Objective:To investigate the relationship between the levels of serum cytokines and chemokines and the prognosis of patients with acute B-ALL after receiving chimeric antigen receptor (CAR)-T cell immunotherapy and acute graft-versus-host disease (aGVHD) in patients after bridging allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:According to the case-control principle, Forty-two patients with B-ALL who received CD19-CAR-T cell immunotherapy bridged to allo-HSCT at Heibei Yanda Ludaopei Hospital from September 18, 2019 to May 9, 2022 were enrolled. Mann-Whitney U test was used to compare the changes of aGVHD-related cytokines and chemokine levels between CAR-T cell immunotherapy and bridging transplantation in different patients at the same time. Their plasma levels of cytokines and chemokines related to aGVHD were monitored at the day before CAR-T therapy and after CAR-T treatment at day 4, 7,14,21,28. The receiver operating characteristic curve was drawn to evaluate the predictive value of cytokines and chemokines in predicting the occurrence and the death of aGVHD patients. Kaplan-Meier method and Log-rank tests were used for Overall survival (OS) analysis. Results:Twenty-four of total 42 patients had aGVHD, of which 11 patients died and 31 patients survived. There was no significant difference in cytokines and chemokines between the aGVHD group and the non-aGVHD group on the day before CAR-T cell treatment. According to statistical analysis, the serum Elafin levels of aGVHD group was higher than that of non-aGVHD group at the 21st day [4 482 (2 811, 6 061) ng/L vs 2 466 (1 948, 3 375) ng/L, Z=3.145, P=0.001] and the 28st day [4 391 (2 808, 5594) ng/L vs 2 463 (1 658, 2 830) ng/L, Z=2.038, P=0.048] separately. At the 14th day, serum cytokines and chemokines levels between the two group were as follows,MIP-1 α [21.02 (12.36, 30.35) ng/L vs 5.56 (3.64, 10.79) ng/L], sCD25 [422.47 (257.99, 1 233.78) IU/ml vs 216.11 (133.75,457.39) IU/ml], Elafin [4 101 (2 393, 5 006) ng/L vs 2 155 (1 781, 3 033) ng/L], IL-6 [119.08 (23.97, 183.43) ng/L vs 8.39 (2.91, 17.42) ng/L] and IL-8 [13.56 (12.50, 24.52) ng/L vs 2.83 (1.73,6.87) ng/L] were at higher levels ( Z=2.653, P=0.007; Z=2.176, P=0. 030; Z=2.058, P=0.041; Z=3.329, P<0.001; Z=3.162, P=0.001). The KM survival curve showed that the cumulative survival rates of patients with higher serum levels of MIP-1α, sCD25, Elafin, IL-6 and IL-8 were lower than those with low levels at day 14, and the difference was statistically significant (χ 2=12.353, 4.890, 6.551, 10.563, 20.755, P<0.05). Conclusion:The outcomes of patients treated with CAR-T cell therapy bridged to allo-HSCT was correlated with serum MIP-1α, sCD25, Elafin, IL-6 and IL-8 levels after receiving CAR-T therapy. High concentrations of MIP-1α, sCD25, Elafin, IL-6 and IL-8 suggest poor prognosis and can be used as biomarkers to suggest appropriate clinical selection of therapy.

Objective:To investigate the significance of multicolor flow cytometry (MFC) monitoring of minimal residual disease (MRD) in the course of allogeneic hematopoietic stem cell transplantation (allo-HSCT) after CD19-chimeric antigen receptor(CAR)-T cell immunotherapy for patients with refractory, relapsed B-cell acute lymphoblastic leukemia (r/r B-ALL).Methods:37 patients with r/r B-ALL admitted to Hebei Yanda Lu Daopei Hospital from January to July 2019, aged 15 (6, 19) years old, including 24 males and 13 females, were treated with CD19-CAR-T cell immunotherapy bridging allo-HSCT. MFC with cytoplasmic CD79a antibody to set up B-cell gates was used to monitor patients′ bone marrow (BM), cerebrospinal fluid (CSF), and tissue samples on day 0 (prior to the CAR-T cell immunotherapy), day 15, day 28 post CAR-T cell immunotherapy, and post transplantation.The MRD values of these samples were analyzed to evaluate the residual tumor cells and metastasis. The killing effect of the CAR-T cells was evaluated by the recovery of CD19+B cells before transplantation and the period between the timepoint when CD19+B cells was recovered and the timepoint when CAR-T cells were infused. Peripheral blood CAR-T cells were counted at different time points. Statistic analysis was performed by Kaplan-Meie assay and Log-rank test to analyze the difference of univariate cumulative survival.Results:(1)Among the 37 patients, 8 died and 29 survived. 5 patients relapsed after transplantation, of which 4 relapsed patients died and 1 survived. (2)MFC MRD negative remission rate of the death group was lower than that of the survival group at the following time points: post-CAR-T therapy and prior to transplantation (5/8 vs. 28/29, χ 2=7.540, P=0.006); day 15 of the CAR-T cell reinfusion (3/8 vs. 24/29, χ 2=6.512, P=0.011); day 28 of the reinfusion (3/8 vs. 276/29, χ 2=10.065, P=0.002). The probability of extramedullary MFC MRD positive tumor infiltration in the death group was higher than that in the survival group(7/8 vs. 14/29, χ 2=3.931, P=0.047). After CAR-T cell immunotherapy, the recovery period of CD19-positive cells in the death group, or the time for CAR-T cells to kill CD19-positive cells, was shorter than that in the survival group [42.00 days(30.00,49.00) vs. 55.00 days(41.50,73.50), Z=0.022, P=0.020]. Conclusion:The positive results of MRD by MFC at the following timepoints may predict unfavorable outcomes, such as post-CAR-T therapy and prior to transplantation, day 15 and 28 of the CAR-T cell immunotherapy, which may provide some guidance for clinical management.

