Amin A R, Coico R F, Finkelman F, Siskind G W, Thorbecke G J
Department of Pathology, New York University Medical Center, NY 10016.
Proc Natl Acad Sci U S A. 1988 Dec;85(23):9179-83. doi: 10.1073/pnas.85.23.9179.
Helper T cells with receptors specific for IgD have immunoaugmenting properties. We have now detected soluble IgD-binding factor in cell supernatants immobilized on nitrocellulose paper by their ability to bind 125I-labeled IgD. IgD-binding factor is released by normal splenic T cells stimulated with recombinant interleukin 2, recombinant interleukin 4, or crosslinked IgD in amounts paralleling the induction of IgD receptors on the cells. IgD receptors are constitutively produced by antigen-specific helper T-cell hybridomas 2H10 and A3.4C6. Incubation of these hybridoma cells with recombinant interleukin 2 increases release of IgD-binding factor while reducing expression of IgD receptors. Specificity of the binding factor for IgD is established by (i) competitive inhibition; (ii) the ability of the binding factor to bind radiolabeled IgD and not monoclonal IgE, IgG2a, or polyclonal IgG; and (iii) the removal of the binding factor on passage through an IgD-Sepharose column and recovery in a subsequent acid eluate.
具有针对IgD的特异性受体的辅助性T细胞具有免疫增强特性。我们现在已经通过其结合125I标记的IgD的能力,在固定于硝酸纤维素纸上的细胞上清液中检测到了可溶性IgD结合因子。IgD结合因子由用重组白细胞介素2、重组白细胞介素4或交联的IgD刺激的正常脾T细胞释放,其释放量与细胞上IgD受体的诱导量平行。抗原特异性辅助性T细胞杂交瘤2H10和A3.4C6组成性产生IgD受体。这些杂交瘤细胞与重组白细胞介素2一起孵育会增加IgD结合因子的释放,同时降低IgD受体的表达。IgD结合因子对IgD的特异性通过以下方式确定:(i)竞争性抑制;(ii)结合因子结合放射性标记的IgD而不结合单克隆IgE、IgG2a或多克隆IgG的能力;(iii)结合因子通过IgD-琼脂糖柱后被去除,并在随后的酸洗脱液中回收。