MAPPING OF A NEUTRALIZING ANTIGENIC SITE OF COXSACKIEVIRUS-B4 BY CONSTRUCTION OF AN ANTIGEN CHIMERA
MAPPING OF A NEUTRALIZING ANTIGENIC SITE OF COXSACKIEVIRUS-B4 BY CONSTRUCTION OF AN ANTIGEN CHIMERA
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DOI：
10.1128/jvi.65.7.3475-3480.1991
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发表时间：
1991-07-01
影响因子：
5.4
通讯作者：
KANDOLF, R
中科院分区：
文献类型：
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作者：
REIMANN, BY;ZELL, R;KANDOLF, R
通过构建柯萨奇病毒B3(CVB3)和柯萨奇病毒B4(CVB4)之间的抗原嵌合体，鉴定了柯萨奇病毒B4(CVB4)的中和抗原部位。该嵌合体命名为CVB3/4，通过对感染性重组CVB3基因进行定点突变，将CVB4结构蛋白VP1的BC环的5个氨基酸插入到CVB3的相应环中。该嵌合基因可在允许的宿主细胞中产生感染循环。所产生的嵌合病毒CVB3/4可被CVB4和CVB3血清型特异性多克隆抗血清中和和沉淀，表明它统一了两种柯萨奇病毒血清型的抗原性。此外，嵌合体还能在兔体内产生抗体，能够中和柯萨奇病毒的两个血清型CVB3和CVB4。CVB4特异性抗原位点插入到CVB3的BC环中降低了病毒复制的效率，导致病毒嵌合体的小斑块形态。综上所述，这些数据提供了在CVB4VP1(氨基酸81到89)的BC环中存在血清型特异性中和抗原位点的证据。我们的发现表明，构建不同类型的嵌合体可以作为鉴定柯萨奇病毒抗原部位的工具。所定位的CVB4特异性抗原表位在CVB3中表达时仍具有免疫原性，表明CVB3可作为异源抗原位点的RNA病毒载体。
A neutralizing antigenic site of coxsackievirus B4 (CVB4) was identified by construction of an antigen chimera between coxsackievirus B3 (CVB3) and CVB4. This chimera, designated CVB3/4, was constructed by inserting five amino acids of the putative BC loop of the structural protein VP1 of CVB4 into the corresponding loop of CVB3 by site-directed mutagenesis of infectious recombinant CVB3 cDNA. The chimeric cDNA was capable of inducing an infectious cycle upon transfection of permissive host cells. The resulting chimeric virus CVB3/4 was neutralized and precipitated by CVB4 and CVB3 serotype-specific polyclonal antisera, demonstrating that it unifies antigenic properties of both coxsackievirus serotypes. In addition, the chimera elicited antibodies in rabbits which were capable of neutralizing the two coxsackievirus serotypes CVB3 and CVB4. The insertion of the CVB4-specific antigenic site into the BC loop of CVB3 reduces the efficiency of viral replication, resulting in a small-plaque morphology of the virus chimera. In summary, these data give evidence for the presence of a serotype-specific neutralizing antigenic site in the BC loop of VP1 of CVB4 (amino acids 81 to 89). Our findings suggest that the construction of intertypic chimeras can be used as a tool for the identification of antigenic sites of coxsackieviruses. The retained immunogenicity of the mapped CVB4-specific antigenic epitope, when expressed in CVB3, indicates that CVB3 can be used as a RNA virus vector for heterologous antigenic sites.