The human leucocyte differentiation antigens (HLDA) workshops: the evolv-ing role of antibodies in research, diagnosis and therapy

The 8^th International Workshop on Human Leucocyte Differentiation Antigens （chaired by HZ and managed by BS） was run over a 4-year period and culminated in a conference in December 2004. Here we review the achievements of the HLDA Workshops and provide links to information on CD molecules and antibodies against them, including the 93 new CDs assigned in the 8^th Workshop. We consider what remains to be achieved （including an estimate of the number of leucocyte surface molecules still to be discovered）, and how the field can best move forward.

Blood group type antigens in pancreatic intraductal papillary mucinous neoplasms

BACKGROUND: There are few data on blood group(BG) types and types of pancreatic cancers. The aims of this study were to study BG types and BG-antigens in pancreatic intraductal papillary mucinous neoplasms(IPMNs). METHODS: BG type and tumor BG-antigen(glycoprotein) expression(studied by immunohistochemistry on tissue microarrays) were analyzed with regard to characteristics of 101 surgically resected pancreatic IPMNs. RESULTS: Non-O BG type predicted invasive carcinoma independently from high serum CA19-9 and male gender. BG type A was observed more frequently in women than in men. Chronic pancreatitis was more frequently seen in patients with BG type B or AB. Aberrant tumor expression(with regard to BG type) of loss of A antigen expression type occurred in 15.0% of IPMNs and of loss of B antigen expression type in 62.5% of IPMNs. Intraneoplasm BG-antigen expression was not related to dysplasia grade or invasion. CONCLUSION: The results of the study suggest that in pancreatic IPMN, non-O BG type predicted invasive carcinoma, whereas for intratumor BG-antigen expression no specific patterns were detected with regard to the progression of glandular epithelial dysplasia or invasion.

CD19 CAR-T细胞治疗难治/复发急性B淋巴细胞白血病儿童及青少年患者的疗效及安全性

目的探讨CD19嵌合抗原受体T细胞(CAR-T)治疗对于儿童及青少年难治/复发急性B淋巴细胞白血病(B-ALL)的疗效及安全性。方法回顾性分析2017年6月至2021年3月接受CD19 CAR-T治疗的<25岁难治/复发B-ALL患者的临床资料,评估该疗法的疗效及安全性。结果共纳入64例难治/复发B-ALL患者,男35例、女29例,中位年龄8.5(1.0~17.0)岁。CD 19 CAR-T回输后1个月进行短期疗效评估,64例患者均获得完全缓解(CR)/完全缓解兼部分血细胞计数缓解(CRi),其中有62例患者达骨髓微小残留病灶(MRD)阴性。细胞因子释放综合征(CRS)及免疫效应细胞相关神经毒性综合征(ICANS)发生率分别为78.1%及23.4%。共22例患者复发,中位复发时间10.1个月,4年总生存(OS)率为(66.0±6.0)%,4年无白血病生存(LFS)率为(63.0±6.0)%。长期随访结果显示桥接异基因造血干细胞移植(allo-HSCT)患者的LFS和OS率均优于未桥接移植患者(4年LFS率:81.8%±6.2%对24.0%±9.8%,4年OS率:81.4%±5.9%对44.4%±11.2%;均P<0.01)。结论CD 19 CAR-T可有效治疗难治/复发B-ALL,输注后桥接allo-HSCT能进一步改善患者的长期生存情况。

儿童CD19 CAR-T细胞治疗相关B细胞再生障碍的临床意义和对策

急性B系淋巴细胞白血病(B-ALL)患儿在CD 19 CAR-T细胞治疗后普遍发生B细胞再生障碍(BCA),BCA持续的时间长短对患者的免疫状态及预后会产生影响。对BCA的充分认识有助于临床医师科学、规范、合理地选择治疗策略,减少CAR-T治疗后白血病患儿的感染机会,提高生活质量,改善预后。

