目的探讨 B淋巴细胞相关抗原在急性 B细胞型淋巴细胞白血病 (B- AL L )细胞表面的表达及其与细胞形态和治疗的关系。方法采用 ABC- AP免疫组化法结合选用多种 B淋巴细胞相关抗原的克隆性抗体对 30 1例初发 (B- AL L )患者的白血病细胞...目的探讨 B淋巴细胞相关抗原在急性 B细胞型淋巴细胞白血病 (B- AL L )细胞表面的表达及其与细胞形态和治疗的关系。方法采用 ABC- AP免疫组化法结合选用多种 B淋巴细胞相关抗原的克隆性抗体对 30 1例初发 (B- AL L )患者的白血病细胞进行细胞表面抗原检测。结果 B- AL L患者的白血病细胞 CD19、HL A - DR呈高阳性率表达 ;而 CD2 0的阳性表达率较低。阳性表达率与 FAB分类分型有一定关系。其中 L1 型 CD 10细胞有高表达率近 90 % ;L2 型 CD10细胞表达率为 6 3.5 % ;L3 型细胞则无 CD10阳性表达。结论结果均与 B- AL L中展开更多
目的:探索CD19 CAR-T细胞治疗复发/难治性B-ALL后细胞因子水平的变化及其临床意义。方法:选取2015年8月至2018年2月我院血液科接受CD19 CAR-T细胞治疗的复发/难治性B-ALL患者23例,应用CBA技术和免疫比浊法检测输注前后血清中IL-6、IFN-...目的:探索CD19 CAR-T细胞治疗复发/难治性B-ALL后细胞因子水平的变化及其临床意义。方法:选取2015年8月至2018年2月我院血液科接受CD19 CAR-T细胞治疗的复发/难治性B-ALL患者23例,应用CBA技术和免疫比浊法检测输注前后血清中IL-6、IFN-γ、IL-10、TNF-α和CRP水平。然后根据CTCAE v4.0将发生CRS的患者分为轻度CRS组(1~2级)和重度CRS组(3~5级),比较两组细胞因子水平的差异及治疗反应。结果:①CD19 CAR-T细胞输注前后患者血清中IL-6(2.00~28.50 vs 34.50~112.00)、IFN-γ(2.13~3.12 vs 4.35~10.07)、IL-10(1.00~1.80 vs 4.01~39.92)、TNF-α(2.91~4.60 vs 4.99~40.18)差异均具有统计学意义(P=0.000,0.000,0.001,0.002);②轻度CRS组和重度CRS患者相比,血清中IL-6(28.75~61.25 vs 91.00~1 074.00)、IFN-γ(3.93~7.28 vs 12.53~15.30)、IL-10(3.13~7.47 vs 39.92~84.97)、TNF-α(4.60~15.77 vs 40.18~62.51)和CRP(4.95~46.86 vs 37.81~120.17)差异均具有统计学意义(P=0.004、0.004、0.003、0.000、0.045);③CD19 CAR-T治疗后完全缓解(包括血象未完全恢复患者)组与未缓解组间上述细胞因子与CRP水平均差异无统计学意义;④IL-6显著升高的重度CRS患者,应用IL-6受体拮抗剂治疗后均好转。结论:IL-6、IFN-γ、IL-10、TNF-α细胞因子的水平可作为判断CRS严重程度的实验室参考指标,并可指导临床靶向选择细胞因子拮抗剂,但对CD19 CAR-T疗效预测价值有待进一步验证。展开更多
Objective To detect the minimal residual disease in children with B-ALL and to evaluate its clinical significance by flow cytometry. Methods 58 childhood B-ALL cases were enrolled into this study and 33 MRD analyses w...Objective To detect the minimal residual disease in children with B-ALL and to evaluate its clinical significance by flow cytometry. Methods 58 childhood B-ALL cases were enrolled into this study and 33 MRD analyses were performed after remission induction therapy.Four-color combinations of fluorochrome labeled monoclonal antibodies against lymphocyte lineage related phenotypes were used to analyze leukemic cells with flow cytometry.The cells from normal bone marrow were used as controls.The combinations of phenotypes that reflect the antigen expression differences between leukemic and normal bone marrow cells on flow cytometry were considered to be the effective phenotype combinations in the first step screening.The effective phenotype combinations were then used to monitor MRD during the disease course after therapy began. Results 58 cases of childhood B-ALL were screened for MRD effective phenotype combinations.The effective phenotype combinations were identified in 89.7% of B-ALL cases in this study.Four-color phenotype combinations were composed of CD10/CD34/CD19 plus another effective marker such as CD38,CD58,CD66c,CD21.The senstitivity of this method was 0.01%,much higher than that of microscopic inspection.In 8 cases,their bone marrow microscopic inspection results showed no remaining leukemic cells;but with flow cytometry,the percentage of leukemic cells were 5.66%,0.36%,1.43%,0.069%,1.55%,2.7%,0.028% and 0.015%,respectively.In risk stratification,all these MRD positive cases were classified into high risk group for relapse and 1 case showed early relapse within 6 months. Conclusion The application of flow cytometry in MRD measurement can significantly improve the sensitivity of detection of remained leukemic cells in childhood B-ALL,and can provide more accurate information on disease progression as well as the efficacy of therapy,thus facilitate future treatment decisions and follow ups.展开更多
