方式生成尿路相关淋巴组织（urinary tract-associated lymphoid tissues, UTALTs）。进一步研究表明，XVII型胶原α1链（collagen type XVII alpha 1 chain, COL17A1）异位表达于覆盖尿路相关淋巴组织的移行上皮（transitional epithelium, TE）中，并通过对尿路相关淋巴组织形成细胞的免疫刺激参与移行上皮的发育。我们从6月龄C57BL/6N小鼠体内分离脾脏，经切碎、洗涤、重悬后进行体外培养。将培养得到的细胞分别以重组人COL17A1刺激与不刺激处理3小时，随后收集核糖核酸（RNA）并开展基因表达分析。