Kenealy W, Reed D, Cybulski R, Tribe D, Taylor P, Stevens C, Matthews T, Petteway S
AIDS Res Hum Retroviruses. 1987 Spring;3(1):95-105. doi: 10.1089/aid.1987.3.95.
Recombinant proteins representing gag and env amino acid sequences of the Human Immunodeficiency Virus (HIV) (HTLV-IIIb) were produced in Escherichia coli and used to analyze sera for the presence of antibodies to HIV. ENV-9 is a protein representing the carboxy terminus of gp120 and part of gp41 which is highly immunoreactive. GAG-1 represents 83% and GAG-55 100% of the amino acids of the gag open reading frame. The purified proteins allow sensitive detection by enzyme linked immunosorbent assay (ELISA) of antibodies directed against either env or gag of HIV. We have determined the reactivity of sera from several HIV exposed individuals, either form high risk populations or with clinically defined conditions, in the ENV-9, GAG-55, and GAG-1 assays and found that two major seropositive groups are observed. The quantitative analysis of sera with env and gag antigens by ELISA showed AIDS patients had very low gag reactivity while retaining high env reactivity. Results obtained with authentic p24 viral protein in both ELISA and radioimmunoassay correlated to those from the GAG-55 ELISA. This correlation and the analysis of sera with both the ENV and GAG ELISAs indicate that the antibodies reactive to gag are specifically affected relative to env reactivity and that different levels of antibodies to separate viral components in these sera may correlate with disease state.
代表人类免疫缺陷病毒(HIV)(HTLV-IIIb)gag和env氨基酸序列的重组蛋白在大肠杆菌中产生,并用于分析血清中是否存在抗HIV抗体。ENV-9是一种代表gp120羧基末端和gp41一部分的蛋白,具有高度免疫反应性。GAG-1代表gag开放阅读框83%的氨基酸,GAG-55代表100%。纯化后的蛋白可通过酶联免疫吸附测定(ELISA)灵敏地检测针对HIV env或gag的抗体。我们测定了来自高危人群或患有临床确诊疾病的几名HIV暴露个体血清在ENV-9、GAG-55和GAG-1检测中的反应性,发现观察到两个主要的血清阳性组。通过ELISA对含有env和gag抗原的血清进行定量分析表明,艾滋病患者的gag反应性非常低,而env反应性保持较高。在ELISA和放射免疫测定中使用真实p24病毒蛋白获得的结果与GAG-55 ELISA的结果相关。这种相关性以及同时使用ENV和GAG ELISA对血清的分析表明,相对于env反应性,对gag有反应的抗体受到特异性影响,并且这些血清中针对不同病毒成分的抗体水平不同可能与疾病状态相关。