Ig/TCR基因重排在儿童急性T淋巴细胞白血病中的表达模式特点
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摘要
目的探讨免疫球蛋白(Ig)/T细胞受体(TCR)基因重排在儿童急性T淋巴细胞白血病(T-ALL)中的表达模式特点及其与临床生物学特征的相关性。方法回顾性纳入首都医科大学附属北京儿童医院(我院)2005年1月1日至2008年12月31日收治的初治T-ALL患儿,分析其初诊时骨髓单个核细胞的Ig/TCR基因重排情况,根据重排情况分为阳性组和阴性组,比较不同组别的临床生物学特征。结果①52例儿童T-ALL中男37例(71.2%),入院时中位年龄8.0(1.8～16.0)岁,初诊时WBC中位数为140.5(2.7～667.1)×10~9·L~(-1),中危38例(73.1%)、高危14例(26.9%)。中位随访时间136.3(1.2～171.7)个月,长期完全缓解38例(73.1%)、复发10例(19.2%),其他原因死亡4例(7.7%)。②TCRB、TCRG、TCRD和IgH克隆性基因重排的发生率分别为85%、85%、38%和21%,94%的患儿检出至少1种基因重排,88%的患儿检出至少2种基因重排。TCRB、TCRD、TCRG和IgH重排分别以完全性V_β-(D_β)-J_β、V_δ-J_δ、V_γⅠ-J_γ1.3/2.3和D_H-J_H不完全重排为主。各种重排的胚系片段使用和连接区序列极具多样性。③10例复发患儿中有6例检测了复发时的Ig/TCR基因重排模式,4例与初诊时完全一致,2例发生改变。④SIL-TAL1融合基因阳性率在11例IgH重排阳性患儿中0%(0/14),在41例IgH重排阴性患儿中为34.1%(14/41),P=0.025。⑤TCRB基因重排阳性组高危比例(20.5%,9/44)低于阴性组(62.5%,5/8),P=0.025。TCRB或TCRG基因重排阳性组第33 d缓解率(89.4%,42/44)高于阴性组(10.6%,5/8),P=0.022,第78 d MRD水平≥10~(-3)的比例(10.8%,4/37)低于阴性组(50.0%,3/6),P=0.045。结论儿童T-ALL初诊时Ig/TCR克隆性基因重排的胚系片段使用和连接区序列极具多样性,有助于进一步性MRD检测标志的筛选。
Objective To investigate the characteristics of immunoglobulin(Ig) and T-cell receptor(TCR) gene rearrangement patterns in childhood T-cell acute lymphoblastic leukemia(T-ALL), and its correlation with the clinical and biological characteristics of the patients. Methods The study included 52 newly diagnosed pediatric T-ALL patients enrolled at Beijing Children's Hospital from January 1 st, 2005 to December 31 th, 2008. Analyze the Ig/TCR gene rearrangement of bone marrow mononuclear cells at diagnosis, including the incidence of TCRB, TCRG, TCRD and IgH gene rearrangements and types, germline fragment usage and junctional characteristics.According to the rearrangement of each gene, it was divided into positive and negative groups, and then the clinical and biological characteristics were compared between different groups. Results ① There were 37 males(71.2%) among the 52 children,aged from 1.8 to 16 years, with a median of 8 years,and the median WBC was 140.5(2.7-667.1) ×10~9·L~(-1). There were 38 cases of IR(73.1%), and 14 cases of HR(26.9%). The follow-up time was 1.2 to 171.7 months and the median follow-up time was 136.3 months. There were 38 cases(73.1%) of long-term complete remission, 10 cases(19.2%) of recurrence and 4 cases(7.7%) of death from other cases. ②The incidence of TCRB, TCRG, TCRD and IgH clonal gene rearrangement was 85%, 85%, 38% and 21%, respectively. Ninety-four percent of the children were detected at least one gene rearrangement, and 88% of the children were at least examined two gene rearrangements. The TCRB, TCRD, TCRG, and IgH rearrangements were dominated by complete V_β-(D_β)-J_β, V_δ-J_δ, V_γI-J_γ1.3/2.3, and D_H-J_H incomplete rearrangement, respectively. Different combinations of germline gene segments resulted in the combinatorial diversity, and the deletion and random insertion of nucleotides at the junction sites created an enormous junctional diversity. ③Among the ten relapsed children, six were compared the Ig/TCR gene rearrangement pattern between diagnosis and recurrence,and four patients had recurrence of Ig/TCR gene rearrangement pattern at the time of initial diagnosis and two patients changed. ④The positive rates of SIL-TAL1 fusion gene were 0 in 11 patients with positive IgH rearrangement and 34.1%(14/41) in 41 children with negative IgH rearrangement,P=0.025. ⑤The high-risk ratio of TCRB gene rearrangement positive group(20.5%, 9/44) was lower than that of negative group(62.5%, 5/8), P=0.025. The remission rate at the 33 rd day of the TCRB or TCRG gene rearrangement positive group(89.4%, 42/44) was higher than that of the negative group(10.6%, 5/8), P=0.022, and the ratio of MRD level ≥10~(-3 )on the 78 th day(10.8%, 4/37) was lower than the negative group(50.0%, 3/6), P=0.045. Conclusion The usage of germline fragments of clonal Ig/TCR gene rearrangements and the DNA sequence of their junction regions were diverse, which was helpful for further screening of MRD markers of T-ALL in children.
