The polymerase L528M mutation cooperates with nucleotide binding-site mutations, increasing hepatitis B virus replication and drug resistance
The polymerase L528M mutation cooperates with nucleotide binding-site mutations, increasing hepatitis B virus replication and drug resistance
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DOI：
10.1172/jci11100
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发表时间：
2001-02-01
影响因子：
15.9
通讯作者：
Omata, M
中科院分区：
文献类型：
--
作者：
Ono, SK;Kato, N;Omata, M
在接受拉米夫定治疗慢性乙肝3年后，67-75%的患者在乙肝病毒聚合酶中出现B区L528M、C区M552I或M552V突变，使乙肝病毒(乙肝病毒)产生抗药性。本研究的目的是评估这些突变对病毒复制和抗病毒药物耐药性的影响。我们通过将HBVdna导入肝癌细胞，并通过Southern blotting检测病毒产物，研究了野生型和5株突变株(L528M、M552I、M552V、L528M/M552I和L528M/M552V)对11种化合物[拉米夫定、阿德福韦、恩替卡韦(BMS-200475)(+)-BCH-189(+/-)-FTC(雷西韦)(-)-FTC(恩曲他滨)(+)-FTC、L-D4FC、L-FMAU(环丙氨酸)、D-DAPD和(-)-卡维韦]的复制适合性和敏感性。与野生型相比，单个C结构域突变体M552I和M552V的复制能力明显降低。然而，B结构域突变L528M的加入恢复了复制能力。只有阿德福韦和恩替卡韦对所有5个突变株都有效，与单一突变株相比，这些化合物需要更高的剂量来抑制双突变株。乙肝病毒聚合酶B区突变(L528M)不仅恢复了C区突变株的复制能力，而且增加了对核苷类似物的耐药性。
After receiving lamivudine for 3 years to treat chronic hepatitis B, 67-75% of patients develop B-domain L528M, C-domain M552I, or M552V mutations in the HBV polymerase that render hepatitis B virus (HBV) drug-resistant. The aim of this study was to evaluate the influence of these mutations on viral replication and resistance to antiviral agents. We investigated the replication fitness and susceptibility of the wild-type and five mutant HBVs (L528M, M552I, M552V, L528M/M552I, and L528M/M552V) to 11 compounds [lamivudine, adefovir, entecavir (BMS-200475) (+)-BCH-189 (+/-)-FTC (racivir) (-)-FTC (emtricitabine) (+)-FTC, L-D4FC, L-FMAU (clevudine), D-DAPD, and (-)-carbovir] by transfecting HBV DNA into hepatoma cells and monitoring viral products by Southern blotting. The replication competency of the single C-domain mutants M552I and M552V was markedly decreased compared With that of wild-type HBV. However, addition of the B-domain mutation L528M restored replication competence. Only adefovir and entecavir were effective against all five HBV mutants, and higher doses of these compounds were necessary to inhibit the double mutants compared with the single mutants. The B-domain mutation (L528M) of HBV polymerase not only restores the replication competence of C-domain mutants, but also increases resistance to nucleoside analogues.