Ohno T, Stribley J A, Wu G, Hinrichs S H, Weisenburger D D, Chan W C
Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha 68198-3135, USA.
N Engl J Med. 1997 Aug 14;337(7):459-65. doi: 10.1056/NEJM199708143370704.
There is general agreement that lymphocytic and histiocytic (L&H) cells, the variants of Reed-Sternberg cells in nodular lymphocyte-predominant Hodgkin's disease, belong to the B-cell lineage. However, the clonality of L&H cells remains controversial.
We used complementarity-determining region 3 (CDR3) of the immunoglobulin heavy-chain gene as a clonal marker to study individual L&H cells isolated by micromanipulation from tissue sections of five patients with nodular lymphocyte-predominant Hodgkin's disease. The heavy-chain CDR3 of each cell was amplified by the polymerase chain reaction. The products were analyzed by gel electrophoresis, and representative amplification products from each patient were sequenced.
L&H cells whose heavy-chain CDR3 was related, indicating the presence of a clonal population, were detected in all five patients and were the dominant population in three. In four of the five patients, members of the clone were found in different nodules in the tissue section, different tissue blocks from the same tumor, or different lymph nodes from the same patient. The CDR3 sequences in each clone frequently contained nucleotide substitutions indicative of intraclonal mutation.
Clonal populations of L&H cells occur in nodular lymphocyte-predominant Hodgkin's disease. Intraclonal variation in nucleotide sequences suggests that hypermutation of the heavy-chain CDR3 continues to occur among the clonal progeny.
人们普遍认为，结节性淋巴细胞为主型霍奇金淋巴瘤中里德-施特恩贝格细胞的变异型，即淋巴细胞和组织细胞（L&H）细胞，属于B细胞谱系。然而，L&H细胞的克隆性仍存在争议。
我们使用免疫球蛋白重链基因的互补决定区3（CDR3）作为克隆标志物，来研究从5例结节性淋巴细胞为主型霍奇金淋巴瘤患者的组织切片中通过显微操作分离出的单个L&H细胞。通过聚合酶链反应扩增每个细胞的重链CDR3。对产物进行凝胶电泳分析，并对每位患者的代表性扩增产物进行测序。
在所有5例患者中均检测到重链CDR3相关的L&H细胞，表明存在克隆群体，其中3例患者中该克隆群体占主导。在5例患者中的4例中，在组织切片的不同结节、同一肿瘤的不同组织块或同一患者的不同淋巴结中发现了克隆成员。每个克隆中的CDR3序列经常包含指示克隆内突变的核苷酸替换。
结节性淋巴细胞为主型霍奇金淋巴瘤中存在L&H细胞的克隆群体。核苷酸序列的克隆内变异表明重链CDR3的超突变在克隆后代中持续发生。
