Rui L X, Park Y M, Choi J Y, Kim B S, Jung G
Department of Biology Education, Seoul National University, Korea.
Korean J Intern Med. 1998 Jul;13(2):95-8. doi: 10.3904/kjim.1998.13.2.95.
DNA polymerase (pol) of Hepatitis B virus (HBV) includes 3 different domains such as terminal protein (TP), reverse transcriptase (RT) and RNase H. Humoral immune responses to each of these proteins have not been well documented previously, although antibody to pol was detected in serum of patients with chronic hepatitis B. We have constructed TP (amino acids 1-182), RT (amino acids 346-685) and RNase H (amino acids 690-832).
By ELISA using each protein expressed in E. coli as antigens, the corresponding antibodies were tested in serum from 40 patients with type B viral chronic liver diseases. (20 HBeAg-positive and 20 HBeAg-negative). As negative controls, sera from 3 healthy young men were used. With the mean values of the OD, which were tested 4 times per each test sample and 3 times per each control sample, we considered to be positive if the mean OD of each test sample is 2-fold or higher than that of controls.
Five of 40 sera (12.5%) contained one or two different antibodies detectable by this method: 4 of 20 HbeAg-positive sera (20%) and 1 of 20 HbeAg-negative sera (5%). Anti-TP, anti-RT and anti-RNase H antibodies were detected in 2.5% (1/40), 10% (4/40) and 7.5% (3/40), respectively. Among 4/20 HbeAg-positive ELISA-positive sera, anti-TP, anti-RT and anti-RNase H were positive in 5% (1/20), 20% (4/20) and 10% (2/20), respectively, while 1 HBeAg-negative ELISA-positive sera were positive only for anti-RNase H.
These results suggest that the corresponding antibody responses to individual recombinant peptides derived from 3 domains of DNA polymerase may tend to be detected more frequently in HBeAg-positive sera than in HBeAg-negative sera from various patients with type B viral chronic liver diseases.
乙型肝炎病毒（HBV）的DNA聚合酶（pol）包含3个不同结构域，即末端蛋白（TP）、逆转录酶（RT）和核糖核酸酶H。此前，针对这些蛋白各自的体液免疫反应尚未得到充分记录，尽管在慢性乙型肝炎患者血清中检测到了抗pol抗体。我们构建了TP（氨基酸1 - 182）、RT（氨基酸346 - 685）和核糖核酸酶H（氨基酸690 - 832）。
以大肠杆菌中表达的各蛋白为抗原，通过酶联免疫吸附测定（ELISA），检测40例B型病毒性慢性肝病患者（20例HBeAg阳性和20例HBeAg阴性）血清中的相应抗体。以3名健康年轻男性的血清作为阴性对照。每个测试样品检测4次，每个对照样品检测3次，取OD平均值。若每个测试样品的平均OD值是对照样品的2倍或更高，则认为测试样品为阳性。
40份血清中有5份（12.5%）含有一种或两种可通过该方法检测到的不同抗体：20份HbeAg阳性血清中有4份（20%），20份HbeAg阴性血清中有1份（5%）。抗TP、抗RT和抗核糖核酸酶H抗体的检测率分别为2.5%（1/40）、10%（4/40）和7.5%（3/40）。在20份HbeAg阳性ELISA阳性血清中的4份中，抗TP、抗RT和抗核糖核酸酶H的阳性率分别为5%（1/20）、20%（4/20）和10%（2/20），而1份HBeAg阴性ELISA阳性血清仅抗核糖核酸酶H呈阳性。
这些结果表明，在各种B型病毒性慢性肝病患者中，HBeAg阳性血清中对源自DNA聚合酶3个结构域的单个重组肽的相应抗体反应，可能比HBeAg阴性血清中更常被检测到。
