Stikvoort Arwen, Chen Yang, Rådestad Emelie, Törlén Johan, Lakshmikanth Tadepally, Björklund Andreas, Mikes Jaromir, Achour Adnane, Gertow Jens, Sundberg Berit, Remberger Mats, Sundin Mikael, Mattsson Jonas, Brodin Petter, Uhlin Michael
Department of Oncology-Pathology, Karolinska Institute, Stockholm, Sweden.
Science for Life Laboratory, Department of Medicine, Karolinska Institute, Stockholm, Sweden.
Front Immunol. 2017 Jun 19;8:717. doi: 10.3389/fimmu.2017.00717. eCollection 2017.
Chronic graft-versus-host disease (cGVHD) is a debilitating complication arising in around half of all patients treated with an allogeneic hematopoietic stem cell transplantation. Even though treatment of severe cGVHD has improved during recent years, it remains one of the main causes of morbidity and mortality in affected patients. Biomarkers in blood that could aid in the diagnosis and classification of cGVHD severity are needed for the development of novel treatment strategies that can alleviate symptoms and reduce the need for painful and sometimes complicated tissue biopsies. Methods that comprehensively profile complex biological systems such as the immune system can reveal unanticipated markers when used with the appropriate methods of data analysis. Here, we used mass cytometry, flow cytometry, enzyme-linked immunosorbent assay, and multiplex assays to systematically profile immune cell populations in 68 patients with varying grades of cGVHD. We identified multiple subpopulations across T, B, and NK-cell lineages that distinguished patients with cGVHD from those without cGVHD and which were associated in varying ways with severity of cGVHD. Specifically, initial flow cytometry demonstrated that patients with more severe cGVHD had lower mucosal-associated T cell frequencies, with a concomitant higher level of CD38 expression on T cells. Mass cytometry could identify unique subpopulations specific for cGVHD severity albeit with some seemingly conflicting results. For instance, patients with severe cGVHD had an increased frequency of activated B cells compared to patients with moderate cGVHD while activated B cells were found at a reduced frequency in patients with mild cGVHD compared to patients without cGVHD. Moreover, results indicate it may be possible to validate mass cytometry results with clinically viable, smaller flow cytometry panels. Finally, no differences in levels of blood soluble markers could be identified, with the exception for the semi-soluble combined marker B-cell activating factor/B cell ratio, which was increased in patients with mild cGVHD compared to patients without cGVHD. These findings suggest that interdependencies between such perturbed subpopulations of cells play a role in cGVHD pathogenesis and can serve as future diagnostic and therapeutic targets.
慢性移植物抗宿主病（cGVHD）是一种致残性并发症，发生在接受异基因造血干细胞移植治疗的所有患者中的约一半。尽管近年来重度cGVHD的治疗有所改善，但它仍然是受影响患者发病和死亡的主要原因之一。开发能够缓解症状并减少对痛苦且有时复杂的组织活检需求的新治疗策略，需要血液中的生物标志物来辅助cGVHD严重程度的诊断和分类。当与适当的数据分析方法一起使用时，全面分析复杂生物系统（如免疫系统）的方法可以揭示意想不到的标志物。在这里，我们使用质谱流式细胞术、流式细胞术、酶联免疫吸附测定和多重测定法，系统地分析了68例不同等级cGVHD患者的免疫细胞群体。我们在T、B和NK细胞谱系中鉴定出多个亚群，这些亚群将cGVHD患者与非cGVHD患者区分开来，并以不同方式与cGVHD的严重程度相关。具体而言，最初的流式细胞术表明，cGVHD较严重的患者黏膜相关T细胞频率较低，同时T细胞上CD38表达水平较高。质谱流式细胞术可以识别特定于cGVHD严重程度的独特亚群，尽管结果有些看似矛盾。例如，与中度cGVHD患者相比，重度cGVHD患者活化B细胞频率增加，而与非cGVHD患者相比，轻度cGVHD患者活化B细胞频率降低。此外，结果表明，有可能用临床上可行的较小流式细胞术检测板验证质谱流式细胞术的结果。最后，除了半溶性联合标志物B细胞活化因子/B细胞比率外，未发现血液可溶性标志物水平存在差异，与非cGVHD患者相比，轻度cGVHD患者的该标志物升高。这些发现表明，这些受干扰的细胞亚群之间的相互依赖性在cGVHD发病机制中起作用，并可作为未来的诊断和治疗靶点。
