IDENTIFICATION AND CHARACTERIZATION OF AN ACIDIC MAJOR SURFACE GLYCOPROTEIN FROM PROCYCLIC STAGE TRYPANOSOMA-CONGOLENSE
IDENTIFICATION AND CHARACTERIZATION OF AN ACIDIC MAJOR SURFACE GLYCOPROTEIN FROM PROCYCLIC STAGE TRYPANOSOMA-CONGOLENSE
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DOI：
10.1016/0166-6851(93)90074-8
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发表时间：
1993-10-01
影响因子：
1.5
通讯作者：
PEARSON, TW
中科院分区：
文献类型：
--
作者：
BEECROFT, RP;RODITI, I;PEARSON, TW
单克隆抗体（单克隆抗体）的衍生对前环培养形式的刚果锥虫和14个被选中的结合到表面的活的前环在免疫荧光测定。这些抗体与T. Congolense（savannah型和Kilifi型）和猿猴锥虫的顺周期性，但不与其他锥虫物种的顺周期性、几种锥虫的血流形式或利什曼原虫相关，因此具有生命周期阶段特异性和亚属特异性。用这些抗体进行的免疫活化细胞分选仪分析表明，在从血流转化为前环形式的过程中，表面抗原的表达动力学与T的前环素或前环酸性重复蛋白（PARP）的表达动力学相似。布氏杆菌早在开始转化后8 h就出现在细胞表面。所有14种抗体在全前环裂解物的免疫印迹分析中检测到40-44和28-32 kDa的宽带，并且对碳水化合物表位具有特异性。用阳离子交换层析和凝胶电泳纯化抗原，证明是一种酸性糖蛋白。纯化抗原的氨基酸微量分析显示谷氨酸/谷氨酰胺和丙氨酸的丰度。溴化氰裂解产生的肽序列与Bayne等人[26]随附论文中描述的基因核苷酸序列预测的氨基酸序列相匹配。与T.然而，其阶段和亚属特异性、表面分布、免疫优势、酸性和从血流转化为前环形式过程中的表达动力学表明，该分子是T.布氏杆菌
Monoclonal antibodies (mAbs) were derived against the procyclic culture form of Trypanosoma congolense and 14 were selected which bound to the surface of living procyclics in immunofluorescence assays. These antibodies bound to procyclics and epimastigotes of T. congolense (both savannah-type and Kilifi-type) and procyclics of Trypanosoma simiae, but not to procyclics of other species of trypanosomes, to bloodstream forms of several species of trypanosomes or to Leishmania, and were thus life cycle stage- and subgenus-specific. Fluorescence-activated cell sorter analysis with these antibodies showed that the kinetics of expression of the surface antigen during transformation from bloodstream to procyclic forms was similar to that of procyclin or procyclic acidic repetitive protein (PARP) of T. brucei spp. appearing at the cell surface as early as 8 h after initiating transformation. All fourteen antibodies detected broad bands of 40-44 and 28-32 kDa in immunoblot analysis of whole procyclic lysates and were specific for carbohydrate epitopes. The antigen was purified by cation-exchange chromatography and gel electrophoresis, and was shown to be an acidic glycoprotein. Amino acid microanalysis of the purified antigen showed an abundance of glutamic acid/glutamine and alanine. Sequences of peptides produced by cyanogen bromide cleavage matched amino acid sequences predicted by the nucleotide sequence of a gene described in the accompanying paper by Bayne et al. [26]. No sequence similarity to T. brucei procyclin/PARP or to any other protein was found. However, its stage and subgenus specificity, surface disposition, immunodominance, acidity and kinetics of expression during transformation from bloodstream to procyclic forms indicate that the molecule is an analog of procyclin/PARP described in T. brucei spp.