Decreased neutralizing antigenicity in IBV S1 protein expressed from mammalian cells.
Decreased neutralizing antigenicity in IBV S1 protein expressed from mammalian cells.
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DOI：
10.1016/j.virusres.2015.06.019
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发表时间：
2015-10-02
期刊：
影响因子：
5
通讯作者：
Honda T
中科院分区：
文献类型：
--
作者：
Andoh K;Suenaga K;Sakaguchi M;Yamazaki K;Honda T
重组传染性支气管炎病毒（IBV） S1蛋白高度糖基化，表面附着许多复杂的n -聚糖。重组S1蛋白可诱导抗IBV S1蛋白的抗体，但大多数抗体不能中和IBV。结果表明，重组S1可能由于失去天然构象或被聚糖掩盖而无法显示中和性表位。研究了在哺乳动物细胞中表达的重组传染性支气管炎病毒（IBV） S1蛋白的抗原性。重组S1表达为与三聚基序肽融合的分泌蛋白，然后用Ni Sepharose纯化。纯化蛋白经Western blotting分析后，与油佐剂混合，给药29日龄特异性无病原体鸡。免疫6周后测定抗ibv中和效价和抗s1 ELISA效价；免疫后的鸡经气管接种IBV，观察纤毛活性。结果表明，重组S1蛋白糖基化程度高，其中和抗原性低于灭活病毒。然而，抗S1酶联免疫吸附试验表明，重组S1蛋白诱导了针对S1的抗体。这些结果表明，重组S1可能保留非中和性表位，但具有非自然的糖基化模式和构象，导致缺乏中和性构象表位。综上所述，哺乳动物细胞表达的重组S1蛋白的中和抗原性降低，不足以诱导产生中和抗体。
The recombinant infectious bronchitis virus (IBV) S1 protein was highly glycosylated and many complex N-glycans were attached on the surface. The recombinant S1 protein elicited antibodies against IBV S1 protein, but most of the antibodies could not neutralize IBV. The results indicated that the recombinant S1 may not be able to display neutralizing epitopes by losing native conformation or masking by glycan. We evaluated the antigenicity of recombinant infectious bronchitis virus (IBV) S1 protein expressed in mammalian cells. Recombinant S1 was expressed as a secreted protein fused with a trimerization motif peptide, then purified using Ni Sepharose. The purified protein was analyzed by Western blotting, mixed with oil adjuvant, and administered to 29-day-old specific-pathogen-free chickens. Six weeks after immunization, anti-IBV neutralizing titer and anti-S1 ELISA titer were determined; immunized chickens then were inoculated with IBV via the trachea and ciliary activity was observed. Results showed that the recombinant S1 protein was highly glycosylated, and the neutralizing antigenicity of recombinant S1 protein was lower than that of inactivated virus. However, anti-S1 ELISA indicated that the recombinant S1 protein induced antibodies against S1. These results suggest that the recombinant S1 may retain non-neutralizing epitopes but have unnatural glycosylation pattern and conformation, resulting in lacking neutralizing conformational epitopes. In conclusion, the neutralizing antigenicity of recombinant S1 protein expressed from mammalian cells was decreased, and was not sufficient to induce neutralizing antibodies.
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