Suchánková A, Ritterová L, Krcmár M, Krchnák V, Vágner J, Jochmus I, Gissmann L, Kanka J, Vonka V
Department of Experimental Virology, Institute of Sera and Vaccines, Prague, Czechoslovakia.
J Gen Virol. 1991 Oct;72 ( Pt 10):2577-81. doi: 10.1099/0022-1317-72-10-2577.
A total of 140 sera originating from healthy women and women with either cervical intraepithelial neoplasia or cervical cancer were tested for the presence of IgG antibody against E7 of human papillomavirus type 16 (HPV-16) by ELISA using a synthetic icosapeptide, denoted 16/E7-2, representing amino acids 11 to 30, and by Western blotting (WB) using a genetically engineered HPV-16 E7 fusion protein. Eighteen sera were found positive in either one or the other test. Positive reactions were more frequently detected in cervical carcinoma patients (12 of 34, 35.2%) than in the other individuals (six of 106, 5.7%). Ten children's (1 to 3 years of age) sera reacted in neither ELISA nor WB with HPV-16 E7. A high degree of concordance between the two tests was found suggesting that both tests detect the same or similar activity. To locate the reacting epitopes in the E7 protein, absorption tests were performed with peptides corresponding to various sections of the protein. Based on the results obtained, sera possessing antibody to HPV-16 E7 could be differentiated into those reactive with only the 16/E7-2 peptide and those reactive with other HPV-16 E7 epitopes.
采用合成的20肽(命名为16/E7-2,代表氨基酸11至30),通过酶联免疫吸附测定(ELISA)法,以及使用基因工程人乳头瘤病毒16型(HPV-16)E7融合蛋白,通过蛋白质免疫印迹法(WB),对总共140份来自健康女性以及患有宫颈上皮内瘤变或宫颈癌的女性的血清,检测针对HPV-16 E7的IgG抗体的存在情况。在其中一项或另一项检测中,发现18份血清呈阳性。在宫颈癌患者中(34例中的12例,35.2%)比在其他个体中(106例中的6例,5.7%)更频繁地检测到阳性反应。10名儿童(1至3岁)的血清在ELISA和WB检测中均未与HPV-16 E7发生反应。发现这两种检测之间具有高度一致性,表明这两种检测检测到相同或相似的活性。为了确定E7蛋白中的反应表位,用与该蛋白不同区段相对应的肽进行了吸收试验。根据所得结果,可将具有HPV-16 E7抗体的血清分为仅与16/E7-2肽反应的血清和与其他HPV-16 E7表位反应的血清。