Chimeric antigen receptor (CAR) T cell therapy has exhibited dramatic anti-tumor effi-cacy in clinical trials. In this study,we reported the transcriptome profiles of bone marrow cells in four B cell acute lymphoblast...Chimeric antigen receptor (CAR) T cell therapy has exhibited dramatic anti-tumor effi-cacy in clinical trials. In this study,we reported the transcriptome profiles of bone marrow cells in four B cell acute lymphoblastic leukemia (B-ALL) patients before and after CD19-specific CAR-T therapy. CD19-CAR-T therapy remarkably reduced the number of leukemia cells,and three patients achieved bone marrow remission (minimal residual disease negative). The efficacy of CD19-CAR-T therapy on B-ALL was positively correlated with the abundance of CAR and immune cell subpopulations,e.g.,CD8+T cells and natural killer (NK) cells,in the bone marrow. Additionally,CD19-CAR-T therapy mainly influenced the expression of genes linked to cell cycle and immune response pathways,including the NK cell mediated cytotoxicity and NOD-like recep-tor signaling pathways. The regulatory network analyses revealed that microRNAs (e.g.,miR-148a-3p and miR-375),acting as oncogenes or tumor suppressors,could regulate the crosstalk between the genes encoding transcription factors (TFs,e.g.,JUN and FOS) and histones (e.g.,HIST1H4A and HIST2H4A) involved in CD19-CAR-T therapy. Furthermore,many long non-coding RNAs showed a high degree of co-expression with TFs or histones (e.g.,FOS and HIST1H4B) and were associated with immune processes. These transcriptome analyses provided important clues for fur-ther understanding the gene expression and related mechanisms underlying the efficacy of CAR-T immunotherapy.展开更多
文摘目的:探索CD19 CAR-T细胞治疗复发/难治性B-ALL后细胞因子水平的变化及其临床意义。方法:选取2015年8月至2018年2月我院血液科接受CD19 CAR-T细胞治疗的复发/难治性B-ALL患者23例,应用CBA技术和免疫比浊法检测输注前后血清中IL-6、IFN-γ、IL-10、TNF-α和CRP水平。然后根据CTCAE v4.0将发生CRS的患者分为轻度CRS组(1~2级)和重度CRS组(3~5级),比较两组细胞因子水平的差异及治疗反应。结果:①CD19 CAR-T细胞输注前后患者血清中IL-6(2.00~28.50 vs 34.50~112.00)、IFN-γ(2.13~3.12 vs 4.35~10.07)、IL-10(1.00~1.80 vs 4.01~39.92)、TNF-α(2.91~4.60 vs 4.99~40.18)差异均具有统计学意义(P=0.000,0.000,0.001,0.002);②轻度CRS组和重度CRS患者相比,血清中IL-6(28.75~61.25 vs 91.00~1 074.00)、IFN-γ(3.93~7.28 vs 12.53~15.30)、IL-10(3.13~7.47 vs 39.92~84.97)、TNF-α(4.60~15.77 vs 40.18~62.51)和CRP(4.95~46.86 vs 37.81~120.17)差异均具有统计学意义(P=0.004、0.004、0.003、0.000、0.045);③CD19 CAR-T治疗后完全缓解(包括血象未完全恢复患者)组与未缓解组间上述细胞因子与CRP水平均差异无统计学意义;④IL-6显著升高的重度CRS患者,应用IL-6受体拮抗剂治疗后均好转。结论:IL-6、IFN-γ、IL-10、TNF-α细胞因子的水平可作为判断CRS严重程度的实验室参考指标,并可指导临床靶向选择细胞因子拮抗剂,但对CD19 CAR-T疗效预测价值有待进一步验证。
