Wong W W, Fearon D T
J Immunol. 1985 Jun;134(6):4048-56.
Affinity-purified rabbit antibody to the human C3b receptor (CR1) cross-reacted with an antigen that was expressed on murine splenocytes, lymph node cells, thymocytes, peritoneal macrophages, erythrocytes, and L929 cells, as assessed by flow cytofluorographic analysis of indirectly stained cells. The cell surface antigen recognized by antihuman CR1 had a Mr of 60,000 to 65,000 on each cell type, and the cross-reactive epitope(s) was sensitive to reduction with dithiothreitol but not to oxidation with NaIO4. Analysis by affinity chromatography of murine splenic B lymphocytes identified two cell surface proteins capable of binding to guinea pig C3b (C3bgp). The larger polypeptide had a Mr of 210,000, was not present in L929 cells, and may represent the murine CR1. The smaller polypeptide of 65,000 Mr was also present on L929 cells and was shown to constitute the cross-reactive antigen. Adsorption of detergent lysates of L929 cells with C3bgp-Sepharose depleted by 44% the antigen recognized by anti-human CR1; the C3bgp-binding protein and the cross-reactive antigen exhibited similar patterns on two-dimensional gel electrophoresis; and the isolated C3b-binding protein could be immunoprecipitated with anti-human CR1. Thus, the murine cell surface protein, termed p65, that is antigenically cross-reactive with human CR1 shares a capacity for binding to C3b in its detergent-solubilized form but is distinct from murine CR1 in its lower Mr, wider cellular distribution, and inability to mediate the adherence of C3b-coated particles in its native, membrane-associated form.
通过间接染色细胞的流式细胞荧光分析评估,亲和纯化的抗人C3b受体(CR1)兔抗体与在小鼠脾细胞、淋巴结细胞、胸腺细胞、腹腔巨噬细胞、红细胞和L929细胞上表达的一种抗原发生交叉反应。抗人CR1识别的细胞表面抗原在每种细胞类型上的分子量为60,000至65,000,并且交叉反应表位对二硫苏糖醇还原敏感,但对高碘酸钠氧化不敏感。对小鼠脾脏B淋巴细胞进行亲和层析分析,鉴定出两种能够结合豚鼠C3b(C3bgp)的细胞表面蛋白。较大的多肽分子量为210,000,不存在于L929细胞中,可能代表小鼠CR1。分子量为65,000的较小多肽也存在于L929细胞上,并被证明构成交叉反应抗原。用C3bgp-琼脂糖吸附L929细胞的去污剂裂解物,可使抗人CR1识别的抗原减少44%;C3bgp结合蛋白和交叉反应抗原在二维凝胶电泳上呈现相似的模式;分离的C3b结合蛋白可用抗人CR1进行免疫沉淀。因此,与人类CR1抗原性交叉反应的小鼠细胞表面蛋白p65,在其去污剂溶解形式下具有结合C3b的能力,但在分子量较低、细胞分布更广泛以及无法以其天然的膜相关形式介导C3b包被颗粒的黏附方面与小鼠CR1不同。