Lyon J A, Haynes J D, Diggs C L, Chulay J D, Pratt-Rossiter J M
J Immunol. 1986 Mar 15;136(6):2252-8.
Some immune sera that inhibit erythrocyte invasion by merozoites also agglutinate the merozoites as they emerge from rupturing schizonts. These immune clusters of merozoites (ICM) possess a surface coat that is cross-linked by antibody and is thicker than the surface coat associated with normal merozoites (NM) obtained from cultures containing preimmune serum. Analysis of metabolically labeled ICM and NM performed by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that washed ICM possessed immune complexes containing antigens representative of schizonts and merozoites. Characteristics of the immune complexes included: a) they were not soluble in pH 8 Triton X-100, b) they were soluble at an acid pH, and c) after pH neutralization they were precipitated by using staphylococcal protein A. Merozoite antigens having Mr of 83, 73, and 45 kDa were associated with immune complexes in ICM. The 83 and 73 kDa antigens were recovered in considerably larger quantities from ICM than from NM. Schizont antigens having Mr of 230, 173 (triplet), 152 (doublet), and 31 kDa were associated with immune complexes in ICM, and a 195 kDa antigen(s) from schizonts and merozoites was also present in the immune complexes. In addition, other antigens of Mr 113, 101, 65, and 51 kDa may have been immune complexed. These 15 antigens accounted for less than 30% of the schizont and merozoite antigens recognized by the immune serum. Immune complexes probably formed between antibodies and a) surface antigens of schizont-infected erythrocytes exposed to antibody before schizont rupture, b) surface antigens of merozoites and schizonts exposed during schizont rupture, and c) soluble antigens normally released during schizont rupture. The antibody components of the immune complexes may have prevented rapid degradation or shedding of some antigens from the merozoite surface. Allowing schizonts to rupture in the presence of inhibitory antibodies (to form ICM) is a useful approach to identifying exposed targets of protective immunity against malaria.
一些能抑制裂殖子侵入红细胞的免疫血清,在裂殖子从破裂的裂殖体中释放出来时也会使其凝集。这些裂殖子免疫簇(ICM)具有一层表面被抗体交联的包膜,且比从含有免疫前血清的培养物中获得的正常裂殖子(NM)的表面包膜更厚。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳对代谢标记的ICM和NM进行分析表明,洗涤后的ICM含有免疫复合物,其中包含代表裂殖体和裂殖子的抗原。这些免疫复合物的特征包括:a)它们在pH 8的Triton X - 100中不溶解,b)在酸性pH下可溶解,c)pH中和后可被葡萄球菌蛋白A沉淀。分子量为83、73和45 kDa的裂殖子抗原与ICM中的免疫复合物相关。与NM相比,从ICM中回收的83和73 kDa抗原的量要多得多。分子量为230、173(三重峰)、152(双峰)和31 kDa的裂殖体抗原与ICM中的免疫复合物相关,并且来自裂殖体和裂殖子的一种195 kDa抗原也存在于免疫复合物中。此外,分子量为113、101、65和51 kDa的其他抗原可能也形成了免疫复合物。这15种抗原占免疫血清识别的裂殖体和裂殖子抗原的比例不到30%。免疫复合物可能在抗体与以下物质之间形成:a)裂殖体破裂前暴露于抗体的裂殖体感染红细胞的表面抗原,b)裂殖体破裂期间暴露的裂殖子和裂殖体的表面抗原,以及c)裂殖体破裂时正常释放的可溶性抗原。免疫复合物中的抗体成分可能阻止了某些抗原从裂殖子表面快速降解或脱落。在抑制性抗体存在的情况下使裂殖体破裂(形成ICM)是鉴定针对疟疾的保护性免疫暴露靶点的一种有用方法。