JAK Inhibitor Withdrawal Causes a Transient Proinflammatory Cascade: A Potential Mechanism for Major Adverse Cardiac Events

This is a study of mesenchymal stromal cells (MSCs) from salivary glands. The salivary glands were derived from participants who had dry eye or dry mouth symptoms but did not have evidence of systemic autoimmune disease (sicca controls). The aim of the study was to determine the effects of Type I and Type II Janus kinase (JAK) inhibitors (JAKinibs) withdrawal on these cells in vitro. The MSCs were treated in vitro with a) IFN-gamma (10 ng/mL) for the entire duration, b) IFN-gamma (10 ng/mL) and withdrawal of ruxolitinib (1 µM), c) IFN-gamma (10 ng/mL) and withdrawal CHZ-868 (1 µM), and d) IFN-gamma (10 ng/mL) and ruxolitinib (1 µM) for the entire duration. For the drug (CHZ-868 and ruxolitinib) conditions, after IFN-gamma treatment for 48 hours, we washed the cells with PBS and treated them with standard growth media with 0.01% DMSO, 10 ng/mL IFN-gamma + 0.01% DMSO, or 10 ng/mL IFN-gamma + JAKinib for 48 hours. For withdrawal conditions, we washed the cells twice and then treated them with standard growth media with either 0.01% DMSO or with 10 ng/mL IFN-gamma + 0.01% DMSO for 24 hours before harvest. ]]>
Inclusion: Referred for labial salivary gland biopsy for suspected Sjögren diseaseExclusion: <18 years of age or not eligible for labial salivary gland biopsy]]>

本研究针对唾液腺来源的间充质基质细胞（mesenchymal stromal cells, MSCs）展开。研究所用唾液腺取自存在眼干或口干症状，但未检出全身性自身免疫病的受试者（干燥对照，sicca controls）。本研究旨在明确Ⅰ型与Ⅱ型Janus激酶（JAK）抑制剂（JAKinibs）停药对这类细胞的体外影响。

实验对间充质基质细胞设置如下4种体外处理方案：a) 全程予以干扰素-γ（IFN-gamma，10 ng/mL）处理；b) 予以干扰素-γ（10 ng/mL）并停用鲁索替尼（ruxolitinib，1 µM）；c) 予以干扰素-γ（10 ng/mL）并停用CHZ-868（1 µM）；d) 全程予以干扰素-γ（10 ng/mL）与鲁索替尼（1 µM）处理。针对药物（CHZ-868与鲁索替尼）处理组：先以干扰素-γ处理细胞48小时后，用磷酸盐缓冲液（PBS）洗涤细胞，随后采用含0.01%二甲基亚砜（DMSO）的标准生长培养基、10 ng/mL干扰素-γ+0.01% DMSO，或10 ng/mL干扰素-γ+JAK抑制剂继续培养48小时。针对停药处理组：先将细胞洗涤两次，随后采用含0.01% DMSO或10 ng/mL干扰素-γ+0.01% DMSO的标准生长培养基培养24小时后收获细胞。

纳入标准：因疑似干燥综合征（Sjögren disease）而行唇腺活检的受试者。
排除标准：年龄未满18周岁，或不符合唇腺活检指征。