Shepley M P, Racaniello V R
Department of Microbiology, Columbia University College of Physicians and Surgeons, New York, New York 10032.
J Virol. 1994 Mar;68(3):1301-8. doi: 10.1128/JVI.68.3.1301-1308.1994.
A monoclonal antibody, AF3, was previously shown to specifically inhibit poliovirus binding to HeLa cells and to detect a 100-kDa glycoprotein only in cell lines and tissues permissive for poliovirus infection. These results suggested that the 100-kDa protein may be involved in the pathogenesis of poliomyelitis and the cellular function of the poliovirus receptor site. To study further the role of the 100-kDa protein in poliovirus attachment, immunoaffinity purification, amino acid sequencing, and cDNA cloning were undertaken. The results demonstrate that antibody AF3 reacts with the lymphocyte homing receptor CD44, a multifunctional cell surface glycoprotein involved in the homing of circulating lymphocytes to lymph nodes and the modulation of lymphocyte adhesion and activation. Antibody AF3 reacts with a subset of CD44 molecules (AF3CD44H), which appears to be a small fraction of the heterogeneously glycosylated CD44 molecules expressed on hematopoietic and nonhematopoietic cells. Anti-CD44 monoclonal antibodies, previously reported to induce CD44-mediated modulation of lymphocyte activation and adhesion, compete with 125I-AF3 in binding assays, demonstrating functional overlap among the epitopes. The anti-CD44 monoclonal antibody A3D8, which binds to a greater molecular weight range of CD44 than does AF3, inhibits poliovirus binding to a similar degree. CD44 does not act as a poliovirus receptor, since CD44-expressing mouse L-cell transformants did not bind poliovirus. The poliovirus receptor and AF3CD44H may be noncovalently associated, or they may interact through the cytoskeleton or signal transduction pathways.
一种单克隆抗体AF3,先前已被证明能特异性抑制脊髓灰质炎病毒与HeLa细胞的结合,并且仅在允许脊髓灰质炎病毒感染的细胞系和组织中检测到一种100 kDa的糖蛋白。这些结果表明,100 kDa蛋白可能参与了脊髓灰质炎的发病机制以及脊髓灰质炎病毒受体位点的细胞功能。为了进一步研究100 kDa蛋白在脊髓灰质炎病毒附着中的作用,进行了免疫亲和纯化、氨基酸测序和cDNA克隆。结果表明,抗体AF3与淋巴细胞归巢受体CD44发生反应,CD44是一种多功能细胞表面糖蛋白,参与循环淋巴细胞归巢至淋巴结以及淋巴细胞黏附和激活的调节。抗体AF3与CD44分子的一个亚群(AF3CD44H)发生反应,该亚群似乎只是在造血细胞和非造血细胞上表达的异质性糖基化CD44分子中的一小部分。先前报道能诱导CD44介导的淋巴细胞激活和黏附调节的抗CD44单克隆抗体,在结合试验中与125I-AF3竞争,表明这些表位之间存在功能重叠。抗CD44单克隆抗体A3D8,其结合的CD44分子量范围比AF3更大,能在相似程度上抑制脊髓灰质炎病毒的结合。CD44并不作为脊髓灰质炎病毒受体,因为表达CD44的小鼠L细胞转化体不结合脊髓灰质炎病毒。脊髓灰质炎病毒受体和AF3CD44H可能非共价结合,或者它们可能通过细胞骨架或信号转导途径相互作用。