CD19^(+)CD24^(hi)CD27^(+)调节性B细胞水平与强直性脊柱炎间的关系探讨

目的:探究CD19^(+)CD24^(hi)CD27^(+)调节性B细胞(Bregs)水平与强直性脊柱炎(AS)的关系。方法:将赣州市人民医院2019年1月至2021年12月间收治的80例AS患者纳为观察组,同期体检合格的健康志愿者60例纳为对照组。根据观察组患者疾病临床分期将其分为进展期及强直期,根据患者疾病活动度将其分为活动组、稳定组,检测并比较观察组及对照组、观察组不同分期患者外周血CD19^(+)CD24^(hi)CD27^(+)Breg占CD19^(+)细胞百分比。分析CD19^(+)CD24^(hi)CD27^(+)Bregs百分比与AS患者病程、晨僵时间、巴斯强直性脊柱炎疾病活动指数评分(BASDAI)、IL-10、血沉(ESR)、C反应蛋白(CRP)及骶髂关节X线片分级等临床特点之间的关系。结果:观察组AS患者外周血CD19^(+)CD24^(hi)CD27^(+)Bregs占CD19^(+)B细胞百分比高于健康对照组,强直期AS患者CD19^(+)CD24^(hi)CD27^(+)Breg占CD19^(+)细胞百分比高于进展期患者,且骨性强直期患者外周血CD19^(+)CD24^(hi)CD27^(+)Breg百分比高于纤维性强直期患者,活动组AS患者外周血CD19^(+)CD24^(hi)CD27^(+)Breg占比高于稳定组,以上差异均有统计学意义(P<0.05)。观察组血清IL-10水平低于对照组,ESR及CRP水平高于对照组,差异有统计学意义(P<0.05)。AS患者外周血CD19^(+)CD24^(hi)CD27^(+)Breg占CD19^(+)B细胞百分比与其BASDAI得分及血清IL-10水平呈正相关,与ESR及CRP水平呈负相关,与患者晨僵时间及髂关节X线分级无明显相关性。结论:AS患者外周血CD19^(+)CD24^(hi)CD27^(+)Breg占CD19^(+)B细胞百分比降低,且其水平与患者疾病分期、活动度及实验室指标IL-10、ESR及CRP间具有一定的相关性。

CD19^(+)CD27^(+)IgD^(+)B细胞与甲状腺相关性眼病患者疾病活动度相关性研究

目的:探讨甲状腺相关性眼病(thyroid-associated ophthalmopathy,TAO)患者外周血CD19^(+)CD27^(+)IgD^(+)B细胞比例变化与病情活动度的关系。方法:采用流式细胞术检测TAO患者和健康对照组CD19^(+)CD27^(+)IgD^(+)B细胞比例,分析TAO患者CD19^(+)CD27^(+)IgD^(+)B细胞比例与健康对照组的差异,并分析CD19^(+)CD27^(+)IgD^(+)B细胞比例与TAO疾病活动度之间的相关性。结果:TAO患者CD19^(+)CD27^(+)IgD^(+)B细胞比例明显低于健康对照(P<0.05);活动性TAO患者CD19^(+)CD27^(+)IgD^(+)B细胞比例细胞显著低于非活动性TAO患者;CD19^(+)CD27^(+)IgD^(+)B细胞比例与TAO患者CAS评分呈显著负相关(r=-0.7128,P<0.001)。结论:TAO患者CD19^(+)CD27^(+)IgD^(+)B细胞比例降低,与疾病活动度呈负相关。

CK19、HBME-1、CD56、BRAF V600E、FAM210B在甲状腺乳头状结构诊断中的应用

目的探讨CK19、HBME-1、CD56、BRAF V600E和FAM210B蛋白表达在甲状腺乳头状结构诊断中的作用及FAM210B和BRAF V600E与甲状腺乳头状癌临床病理特征的关系。方法连续收集2017年1月—2020年9月××省第二人民医院PTC患者的病理标本279例,甲状腺乳头状增生标本85例。用免疫组化SP法检测甲状腺乳头状癌(papillary thyroid carcinoma,PTC)和甲状腺乳头状增生中CK19、HBME-1、CD56、BRAF V600E和FAM210B的表达情况,分析FAM210B和BRAF V600E在不同临床病理特征的PTC组织中的表达情况。结果CD56在PTC组织中表达率为10.03%,在甲状腺乳头状增生中表达率为94.12%;CK19、HBME-1、BRAF V600E、FAM210B在PTC组织中的阳性率均高于甲状腺乳头状增生,差异有统计学意义(P<0.01);FAM210B表达与年龄、肿块大小和淋巴结转移情况具有相关性,BRAF V600E表达与性别、年龄、肿块大小和淋巴结转移情况具有相关性,差异有统计学意义(P<0.01)。结论CK19、HBME-1、FAM210B、BRAF V600E联合CD56标记有助于PTC的诊断和鉴别诊断;检测FAM210B和BRAF V600E有助于评估肿瘤的生物学行为及预后,对指导治疗和提高疗效有重要意义。

