目的:慢性移植物抗宿主病(cGVHD)是异基因造血干细胞移植后的重要并发症,通常需要利妥昔单抗(RTX)和环孢素A(CsA)等治疗干预措施。本研究旨在阐明RTX和CsA共同解决cGVHD中B细胞失调的机制。为该病的治疗和预后评估提供了理论基础和科学依据。
方法:通过对受体小鼠进行全身照射,然后注射来自供体小鼠的骨髓细胞和脾细胞的混合悬浮液,建立总共30个cGVHD小鼠模型。从模型建立后的第2天到第29天,小鼠接受皮下施用RTX和CsA。在整个研究过程中,体重,临床cGVHD评分,并监测生存率。通过眶静脉丛收集血样。采用酶联免疫吸附试验(ELISA)检测血清B细胞活化因子(BAFF)和促炎因子水平,通过流式细胞术评估血液样品中调节性B细胞(Bregs)的比例。
结果:患有cGVHD的小鼠体重减轻了14.5%,临床评分升高,与对照组相比,症状更严重。值得注意的是,cGVHD组和对照组的所有小鼠均存活直至研究结束.cGVHD的诱导导致B细胞失调,血清BAFF水平升高和Bregs比例降低证明。然而,RTX联合CsA治疗可改善cGVHD小鼠的B细胞失调并显著降低血清促炎因子水平,TNF-α降低39.78%,IL-6降低37.89%。
结论:RTX和CsA的组合有效缓解cGVHD中的B细胞失调,从而降低疾病的严重程度和进展。
OBJECTIVE: Chronic graft-versus-host disease (cGVHD) is a significant complication following allogenic hematopoietic stem cell transplantation, often necessitating therapeutic interventions such as rituximab (RTX) and cyclosporin A (CsA). This study aims to elucidate the mechanisms by which RTX and CsA jointly address B-cell dysregulation in cGVHD, providing a theoretical foundation and scientific rationale for the treatment and prognostic evaluation of this condition.
METHODS: A total of 30 cGVHD mouse models were established by subjecting recipient mice to total body irradiation followed by injection of a mixed suspension of bone marrow cells and splenocytes from donor mice. From Day 2 to Day 29 post-model establishment, the mice received subcutaneous administration of RTX and CsA. Throughout the study, body weight, clinical cGVHD scores, and survival rates were monitored. Blood samples were collected via the orbital venous plexus. Serum levels of B-cell activating factor (BAFF) and
pro-inflammatory factors were measured using enzyme-linked immunosorbent assay (ELISA), and the ratio of regulatory B cells (Bregs) in the blood sample was assessed via flow cytometry.
RESULTS: Mice with cGVHD exhibited a 14.5% decrease in body weight, elevated clinical scores, and more severe symptoms compared to the control group. Notably, all mice in both the cGVHD and control groups survived until the conclusion of the study. Induction of cGVHD resulted in B-cell dysregulation, evidenced by elevated serum BAFF levels and a decreased proportion of Bregs. However, treatment with RTX combined with CsA ameliorated B-cell dysregulation and significantly reduced serum levels of
pro-inflammatory factors in cGVHD mice, with decreases of 39.78% in TNF-α and 37.89% in IL-6.
CONCLUSIONS: The combination of RTX and CsA effectively mitigates B-cell dysregulation in cGVHD, thereby reducing the severity and progression of the disease.