ELECTROPHORETIC ANALYSIS OF POLYPEPTIDES IMMUNE PRECIPITATED FROM CYTOMEGALOVIRUS-INFECTED CELL-EXTRACTS BY HUMAN-SERA
ELECTROPHORETIC ANALYSIS OF POLYPEPTIDES IMMUNE PRECIPITATED FROM CYTOMEGALOVIRUS-INFECTED CELL-EXTRACTS BY HUMAN-SERA
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DOI：
10.1128/iai.36.3.933-942.1982
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发表时间：
1982-01-01
影响因子：
3.1
通讯作者：
CREMER, N
中科院分区：
文献类型：
--
作者：
PEREIRA, L;HOFFMAN, M;CREMER, N
补体固定试验对巨细胞病毒（CMV）感染的血清诊断依赖于急性期至恢复期血清抗体滴度上升4倍。冻融和甘氨酸提取的感染细胞[人胎儿肺HFDL]培养抗原用于这些试验，在与相同的血清反应中具有明显不同的滴度。对冻融后的cmv感染细胞多肽和甘氨酸提取抗原进行了鉴定，并对23对人急性和恢复期血清沉淀的蛋白进行了鉴定。首先，用[35S]蛋氨酸对感染细胞制备的冻融和甘氨酸提取抗原进行放射性标记，并在十二酰硫酸钠-聚丙烯酰胺凝胶中进行电泳，得到类似的图案，大部分标记包含在晚期结构蛋白和糖蛋白中。甘氨酸提取制剂比冻融抗原含有更高比例的可溶性66,000和50,000-MW蛋白。其次，恢复期血清沉淀蛋白迁移，表观分子量分别为150,000、130,000、110,000、96,000、74,000、66,000、50,000、34,000、32,000和25,000。其中，130,000,110,000,96,000,66,000,50,000和25,000 MW的蛋白质与氨基标记的多肽相结合。人血清中的IgG和IgM抗体都能从cmv感染的细胞制剂中沉淀出这些蛋白。结果对巨细胞病毒感染的血清诊断意义进行了讨论。
Serodiagnosis of cytomegalovirus (CMV) infection by complement fixation tests depends on showing a 4-fold rise in antibody titer from acute- to convalescent-phase sera. Freeze-thaw and glycine-extracted, infected cell [human fetal lung HFDL] culture antigens used for these tests give markedly different titers in reactions with the same sera. The CMV-infected cell polypeptides contained in freeze-thaw and glycine-extracted antigens were characterized, and the proteins precipitated by 23 pairs of human acute and convalescent sera were identified. First, freeze-thaw and glycine-extracted antigens prepared from infected cells radiolabeled with [35S]methionine and subjected to electrophoresis in sodium dodecyl sulfate-polyacrylamide gels yielded similar patterns, and the bulk of the label was contained in late structural proteins and glycoproteins. Glycine-extracted preparations contained a greater proportion of soluble 66,000- and 50,000-MW proteins than did freeze-thaw antigens. Second, convalescent sera precipitated proteins migrating with apparent MW of 150,000, 130,000, 110,000, 96,000, 74,000, 66,000, 50,000, 34,000, 32,000 and 25,000. Of these, the 130,000-, 110,000-, 96,000-, 66,000-, 50,000- and 25,000 MW proteins comigraated with glucosamine-labeled polypeptides. Both IgG and IgM antibodies in human sera precipitated these proteins from CMV-infected cell preparations. Implications of the results for serodiagnosis of CMV infections are discussed.