系统性红斑狼疮患者妊娠期血清CCL19和sCD163水平及其对母婴结局的影响

目的:探讨外周血趋化因子配体19(CCL19)和可溶性CD163(sCD163)在系统性红斑狼疮(SLE)患者妊娠期血清中的水平变化,并阐明其对母婴结局的影响。方法:选取180例妊娠期SLE患者作为SLE组,根据母婴结局分为妊娠成功组(n=132)和妊娠失败组(n=48);另外随机选取同期产检的180例健康孕妇作为对照组。收集2组研究对象一般资料,试剂盒检测2组研究对象血清中CCL19和sCD163水平及相关血清因子水平。采用多因素Logistic回归分析检测SLE患者妊娠失败的影响因素,受试者工作特征(ROC)曲线分析SLE组血清中CCL19和sCD163水平预测妊娠结局的效果。结果:与对照组比较,SLE组患者血清中补体C3和补体C4水平明显降低(P<0.05),血清中红细胞沉降率(ESR)、肌酐(CR)、抗心磷脂抗体(ACA)-IgG、抗β2糖蛋白Ⅰ(anti-β2GPⅠ)、CCL19和sCD163水平明显升高(P<0.01)。与妊娠成功组比较,妊娠失败组患者血清中补体C3和补体C4水平明显降低(P<0.01),血清中ESR、CR、ACA-IgG、anti-β2GPⅠ、CCL19和sCD163水平均明显升高(P<0.01)。血清中CCL19、sCD163、ESR、CR、ACA-IgG和anti-β2GPⅠ水平是妊娠期SLE患者妊娠失败的危险因素(P<0.05或P<0.01),补体C3和补体C4水平是妊娠期SLE患者妊娠失败的保护因素(P<0.01)。血清CCL19水平预测妊娠期SLE患者妊娠失败的ROC曲线下面积(AUC)为0.726,血清sCD163水平预测妊娠期SLE患者妊娠失败预后的AUC为0.789,二者联合预测妊娠期SLE患者妊娠失败的AUC为0.835。二者联合预测妊娠期SLE患者妊娠失败的效能优于CCL19和sCD163水平各自单独预测(Z_(联合检测-CCL19)=3.066,P=0.002;Z_(联合检测-sCD163)=2.087,P=0.037)。结论:SLE患者妊娠期血清中CCL19和sCD163水平明显升高,可能导致患者母婴结局不良。

超声联合Galectin-3、CK19、CD56在甲状腺乳头状癌颈部淋巴结转移中的诊断价值

目的 回顾性分析超声联合半乳凝素-3(Galectin-3)、细胞角蛋白19(CK19)、CD56在甲状腺乳头状癌(PTC)颈部淋巴结转移(CLNM)中的诊断价值。方法 回顾性选取2022年1月至2023年11月于安庆市立医院行手术治疗的98例PTC患者的临床资料。按照是否发生CLNM将98例PTC患者分为CLNM组(n=25)、非CLNM组(n=73)。所有患者均行超声及Galectin-3、CK19、CD56诊断,观察两组性别、年龄及超声特征;PTC患者Galectin-3、CK19、CD56表达情况;两组Galectin-3、CK19、CD56表达情况;采用受试者操作特征(ROC)曲线分析超声联合Galectin-3、CK19、CD56对PTC CLNM的诊断价值;经多因素Logistic回归分析法分析导致PTC CLNM的因素。结果 两组性别、年龄、病灶部位、边缘规则情况、回声高低、回声均匀及病灶内部成分比较,差异均无统计学意义(P>0.05);两组病灶数目、病灶最大径、被膜侵犯、纵横比及微钙化比较,差异均有统计学意义(P<0.05)。经免疫组织化学染色法检测,PTC患者Galectin-3、CK19及CD56的阳性表达率分别为84.69%、82.65%、38.78%。CLNM组患者Galectin-3、CK19、CD56阳性表达率分别为100.00%、100.00%、12.00%,其中Galectin-3、CK19阳性表达率均高于非CLNM组(79.45%、76.71%),CD56阳性表达率低于非CLNM组(47.95%),差异均有统计学意义(P<0.05)。经ROC曲线分析,超声联合Galectin-3、CK19、CD56对PTC CLNM的诊断价值高于四者单独诊断PTC CLNM的诊断价值。经多因素Logistic回归分析,年龄<30、病灶最大径>10 mm、被膜侵犯、纵横比>1、微钙化及Galectin-3、CK19是导致PTC CLNM的危险因素(P<0.05),CD56阳性是PTC CLNM的保护因素(P<0.05)。结论 超声联合Galectin-3、CK19、CD56对PTC CLNM具有较高的诊断价值。

