背景:流式细胞术在急性淋巴细胞白血病(ALL)的诊断中以及在需要抗原特异性免疫疗法时具有重要意义。我们研究了泼尼松龙的作用,长春新碱,柔红霉素,天冬酰胺酶和甲氨蝶呤对母细胞上抗原表达的影响可能会影响抗原特异性治疗的计划以及基于风险的治疗分配。
方法:将年龄≤17岁的从头B细胞ALL(B-ALL)患者纳入研究。分离Blast细胞并在体外暴露于对数增加浓度的5种单独的细胞毒性药物。然后,通过定量流式细胞术检测CD10,CD19,CD20,CD27,CD34,CD45,CD58,CD66c和CD137抗原的表达。
结果:细胞毒性药物引起抗原表达的剂量依赖性或剂量非依赖性调节。柔红霉素引起CD10、CD19、CD34、CD45和CD58的剂量依赖性下调和CD137的上调。长春新碱引起CD19和CD58的剂量依赖性下调和CD45的上调。柔红霉素还引起CD27的剂量依赖性下调和CD10、CD19、CD27、CD34和CD58的泼尼松龙下调。仅检测到与柔红霉素的特定剂量有关的CD20的下调。
结论:研究结果表明,细胞毒性药物可以改变对免疫治疗重要的抗原的表达。重要的是,柔红霉素,泼尼松龙和长春新碱引起CD19和CD58的下调,这表明在双特异性抗体或CAR-T细胞治疗之前的桥接治疗中,更好地避免使用这些药物.此外,不同药物诱导的母细胞免疫表型变化也可能影响基于风险的治疗分配。
BACKGROUND: Flow cytometry plays is important in the diagnosis of acute lymphoblastic leukaemia (ALL) and when antigen-specific immunotherapy is indicated. We have investigated the effects of prednisolone, vincristine,
daunorubicin, asparaginase and methotrexate on the antigen expression on blast cells that could influence the planning of antigen-specific therapy as well as risk-based treatment assignment.
METHODS: Patients aged ≤ 17 years with de novo B-cell ALL (B-ALL) were enrolled in the study. Blast cells were isolated and exposed in vitro to 5 individual cytotoxic drugs in logarithmically increasing concentrations. Then, the expression of CD10, CD19, CD20, CD27, CD34, CD45, CD58, CD66c and CD137 antigens was determined by quantitative flow cytometry.
RESULTS: Cytotoxic drugs caused dose-dependent or dose-independent modulation of antigen expression.
Daunorubicin caused a dose-dependent down-modulation of CD10, CD19, CD34, CD45 and CD58 and an up-modulation of CD137. Vincristine caused a dose-dependent down-modulation of CD19 and CD58 and an up-modulation of CD45.
Daunorubicin also caused dose-independent down-modulation of CD27 and prednisolone down-modulation of CD10, CD19, CD27, CD34 and CD58. Down-modulation of CD20 was detected only in relation to the specific dose of
daunorubicin.
CONCLUSIONS: The results of the study have shown that cytotoxic drugs can alter the expression of antigens that are important for immunotherapy. Importantly,
daunorubicin, prednisolone and vincristine caused down-modulation of CD19 and CD58, suggesting that these drugs are better avoided during bridging therapy prior to bispecific antibodies or CAR-T cell therapy. In addition, immunophenotypic changes on blast cells induced by different drugs could also influence risk-based treatment assignment.