Monoclonal antibody resistant mutant of Peste des petits ruminants vaccine virus.
Monoclonal antibody resistant mutant of Peste des petits ruminants vaccine virus.
复制标题
小反刍兽疫疫苗病毒的单克隆抗体抗性突变体。
DOI：
10.1007/s13337-018-0483-z
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Getachew B
中科院分区：
文献类型：
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作者：
Getachew B
用于控制小反刍兽疫的现有疫苗不利于区分感染动物和接种动物（DIVA）。因此，本研究的目的是分离和表征的单克隆抗体抗性突变体的印度株疫苗病毒“PPRV-Sungri/96”的选择压力下的病毒中和单克隆抗体“4 B11”特异性的血凝素（H）蛋白。我们成功地分离了五个单克隆抗体抗性（mAr）突变体（PPRV-RM 5、PPRV-RM 6、PPRV-RM 7、PPRV-E6和PPRV-E7）。在间接ELISA、细胞ELISA和间接免疫荧光试验中，mAr突变株与抗HmAb 4 B11不发生反应，而与对照抗核蛋白mAb 4G 6发生反应，与亲本疫苗病毒“PPRV-Sungri/96”相似。mAr突变体的细胞计数分析显示与mAb 4 B11结合的丧失，同时保持与mAb 4G 6的结合，或多或少类似于“PPRV-Sungri/96”。mAr突变体的H蛋白基因的序列分析导致鉴定出导致H蛋白的位置263和502（L263 P和R502 P）处的氨基酸取代的两个核苷酸变化，表明mAb 4 B11的表位在性质上可能是构象的。尽管mAr突变体生长到与亲本疫苗病毒（PPRV-Sungri/96）相似的滴度，但在山羊体内研究mAr突变体作为可能的阴性标记疫苗候选物的工作不能成功地用基于mAb 4 B11的竞争性ELISA证明。然而，在位置788处的核苷酸变化（T-C）之一（mAr突变体病毒所特有的）导致限制性酶识别位点（BglII）的消除。这可以用来区分mAr突变疫苗病毒从其他可用的疫苗和现场使用限制性片段长度多态性的菌株。然而，mAr突变体PPRV-E6不能用作DIVA疫苗的候选毒株，因为基于血清学不能区分针对它的免疫应答。
The available vaccines for control ofPeste des petits ruminantsdo not favour differentiation of infected and vaccinated animals (DIVA). Hence, the present study was aimed to isolate and characterize monoclonal antibody resistant mutant of an Indian strain of vaccine virus “PPRV-Sungri/96” under selection pressure of virus neutralizing monoclonal antibody ‘4B11’ specific to haemagglutinin (H) protein. We successfully isolated five monoclonal antibody resistant (mAr) mutants (PPRV-RM5, PPRV-RM6, PPRV-RM7, PPRV- E6 and PPRV- E7). The mAr mutants did not react with the anti-H mAb 4B11 whereas reacted with control anti-nucleoprotein mAb 4G6, similar to the parent vaccine virus “PPRV-Sungri/96” in indirect ELISA, cell ELISA and indirect immunofluorescence test. Cytometry analysis of mAr mutants revealed loss of binding to mAb 4B11 while maintaining binding to mAb 4G6, more or less similar to “PPRV-Sungri/96”. The sequence analysis of the H-protein gene of the mAr mutants resulted in identification of two nucleotide changes leading to amino acid substitutions at position 263 and 502 (L263P and R502P) of the H protein indicating that the epitope of mAb 4B11 could be conformational in nature. Though, mAr mutant grew to a similar titre as parent vaccine virus (PPRV-Sungri/96), the in vivo work in goats to study the mAr mutant as possible negative marker vaccine candidate could not be successfully proved with mAb 4B11 based competitive ELISA. However, one of the nucleotide change (T-C) at position 788, unique to mAr mutant virus resulted in abolition of a restriction enzyme recognition site (BglII). This could be used to differentiate mAr mutant vaccine virus from other available vaccine and field strains using restriction fragment length polymorphism. However, the mAr mutant PPRV-E6 cannot be used as a candidate strain for DIVA vaccine as immune response against it cannot be differentiated based on serology.