系统性红斑狼疮患者外周血CD19^(+)CD25^(+)Breg比例和细胞表面PD-1和PD-L1的表达及其与临床指标的相关性

目的 研究程序性死亡蛋白1(PD-1)与其配体PD-L1在系统性红斑狼疮(SLE)患者外周血CD19^(+)CD25^(+)调节性B细胞(Breg)的表达及临床意义。方法收集50例SLE患者和41例健康人(HC)外周血标本,采用流式细胞仪检测外周血中CD19^(+)CD25^(+)Breg的比例,以及CD19^(+)CD25^(+)Breg的PD-1、PD-L1表达,同时采集患者的临床表现和实验室指标等临床信息。使用免疫磁珠分选CD4^(+)T细胞与CD19^(+)B细胞,体外细胞共培养,检测Breg的分化。结果SLE患者活动组外周血中CD19^(+)CD25^(+)Breg比例低于HC,CD25^(+)B细胞PD-1、PD-L1的表达高于HC;SLE患者有胸腔积液、关节炎、C反应蛋白(CRP)上升的患者Breg频率高于对应阴性组;SLE患者有IgM下降、抗核糖核蛋白(RNP)抗体阳性的患者Breg频率低于对应阴性组;SLE患者有感染、发热、关节炎、IgA上升的患者CD19^(+)CD25^(+)PD-1^(+)细胞频率高于对应阴性组;SLE患者有感染、发热、IgA上升的患者CD19^(+)CD25^(+)PD-L1^(+)细胞频率高于对应阴性组;活化的CD4^(+)T细胞有利于CD19^(+)B细胞表面CD25的表达。结论SLE患者外周血中CD19^(+)CD25^(+)Breg降低,但细胞表面PD-1与PD-L1的表达增加,并且与相关临床表现及实验室参数存在一定的关系。活化的CD4^(+)T细胞促进Breg的分化。

Serum tumor markers (carcinoembryonic antigen, carbohydrate antigen 19-9, carbohydrate antigen 72-4, carbohydrate antigen 24-2, ferritin) and gastric cancer prognosis correlation

BACKGROUND Gastric cancer is a kind of malignant tumor which is prevalent all over the world.Although some progress has been made in the treatment of gastric cancer,its prognosis is still not optimistic,so it is of great significance to find reliable prog-nostic indicators to guide the treatment and management of patients with gastric cancer.AIM To explore the relationship between serum levels of five biomarkers[carcinoem-bryonic antigen(CEA),carbohydrate antigen(CA)19-9,CA72-4,CA24-2,and ferritin]and prognosis in patients with gastric cancer.METHODS This study included 200 patients with gastric adenocarcinoma,and conducted an in-depth analysis of their baseline characteristics,relationship between tumor markers and staging,and prognosis.The study found that CA19-9 has a signi-ficant correlation with tumor stage,the average levels of CA24-2,CEA,CA72-4 and ferritin were slightly increased disregarding the stage of tumor.Survival analysis showed that increases in CEA,CA19-9,CA24-2,and ferritin were all associated with shortened overall survival of patients.Further multivariate ana-lysis revealed that elevated serum CA72-4 levels were an inde-pendent adverse prognostic factor.RESULTS This study reveals that there is a significant correlation between the expression levels of serum tumor markers CEA,CA19-9,CA72-4,CA24-2 and ferritin in patients with gastric cancer and prognosis,and can be used as important indicators for prognostic evaluation of gastric cancer.In particular,markers that appear abnormally elevated initially may help identify gastric cancer patients with poor prognosis.CONCLUSION Serum CEA and CA19-9 play an important role in the prognosis assessment of gastric cancer,and are effective tools to guide clinical practice and optimize individualized treatment strategies for gastric cancer patients.

