急性B淋巴细胞白血病免疫球蛋白重链可变区基因的分子特.docVIP

急性B淋巴细胞白血病免疫球蛋白重链可变区基因的分子特征 刘英1 朱平2 胡亚美1 [摘要] 目的 探讨急性B淋巴细胞白血病（B-ALL） PCR扩增108例ALL不同免疫球蛋白重链可变区（IgHV）亚家族的重排基因；PCR产物直接测序并翻译成氨基酸序列；利用生物信息资源分析B-ALL IgHV基因的重排类型、胚系基因片段利用及体细胞突变并预测T细胞表位。结果 66%ALL检测到完整的VHDJH重排克隆，其中34例单等位基因重排（47.89%）30例双等位基因重排（42.25%）7例（9.86%）寡克隆基因重排。40份B-ALL IgHV基因序13份读框内（in frame）重排5份含终止密码子27份读框移位（frameshift）功能性重排。重排使用最多的VH家族为VH3（11/40）VH4（11/40）和 VH1（8/40），占75%VH4-59（5/40）和VH4-34（4/40）VH4家族中频率最高优先利用的D家族为D3（35%）和D2（27.5%），D7-27的利用率（15%）明显高于正常PBL（p= 1.29E-12）JH6是B-ALL利用最多的JH基因片段（42.50%），JH4（30.00%）次之；20%（8/40）的B-ALL在DJHN核苷酸的插入 高于正常PBL (p=0.017)。22.%B-ALL IgHV基因发生体细胞突变，突变的碱基散布于IgHV全长，CDR区替代突变与静寂突变的比例(1；26例B-ALL重链可变区的HLA I类分子结合肽80%以上位于框架区,同一VH家族的B-ALL有相同的1~2条九肽。 结论 B-ALL起源于未受抗原刺激的祖B或前B细胞，它们的IgHV基因具有胚系特征；白血病的恶性转化往往发生于胚胎期；生物信息分析B-ALL的T细胞表位主要位于框架1和框架3区。 [关键词] 基因，免疫球蛋白；表位，T细胞；白血病，淋巴细胞性；聚合酶链反应；生物信息资源 Characteristics of immunoglobulin heavy chain variable region genes in B-cell acute lymphoblastic leukemia LIU Ying *, ZHU Ping , Hu Yamei .*Beijing Children(s Hospital Affiliated to Capital University of Medical Sciences , Beijing 100045 . [Abstract] Objective To investigate the origin and T-cell epitopes of B-cell acute lymphoblastic leukemia. Methods Rearrangement genes of different VH families were amplified by PCR and directly sequenced for 108 children with acute lymphoblastic leukemia. Nucleotide Sequences available were translated into amino acid sequences. Bioinformatics were applied for analyzing recombination patterns，somatic mutations and usage of germline gene segments and predicting T cell epitopes. Results Complete VHDJH rearrangements were identified in 71(66%)out of 108 cases with 34（47.89%））9.86%）oligoclonal rearrangements. Among 40 B-ALL IgHV gene sequences available ,13 were in frame, whereas 27 were frameshifts; 5 in frame joinings contained stop codons, leaving 8(20%) potentially functional rearrangements. VH3（11/40）VH4（11/40）and VH1（8/40）VH families in B-ALL; VH4-59 and VH4-34 were the most frequently rearranged VH4 family gene segm