Blokland Sofie L M, van Vliet-Moret Fréderique M, Hillen Maarten R, Pandit Aridaman, Goldschmeding Roel, Kruize Aike A, Bouma Gerben, van Maurik André, Olek Sven, Hoffmueller Ulrich, van Roon Joel A G, Radstake Timothy R D J
Department of Immunology, Laboratory of Translational Immunology, Utrecht University, Utrecht, The Netherlands.
Department of Rheumatology & Clinical Immunology, Utrecht University, Utrecht, The Netherlands.
Rheumatology (Oxford). 2020 Feb 1;59(2):335-343. doi: 10.1093/rheumatology/kez268.
To investigate whether epigenetic cell counting represents a novel method to quantify immune cells in salivary glands of patients with different forms of Sjögren's and sicca syndrome and to capture immunopathology and potentially aid in diagnosis.
DNA from frozen salivary gland tissue sections of sicca patients was used for bisulphite conversion of demethylated DNA cytosine residues, followed by cell-specific quantitative PCR to calculate cell percentages in relation to total tissue cell numbers as quantified by housekeeping gene demethylation. The percentages of epigenetically quantified cells were correlated to RNA expression of matched salivary gland tissue and histological and clinical parameters.
The percentages of epigenetically quantified CD3, CD4, CD8, T follicular helper (Tfh) cells, FoxP3+ regulatory T cells and B cells were significantly increased in the salivary glands of patients with SS. Unsupervised clustering using these percentages identified patient subsets with an increased lymphocytic focus score and local B cell hyperactivity and classifies patients different from conventional classification criteria. In particular, Tfh cells were shown to strongly correlate with the expression of CXCL13, lymphocytic focus scores, local B cell hyperactivity and anti-SSA positivity.
Epigenetic cell counting is a promising novel tool to objectively and easily quantify immune cells in the labial salivary gland of sicca patients, with a relatively small amount of tissue needed. In view of the potential of this technique to include a huge number of (cell-specific) biomarkers, this opens up new standardized ways of salivary gland analysis with high relevance for patient classification, understanding of immunopathology and monitoring of drug responses in clinical trials.
研究表观遗传学细胞计数是否代表一种新方法，用于量化不同形式干燥综合征和口干燥症患者唾液腺中的免疫细胞，并捕捉免疫病理学特征， potentially aid in diagnosis.
使用干燥症患者冷冻唾液腺组织切片的DNA进行去甲基化DNA胞嘧啶残基的亚硫酸氢盐转化，随后进行细胞特异性定量PCR，以计算相对于管家基因去甲基化定量的总组织细胞数的细胞百分比。表观遗传学定量细胞的百分比与匹配唾液腺组织的RNA表达、组织学和临床参数相关。
干燥综合征患者唾液腺中表观遗传学定量的CD3、CD4、CD8、滤泡辅助性T细胞（Tfh）、FoxP3+调节性T细胞和B细胞的百分比显著增加。使用这些百分比进行无监督聚类可识别出淋巴细胞聚集评分增加和局部B细胞活性亢进的患者亚组，并对不同于传统分类标准的患者进行分类。特别是，Tfh细胞与CXCL13的表达、淋巴细胞聚集评分、局部B细胞活性亢进和抗SSA阳性密切相关。
表观遗传学细胞计数是一种有前景的新工具，可客观、轻松地量化干燥症患者唇腺中的免疫细胞，所需组织量相对较少。鉴于该技术包含大量（细胞特异性）生物标志物的潜力，这为唾液腺分析开辟了新的标准化方法，对患者分类、免疫病理学理解和临床试验中药物反应监测具有高度相关性。
