Chronic lymphocytic leukemia (CLL) is a strikingly heterogeneous disease both at the molecular level and clinical disease course. The key proliferation and survival signals malignant B cells obtain within lymph nodes or bone marrow. Moreover, CLL B cells and tumor microenvironment dynamically co-evolve organizing inflammatory and immunosuppressive microenvironment by direct contact with surrounding cells and secretion of cytokines, growth factors, or extracellular vesicles. Finding a way to weaken obtaining by malignant B cells supportive signals may improve CLL outcome and optimize treatment strategies. The aim of this study was to evaluate whether CD150 and CD180 cell surface receptors could be involved in the regulation of CLL B cells-derived cytokines (CCL3, CCL4, IL-6, and IL-10). The study was performed on malignant B cells isolated from peripheral blood of primary CLL patients. Flow cytometry, qPCR, ELISA, western blot, ex vivo cell surface ligation assay, ex vivo drug sensitivity assay, and cell viability assay were used in this study. The CCL3, CCL4, IL-6, and IL-10 mRNA expression levels were heterogeneously presented among studied CLL cases. The elevated CCL3/CCL4 and decreased IL-6/IL-10 expression level are specific features of CLL B cells with positive CD150 and CD180 expression status. Ligation of CD150 and CD180 receptors did not affect CCL3/CCL4 mRNA expression level in CLL B cells, while IL-6 and IL-10 were significantly decreased. After malignant B cells stimulation via CD150 and CD180 observed reduced IL-10 but not IL-6 level in culture supernatants. Ligation of CD150 and CD180 was linked to the classical NF-κB pathway via regulation of phosphorylation level of NF-κB inhibitor IκBα. We found several correlations between basal mRNA expression levels of CLL3, CCL4, IL-6, and IL-10 in CLL B cells and their sensitivity to chemotherapeutic drugs ex vivo. High CCL3/CCL4 and low IL-10 mRNA expression levels are associated with malignant B cells` sensitivity to BEN, while high IL-6 levels are a sign of CLL B cells` resistance to FC. The revealed involvement of CD150 and CD180 in cytokine regulation expands our knowledge of the role of CD150 and CD180 in the pathobiology of CLL and their contribution to a favorable clinical outcome. Determining the cytokines expression levels together with CD150 and CD180 expression status may help to predict the responsiveness of CLL B cells to chemotherapeutic drugs and optimize personalized chemotherapy scheme.

中文翻译：

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CD150 和 CD180 是慢性淋巴细胞白血病 B 细胞中 IL-10 表达和分泌的负调节因子。

慢性淋巴细胞白血病 (CLL) 在分子水平和临床病程上都是一种异质性显着的疾病。关键的增殖和存活信号恶性 B 细胞在淋巴结或骨髓内获得。此外，CLL B 细胞和肿瘤微环境通过与周围细胞直接接触和分泌细胞因子、生长因子或细胞外囊泡，动态地共同进化组织炎症和免疫抑制微环境。寻找一种方法来削弱恶性 B 细胞获得的支持信号可能会改善 CLL 的结果并优化治疗策略。本研究的目的是评估 CD150 和 CD180 细胞表面受体是否可能参与调节 CLL B 细胞衍生的细胞因子（CCL3、CCL4、IL-6 和 IL-10）。该研究是对从原发性 CLL 患者外周血中分离的恶性 B 细胞进行的。本研究使用流式细胞术、qPCR、ELISA、蛋白质印迹、离体细胞表面连接测定、离体药物敏感性测定和细胞活力测定。在研究的 CLL 病例中，CCL3、CCL4、IL-6 和 IL-10 mRNA 表达水平呈异质性。CCL3/CCL4 升高和 IL-6/IL-10 表达水平降低是 CD150 和 CD180 表达状态呈阳性的 CLL B 细胞的特定特征。CD150和CD180受体的连接不影响CLL B细胞中CCL3/CCL4 mRNA的表达水平，而IL-6和IL-10显着降低。在通过 CD150 和 CD180 刺激恶性 B 细胞后，在培养上清液中观察到 IL-10 水平降低，但 IL-6 水平没有降低。CD150 和 CD180 的连接通过调节 NF-κB 抑制剂 IκBα 的磷酸化水平与经典的 NF-κB 途径相关联。我们发现 CLL B 细胞中 CLL3、CCL4、IL-6 和 IL-10 的基础 mRNA 表达水平与其离体对化疗药物的敏感性之间存在一些相关性。高 CCL3/CCL4 和低 IL-10 mRNA 表达水平与恶性 B 细胞对 BEN 的敏感性有关，而高 IL-6 水平是 CLL B 细胞对 FC 抗性的标志。CD150 和 CD180 参与细胞因子调节的揭示扩大了我们对 CD150 和 CD180 在 CLL 病理生物学中的作用及其对良好临床结果的贡献的认识。