Conjoint Kidney Research Laboratory, Pathology Queensland, Queensland Institute of Medical Research, Level 9, Bancroft Centre, Herston 4006, Queensland, Australia.
Am J Physiol Renal Physiol. 2013 Nov 15;305(10):F1391-401. doi: 10.1152/ajprenal.00318.2013. Epub 2013 Sep 18.
Dendritic cells (DCs) play critical roles in immune-mediated kidney diseases. Little is known, however, about DC subsets in human chronic kidney disease, with previous studies restricted to a limited set of pathologies and to using immunohistochemical methods. In this study, we developed novel protocols for extracting renal DC subsets from diseased human kidneys and identified, enumerated, and phenotyped them by multicolor flow cytometry. We detected significantly greater numbers of total DCs as well as CD141(hi) and CD1c(+) myeloid DC (mDCs) subsets in diseased biopsies with interstitial fibrosis than diseased biopsies without fibrosis or healthy kidney tissue. In contrast, plasmacytoid DC numbers were significantly higher in the fibrotic group compared with healthy tissue only. Numbers of all DC subsets correlated with loss of kidney function, recorded as estimated glomerular filtration rate. CD141(hi) DCs expressed C-type lectin domain family 9 member A (CLEC9A), whereas the majority of CD1c(+) DCs lacked the expression of CD1a and DC-specific ICAM-3-grabbing nonintegrin (DC-SIGN), suggesting these mDC subsets may be circulating CD141(hi) and CD1c(+) blood DCs infiltrating kidney tissue. Our analysis revealed CLEC9A(+) and CD1c(+) cells were restricted to the tubulointerstitium. Notably, DC expression of the costimulatory and maturation molecule CD86 was significantly increased in both diseased cohorts compared with healthy tissue. Transforming growth factor-β levels in dissociated tissue supernatants were significantly elevated in diseased biopsies with fibrosis compared with nonfibrotic biopsies, with mDCs identified as a major source of this profibrotic cytokine. Collectively, our data indicate that activated mDC subsets, likely recruited into the tubulointerstitium, are positioned to play a role in the development of fibrosis and, thus, progression to chronic kidney disease.
树突状细胞 (DCs) 在免疫介导的肾脏疾病中发挥关键作用。然而，人们对人类慢性肾脏病中的 DC 亚群知之甚少，以前的研究仅限于有限的一组病理学，并使用免疫组织化学方法。在这项研究中，我们开发了从患病人类肾脏中提取肾 DC 亚群的新方案，并通过多色流式细胞术对其进行鉴定、计数和表型分析。我们发现，与无纤维化的患病活检或健康肾组织相比，具有间质纤维化的患病活检中的总 DC 以及 CD141(hi)和 CD1c(+)髓样 DC (mDC)亚群数量显著增加。相比之下，与健康组织相比，成纤维细胞组中的浆细胞样 DC 数量显著更高。所有 DC 亚群的数量与肾功能丧失相关，记录为估计肾小球滤过率。CD141(hi) DC 表达 C 型凝集素结构域家族 9 成员 A (CLEC9A)，而大多数 CD1c(+) DC 缺乏 CD1a 和 DC 特异性 ICAM-3 抓取非整合素 (DC-SIGN) 的表达，表明这些 mDC 亚群可能是循环 CD141(hi)和 CD1c(+)血液 DC 浸润肾脏组织。我们的分析表明，CLEC9A(+)和 CD1c(+)细胞仅限于肾小管间质。值得注意的是，与健康组织相比，两组患病组织中 DC 表达的共刺激和成熟分子 CD86 显著增加。与非纤维化活检相比，具有纤维化的患病活检中分离组织上清液中的转化生长因子-β水平显著升高，mDC 被鉴定为这种促纤维化细胞因子的主要来源。总的来说，我们的数据表明，激活的 mDC 亚群可能被募集到肾小管间质中，在纤维化的发展中发挥作用，从而导致慢性肾脏病的进展。