Objective To investigate the characteristics of immunoglobulin(Ig) and T-cell receptor(TCR) gene rearrangement patterns in childhood T-cell acute lymphoblastic leukemia(T-ALL), and its correlation with the clinical and biological characteristics of the patients. Methods The study included 52 newly diagnosed pediatric T-ALL patients enrolled at Beijing Children's Hospital from January 1 st, 2005 to December 31 th, 2008. Analyze the Ig/TCR gene rearrangement of bone marrow mononuclear cells at diagnosis, including the incidence of TCRB, TCRG, TCRD and IgH gene rearrangements and types, germline fragment usage and junctional characteristics.According to the rearrangement of each gene, it was divided into positive and negative groups, and then the clinical and biological characteristics were compared between different groups. Results ① There were 37 males(71.2%) among the 52 children,aged from 1.8 to 16 years, with a median of 8 years,and the median WBC was 140.5(2.7-667.1) ×10~9·L~(-1). There were 38 cases of IR(73.1%), and 14 cases of HR(26.9%). The follow-up time was 1.2 to 171.7 months and the median follow-up time was 136.3 months. There were 38 cases(73.1%) of long-term complete remission, 10 cases(19.2%) of recurrence and 4 cases(7.7%) of death from other cases. ②The incidence of TCRB, TCRG, TCRD and IgH clonal gene rearrangement was 85%, 85%, 38% and 21%, respectively. Ninety-four percent of the children were detected at least one gene rearrangement, and 88% of the children were at least examined two gene rearrangements. The TCRB, TCRD, TCRG, and IgH rearrangements were dominated by complete V_β-(D_β)-J_β, V_δ-J_δ, V_γI-J_γ1.3/2.3, and D_H-J_H incomplete rearrangement, respectively. Different combinations of germline gene segments resulted in the combinatorial diversity, and the deletion and random insertion of nucleotides at the junction sites created an enormous junctional diversity. ③Among the ten relapsed children, six were compared the Ig/TCR gene rearrangement pattern between diagnosis and recurrence,and four patients had recurrence of Ig/TCR gene rearrangement pattern at the time of initial diagnosis and two patients changed. ④The positive rates of SIL-TAL1 fusion gene were 0 in 11 patients with positive IgH rearrangement and 34.1%(14/41) in 41 children with negative IgH rearrangement,P=0.025. ⑤The high-risk ratio of TCRB gene rearrangement positive group(20.5%, 9/44) was lower than that of negative group(62.5%, 5/8), P=0.025. The remission rate at the 33 rd day of the TCRB or TCRG gene rearrangement positive group(89.4%, 42/44) was higher than that of the negative group(10.6%, 5/8), P=0.022, and the ratio of MRD level ≥10~(-3 )on the 78 th day(10.8%, 4/37) was lower than the negative group(50.0%, 3/6), P=0.045. Conclusion The usage of germline fragments of clonal Ig/TCR gene rearrangements and the DNA sequence of their junction regions were diverse, which was helpful for further screening of MRD markers of T-ALL in children.
