Subcellular distribution of particle-associated antigens in Trypanosoma rhodesiense.
Subcellular distribution of particle-associated antigens in Trypanosoma rhodesiense.
复制标题
罗得西亚锥虫颗粒相关抗原的亚细胞分布。
DOI：
10.4049/jimmunol.128.6.2656
复制
发表时间：
1982
影响因子：
4.4
通讯作者：
J. McLaughlin
中科院分区：
文献类型：
--
作者：
J. McLaughlin
非洲锥虫罗得西亚锥虫的血流形式受到分析细胞分离程序使用等重蔗糖梯度离心。采用融合火箭免疫电泳对所得组分进行分析，结果显示，在密度增量为1.10 ~ 1.15和1.20 ~ 1.22之间，与颗粒沉积相关的抗原成分最多。从各种标记酶和荧光标记荧光胺的分布来看，这些粒子群分别来源于鞭毛袋和鞭毛表面膜。假设的鞭毛口袋和表面膜部分的交叉免疫电泳分别显示了至少7种和10种不同的抗原，只有有限的交叉反应性。在交叉亲和免疫电泳的基础上，每种膜抗原中有四到五个是糖蛋白，沉淀在含有豆豆蛋白a的凝胶中。在使用的各种洗涤剂处理中，发现在0.4% Triton X-100存在的情况下，使用两性离子洗涤剂Zwittergent 3-12（0.1%）可获得最佳的糖蛋白抗原提取。融合火箭免疫电泳还发现了一个与平衡密度约为1.180的粒子群相关的单一突出抗原，根据标记酶的分布确定为溶酶体。其他细胞内位点的抗原性不明显。
Bloodstream forms of the African trypanosome Trypanosoma rhodesiense were subjected to analytic cell fractionation procedures using isopycnic sucrose gradient centrifugation. The fractions obtained were analyzed using fused rocket immunoelectrophoresis, which revealed the most antigenic components to be associated with particles sedimenting between density increments of 1.10 to 1.15 and 1.20 and 1.22. From the distribution of various marker enzymes and the fluorogenic label fluorescamine, these particle populations were identified as derived from flagella pocket and surface membrane, respectively. Crossed immunoelectrophoresis of postulated flagella pocket and surface membrane fractions demonstrated at least seven and 10 distinct antigens, respectively, with only limited cross-reactivity. On the basis of crossed affini-immunoelectrophoresis four to five of each of these membrane antigens were glycoproteins, being precipitated in a concanavalin A-containing gel. Of the various detergent treatments used, it was found that optimum extraction of these glycoprotein antigens was achieved using the zwitterionic detergent Zwittergent 3-12 (0.1%) in the presence of 0.4% Triton X-100. Fused rocket immunoelectrophoresis also revealed a single prominent antigen associated with a particle population having an equilibrium density of approximately 1.180, which was identified as lysosomal from the distribution of marker enzymes. Other intracellular sites were not significantly antigenic.