Carroll W L, Thielemans K, Dilley J, Levy R
J Immunol Methods. 1986 May 1;89(1):61-72. doi: 10.1016/0022-1759(86)90032-3.
Surface idiotype (Id) of B cell malignancies is an excellent tumor-specific marker. We have, however, recently described heterogeneity of tumor Id in some cases. We therefore sought a way to isolate, reliably and efficiently, different species of idiotype from a potentially heterogeneous population. In this report we demonstrate our success using a series of mouse X human heterohybridomas as fusion partners with human B cell tumors. Three lines (K6H6/B5, K6H9/G12, SBC/H20) demonstrated excellent fusion efficiency with 75%-85% of wells plated containing hybrids. Two cell lines, K6H9/G12 and SBC/H20 had a tendency to secrete a single Ig chain (heavy or light chain), whereas the K6H6/B5 cell line secreted whole immunoglobulin (Ig) in greater than 80% of the hybrids. This line secreted significant amounts of Ig (2.73 micrograms/ml/10(6) cells) and was relatively stable in culture. Since this line has such a high fusion efficiency the products of normal B cells admixed with tumor may be recovered, allowing the opportunity of isolating host anti-tumor antibodies. In order to prove that hybrids were derived from the tumor, Southern blot analysis of rearranged DNA was performed in selected cases. Fusions with this line provide the potential for recovering many different species of idiotype in a mixed population. This will facilitate the production of mouse monoclonal anti-idiotype antibodies against many variants and against different idiotopes.
B细胞恶性肿瘤的表面独特型(Id)是一种出色的肿瘤特异性标志物。然而,我们最近在某些病例中描述了肿瘤Id的异质性。因此,我们寻求一种方法,能够从潜在的异质群体中可靠且高效地分离出不同种类的独特型。在本报告中,我们展示了使用一系列小鼠X人异源杂交瘤作为与人B细胞肿瘤融合伙伴的成功经验。三个细胞系(K6H6/B5、K6H9/G12、SBC/H20)表现出优异的融合效率,接种的孔中有75%-85%含有杂交细胞。两个细胞系K6H9/G12和SBC/H20倾向于分泌单一的Ig链(重链或轻链),而K6H6/B5细胞系在超过80%的杂交细胞中分泌完整的免疫球蛋白(Ig)。该细胞系分泌大量的Ig(2.73微克/毫升/10⁶个细胞),并且在培养中相对稳定。由于该细胞系具有如此高的融合效率,与肿瘤混合的正常B细胞产物可能被回收,从而有机会分离宿主抗肿瘤抗体。为了证明杂交细胞源自肿瘤,在选定的病例中进行了重排DNA的Southern印迹分析。与该细胞系的融合为在混合群体中回收许多不同种类的独特型提供了可能性。这将有助于生产针对许多变体和不同独特位的小鼠单克隆抗独特型抗体。