Wrin T, Loh T P, Vennari J C, Schuitemaker H, Nunberg J H
Department of Immunology, Research Virology Laboratory, Genentech Inc., South San Francisco, California 94080.
J Virol. 1995 Jan;69(1):39-48. doi: 10.1128/JVI.69.1.39-48.1995.
Seven diverse primary isolates of human immunodeficiency virus type 1 (HIV-1) were examined and found to be refractory to neutralization by antisera to recombinant gp120 (rgp120) protein from HIV-1 MN. This stands in marked contrast to the sensitivity exhibited by certain laboratory-adapted viruses. To understand the difference between primary and laboratory-adapted viruses, we adapted the primary virus ACH 168.10 to growth in the FDA/H9 cell line. ACH 168.10 was chosen because the V3 region of gp120 closely matches that of MN. After 4 weeks, infection became evident. The virus (168A) replicated in FDA/H9 cells with extensive cytopathic effect but was unchanged in sensitivity to antibody-mediated neutralization. Thus, growth in cell lines is not sufficient to render primary virus sensitive to neutralization. The 168A virus was, however, partially sensitive to CD4 immunoadhesin (CD4-Ig). Adaptation was continued to produce a persistently infected FDA/H9 culture that displayed minimal cytopathic effect. The virus (168C) was now sensitive to neutralization by MN rgp120 vaccine sera and by MN-specific monoclonal antibodies and showed increased sensitivity to HIVIG and CD4-Ig. 168C encoded three amino acid changes in gp120, including one within the V3 loop (I-166-->R, I-282-->N, G-318-->R). MN-specific monoclonal antibodies bound equally to the surface of cells infected with either neutralization-resistant or -sensitive virus. The coincidence of changes in neutralization sensitivity with changes in cell tropism and cytopathic effect suggests a common underlying mechanism(s) acting through the whole of the envelope protein complex.
对7种不同的人类免疫缺陷病毒1型(HIV-1)原代分离株进行了检测,发现它们对来自HIV-1 MN的重组gp120(rgp120)蛋白的抗血清中和作用具有抗性。这与某些实验室适应病毒所表现出的敏感性形成了显著对比。为了了解原代病毒和实验室适应病毒之间的差异,我们将原代病毒ACH 168.10适应于在FDA/H9细胞系中生长。选择ACH 168.10是因为gp120的V3区域与MN的V3区域紧密匹配。4周后,感染变得明显。该病毒(168A)在FDA/H9细胞中复制,具有广泛的细胞病变效应,但对抗体介导的中和作用的敏感性没有改变。因此,在细胞系中生长不足以使原代病毒对中和作用敏感。然而,168A病毒对CD4免疫粘附素(CD4-Ig)部分敏感。继续进行适应,以产生一个持续感染的FDA/H9培养物,该培养物显示出最小的细胞病变效应。该病毒(168C)现在对MN rgp120疫苗血清和MN特异性单克隆抗体的中和作用敏感,并对HIVIG和CD4-Ig表现出更高的敏感性。168C在gp120中编码了三个氨基酸变化,包括V3环内的一个变化(I-166→R,I-282→N,G-318→R)。MN特异性单克隆抗体与感染了中和抗性或敏感病毒的细胞表面的结合程度相同。中和敏感性的变化与细胞嗜性和细胞病变效应的变化同时出现,表明存在一种共同的潜在机制,通过整个包膜蛋白复合物起作用。