IDENTIFICATION OF DISTINCT ANTIGENIC DETERMINANTS ON DENGUE-2 VIRUS USING MONOCLONAL-ANTIBODIES
IDENTIFICATION OF DISTINCT ANTIGENIC DETERMINANTS ON DENGUE-2 VIRUS USING MONOCLONAL-ANTIBODIES
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DOI：
10.4269/ajtmh.1982.31.548
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发表时间：
1982-01-01
影响因子：
3.3
通讯作者：
DALRYMPLE, JM
中科院分区：
文献类型：
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作者：
GENTRY, MK;HENCHAL, EA;DALRYMPLE, JM
从免疫小鼠淋巴细胞与小鼠骨髓瘤细胞融合产生的淋巴细胞杂交瘤细胞中获得针对登革2型病毒新几内亚C株抗原决定簇的单抗。杂交瘤细胞培养上清采用洗涤剂溶解的登革2型感染细胞抗原的放射免疫分析法筛选。22株杂交瘤细胞诱导的腹水中的单抗经血凝抑制试验、空斑减少中和试验、免疫荧光试验和补体结合试验鉴定。在两组中都观察到了类型特异性抗体和广泛交叉反应抗体，两组中都有Ig亚类IgG1和IgG2a的表达。使用这些抗体确定了病毒粒子上至少3个不同的抗原决定簇。一株杂交瘤产生的抗体能识别登革2型病毒的特异性决定簇，并表现出高滴度的中和，但通过血凝抑制产生低滴度的抗体。四种制剂与一种特定类型的决定簇发生反应，并表现出血凝抑制，但没有中和。杂交瘤(17)产生的抗体在所有测试中都具有广泛的交叉反应。只有2种制剂与登革热-2抗原发生补体结合反应；两种制剂都有交叉反应。免疫荧光特异性或交叉反应与中和和/或血凝抑制相关。用于免疫荧光鉴定登革2型病毒感染细胞的登革2型病毒特异性抗体保存在美国类型培养库杂交瘤细胞库中。
Monoclonal antibodies directed against antigenic determinants of the New Guinea C strain of dengue-2 virus were obtained from lymphocyte hybridomas produced by fusing immune mouse lymphocytes with mouse myeloma cells. Hybridoma cell culture supernatants were screened by using a radioimmunoassay employing detergent-solubilized dengue-2 infected cell antigens. Monoclonal antibodies in ascitic fluids induced by 22 selected hybridomas were characterized by the hemagglutination-inhibition, plaque reduction neutralization, immunofluorescence and complement-fixation tests. Both type-specific and broadly cross-reactive antibodies were observed, and Ig subclasses IgG1 and IgG2a were represented in both groups. At least 3 distinct antigenic determinants on the virion were defined using these antibodies. A single hybridoma produced antibody which recognized a dengue-2 virus type-specific determinant and exhibited high titered neutralization but had a low titer by hemagglutination inhibition. Four preparations reacted with a type-specific determinant and exhibited hemagglutination inhibition but did not neutralize. Hybridomas (17) produced antibodies which were broadly cross reactive in all tests. Only 2 preparations reacted by complement fixation with dengue-2 antigens; both were cross reactive. Immunofluorescence specificity or cross reactivity correlated with neutralization and/or hemagglutination-inhibition. The dengue-2 virus type-specific antibody useful for identification of dengue-2 infected cells by immunofluorescence was deposited in the Hybridoma Cell Bank of the American Type Culture Collection.