Objective To study the effect of peripheral blood cell paremeters in the patients with refractory/relapsed acute B-lymphoblastic leukemia treated with CD19 chimeric antigen receptor T cells (CD19-CAR-T). Methods We retrospectively analyzed the clinical data of patients with refractory/relapsed acute B-lymphoblastic leukemia treated with CD19-CAR-T cells in Beijing Ludaopei hospital from May 2017 to May 2020 . The peripheral blood cell parameters were compared between 46 patients with minimal residual disease (MRD) negative (MRD negative group) and 38 patients with minimal residual disease positive (MRD positive group) after CAR-T cells therapy. Results The mean values of leukocytes, hemoglobin, platelets and neutrophils(LHPN0) in the MRD negative group were signifi cantly higher than those in the MRD positive group on the day of CAR-T cells transfusion, and the diff erencewas statistically signifi cant (P＜0.05). The mean platelet values on the 7th, 14th and 21st day (PLT7, PLT14, PLT21)after CD19-CAR-T cells transfusion were signifi cantly higher in the MRD negative group than those in the MRD positive group, respectively, and the diff erencewas statistically signifi cant (P＜0.05). Conclusion The higher value of LHPN0, PLT7, PLT14, PLT21 may indicate that CD19-CAR-T cell treatment may be eff ective. The hematopoietic recovery is faster after CD19-CAR-T treatment than after chemotherapy.

目的:探究自体外周血造血干细胞移植(APBSCT)治疗T细胞淋巴瘤的治疗效果.方法:回顾性分析2014年6月-2018年6月期间接受APBSCT治疗的本病患者50例的临床资料、治疗方法以及随访结果.结果:患者输注数量为单个核细胞(1.4-6.3)×108/kg;中性粒细胞的植入时间为8-25d,中位时间11d;血小板植入时间为8-45d,中位时间为15.4d.所有患者均顺利完成移植,未见预处理毒性造成非复发死亡.中位随访时间为13.2个月,患者2年预期PFS率68.00%(34/50);2年OS率70.00%(35/50).结论:T细胞淋巴瘤患者采用APBSCT治疗的效果可靠,并发症少,其应用价值较高,值得推广.

目的:评价抗CD19嵌合抗原受体T(chimeric antigen receptor T,CAR-T)细胞IM19在复发难治B细胞血液系统恶性肿瘤中的疗效及安全性.方法:12例复发难治B细胞血液系统恶性肿瘤患者接受IM19治疗,包括6例B细胞非霍奇金淋巴瘤和6例急性B淋巴细胞白血病患者.患者接受3×105～10×105cells·kg-1 CAR-T细胞输注.细胞回输后1和3个月评估疗效,并监测不良反应事件的发生情况,同时检测细胞扩增以及细胞因子释放.结果:12例患者中有1 1例达到完全缓解,患者体内可以检测到IM19扩增,以及白介素-6和白介素-10水平升高.没有患者发生≥3级的细胞因子释放综合征和CAR-T细胞相关的神经系统毒性.结论:IM19治疗复发难治B细胞血液系统恶性肿瘤安全有效.

