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目的 比较人类胚胎发育期的淋巴细胞克隆 ,儿童急性淋巴细胞白血病 (ALL)和多发性骨髓瘤 (MM)的免疫球蛋白重链 (IgH)基因 ,确定与白血病恶性克隆有关的基因特征。方法 利用IgH可变区各个基因家族 (VH1 VH6 )和结合区 (J)的特异序列做引物 ,RT PCR扩增IgH重排的CDR3区基因 ,然后用变性胶长电泳的方法 ,获得不同发育期胚胎的肝 ,脾 ,胸腺和骨髓B细胞克隆的基因指纹图谱 ,同样也获得 15例ALL和 1例MMB细胞基因指纹图谱 ,测序对比分析ALL及MM病人恶性细胞IgH基因的序列特征。结果 以IgHVH1 VH6基因家族为标志的各个B细胞克隆出现有一定规律。肝脏VH6基因是占优势的家族 ,而脾脏中VH3为优势家族 ,骨髓更多利用的也是VH3。ALL 9/15例表现为B淋巴细胞单克隆的增生 ,3例属VH1家族 ,3例属VH6家族 ,2例同时有VH3和VH4家族基因。患者其他基因家族表现为正常的均匀分布的梯度状的多克隆谱系或者受到抑制。 1例ALL的VH1、VH3和VH6家族呈寡克隆现象。 6例ALL呈正常的多克隆分布。克隆并测序分析 1例ALL单克隆VH3序列 ,发现与胚胎 13周的序列相似并具胚胎期特点。 1例成人MM的IgH序列特点则完全不同 ,出现体细胞基因的高突变。结论 ALL的IgH基因重排可以有不同的VH1,VH3,VH4 ,VH6重排的克隆类型 ;儿童白血病发病可
Objective To compare the immunoglobulin heavy chain (IgH) genes of lymphocyte clones in human embryonic development, childhood acute lymphoblastic leukemia (ALL) and multiple myeloma (MM), and to determine the genetic characteristics associated with malignant leukemia clones. Methods The primers were used as the primers for the specific sequences of VH1 VH6 and the binding region of IgH variable region. RT PCR was used to amplify the IgH rearranged CDR3 region genes, and then the different developmental stages were obtained by long gel electrophoresis. The gene fingerprints of embryonic liver, spleen, thymus, and bone marrow B cell clones were also obtained from the DNA fingerprinting of 15 ALL and 1 MMB cells. The sequence characteristics of malignant IgH genes in ALL and MM patients were analyzed by sequencing. RESULTS: The B cell clones marked with the IgHVH1 VH6 gene family showed certain patterns. Liver VH6 gene is the predominant family, while VH3 is the dominant family in the spleen and VH3 is the more abundant bone marrow. ALL 9/15 cases showed the proliferation of B lymphocyte monoclones, 3 cases belonged to VH1 family, 3 cases belonged to VH6 family, 2 cases also had VH3 and VH4 family genes. The patient’s other gene family showed normal, evenly distributed, gradient-like polyclonal lines or was suppressed. In 1 case of ALL, the VH1, VH3, and VH6 families were oligoclonal. Six cases of ALL showed a normal polyclonal distribution. Clone and sequencing analysis of a single ALL VH3 sequence was found to be similar to the embryonic 13-week sequence and embryonic stage characteristics. The IgH sequence characteristics of an adult MM were completely different, and there was a high mutation in the somatic gene. Conclusion The IgH gene rearrangement of ALL may have different clone types of VH1, VH3, VH4, and VH6 rearrangements;