文摘Objective To detect the minimal residual disease in children with B-ALL and to evaluate its clinical significance by flow cytometry. Methods 58 childhood B-ALL cases were enrolled into this study and 33 MRD analyses were performed after remission induction therapy.Four-color combinations of fluorochrome labeled monoclonal antibodies against lymphocyte lineage related phenotypes were used to analyze leukemic cells with flow cytometry.The cells from normal bone marrow were used as controls.The combinations of phenotypes that reflect the antigen expression differences between leukemic and normal bone marrow cells on flow cytometry were considered to be the effective phenotype combinations in the first step screening.The effective phenotype combinations were then used to monitor MRD during the disease course after therapy began. Results 58 cases of childhood B-ALL were screened for MRD effective phenotype combinations.The effective phenotype combinations were identified in 89.7% of B-ALL cases in this study.Four-color phenotype combinations were composed of CD10/CD34/CD19 plus another effective marker such as CD38,CD58,CD66c,CD21.The senstitivity of this method was 0.01%,much higher than that of microscopic inspection.In 8 cases,their bone marrow microscopic inspection results showed no remaining leukemic cells;but with flow cytometry,the percentage of leukemic cells were 5.66%,0.36%,1.43%,0.069%,1.55%,2.7%,0.028% and 0.015%,respectively.In risk stratification,all these MRD positive cases were classified into high risk group for relapse and 1 case showed early relapse within 6 months. Conclusion The application of flow cytometry in MRD measurement can significantly improve the sensitivity of detection of remained leukemic cells in childhood B-ALL,and can provide more accurate information on disease progression as well as the efficacy of therapy,thus facilitate future treatment decisions and follow ups.
基金the National Natural Science Foundation of China (Grant Nos. 31822030, 31801113, and 31771458)the National Key R&D Program of China (Grant No. 2017YFA0700403)China Postdoctoral Science Foundation (Grant No. 2018M632830)
文摘Chimeric antigen receptor (CAR) T cell therapy has exhibited dramatic anti-tumor effi-cacy in clinical trials. In this study,we reported the transcriptome profiles of bone marrow cells in four B cell acute lymphoblastic leukemia (B-ALL) patients before and after CD19-specific CAR-T therapy. CD19-CAR-T therapy remarkably reduced the number of leukemia cells,and three patients achieved bone marrow remission (minimal residual disease negative). The efficacy of CD19-CAR-T therapy on B-ALL was positively correlated with the abundance of CAR and immune cell subpopulations,e.g.,CD8+T cells and natural killer (NK) cells,in the bone marrow. Additionally,CD19-CAR-T therapy mainly influenced the expression of genes linked to cell cycle and immune response pathways,including the NK cell mediated cytotoxicity and NOD-like recep-tor signaling pathways. The regulatory network analyses revealed that microRNAs (e.g.,miR-148a-3p and miR-375),acting as oncogenes or tumor suppressors,could regulate the crosstalk between the genes encoding transcription factors (TFs,e.g.,JUN and FOS) and histones (e.g.,HIST1H4A and HIST2H4A) involved in CD19-CAR-T therapy. Furthermore,many long non-coding RNAs showed a high degree of co-expression with TFs or histones (e.g.,FOS and HIST1H4B) and were associated with immune processes. These transcriptome analyses provided important clues for fur-ther understanding the gene expression and related mechanisms underlying the efficacy of CAR-T immunotherapy.