引文
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[8] 李志刚,吴敏媛,朱平,等.白血病29种染色体畸变形成的融合基因分析.中华儿科杂志,2001,39(11):682-685
[9] Spits H.Development of alphabeta T cells in the human thymus.Nat Rev Immunol,2002,2(10):760-772
[10] Dik WA,Pike-Overzet K,Weerkamp F,et al.New insights on human T cell development by quantitative T cell receptor gene rearrangement studies and gene expression profiling.J Exp Med,2005,201(11):1715-1723
[11] Kolenova A,Hikkel I,Ilencikova D,et al.Minimal residual disease detection using real-time quantitative PCR analysis of immunoglobulin and T-cell receptor gene rearrangements in the non-MRD-based ALL IC-BFM 2002 protocol for childhood ALL:Slovak experience.Neoplasma,2010,57(6):552-561
[12] Szczepański T,Pongers-Willemse MJ,Langerak AW,et al.Ig heavy chain gene rearrangements in T-cell acute lymphoblastic leukemia exhibit predominant DH6-19 and DH7-27 gene usage,can result in complete V-D-J rearrangements,and are rare in T-cell receptor alpha beta lineage.Blood,1999,93(12):4079-4085
[13] Flohr T,Schrauder A,Cazzaniga G,et al.Minimal residual disease-directed risk stratification using real-time quantitative PCR analysis of immunoglobulin and T-cell receptor gene rearrangements in the international multicenter trial AIEOP-BFM ALL 2000 for childhood acute lymphoblastic leukemia.Leukemia,2008,22(4):771-782
[14] 刘婕好,李志刚,高超,等.儿童急性T淋巴细胞白血病T细胞受体β链基因重排的特点及其在微小残留病定量检测中的意义.中华儿科杂志,2008,46(z1):18-24
[15] Sudhakar N,Nancy NK,Rajalekshmy KR,et al.T-cell receptor gamma and delta gene rearrangements and junctional region characteristics in south Indian patients with T-cell acute lymphoblastic leukemia.Am J Hematol,2007,82(3):215-221
[16] Meleshko AN,Lipay NV,Stasevich IV,et al.Rearrangements of IgH,TCRD and TCRG genes as clonality marker of childhood acute lymphoblastic leukemia.Exp Oncol,2005,27(4):319-324
[17] Szczepański T,Langerak AW,Willemse MJ,et al.T cell receptor gamma (TCRG) gene rearrangements in T cell acute lymphoblastic leukemia refelct 'end-stage' recombinations:implications for minimal residual disease monitoring.Leukemia,2000,14(7):1208-1214
[18] van der Velden VH,Wijkhuijs JM,Jacobs DC,et al.T cell receptor gamma gene rearrangements as targets for detection of minimal residual disease in acute lymphoblastic leukemia by real-time quantitative PCR analysis.Leukemia,2002,16(7):1372-1380
[19]Szczepański T,van der Velden VH,Raff T,et al.Comparative analysis of T-cell receptor gene rearrangements at diagnosis and relapse of T-cell acute lymphoblastic leukemia (T-ALL) shows high stability of clonal markers for monitoring of minimal residual disease and reveals the occurrence of second T-ALL.Leukemia,2003,17(11):2149-2156
[2] Borowitz MJ,Wood BL,Devidas M,et al.Prognostic significance of minimal residual disease in high risk B-ALL:a report from Children's Oncology Group study AALL0232.Blood,2015,126(8):964-971
[3] Flohr T,Schrauder A,Cazzaniga G,et al.Minimal residual disease-directed risk stratification using real-time quantitative PCR analysis of immunoglobulin and T-cell receptor gene rearrangements in the international multicenter trial AIEOP-BFM ALL 2000 for childhood acute lymphoblastic leukemia.Leukemia,2008,22(4):771-782
