Maeno K, Kilbourne E D
J Virol. 1970 Feb;5(2):153-64. doi: 10.1128/JVI.5.2.153-164.1970.
Specific antisera for hemagglutinin (HA) and neuraminidase antigens of influenza A(2) virus (A(2)E) were produced through the segregation of the two proteins in reciprocal viral recombinants of A(2)E and A(0)e viruses. Gamma globulin fractions of these specific antisera and of antiserum specific for the nucleoprotein (NP) antigen of A(0)e virus were conjugated with fluorescein isothiocyanate and employed to follow the synthesis of the three structural proteins in clone 1-5C-4 human aneuploid cells, with parallel measurement of serological and biological activity of the antigens by other techniques. In this system, NP antigen appeared first (at 3 hr) in the cell nucleus, whereas HA and neuraminidase appeared coincidentally, at 4 hr after infection, in the cytoplasm. The initial detectability of biological or complement-fixing activity of the proteins coincided with their demonstrability as stainable antigens. Late in infection, all three antigens were detected at the cell surface. Antibody specific for HA partially blocked the intracellular staining of neuraminidase and inhibited the enzymatic activity of both extracted and intact extracellular virus. These observations suggest the close intracytoplasmic proximity of the two envelope antigens and perhaps their initial association in a larger protein.
通过在甲型流感病毒（A(2)E）和A(0)e病毒的相互重组病毒中分离这两种蛋白质，制备了针对甲型流感病毒（A(2)）血凝素（HA）和神经氨酸酶抗原的特异性抗血清。将这些特异性抗血清的γ球蛋白组分以及针对A(0)e病毒核蛋白（NP）抗原的特异性抗血清与异硫氰酸荧光素偶联，并用于追踪1-5C-4人非整倍体细胞中三种结构蛋白的合成，同时用其他技术平行测定抗原的血清学和生物学活性。在这个系统中，NP抗原首先（在3小时时）出现在细胞核中，而HA和神经氨酸酶在感染后4小时时同时出现在细胞质中。蛋白质的生物学或补体结合活性的最初可检测性与其作为可染色抗原的可证明性一致。在感染后期，所有三种抗原都在细胞表面被检测到。针对HA的抗体部分阻断了神经氨酸酶的细胞内染色，并抑制了提取的和完整的细胞外病毒的酶活性。这些观察结果表明两种包膜抗原在细胞质内紧密相邻，也许它们最初在一种更大的蛋白质中结合在一起。