The combined detection of carcinoembryonic antigen,carcinogenic antigen 125,and carcinogenic antigen 19-9 in colorectal cancer patients

BACKGROUND Hepatic metastases are common and difficult to treat after colorectal cancer(CRC)surgery.The predictive value of carcinoembryonic antigen(CEA),cancer antigen(CA)125 and CA19-9 combined tests for liver metastasis is unclear.AIM To evaluate predictive value of combined tests for CEA,CA125,and CA19-9 levels in patients with liver metastases of CRC.METHODS The retrospective study included patients with CRC alone(50 cases)and patients with CRC combined with liver metastases(50 cases)who were hospitalized between January 2021 and January 2023.Serum CEA,CA125 and CA19-9 levels were compared between the two groups,and binary logistic regression was used to analyze the predictive value of the combination of these tumor markers in liver metastasis.In addition,we performed receiver operating characteristic(ROC)curve analysis to assess its diagnostic accuracy.RESULTS The results showed that the serum CEA,CA125 and CA19-9 levels in the CRC with liver metastasis group were significantly higher than those in the CRC alone group.Specifically,the average serum CEA level in the CRC with liver metastasis group was 162.03±810.01 ng/mL,while that in the CRC alone group was 5.71±9.76 ng/mL;the average serum CA125 levels were 43.47±83.52 U/mL respectively.and 13.5±19.68 U/mL;the average serum CA19-9 levels were 184.46±473.13 U/mL and 26.55±43.96 U/mL respectively.In addition,binary logistic regression analysis showed that CA125 was significant in predicting CRC liver metastasis(P<0.05).ROC curve analysis results showed that the areas under the ROC curves of CEA,CA125 and CA19-9 were 0.607,0.692 and 0.586.CONCLUSION These results suggest that combined detection of these tumor markers may help early detection and intervention of CRC liver metastasis,thereby improving patient prognosis.

Combining prognostic value of serum carbohydrate antigen 19-9 and tumor size reduction ratio in pancreatic ductal adenocarcinoma

BACKGROUND Pancreatic ductal adenocarcinoma(PDAC)is a common cancer with increasing morbidity and mortality due to changes of social environment.AIM To evaluate the significance of serum carbohydrate antigen 19-9(CA19-9)and tumor size changes pre-and post-neoadjuvant therapy(NAT).METHODS This retrospective study was conducted at the Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment,Chongqing University Cancer Hospital.This study specifically assessed CA19-9 levels and tumor size before and after NAT.RESULTS A total of 156 patients who completed NAT and subsequently underwent tumor resection were included in this study.The average age was 65.4±10.6 years and 72(46.2%)patients were female.Before survival analysis,we defined the post-NAT serum CA19-9 level/pre-NAT serum CA19-9 level as the CA19-9 ratio(CR).The patients were divided into three groups:CR<0.5,CR>0.5 and<1 and CR>1.With regard to tumor size measured by both computed tomography and magnetic resonance imaging,we defined the post-NAT tumor size/pre-NAT tumor size as the tumor size ratio(TR).The patients were then divided into three groups:TR<0.5,TR>0.5 and<1 and TR>1.Based on these groups divided according to CR and TR,we performed both overall survival(OS)and disease-free survival(DFS)analyses.Log-rank tests showed that both OS and DFS were significantly different among the groups according to CR and TR(P<0.05).CR and TR after NAT were associated with increased odds of achieving a complete or near-complete pathologic response.Moreover,CR(hazard ratio:1.721,95%CI:1.373-3.762;P=0.006),and TR(hazard ratio:1.435,95%CI:1.275-4.363;P=0.014)were identified as independent factors associated with OS.CONCLUSION This study demonstrated that post-NAT serum CA19-9 level/pre-NAT serum CA19-9 level and post-NAT tumor size/pre-NAT tumor size were independent factors associated with OS in patients with PDAC who received NAT and subsequent surgical resection.

