LOSS OF HAEMAGGLUTINATION ACTIVITY OF AN INFECTIOUS BRONCHITIS VIRUS AFTER ATTENUATION

Yu-Ting Liu, Ching-Ho Wang
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Abstract

Avian infectious bronchitis virus (IBV) has been reported to acquire haemagglutination (HA) activity after treatment with neuraminidase, which depends on the IBV S1 protein. The purpose of this study was to test the IBV HA activity and its relationship to the S1 sequences. A wild strain of IBV, 2575/98 possessed HA activity after neuraminidase treatment. On the contrary, its attenuated strain through 75 chicken embryo passages did not. The nucleotide sequence differences in S1 genes before and after attenuation were C166A and G280T, resulting in amino acid changes of P56T and A94S. The S1 and S genes of the wild strain were cloned and expressed in a baculovirus (B) expression system. To test the relationship between HA activity and the S sequence changes after attenuation, the two nucleotide residues were mutated. The HA activity of those recombinant baculoviruses (rBs) was tested. The results showed that rB containing S gene from wild strain possesses the HA activity, that containing S gene with C166A mutation shows partial HA but that containing G280T mutation shows no any HA activity. Thus, a Taiwan IBV strain 2575/98 loses its HA activity after attenuation and this loss might be related to amino acid changes of P56T and A94S in the IBV S protein.
传染性支气管炎病毒衰减后血凝活性的丧失
据报道,禽类传染性支气管炎病毒(IBV)在用神经氨酸酶治疗后获得血凝(HA)活性,这取决于IBV S1蛋白。本研究的目的是检测IBV HA活性及其与S1序列的关系。IBV野生菌株2575/98经神经氨酸酶处理后具有HA活性。相反,其减毒株通过75个鸡胚传代却没有。衰减前后S1基因的核苷酸序列差异为C166A和G280T,导致P56T和A94S氨基酸发生变化。克隆了野生菌株的S1和S基因,并在杆状病毒(B)表达系统中表达。为了检验HA活性与衰减后S序列变化的关系,对两个核苷酸残基进行了突变。检测了重组杆状病毒(rBs)的HA活性。结果表明,野生菌株含有S基因的rB具有HA活性,含有C166A突变的S基因的rB具有部分HA活性,而含有G280T突变的rB没有HA活性。因此,台湾IBV 2575/98菌株HA活性衰减后丧失,这种丧失可能与IBV S蛋白中P56T和A94S氨基酸的变化有关。
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