Huff T F, Ishizaka K
Proc Natl Acad Sci U S A. 1984 Mar;81(5):1514-8. doi: 10.1073/pnas.81.5.1514.
Normal human T cells that proliferated in the presence of interleukin 2 (IL-2) formed IgE-binding factors when incubated with human IgE. These cells were then fused with a mutant of the human T-cell line CEM. Incubation of five hybridomas with human IgE or culture of the cells in IgE-coated wells resulted in the formation of IgE-binding factors. One hour of incubation with 10 micrograms of human IgE per ml was sufficient to induce the hybridomas to form IgE-binding factors. Polymerized IgE was much more efficient than monomeric IgE for the induction of the factor formation. As little as 10 ng of IgE dimer per ml was sufficient to induce factor formation. The IgE-binding factors produced by the hybridomas bound to human IgE-coated Sepharose and were recovered from the beads by elution at acid pH. The factors had low affinity for rat IgE but failed to bind to human IgG. The IgE-binding factors formed by four hybridomas had a Mr between 25,000 and 30,000, whereas one hybridoma formed IgE-binding factors of Mr 30,000 and Mr 15,000. The IgE-binding formed by all of the hybridomas had affinity for concanavalin A, indicating that the factors are glycoproteins.
在白细胞介素2（IL-2）存在下增殖的正常人T细胞，与人IgE一起孵育时会形成IgE结合因子。然后将这些细胞与人类T细胞系CEM的一个突变体进行融合。用人类IgE孵育五个杂交瘤，或将细胞在包被有IgE的孔中培养，都会导致IgE结合因子的形成。每毫升用10微克人类IgE孵育1小时就足以诱导杂交瘤形成IgE结合因子。聚合IgE比单体IgE在诱导因子形成方面效率更高。每毫升低至10纳克的IgE二聚体就足以诱导因子形成。杂交瘤产生的IgE结合因子与包被有人类IgE的琼脂糖结合，并通过在酸性pH下洗脱从珠子上回收。这些因子对大鼠IgE亲和力低，但不与人IgG结合。四个杂交瘤形成的IgE结合因子的分子量在25,000至30,000之间，而一个杂交瘤形成的IgE结合因子的分子量为30,000和15,000。所有杂交瘤形成的IgE结合物对伴刀豆球蛋白A有亲和力，表明这些因子是糖蛋白。
