Relation of circulating levels of human immunodeficiency virus (HIV) antigen, antibody to p24, and HIV-containing immune complexes in HIV-infected patients.
Relation of circulating levels of human immunodeficiency virus (HIV) antigen, antibody to p24, and HIV-containing immune complexes in HIV-infected patients.
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HIV 感染者体内人类免疫缺陷病毒 (HIV) 抗原、p24 抗体和含 HIV 免疫复合物循环水平的关系。
DOI：
10.1093/infdis/158.5.1088
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Hollander,H
中科院分区：
文献类型：
--
作者：
McHugh,TM;Stites,DP;Busch,MP;Krowka,JF;Stricker,RB;Hollander,H
材料和方法来自患者的样本。血清采集自 30 名健康志愿者（作为对照对象）和 53 名患有各种风湿性疾病的患者。通过商业酶免疫测定（雅培实验室，北芝加哥，111）和间接免疫荧光测定[10]测试时，对照受试者和风湿病患者的 HIV 抗体呈阴性。收集了 73 名 HIV 感染者的血清；我们中的一位 (HH) 将这些患者分类为无症状、轻度症状或患有艾滋病。所有这些患者的艾滋病毒抗体均呈阳性，并且均为男性同性恋。无症状类别的患者没有提示 HIV 感染的症状，包括反复发烧、体重减轻或淋巴结肿大。轻度症状组包括具有任何提示 HIV 感染症状但不符合疾病控制中心 (CDC) 艾滋病诊断标准的患者。所有艾滋病患者均符合疾病预防控制中心的诊断标准。免疫复合物测定。如前所述 [9]，使用固相 Clq 微球免疫测定法通过流式细胞术测量免疫复合物。通过在 FACS 分析仪中使用链霉亲和素-生物素扩增程序，并在 546 nm 处最大激发藻红蛋白 (PE)，获得了更高的灵敏度 [11]（Becton Dickinson 免疫细胞计数系统，山景城，加利福尼亚州）。为了检测免疫复合物中的 HIV，我们使用了针对 HIV 蛋白的小鼠单克隆抗体库。该库包含针对pi 8的抗体和针对gp 110的抗体（由西雅图Genetic Systems的K.Shriver博士提供）以及针对p24的抗体（由加州大学戴维斯分校的J.Carlson博士提供）。抗体库与结合至固相C1q的测试血清反应，随后与针对小鼠IgG的生物素化山羊F(ab')2抗体(Tago，Burlingame，CA)反应。然后添加链霉亲和素-PE（由 Becton Dickinson N. Warner 博士提供）作为荧光标记。含有 IgG 的免疫复合物的总量也使用生物素化的山羊 F (ab,) 2 抗人 IgG 抗体 (Tago) 进行定量，然后使用链霉亲和素-PE。然后通过流式细胞术测量微球相关荧光，结果以标准曲线中聚集的人 IgG/mL 的微克当量 (ng Eq) 表示，如前所述 [9]。
Materials and MethodsSamples from patients. Sera were collected from 30 healthy volunteers who were used as control subjects and from 53 patients with various rheumatic diseases. The control subjects and the patients with rheumatic diseases were negative for HIV antibody when tested by a commercial enzyme immunoassay (Abbott Laboratories, North Chicago, 111) and an indirect immunofluorescence assay [10]. Sera from 73 HIV-infected subjects were collected; these patients were classified, by one of us (HH), as being asymptomatic, mildly symptomatic, or having AIDS. All of these patients were positive for HIV antibody and were homosexual men. The patients in the asymptomatic category were without symptoms suggestive of HIV in-fection, including recurrent fevers, weight loss, or lymph-adenopathy. The mildly symptomatic group included patients with any symptoms suggestive of HIV infection without meeting the Centers for Disease Control's (CDC) criteria for the diagnosis of AIDS. All of the patients with AIDS met the CDC criteria for such a diagnosis. Immune complex assay. Immune complexes were measured by flow cytometry using a solid-phase, Clq micro-sphere immunoassay, as previously described [9]. Increased sensitivity was obtained by using a streptavidin-biotin amplification procedure with maximum excitation of phycoerythrin (PE) at 546 nm in a FACS Analyzer [11](Becton Dickinson Immunocytometry Systems, Mountain View, Calif).To detect HIV in immune complexes, we used a pool of mouse monoclonal antibodies to HIV proteins. This pool contained antibody to pi 8 and antibody to gp 110 (provided by Dr. K. Shriver, Genetic Systems, Seattle) and antibody to p24 (provided by Dr. J. Carlson, University of California at Davis). The antibody pool was reacted with test serum bound to solid-phase Clq, followed by reac-tion with a biotinylated goat F (ab') 2 antibody to mouse IgG (Tago, Burlingame, Calif). Streptavidin-PE (provided by Dr. N. Warner, Becton Dickinson) was then added as the fluorescent label. The total amount of immune com-plexes containing IgG was also quantitated using biotinylated goat F (ab,) 2 antibody to human IgG (Tago), followed by streptavidin-PE. Microsphere-associated fluorescence was then measured by flow cytometry, and the results were expressed in microgram equivalents (ng Eq) of aggregated human IgG/mL from a standard curve, as previously described [9].