Rudders R A, Jespersen D L, Zacks J, Sikorski A, DeLellis R A, Krontiris T
Department of Medicine, New England Medical Center Hospital, Boston, MA 02111.
J Immunol. 1990 Jan 1;144(1):396-407.
Secretory heterohybrid clones from seven pristine human B cell lymphomas of diverse histologic types were established to investigate the question of tumor clonal diversity. We found that in six tumors, heterohybrid-derived Ig showed similar band patterns in IEF; families of anti-Id prepared from tumor Ig reacted uniformly with individual heterohybrids and original tumor; and the V gene loci displayed little variation on Southern analysis. In one patient who was followed with serial multiple site biopsies over a 14-mo period, clonal Id was preserved until the final stage of his disease, in spite of cytotoxic treatment. In a single follicular tumor (J.M.), each of the anti-Id reacted uniformly with the parent tumor and the individual heterohybrids, except that three of six clones failed to react with a single anti-Id family member. A Southern analysis of the VH gene locus revealed an identical gene rearrangement that was shared by the parent tumor and each heterohybrid. However, there was considerable heterogeneity of J.M. heterohybrid Ig in IEF gels, and we demonstrated the production of variant lambda L chains by the heterohybrid clones. One type of lambda L chain had a normal mobility in SDS-PAGE gels but larger lambda variants were produced by four of six heterohybrids. A Southern analysis of the VL gene displayed considerable variation in the type of lambda rearrangement present in the various heterohybrids, suggesting extensive diversity at the VL gene locus. In a second tumor (S.C.) that exhibited uniform anti-Id tumor reactivity we were also able to demonstrate the presence of a second minor tumor cell population (a biclonal tumor). Our data suggest that intraclonal VH variation may vary considerably with lymphoma subtype and mutagenic exposure and that an additional mechanism for generating spontaneous intraclonal heterogeneity is genetic variation at the VL locus.
建立了来自七种不同组织学类型的原始人类B细胞淋巴瘤的分泌性异源杂交克隆，以研究肿瘤克隆多样性问题。我们发现，在六个肿瘤中，异源杂交衍生的Ig在IEF中显示出相似的条带模式；从肿瘤Ig制备的抗独特型抗体家族与单个异源杂交体和原始肿瘤均能均匀反应；并且Southern分析显示V基因座几乎没有变异。在一名患者中，在14个月的时间里进行了多次多部位连续活检，尽管进行了细胞毒性治疗，但克隆性独特型在其疾病的最后阶段仍得以保留。在一个单一的滤泡性肿瘤（J.M.）中，除了六个克隆中有三个未能与一个抗独特型抗体家族成员反应外，每个抗独特型抗体与亲本肿瘤和单个异源杂交体均能均匀反应。对VH基因座的Southern分析显示，亲本肿瘤和每个异源杂交体共享相同的基因重排。然而，J.M.异源杂交Ig在IEF凝胶中存在相当大的异质性，并且我们证明了异源杂交克隆产生了变异的λ轻链。一种类型的λ轻链在SDS-PAGE凝胶中具有正常的迁移率，但六个异源杂交体中有四个产生了更大的λ变体。对VL基因的Southern分析显示，各种异源杂交体中存在的λ重排类型有很大差异，表明VL基因座存在广泛的多样性。在第二个表现出均匀抗独特型肿瘤反应性的肿瘤（S.C.）中，我们也能够证明存在第二个次要肿瘤细胞群体（双克隆肿瘤）。我们的数据表明，克隆内VH变异可能因淋巴瘤亚型和诱变暴露而有很大差异，并且产生自发克隆内异质性的另一种机制是VL基因座的遗传变异。
