Blatt N B, Bill R M, Glick G D
University of Michigan, Department of Chemistry, Ann Arbor 48109-1055, USA.
Hybridoma. 1998 Feb;17(1):33-9. doi: 10.1089/hyb.1998.17.33.
We have previously described the isolation and in vitro binding properties of eight anti-DNA monoclonal antibodies (MAbs) from an MRL-lpr mouse. In light of recent reports that have indicated it is possible to isolate multiple MAbs from a single hybridoma, our pathogenic hybridoma, 11F8, was examined for evidence of similar phenomena. Chromosome counting suggested that 11F8 cells are unusual and might indeed be expressing multiple heavy and/or light chains. PCR, cloning, and sequencing of immunoglobulin heavy and light chains indicate that 11F8 displays expression of both gamma 2a and gamma 3 heavy chains at the DNA level. Flow cytometry and amino acid sequencing reveals that expression of multiple isotypes also occurs at the protein level but only a single heavy- and light-chain sequence is able to bind DNA. Based on these results, we conclude that 11F8 is an unusual hybridoma that secretes two distinct heavy and at least one light chain from a single cell, and may represent a trioma, a stable three-cell fusion.
我们之前描述过从一只MRL - lpr小鼠中分离出的8种抗DNA单克隆抗体（MAb）及其体外结合特性。鉴于最近有报道表明有可能从单个杂交瘤中分离出多种MAb，我们对致病性杂交瘤11F8进行了检测，以寻找类似现象的证据。染色体计数表明11F8细胞不同寻常，可能确实在表达多种重链和/或轻链。免疫球蛋白重链和轻链的聚合酶链反应（PCR）、克隆及测序表明，11F8在DNA水平上显示出γ2a和γ3重链的表达。流式细胞术和氨基酸测序显示，多种同种型的表达在蛋白质水平上也会发生，但只有单一的重链和轻链序列能够结合DNA。基于这些结果，我们得出结论，11F8是一种不同寻常的杂交瘤，它从单个细胞中分泌出两种不同的重链和至少一种轻链，可能代表一种三瘤细胞，即一种稳定的三细胞融合体。
