A syngeneic monoclonal antibody to murine Meth-A sarcoma (HepSS-1) recognizes heparan sulfate glycosaminoglycan (HS-GAG): cell density and transformation dependent alteration in cell surface HS-GAG defined by HepSS-1.
A syngeneic monoclonal antibody to murine Meth-A sarcoma (HepSS-1) recognizes heparan sulfate glycosaminoglycan (HS-GAG): cell density and transformation dependent alteration in cell surface HS-GAG defined by HepSS-1.
复制标题
鼠 Meth-A 肉瘤 (HepSS-1) 的同源单克隆抗体可识别硫酸乙酰肝素糖胺聚糖 (HS-GAG)：HepSS-1 定义的细胞表面 HS-GAG 中的细胞密度和转化依赖性改变。
作者：
S. Kure;O. Yoshie
我们已经分离出一个同源的单克隆抗体（HepSS-1）反应的小鼠甲基胆蒽诱导的纤维肉瘤，Meth-A。HepSS-1还与多种建立的和新鲜的正常细胞结合，这些细胞不仅来自小鼠，而且来自其他物种，如人、猴、大鼠、仓鼠和鸡。用HepSS-1免疫沉淀表面碘化的Meth-A细胞提取物，以及Meth-A细胞提取物的Sepharose 4 B凝胶层析和通过放射免疫测定法检测HepSS-1识别的抗原，揭示了HepSS-1抗原由几种分子种类组成，其中一种大至约10（6）道尔顿。以下证据表明HepSS-1特异性识别硫酸乙酰肝素糖胺聚糖（HS-GAG）中存在的表位。首先，用肝素酶或肝素酶处理Meth-A细胞，但不用软骨素酶ABC或透明质酸酶处理Meth-A细胞，导致HepSS-1结合丧失。第二，HS-GAG而不是七种其他类型的GAG（透明质酸、肝素、软骨素、4-硫酸软骨素、6-硫酸软骨素、硫酸皮肤素和硫酸角质素）抑制HepSS-1与Meth-A细胞的结合。第三，HepSS-1与HS-GAG结合，但不与其他七种类型的GAG结合。从HepSS-1与各种修饰的HS-GAG和鲸ω-肝素的结合分析，另外表明HepSS-1识别与O-硫酸化和N-乙酰化葡糖胺密切相关的表位。我们发现NIH 3 T3细胞在低细胞密度时比汇合时表达更多的HepSS-1表位，并且在G2 + M中比G1中表达更多的HepSS-1表位，而用Kirsten-ras癌基因或SV-40转化的NIH 3 T3细胞表达高水平的HepSS-1表位，并且不再显示HepSS-1表位数量的密度依赖性变化。这些观察结果也通过使用NIH 3 T3细胞重现，NIH 3 T3细胞用温度敏感性Kirsten鼠肉瘤病毒转化，保持在允许和非允许温度下。因此，HepSS-1是第一个单克隆抗体HS-GAG，似乎是有用的，以阐明细胞表面HS-GAG在正常细胞生长和细胞转化的变化。
We have isolated a syngeneic monoclonal antibody (HepSS-1) reactive to a murine methylcholanthrene-induced fibrosarcoma, Meth-A. HepSS-1 also bound to a wide variety of established and fresh normal cells derived from not only mice but also other species such as human, monkey, rat, hamster, and chicken. Immunoprecipitation of surface iodinated Meth-A cell extract with HepSS-1, as well as Sepharose 4B gel chromatography of Meth-A cell extract and detection of antigens recognized by HepSS-1 by a sandwich-type radioimmunoassay revealed that the HepSS-1 antigens were composed of several molecular species, with one as large as approximately 10(6) daltons. The following evidence indicates that HepSS-1 specifically recognizes an epitope present in heparan sulfate glycosaminoglycan (HS-GAG). First, treatment of Meth-A cells with heparitinase or heparinase, but not with chondroitinase ABC or hyaluronidase, resulted in the loss of HepSS-1 binding. Second, HS-GAG but not seven other types of GAG (hyaluronic acid, heparin, chondroitin, chondroitin 4-sulfate, chondroitin 6-sulfate, dermatan sulfate, and keratan sulfate) inhibited HepSS-1 binding to Meth-A cells. Third, HepSS-1 bound with HS-GAG but not with the seven other types of GAG. From the binding analysis of HepSS-1 to various modified HS-GAG and whale omega-heparin, it is additionally suggested that HepSS-1 recognizes an epitope closely related to O-sulfated and N-acetylated glucosamine. We found that NIH 3T3 cells expressed more HepSS-1 epitopes at a low cell density than at confluency and in G2 + M than in G1, whereas NIH 3T3 cells transformed with Kirsten-ras oncogene or SV-40 expressed high levels of HepSS-1 epitopes and ceased to show the density-dependent change in the amount of HepSS-1 epitopes. These observations were also reproduced by using NIH 3T3 cells transformed with a temperature sensitive Kirsten murine sarcoma virus maintained at permissive and non-permissive temperatures. Thus HepSS-1 is a first monoclonal antibody to HS-GAG and seems to be useful to elucidate changes in cell surface HS-GAG in normal cell growth and cell transformation.