Krause S W, Rehli M, Heinz S, Ebner R, Andreesen R
Department of Haematology, University of Regensburg, D-93042 Regensburg, Germany.
Biochem J. 2000 Mar 15;346 Pt 3(Pt 3):729-36.
MAX.3 is a monoclonal antibody that preferentially reacts with mature macrophages (MAC), monocyte-derived dendritic cells, megakaryocytes and platelets. In this study, we describe the characterization, purification and identification of the MAX.3 antigen. Immunoprecipitation and SDS/PAGE revealed different molecular masses of MAX.3 antigen in MAC (60-90 kDa) and platelets (58-64 kDa), whereas a similar size (45 kDa) was observed in both cell types after digestion with N-glycosidase F. Lectin affinity and sequential treatment with different glycosidases suggests complex type glycosylation of MAX.3 antigen in MAC and hybrid type glycosylation in platelets. Amino acid sequencing led to the identification of a corresponding cDNA clone and showed its identity to the sequence of the CD84 antigen, a member of the CD2 family of cell surface molecules. MAX.3/CD84 was further studied by immunohistochemistry and a variable expression was found on tissue MAC, confirming this antigen to be mainly a marker for MAC in situ.
MAX.3是一种单克隆抗体,它优先与成熟巨噬细胞(MAC)、单核细胞衍生的树突状细胞、巨核细胞和血小板发生反应。在本研究中,我们描述了MAX.3抗原的特性、纯化和鉴定。免疫沉淀和SDS/聚丙烯酰胺凝胶电泳显示,MAX.3抗原在MAC(60 - 90 kDa)和血小板(58 - 64 kDa)中的分子量不同,而在用N-糖苷酶F消化后,两种细胞类型中观察到相似大小(45 kDa)。凝集素亲和性以及用不同糖苷酶的顺序处理表明,MAX.3抗原在MAC中为复合型糖基化,在血小板中为杂合型糖基化。氨基酸测序导致鉴定出一个相应的cDNA克隆,并显示其与细胞表面分子CD2家族成员CD84抗原的序列相同。通过免疫组织化学对MAX.3/CD84进行了进一步研究,发现在组织MAC上有可变表达,证实该抗原主要是原位MAC的标志物。