NEUTRALIZATION OF CRPV INFECTIVITY BY MONOCLONAL-ANTIBODIES THAT IDENTIFY CONFORMATIONAL EPITOPES ON INTACT VIRIONS
NEUTRALIZATION OF CRPV INFECTIVITY BY MONOCLONAL-ANTIBODIES THAT IDENTIFY CONFORMATIONAL EPITOPES ON INTACT VIRIONS
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DOI：
10.1016/0168-1702(91)90031-p
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发表时间：
1991-11-01
期刊：
影响因子：
5
通讯作者：
KREIDER, JW
中科院分区：
文献类型：
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作者：
CHRISTENSEN, ND;KREIDER, JW
制备了抗棉尾兔乳头瘤病毒(CRPV)的单抗，并在家兔NZW上进行了CRPV诱导的乳头状瘤的中和试验。将完整的CRPV用CsCl半提纯后免疫BALB/c小鼠。通过与淋巴结细胞融合制备杂交瘤细胞，用酶联免疫吸附试验(ELISA)分析杂交瘤细胞培养上清液对完整和破坏的CRPV病毒抗原的反应性。最初选择了22种对CRPV有反应的培养上清。10例仅对完整CRPV呈阳性反应，4例仅对破碎型CRPV呈阳性反应，8例同时对完整和破碎型CRPV病毒抗原呈阳性反应。这些上清液中没有一种抗体能识别免疫印迹法分离的CRPV衣壳蛋白(L1和L2)上的表位。选择了5株产生与完整CRPV结合的抗体，而对完整的HPV-11或BPV-1不起反应的杂交瘤细胞，并进行了抗体介导的CRPV感染性中和试验。5株单抗均为中和抗体，并确定了CRPV病毒粒子上的表位，这些表位在本质上是非线性和构象的。竞争酶联免疫吸附试验显示，5株中和单抗在完整的CRPV病毒粒子上识别出相似的表位或表位簇。
Monoclonal antibodies were generated against cottontail rabbit papillomavirus (CRPV) and tested for neutralization of CRPV-induced papillomas on domestic NZW rabbits. Intact CRPV was semi-purified on CsCl gradients and used to immunize BALB/c mice. Hybridomas were prepared from a fusion with lymph node cells, and supernatants from growing hybridomas were analyzed by enzyme-linked immunosorbent assay (ELISA) for reactivity to both intact and disrupted CRPV virion antigen. Supernatants from 22 cultures were initially selected that were responsive to CRPV. Ten were reactive to intact CRPV alone, 4 were reactive only to disrupted CRPV, and 8 were reactive to both intact and disrupted CRPV virion antigen. None of these supernatants contained antibodies which recognized epitopes on CRPV capsid proteins (LI and L2) that were separated on Western blots. Five hybridomas which produced antibodies that bound to intact CRPV, and did not react to intact HPV-11 or BPV-1 were selected and tested for antibody-mediated neutralization of CRPV infectivity. All five monoclonal antibodies were neutralizing, and identified epitopes on intact CRPV virions which were non-linear and conformational in nature. The five neutralizing monoclonal antibodies appeared to recognize a similar epitope or epitope cluster on the intact CRPV virion as determined by competition ELISA.