@article{oai:twinkle.repo.nii.ac.jp:00018348,
 author = {ISHIKAWA, Kayo and HASEGAWA, Kiyoshi and NARITOMI, Takuma and IIZUKA, Aiko and HAYASHI, Naoaki},
 issue = {4},
 journal = {東京女子医科大学雑誌},
 month = {Apr},
 note = {[目的] 劇症肝炎を含む,急性B型肝炎のcore promoterの塩基配列を解析し,さらに,肝炎の重症度と変異株のpromoter活性の相関につき検討を行った.[対象と方法] 劇症肝炎(FH)7例,重症急性肝炎(SAH)5例,急性肝炎(AH)6例を対象とし,core promoter,precore/coreを含む領域をPCR法で増幅したのち,増幅産物をpGEM-5Zへサブクローニングし,最低5クローンの塩基配列を調べた.また,すべての症例のcore promoterを含む領域のPCR産物をCAT enhancer vector(Promega)に挿入し,リポフェクチン法で培養肝細胞(Huh7)にトランスフェクションした後,QUAN-T CAT(Amarsham)を用いてpromoter活性を測定した.[結果] core promoter領域の変異はFHで平均4.6個,SAHで3.4個,AHで0.5個と,重症化に伴い頻度が増加した.nt 1762とnt 1764のA→T,G→Aのいわゆるdouble mutation(MT62, MT64)は,FH 7例中2例,SAH 5例中3例に認めた.またFH 5例,SAH 2例にnt 1773のC→T(MT73)の変異を認めたが,FHあるいはSAHの全例に共通する変異はなかった.次にcore promoterの変異株のpromoter活性をCAT assayで調べたところ,野生株(WT)で3202 ± 122cpm,MT62,64,73 : 3625 ± 746cpm,MT62,64:3699 ± 122cpm,MT73 : 3027 ± 570cpmであり,そのほかの変異のパターンを持つ症例でも,WTと比較しpromoter活性の低下,あるいは亢進のいずれも認められなかった.[考察] core promoter領域の変異は,重症度と相関することから,肝炎の重症化との関連が示唆された.しかし,我々の用いたアッセイ系ではpromoter活性に差がみられず,重症化の要因は転写活性の変化とは別の機序によると考えられた., This study sought to identify the contribution of mutations in the core promoter region of the hepatitis B virus (HBV) to the pathogenesis of severe acute onset hepatitis B. Eighteen patients were enrolled in the study: 7 with fulminant hepatitis (FH), 5 with severe acute hepatitis (SAH), and 6 with acute self-limited hepatitis (AH). Sequences and transcriptional activity of core promoter region of HBV from the patients were studied. The mean number of base changes increased with the severity of the hepatitis: 4.6 ± 3.1 in FH, 3.4 ± 1.7 in SAH, and 0.5 ± 0.5 in AH. A double mutation, A (1762)→T and G (1764)→A, was found in 2 of FH, 3 of SAH, and none of AH. AC (1773)→T mutation was found in 5 of FH, 2 of SAH, and none of AH. There was no significant difference in chloramphenicol acetyl transferase activity between different mutated bases care promotor sequences. This finding suggests that the number of mutations in core promoter region was correlated with severity of liver disease, but development of fulminant hepatitis was not a consequence of enhanced core promoter activity.},
 pages = {207--214},
 title = {Comparison of Core Promoter Sequence and Activity Between Naturally Occurring Hepatitis B Virus Mutants from Patients with Acute Hepatitis B},
 volume = {71},
 year = {2001}
}