Neuroanatomy, University Medical Center Göttingen, Göttingen, Germany.
Pediatric Infectious Diseases, University Children's Hospital Mannheim, Heidelberg University, Heidelberg, Germany.
Immunol Res. 2017 Dec;65(6):1110-1123. doi: 10.1007/s12026-017-8952-9.
Chlamydia trachomatis (CT) and the Chlamydophila species (CS) Chlamydophila pneumoniae (CPn), and Chlamydophila psittaci (CPs) are suggested to induce autoantibodies causative of several human autoimmune disorders like rheumatoid arthritis and systemic lupus erythematosus (SLE). The aim of the present study was therefore to identify cellular protein interaction partners with antisera to CT (α-CT) or CS (α-CS) and to identify functional consequences of such interaction in vitro. As detected with a commercial first trimester human prenatal brain multiprotein array (hEXselect, Engine, Germany), the most frequent interaction partner with both α-CT and α-CS was the ribosomal small subunit protein RPS27a. This could be confirmed by Western blot analysis with a recombinant RPS27a sample. In addition, immunocytochemistry with both antisera in the human choroid plexus papilloma cell line HIBCPP revealed a granular cytoplasmic staining, and Western blot analysis with whole-cell protein samples of HIBCPP cells revealed both antisera to label protein bands of different molecular weights and intensity. By 2D Western blot analysis and mass spectrometry, one of the protein spots interacting with α-CT could be identified as the RPS27a. Finally, two different methods for the detection of protein synthesis activity, the SUnSET technique and an HPG fluorescence assay revealed both antisera to cause reduced translational activity in HIBCPP cells. Together with previous findings of RPS27a as an autoimmune target in a mouse model of systemic lupus erythematosus (SLE), these results suggest that infections with CT and/or CS could induce SLE-associated immune modifications. However, direct evidence for a pathogenic role of these interactions for SLE demands further investigations.
沙眼衣原体（CT）和衣原体属（CS）肺炎衣原体（CPn）和鹦鹉热衣原体（CPs）被认为会诱导自身抗体，从而导致几种人类自身免疫性疾病，如类风湿关节炎和系统性红斑狼疮（SLE）。因此，本研究的目的是鉴定与 CT（α-CT）或 CS（α-CS）抗血清发生相互作用的细胞蛋白相互作用伙伴，并鉴定体外这种相互作用的功能后果。如商业性的用于检测人早孕期胎儿大脑多蛋白阵列（hEXselect，Engine，德国）检测到的，与α-CT 和 α-CS 最频繁相互作用的是核糖体小亚基蛋白 RPS27a。这可以通过使用重组 RPS27a 样本的 Western blot 分析来确认。此外，在人脉络丛乳头状瘤细胞系 HIBCPP 中用两种抗血清进行免疫细胞化学染色，显示出颗粒状细胞质染色，用 HIBCPP 细胞的全细胞蛋白样本进行 Western blot 分析显示，两种抗血清都可以标记不同分子量和强度的蛋白条带。通过 2D Western blot 分析和质谱分析，与α-CT 相互作用的一个蛋白点被鉴定为 RPS27a。最后，使用两种不同的检测蛋白合成活性的方法，SUnSET 技术和 HPG 荧光测定法，发现两种抗血清都导致 HIBCPP 细胞的翻译活性降低。结合之前在系统性红斑狼疮（SLE）小鼠模型中发现 RPS27a 是自身免疫靶标的研究结果，这些结果表明，CT 和/或 CS 的感染可能会引起与 SLE 相关的免疫改变。然而，这些相互作用对 SLE 的致病作用的确切证据需要进一步研究。
