Funkhouser A, Clements M L, Slome S, Clayman B, Viscidi R
Center for Immunization Research, Johns Hopkins University School of Hygiene and Public Health, Baltimore, Maryland 21205.
AIDS Res Hum Retroviruses. 1993 Jul;9(7):627-32. doi: 10.1089/aid.1993.9.627.
An enzyme immunoassay (EIA) was developed to detect secretory IgA (sIgA) antibodies to HIV-1 envelope glycoprotein, using a mouse monoclonal antibody and a highly purified, baculovirus-expressed recombinant gp160 (rgp160) as antigen. Detection of sIgA was enhanced by prior immunoprecipitation of IgG. IgG and sIgA rgp160 antibodies were measured in parotid saliva and nasal wash samples of 18 HIV-1-seropositive volunteers and 14 HIV-1-seronegative adult volunteers immunized 3 times with HIV-1 IIIB rgp160 vaccine at 1 of 4 dosage levels: 40 micrograms (N = 3), 80 micrograms (N = 3), 160 micrograms (N = 4), and 640 micrograms (N = 4). We detected rgp160-specific IgG antibody in the nasal wash samples of all HIV-1-seropositive volunteers and 4/8 vaccinees (50%) immunized with the two highest doses of rgp160 vaccine. All infected volunteers tested had rgp160-specific sIgA in their nasal wash samples. None of the vaccinees and very few of infected volunteer specimens had detectable antibody in the parotid saliva samples (5/8 had IgG and 1/8 had sIgA). We also detected IgG antibody to rgp160 in the sera of all infected volunteers and 13/14 vaccinees (93%). With this EIA, sIgA antibody can be measured in mucosal secretions of recipients of appropriate candidate HIV-1 vaccines.
开发了一种酶免疫测定法(EIA)来检测针对HIV-1包膜糖蛋白的分泌型IgA(sIgA)抗体,使用小鼠单克隆抗体和高度纯化的、杆状病毒表达的重组gp160(rgp160)作为抗原。通过预先对IgG进行免疫沉淀来增强sIgA的检测。在18名HIV-1血清阳性志愿者和14名HIV-1血清阴性成年志愿者的腮腺唾液和鼻腔冲洗样本中测量了IgG和sIgA rgp160抗体,这些志愿者以4种剂量水平之一用HIV-1 IIIB rgp160疫苗进行了3次免疫:40微克(N = 3)、80微克(N = 3)、160微克(N = 4)和640微克(N = 4)。我们在所有HIV-1血清阳性志愿者以及4/8(50%)接受两种最高剂量rgp160疫苗免疫的接种者的鼻腔冲洗样本中检测到了rgp160特异性IgG抗体。所有检测的感染志愿者的鼻腔冲洗样本中都有rgp160特异性sIgA。在腮腺唾液样本中,没有接种者以及极少感染志愿者的样本能检测到抗体(5/8有IgG,1/8有sIgA)。我们还在所有感染志愿者和13/14(93%)接种者的血清中检测到了针对rgp160的IgG抗体。通过这种EIA,可以在合适的候选HIV-1疫苗接种者的黏膜分泌物中测量sIgA抗体。