Gao Jingjing, Li Chuntuan, Lin Xingzhi, Zhuang Yanling, Wang Mingquan, Lin Hongjun, Zhu Xiongpeng
Department of Blood transfusion, Quanzhou First Hospital affiliated to Fujian Medical University, Quanzhou, Fujian, 362000, China.
Department of Hematology, Quanzhou First Hospital affiliated to Fujian Medical University, Quanzhou, Fujian, 362000, China.
Ann Hematol. 2025 Apr;104(4):2449-2459. doi: 10.1007/s00277-025-06273-6. Epub 2025 Apr 10.
Autoimmune haemolytic anaemia (AIHA) is common secondary to diffuse large B-cell lymphoma (DLBCL). However, there are no reports on tumour B-cells differentiating into plasmablasts in DLBCL secondary AIHA. To state impact of the interferon regulatory factor 4 (IRF4) on DLBCL and explore the mechanism of IRF4 on plasmablast differentiation.We analysed the expression of immunity-related genes from the Gene Expression Omnibus and correlated predictors from clinical and laboratory data using R package and various statistical tools. Western blotting (WB) was used to detect protein levels in DLBCL cell lines of different subtypes to investigate the plasmablast and activation of mTORC1. To furtherly validate mTOR regulation of plasmablast differentiation, mTOR-activated and -inhibited cell models were constructed by CCK8 and flow cytometry (FCM) was used to assess the proportion of CD38 positive cells. We found that IRF4 was highly expressed in activated B-cell-like (ABC) DLBCL vs. germinal centre B-cell-like (GCB) DLBCL. Positive MUM-1and low haemoglobin values were corrected to non-CGB patients. Plasmablast indictors (BLIMP-1, ARF4, IRE1α, and IRF4) and mTORC1 pathway indictors (mTOR, p70S6K and phosphorylated-p70S6K) were different level in ABC cell lines. After successfully constructing cell models, the proportion of CD38 cells changed in mTOR-activated and -inhibited ABC-DLBCL cell models. We first pointed out that the role of the IRF4 invovling in DLBCL cell plasmablast differentiation via mTORC1 pathway. These findings could be extended to provide experimental evidence for novel treatments of secondary AIHA in ABC-DLBCL.
自身免疫性溶血性贫血（AIHA）是弥漫性大B细胞淋巴瘤（DLBCL）常见的继发病症。然而，尚无关于DLBCL继发AIHA中肿瘤B细胞分化为浆母细胞的报道。为阐明干扰素调节因子4（IRF4）对DLBCL的影响并探究IRF4在浆母细胞分化中的机制。我们从基因表达综合数据库分析了免疫相关基因的表达，并使用R软件包和各种统计工具对临床和实验室数据的相关预测指标进行了分析。采用蛋白质免疫印迹法（WB）检测不同亚型DLBCL细胞系中的蛋白水平，以研究浆母细胞及雷帕霉素靶蛋白复合体1（mTORC1）的激活情况。为进一步验证mTOR对浆母细胞分化的调控作用，通过细胞计数试剂盒8（CCK8）构建mTOR激活和抑制细胞模型，并用流式细胞术（FCM）评估CD38阳性细胞的比例。我们发现，与生发中心B细胞样（GCB）DLBCL相比，IRF4在活化B细胞样（ABC）DLBCL中高表达。将MUM-1阳性和血红蛋白值低的患者校正为非GCB患者。浆母细胞指标（B淋巴细胞诱导成熟蛋白1、ARF4、肌醇需求酶1α和IRF4）和mTORC1通路指标（mTOR、核糖体蛋白S6激酶p70S6K和磷酸化-p70S6K）在ABC细胞系中的表达水平各异。成功构建细胞模型后，mTOR激活和抑制的ABC-DLBCL细胞模型中CD38细胞的比例发生了变化。我们首次指出，IRF4通过mTORC1途径参与DLBCL细胞浆母细胞分化。这些发现可为ABC-DLBCL继发AIHA的新治疗方法提供实验依据。
