Patterns of circulating hepatitis B virus serum nucleic acids during lamivudine therapy
Patterns of circulating hepatitis B virus serum nucleic acids during lamivudine therapy
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DOI：
10.1196/annals.1318.042
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发表时间：
2004-01-01
期刊：
影响因子：
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通讯作者：
Schröder, CH
中科院分区：
文献类型：
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作者：
Hacker, HJ;Zhang, W;Schröder, CH
拉米夫定治疗慢性乙肝病毒感染者可导致乙肝病毒血清DNA迅速下降。因为停药后，乙肝病毒的复制迅速达到治疗前的值，我们解决了这样一个问题：在治疗过程中，可辨别的循环病毒DNA和RNA的组成和数量的变化是否可以为这种现象提供解释。从两名慢性感染患者的系列血清样本中提取核酸。第一名患者接受拉米夫定治疗14周，而第二名患者显示为核心基因突变的乙肝病毒，接受拉米夫定治疗10周。应用竞争性聚合酶链式反应和逆转录酶/聚合酶链式反应对复制过程中先后合成的3个序列片段，即X、C和X-PreC进行分析。还分析了乙肝病毒转录本中的差异多聚腺苷酸化。在治疗开始时，在第一个患者的所有三个片段中发现了相同的DNA拷贝数(10(9)/毫升)。C节段DNA出现了预期的快速下降。X-prec，一个仅在正链DNA上连续的靶点，表现出类似的行为。相反，治疗结束时X片段DNA COP数下降不明显，保持在高于C和X-Prec片段的值(10(7)/毫升)(均约2×10(5)/毫升)。X片段RNA的持续拷贝数约为10(7)个/m L，而C和X-prec RNA的拷贝数减少到约10(5)个/m L。多腺苷化的HBVRNA，包括全长和截短的，最初保持在10(5)拷贝/毫升，但在治疗结束时下降到10(4)到10(3)拷贝/毫升。正如预期的那样，在第二个患者中没有检测到C节段DNA和RNA，而X和X-prec片段显示出与第一个患者基本相同的模式，尽管水平略低。我们的结论是：(1)在拉米夫定治疗期间，实际的乙肝病毒基因组等价物的数量只能通过X片段DNA来评估，因为它首先是反转录的；(2)拉米夫定在类似水平上诱导C和X片段的DNA和RNA共存，表明逆转录HBVDNA减链的药物滞留中间体；以及(3)包装的HBVRNA缺乏聚(A)尾巴，而多腺化的RNA可能没有包装。
Lamivudine treatment of individuals with chronic HBV infection leads to a rapid decline of hepatitis B virus (HBV) serum DNA. Because HBV replication quickly reaches pretreatment values following cessation of the drug, we addressed the question of whether changes during therapy in composition and amount of discernible circulating viral DNA and RNA might provide an explanation for this phenomenon. Nucleic acids were extracted from serial serum samples of two chronically infected patients. The first patient was treated with lamivudine for 14 weeks, whereas the second one, who displayed an HBV virus with a core gene mutation, received lamivudine for 10 weeks. Three sequence segments of the HBV genome synthesized successively during replication, namely, X, C, and X-preC, were analyzed via competitive polymerase chain reaction (PCR) and reverse transcriptase (RT)/PCR. HBV transcripts were also analyzed for differential polyadenylation. At the start of treatment, identical DNA copy numbers (10(9)/mL) were found for all three segments in the first patient. C segment DNA displayed the expected rapid decline. X-preC, a target contiguous only on plus-strand DNA, behaved similarly. In contrast, the X segment DNA cop numbers showed a less pronounced decrease, remaining at higher values (10(7)/mL) than the C and X-preC segments (both about 2 x 10(5)/mL) at the end of therapy. X segment RNA displayed a persisting copy number of about 10(7)/mL, whereas C and X-preC RNA decreased to about 10(5) copies/mL. Polyadenylated HBV RNA, both full-length and truncated, initially persisted at 10(5) but decreased to 10(4) to 10(3) copies/mL at the end of treatment. As expected, C segment DNA and RNA were not detected in the second patient, whereas X and X-preC segments showed essentially the same pattern as the first patient, although at a slightly lower level. We conclude that: (1) actual numbers of HBV genome equivalents during lamivudine therapy can be assessed only via X segment DNA, because it is reverse transcribed first; (2) lamivudine induces coexistence of DNA and RNA for the C and X segments at similar levels, indicating drug-arrested intermediates of reverse-transcribed HBV DNA minus-strand; and (3) packaged HBV RNA lacks a poly(A) tail, whereas polyadenylated RNA is likely not packaged.