典型和不典型免疫表型慢性淋巴细胞白血病遗传学和分子生物学特征研究

Study of cytogenetics and molecular biology in typical and atypical immunophenotypic chronic lymphocytic leukemia

目的探讨典型和不典型免疫表型慢性淋巴细胞白血病(CLL)在免疫表型、遗传学及分子生物学方面的差异及遗传学异常与基因突变的相关性.方法依据英国马斯登皇家医院免疫分型积分系统对2014年11月至2021年5月期间南京医科大学第一附属医院收治的488例初诊CLL患者进行分类,其中积分4~5分为典型免疫表型CLL(tCLL)(382例),积分3分为不典型免疫表型CLL(aCLL)(106例).采用多参数流式细胞术对所有CLL患者外周血标本进行免疫表型检测,荧光原位杂交(FISH)技术检测359例CLL患者的遗传学异常,二代测序(NGS)技术检测330例CLL患者基因突变情况.结果aCLL患者CD10、CD22、CD49d、CD81和FMC7阳性表达率显著高于tCLL患者(P值分别为0.020、<0.001、<0.001、0.027和<0.001),CD5、CD23、CD148和CD200的阳性表达率显著低于tCLL患者(P值分别为<0.001、0.017、0.041和<0.001).aCLL患者+12阳性率显著高于tCLL患者(P<0.001),del(13q14)阳性率显著低于tCLL患者(P<0.001),同时aCLL患者NOTCHl突变发生率高于tCLL患者(P=0.038),其余基因突变发生率在两组间的差异均无统计学意义(P值均>0.05).生存分析显示,tCLL和aCLL患者总生存(OS)率与无治疗生存(TFS)率的差异无统计学意义(P值均>0.05).结论tCLL与aCLL患者的抗原表达特征、细胞遗传学及体细胞突变均存在差异,有助于tCLL与aCLL的诊断和鉴别诊断.

Objective To analyze the differences in immunophenotype,cytogenetics,and molecular biology between typical and atypical immunophenotype chronic lymphocytic leukemia(CLL),and explore the correlation of cytogenetic anomalies with gene mutations.Methods This study included 488 patients diagnosed in the First Affiliated Hospital of Nanjing Medical University between November 2014 and May 2021.Of these,382 patients scored 4-5 points,which was typical CLL(tCLL),and 106 scored 3 points,which was atypical CLL(aCLL)as per the Royal Marsden Hospital Immunomarker Integral System.Peripheral blood cells were collected for immunophenotype by multiparameter flow cytometry in 488 patients,fluorescence in situ hybridization(FISH)was employed to detect cytogenetic anomalies in 359 patients,and gene mutations were detected by next-generation sequencing(NGS)in 330 patients.Results The positive rates of CD10,CD22,CD49d,CD81,and FMC7 were significantly higher in the aCLL compared with the tCLL group(P=0.020,P<0.001,P<0.001,P=0.027,and P<0.001,respectively),while the positive rates of CD5,CD23,CD148,and CD200 were lower in the former compared to the latter(P<0.001,P=0.017,P=0.041,and P<0.001,respectively).aCLL exhibited a higher frequency of trisomy 12 and lower frequency of del(13q14)compared to the tCLL group(P<0.001 and P<0.001,respectively).Moreover,aCLL patients also showed a higher incidence of NOTCH1 mutations than the tCLL patients(P=0.038),while no statistically significant differences in other gene mutations occurred between the two groups.No significant differences in overall survival(OS)and treatment-free survival(TFS)occurred between aCLL and tCLL using Kaplan-Meier analysis(P>0.05).Conclusion aCLL has characteristic immunophenotype,cytogenetic,and somatic mutation that differ from tCLL,and this can provide reliable information for the diagnosis and differential diagnosis between the two groups.