Werkmeister J A, Shukla D D
CSIRO, Division of Biomolecular Engineering, Parkville, Victoria, Australia.
J Virol Methods. 1991 Sep;34(1):71-9. doi: 10.1016/0166-0934(91)90122-g.
Polyclonal antisera to potyviruses contain virus-specific as well as cross-reacting antibodies. The virus-specific antibodies are directed to the surface-located, N-terminal region of the coat protein, whereas cross-reacting antibodies are produced against multiple epitopes within the core region of the coat protein (minus N and C termini), which displays extensive sequence homology among distinct potyviruses. In the present study, immunological tolerance was induced in mice against the cross-reactive central core region of bean yellow mosaic virus (BYMV) using a rat monoclonal antibody (mAb) to the L3T4 molecule (the mouse equivalent of CD4). Generation of specific antisera reactive to the N terminus of BYMV was attained in tolerized mice by secondary immunization with whole viral coat protein from BYMV. This approach appears to be ideally suited to potyviruses where a two-third of the coat protein molecule contains immunogenic epitopes which can result in cross-reacting antibodies.
针对马铃薯Y病毒属病毒的多克隆抗血清包含病毒特异性抗体以及交叉反应抗体。病毒特异性抗体针对外壳蛋白位于表面的N端区域,而交叉反应抗体则针对外壳蛋白核心区域(不含N端和C端)内的多个表位产生,该核心区域在不同的马铃薯Y病毒属病毒之间显示出广泛的序列同源性。在本研究中,使用针对L3T4分子(小鼠中的CD4等同物)的大鼠单克隆抗体在小鼠中诱导针对菜豆黄花叶病毒(BYMV)交叉反应性中央核心区域的免疫耐受性。通过用来自BYMV的完整病毒外壳蛋白进行二次免疫,在耐受小鼠中产生了对BYMV N端具有反应性的特异性抗血清。这种方法似乎非常适合于马铃薯Y病毒属病毒,其中三分之二的外壳蛋白分子包含可导致交叉反应抗体的免疫原性表位。