表达PD-1 shRNA增强靶向CD19 CAR-T细胞对肿瘤细胞的杀伤能力

目的:设计和构建表达PD-1 shRNA的靶向CD19 CAR-T细胞并验证其体外肿瘤细胞杀伤能力。方法:设计并构建表达PD-1 shRNA的CD19 CAR分子基因,将其包装成逆转录病毒载体,通过qPCR法检测病毒载体拷贝数。将慢病毒转导人原代T细胞,获得三种CAR-T细胞,分别为RNAU6-CD19 CAR-T、PD-1 shRNA1-CD19 CAR-T、PD-1 shRNA2-CD19 CAR-T细胞。采用qPCR法检测三种CAR-T细胞中PD-1 mRNA的表达水平,流式细胞术检测三种CAR-T细胞中PD-1表达水平,萤光素酶报告基因实验、流式细胞术检测在不同效靶比时CAR-T细胞对CD19阳性靶细胞(人淋巴瘤daudi细胞)的杀伤功能。结果:RNAU6-CD19 CAR、PD-1 shRNA1-CD19 CAR、PD-1 shRNA2-CD19 CAR三种CAR分子成功包装成逆转录病毒载体,病毒载体拷贝数均高于1×10^(7)拷贝/mL,转导人原代T细胞获得CAR-T细胞,RNAU6-CD19 CAR-T、PD-1 shRNA1-CD19 CAR-T、PD-1 shRNA2-CD19 CAR-T细胞转导效率分别为43.1%、55.1%、41.7%。与RNAU6-CD19 CAR-T细胞相比,PD-1 shRNA1-CD19 CAR-T、PD-1 shRNA2-CD19 CAR-T细胞中PD-1 mRNA表达水平均显著降低(均P<0.01)、细胞表面PD-1表达水平更低(均P<0.01)、体外对daudi细胞的杀伤率更高(P<0.05或P<0.01)。结论:成功构建表达PD-1 shRNA的靶向CD19 CAR-T细胞,其对CD19阳性靶细胞的杀伤率显著提高,PD-1 mRNA及其翻译产物PD-1的表达减少,CAR-T细胞的耗竭减缓。

Preclinical evaluation of cyclophosphamide and fludarabine combined with CD19 CAR-T in the treatment of B-cell hematologic malignancies in vivo

Background:Chimeric antigen receptor T(CAR-T)cell therapy has achieved marked therapeutic success in ameliorating hematological malignancies.However,there is an extant void in the clinical guidelines concerning the most effective chemotherapy regimen prior to chimeric antigen receptor T(CAR-T)cell therapy,as well as the optimal timing for CAR-T cell infusion post-chemotherapy.Materials and Methods:We employed cell-derived tumor xenograft(CDX)murine models to delineate the optimal pre-conditioning chemotherapy regimen and timing for CAR-T cell treatment.Furthermore,transcriptome sequencing was implemented to identify the therapeutic targets and elucidate the underlying mechanisms governing the treatment regimen.Results:Our preclinical in vivo evaluation determined that a combination of cyclophosphamide and fludarabine,followed by the infusion of CD19 CAR-T cells five days subsequent to the chemotherapy,exerts the most efficacious therapeutic effect in B-cell hematological malignancies.Concurrently,RNA-seq data indicated that the therapeutic efficacy predominantly perturbs tumor cell metabolism,primarily through the inhibition of key mitochondrial targets,such as C-Jun Kinase enzyme(C-JUN).Conclusion:In summary,the present study offers critical clinical guidance and serves as an authoritative reference for the deployment of CD19 CAR-T cell therapy in the treatment of B-cell hematological malignancies.

C57BL/6J小鼠背景的CD19嵌合抗原受体T细胞的制备及优化

目的:探索C57BL/6J小鼠CD3^(+)T细胞体外刺激条件、最佳培养和感染时间,以期提高CD19嵌合抗原受体T细胞(m CD19 CAR-T)的感染效率。方法:将纯化的C57BL/6J小鼠CD3^(+)T细胞在包被CD3/CD28抗体(anti-CD3/CD28 coated)、包被CD3抗体+可溶性CD28抗体(anti-CD3 coated+soluble anti-CD28)和包被CD3抗体(anti-CD3 coated)3种不同条件下分别刺激12 h和24 h,后续培养24 h、48 h和72 h并记录细胞克隆数。在上述3种条件下分别刺激C57BL/6J小鼠CD3^(+)T细胞12、24和36 h,然后加入白介素(IL)-2(100 U/ml),镜下记录培养24 h、48 h和72 h的细胞克隆数;此条件下取刺激24 h的CD3^(+)T细胞,感染m CD19 CAR-T逆转录病毒,制备m CD19 CAR-T细胞,流式检测48 h时3组中GFP+CAR-T细胞的百分率。结果:利用获得BALB/c小鼠m CD19 CAR-T细胞的最优化条件,制备C57BL/6J小鼠的m CD19 CAR-T细胞感染效率仅5.23%;anti-CD3^(+)soluble anti-CD28 coated刺激C57BL/6J小鼠CD3^(+)T细胞24 h,终止刺激继续培养至48 h时细胞克隆数最高;在上述条件刺激24 h后加入IL-2培养48 h,anti-CD3^(+)soluble anti-CD28 coated组T细胞增殖克隆数量显著增加,且CAR-T细胞感染效率为17.63%±4.17%。结论:C57BL/6J小鼠来源T细胞制备CAR-T细胞的最佳条件与BABL/c小鼠不同;anti-CD3^(+)soluble anti-CD28 coated+IL-2刺激条件下诱导的T细胞感染逆转录病毒后可获得最高效率的m CD19 CAR-T细胞。