目的 观察西达本胺联合化疗治疗复发、难治性急性白血病的临床疗效.方法 回顾性分析2015年5月至2017年10月在本院治疗的复发、难治性急性白血病患者38例,其中急性髓系白血病(AML)及骨髓异常增生综合征EB2期(MDS-EB2)患者24例,急性淋巴细胞白血病(ALL)患者14例.≤14岁患者13例,＞ 14岁患者25例.所有患者给予二线化疗方案联合西达本胺治疗,成人给予西达本胺20 mg,患儿按实际体重/50 kg×20 mg给药,均口服,每周2次,连续给药1～3个月.计算患者总反应率(ORR)、完全缓解(CR)率和部分缓解(PR)率,并分析患者的年龄、性别、疾病种类等一般资料和染色体、基因的不同对疗效的影响.结果 所有患者ORR为55％,CR率为37％.≤14岁患者ORR和CR率分别为69％和46％,均高于＞14岁患者(48％和32％),差异有显著意义(P＜0.05).不同性别和病种的患者,ORR和CR率均无显著差异(P＞0.05).病程≤2个疗程未缓解(NR)的患者,其ORR和CR率分别为80％和53％,均高于病程＞2个疗程NR/复发患者(39％和26％),差异有显著意义(P＜0.05).MLL-PTD基因突变患者10例,ORR为60％,CR率为30％.染色体不同、基因突变或无突变患者的ORR和CR率均无显著差异(P＞0.05).结论 西达本胺联合化疗能有效改善复发、难治性急性白血病的ORR和CR率,在儿童中应用效果优于成人,早期应用效果优于晚期,在MLL-PTD突变患者中应用疗效良好.

急性红白血病的发病率较低,且不同患者个体间的疾病演变及预后差异较大,以下通过回顾分析2014年以前就诊的4例,应用WHO旧版分类方法诊断的急性红白血病患者,讨论该类疾病的骨髓形态学、免疫分型、细胞遗传学、分子生物学特点、病程演变及转归,分析影响患者预后的因素及治疗方法.

目的:研究儿童外周血造血干细胞的采集过程以及外周血造血干细胞采集对儿童供者健康的影响,以评估儿童作为异基因造血干细胞移植供者,特别是当干细胞受者为成人时的安全性和有效性.方法:回顾性分析14例儿童作为供者进行外周血造血干细胞的动员、采集过程和采集物的相关临床资料.结果:14例儿童供者中位年龄6(2～10)岁.干细胞采集物用于半相同移植4例,同胞全合移植10例.骨髓联合外周血造血干细胞移植6例,单纯外周血造血干细胞移植8例.供者外周血干细胞采集总次数分别为1次4例,2次5例,3次3例,4次2例.除1例植入失败以外,其他受者均顺利植活.儿童在整个外周血造血干细胞采集过程中无明显不良反应.结论:儿童供者通过分次动员和采集可以采集出足量的造血干细胞用于异基因造血干细胞移植.儿童采集外周血造血干细胞是安全的,儿童供者所采集的外周血造血干细胞能够满足单纯外周血干细胞移植或骨髓联合外周血干细胞移植的需要.

OBJECTIVE:To study the predictable value of monitoring minimal residual disease (MRD) regularly by flow cytometry (FCM) in patients with acute leukemia (AL) in the first complete remission (CR(1)).METHODS:From April 2005 to July 2009, AL patients who had got CR(1) after chemotherapy were regularly monitored for MRD in bone marrow by FCM to relapse or to July 2010 in Beijing Daopei Hospital (not including those received stem cell transplantation). The special antibody combinations were employed for each patient according to aberrant expression of leukemia cells. MRD(+) was defined as the aberrant cells more than 0.01%. The probability of continuous CR (CCR) was calculated by Kaplan-Meier formula, and the statistical difference between two CCR probabilities was evaluated by log-rank test.RESULTS:A total of 163 AL patients in CR(1) were monitored to relapse or to July 2010. Among 89 AML patients referred to our hospital within 1 year after diagnosis, 30 cases were in MRD(+) and 59 cases MRD(-) till 12 months following chemotherapy, 3/30 patients in MRD(+) and 47/59 remained in CCR to July 2010. The probability of CCR at 24, 36 months was 13%, 13%in MRD(+) group, 94%, 78% in MRD(-) group respectively, the difference between them was statistically significant (P < 0.01). Among 35 ALL referred to our hospital within 5 months after diagnosis, 13 cases were MRD(+) and 22 cases MRD(-) till 5 months following chemotherapy, 0/13 patients in MRD(+) and 20/22 patients in MRD(-) remained in CCR to July 2010. The probability of CCR at 24, 36 months was 0% in MRD(+) group, 96%, 96% in MRD(-) group respectively, the difference between them was statistically significant (P < 0.01). Over the time point above, all patients with MRD(+) or their MRD from negative to positive relapsed finally, and most patients with MRD(-) remained CCR to July 2010.CONCLUSION:It had a clinical prognostic value to monitor MRD regularly by FCM in the patients with AL after CR(1).