[4] O'Connor D1,Enshaei A,Bartram J,et al.Genotype-specific minimal residual disease interpretation improves stratification in pediatric acute lymphoblastic leukemia.J Clin Oncol,2018,36(1):34-43
[5] van Krieken JH,Langerak AW,Macintyre EA,et al.Improved reliability of lymphoma diagnostics via PCR-based clonality testing:report of the BIOMED-2 Concerted Action BHM4-CT98-3936.Leukemia,2007,21(2):201-206
[6] Langerak AW,Groenen PJ,Brüggemann M,et al.EuroClonality/BIOMED-2 guidelines for interpretation and reporting of Ig/TCR clonality testing in suspected lymphoproliferations.Leukemia,2012,26(10):2159-2171
[7]中华医学会儿科学分会血液学组,中华儿科杂志编辑委员会.儿童急性淋巴细胞白血病诊疗建议(第四次修订).中华儿科杂志,2014,52(9):641-644
[8] 李志刚,吴敏媛,朱平,等.白血病29种染色体畸变形成的融合基因分析.中华儿科杂志,2001,39(11):682-685
[9] Spits H.Development of alphabeta T cells in the human thymus.Nat Rev Immunol,2002,2(10):760-772
[10] Dik WA,Pike-Overzet K,Weerkamp F,et al.New insights on human T cell development by quantitative T cell receptor gene rearrangement studies and gene expression profiling.J Exp Med,2005,201(11):1715-1723
[11] Kolenova A,Hikkel I,Ilencikova D,et al.Minimal residual disease detection using real-time quantitative PCR analysis of immunoglobulin and T-cell receptor gene rearrangements in the non-MRD-based ALL IC-BFM 2002 protocol for childhood ALL:Slovak experience.Neoplasma,2010,57(6):552-561
[12] Szczepański T,Pongers-Willemse MJ,Langerak AW,et al.Ig heavy chain gene rearrangements in T-cell acute lymphoblastic leukemia exhibit predominant DH6-19 and DH7-27 gene usage,can result in complete V-D-J rearrangements,and are rare in T-cell receptor alpha beta lineage.Blood,1999,93(12):4079-4085
[13] Flohr T,Schrauder A,Cazzaniga G,et al.Minimal residual disease-directed risk stratification using real-time quantitative PCR analysis of immunoglobulin and T-cell receptor gene rearrangements in the international multicenter trial AIEOP-BFM ALL 2000 for childhood acute lymphoblastic leukemia.Leukemia,2008,22(4):771-782
[14] 刘婕好,李志刚,高超,等.儿童急性T淋巴细胞白血病T细胞受体β链基因重排的特点及其在微小残留病定量检测中的意义.中华儿科杂志,2008,46(z1):18-24
[15] Sudhakar N,Nancy NK,Rajalekshmy KR,et al.T-cell receptor gamma and delta gene rearrangements and junctional region characteristics in south Indian patients with T-cell acute lymphoblastic leukemia.Am J Hematol,2007,82(3):215-221
[16] Meleshko AN,Lipay NV,Stasevich IV,et al.Rearrangements of IgH,TCRD and TCRG genes as clonality marker of childhood acute lymphoblastic leukemia.Exp Oncol,2005,27(4):319-324
[17] Szczepański T,Langerak AW,Willemse MJ,et al.T cell receptor gamma (TCRG) gene rearrangements in T cell acute lymphoblastic leukemia refelct 'end-stage' recombinations:implications for minimal residual disease monitoring.Leukemia,2000,14(7):1208-1214
[18] van der Velden VH,Wijkhuijs JM,Jacobs DC,et al.T cell receptor gamma gene rearrangements as targets for detection of minimal residual disease in acute lymphoblastic leukemia by real-time quantitative PCR analysis.Leukemia,2002,16(7):1372-1380
[19]Szczepański T,van der Velden VH,Raff T,et al.Comparative analysis of T-cell receptor gene rearrangements at diagnosis and relapse of T-cell acute lymphoblastic leukemia (T-ALL) shows high stability of clonal markers for monitoring of minimal residual disease and reveals the occurrence of second T-ALL.Leukemia,2003,17(11):2149-2156