Kinetics of the antibody response to BamHI-K nuclear antigen in uncomplicated infectious mononucleosis.
Kinetics of the antibody response to BamHI-K nuclear antigen in uncomplicated infectious mononucleosis.
复制标题
无并发症的传染性单核细胞增多症中抗体对 BamHI-K 核抗原反应的动力学。
DOI：
10.1093/infdis/154.2.346
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Miller,G
中科院分区：
文献类型：
--
作者：
Niederman,JC;Miller,G
材料与方法从28名男性和15名女性大学生（17 - 29岁）与嗜异性抗体阳性IM获得220份血清。血清学技术。VCA的IgG和IgM抗体通过先前建立的方法测量。通过Raji细胞上的抗补体免疫荧光检测对EBNA的反应性;补体来源是缺乏EBV抗体的新鲜冷冻人血清。用EBV DNA的BamHI-K片段和单纯疱疹病毒胸苷激酶基因共转化小鼠LTK细胞，使其表达BamHI-K核抗原。胰蛋白酶消化后，将细胞在Microtekr载玻片上的六孔室中生长两至三天。然后将载玻片在室温下在丙酮和甲醇的2：1混合物中固定10分钟。首先通过抗补体免疫荧光法在含有BamHI-K核抗原的转化L细胞上以1：5稀释度筛选试验血清，随后进行滴定[12]。标本咽喉冲洗液包括20 ml含漱液，含RPMI 1640培养基，补充10%胎牛血清和抗生素。将洗涤液储存在-70 ℃下直至进行EBV测试.转变
Materials and MethodsSera. Two hundred twenty sera were obtained from 28 male and 15 female university students (17-29 years of age) with heterophile antibody-positive IM. Serological techniques. IgG and IgM antibodies to VCA were measured by previously established methods. Reactivity to EBNA was detected by anticomplement immunofluorescence on Raji cells; the complement source was a fresh, frozen human serum lacking antibody to EBV. BamHI-K nuclear antigen was expressed in mouse LTK-cells that had been cotransformed with the BamHI-K frag-ment of EBV DNA and the thymidine-kinase gene of herpes simplex virus. Cells were grown in six-well chambers on Microtekr slides for two to three days after trypsinization. The slides were then fixed for 10 min at room tem-perature in a 2: 1 mixture of acetone and methanol. Test sera were initially screened at a 1: 5 dilution by anticomplement immunofluorescence on transformed L cells contain-ing BamHI-K nuclear antigen and were later titered [12]. Specimens. Throat washings consisted of 20-ml gargles of RPMI 1640 culture medium supplemented with 10% fetal calf serum and antibiotics. The washings were stored at-70 C until tested for EBV. Transformation of