CD19 CAR-T细胞治疗的安全性:基于FAERS数据库的不良反应事件信号挖掘与分析

目的分析目前已上市的CD19嵌合抗原受体T(chimeric antigen receptor T,CAR-T)细胞药物在治疗中出现的不良反应事件(adverse effect,AE),为临床用药提供依据。方法使用美国食品药品监督管理局不良事件报告系统(FDA adverse event reporting system,FAERS)数据库,经过数据检索、筛选、去重、整理和分析等处理后,获取4种已上市的CD19 CAR-T药物阿基仑赛(Axicabtagene ciloleucel,Axi-cel)、替沙仑赛(Tisagenleucel,Tisa-cel)、布瑞基奥仑赛(Brexucabtagene autoleucel,Brexu-cel)和瑞基奥仑赛(Lisocabtagene maraleucel,Liso-cel)2017年第1季度至2023年第4季度的基线资料和AE结果。采用报告比值比(reporting odds ratio,ROR)法和比例报告比值比(proportional ADR reporting ratio,PRR)法分析和比较其AE。结果本研究利用FAERS数据库,共得到4种已上市的CD19 CAR-T药物的AE报告5335份。4种CD19 CAR-T药物AE的男性占比均高于女性;年龄分布均在18岁以上;体重则多数集中在50~100 kg。CD19 CAR-T药物AE共累及到23类系统器官分类(system organ classification,SOC),主要集中在神经系统疾病和免疫系统疾病等,其中以细胞因子释放综合征(cytokine release syndrome,CRS)、免疫效应细胞相关性神经毒性综合征(immune effector cell-associated neurotoxicity syndrome,ICANS)最为常见。此外,还发现一些说明书中未列出的AE,如骨髓异常增生综合征(myelodysplastic syndromes,MDS)、急性肾损伤(acute kidney injury,AKI)和脓毒症等。结论在CD 19 CAR-T药物的临床实践中,除需关注药物说明书中提及的AE外,还应注意可能发生的未提及的AE,如MDS、AKI和脓毒症等。

FDA调查接受靶向BCMA或CD19的自体嵌合抗原受体(CAR)-T细胞免疫治疗后发生T细胞恶性肿瘤的严重风险

美国食品药品管理局(FDA)发布消息,已收到接受靶向BCMA或CD19的自体CAR-T细胞免疫治疗后患者出现T细胞恶性肿瘤(包括嵌合抗原受体CAR阳性淋巴瘤)的报告。这些报告来自临床试验和/或上市后不良事件(AE)。

FDA调查接受靶向BCMA或CD19的自体嵌合抗原受体(CAR)-T细胞免疫治疗后发生T细胞恶性肿瘤的严重风险

美国食品药品监督管理局(FDA)发布消息,已收到接受靶向BCMA或CD19的自体CAR-T细胞免疫治疗后患者出现T细胞恶性肿瘤(包括嵌合抗原受体CAR阳性淋巴瘤)的报告。这些报告来自临床试验和/或上市后不良事件(AE)。FDA已确定,T细胞恶性肿瘤的风险适用于目前批准的所有经基因修饰的靶向BCMA和靶向CD19的自体CAR-T细胞免疫疗法。接受过多种同类产品治疗的患者发生了T细胞恶性肿瘤。

美国FDA向靶向BCMA或CD19的自体嵌合抗原受体(CAR)-T细胞免疫治疗类产品相关企业发送标签安全性内容变更通知函

美国食品药品管理局(FDA)发函要求企业修改靶向BCMA或CD19的自体嵌合抗原受体(CAR)-T细胞免疫治疗类产品标签的安全性内容,增加继发性恶性肿瘤相关风险提示。此类产品标签信息应包含以下内容:1.黑框警告,处方信息要点接受靶向BCMA或CD19的自体CAR-T细胞免疫治疗后出现了T细胞恶性肿瘤。