放射性碘化杂交重组干扰素α A/D与小鼠和人类淋巴细胞系结合的特性分析。

Characterization of the binding of radioiodinated hybrid recombinant IFN-alpha A/D to murine and human lymphoid cell lines.

作者信息

Faltynek C R, Princler G L, Schwabe M, Shata M T, Lewis G K, Kamin-Lewis R M

机构信息

Department of Immunopharmacology, Sterling Research Group, Malvern, PA 19355.

出版信息

J Interferon Res. 1990 Feb;10(1):55-64. doi: 10.1089/jir.1990.10.55.

DOI:10.1089/jir.1990.10.55

PMID:2139461

Abstract

The hybrid recombinant human interferon (IFN) rIFN-alpha A/D was radioiodinated. Specific binding of [125I]rIFN-alpha A/D was observed with both human and murine cell lines. The binding of [125I]rIFN-alpha A/D to human Daudi cells had similar characteristics to the previously described binding of [125I]rIFN-alpha A or -alpha 2. The following lines of evidence demonstrated that [125I]rIFN-alpha A/D bound with high affinity to the same receptor on murine cells as murine IFN-alpha and -beta: (i) the binding of [125I]rIFN-alpha A/D to murine LBRM cells was inhibited to a similar extent by natural murine IFN-alpha, natural murine IFN-beta, and rIFN-A/D; (ii) the Kd (approximately 2 X 10(-10) M) obtained from both competition experiments and saturation binding experiments with [125I]rIFN-alpha A/D was comparable to the previously reported Kd for the binding of natural murine IFN-alpha and -beta to other murine cell lines; (iii) the size of the cross-linked [125I]rIFN-alpha A/D receptor complex formed on murine LBRM cells was similar to the previously reported cross-linked complex formed after binding radioiodinated natural murine IFN-beta to other murine cell lines. Due to the current lack of readily available recombinant murine IFN-alpha or -beta for radiolabeling and the previously demonstrated biological activity of rIFN-alpha A/D on murine cells, [125I]rIFN-alpha A/D should prove to be a useful reagent for further studies of murine IFN receptors.

摘要

对杂交重组人干扰素（IFN）rIFN-α A/D进行了放射性碘标记。在人和鼠细胞系中均观察到了[125I]rIFN-α A/D的特异性结合。[125I]rIFN-α A/D与人Daudi细胞的结合特性与先前描述的[125I]rIFN-α A或-α 2的结合特性相似。以下一系列证据表明，[125I]rIFN-α A/D与鼠IFN-α和-β以高亲和力结合于鼠细胞上的同一受体：（i）天然鼠IFN-α、天然鼠IFN-β和rIFN-A/D对[125I]rIFN-α A/D与鼠LBRM细胞的结合抑制程度相似；（ii）通过[125I]rIFN-α A/D的竞争实验和饱和结合实验获得的Kd（约2×10-10 M）与先前报道的天然鼠IFN-α和-β与其他鼠细胞系结合的Kd相当；（iii）在鼠LBRM细胞上形成的交联[125I]rIFN-α A/D受体复合物的大小与先前报道的放射性碘标记的天然鼠IFN-β与其他鼠细胞系结合后形成的交联复合物相似。由于目前缺乏易于获得的用于放射性标记的重组鼠IFN-α或-β，且先前已证明rIFN-α A/D对鼠细胞具有生物学活性，[125I]rIFN-α A/D应被证明是进一步研究鼠IFN受体的有用试剂。