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C 型肝炎病毒是一正單股 RNA病毒,常會引起持續性的感染而導致慢性肝
臟疾病和肝癌.C型肝炎病毒的核心蛋白共包含有191個胺基酸,在整個核心
蛋白 前半段靠近N端,已證實是最能引起免疫反應的區域.在本論文中,兩
個能表現出C型肝炎核心蛋白全長1到190個胺基酸及部份1到98個胺基酸,
都分別接在E.coli表現質體pRSET-B上面.在pHBcPreS1PrS2質體,能表現B
型肝炎病毒核心抗原融合蛋白 質粒子 (3-144胺基酸)及其所攜帶的B型肝
炎病毒表面抗原PreS1(1-20胺基酸)和PreS2(1-26胺基酸)片段 ,如將含有
C型肝炎病毒核心蛋白N端98個胺基酸的片段也接在此質體上面,一樣也能
在E.coli內表現. 透過西方墨點法分析 ,證實C型肝炎病毒全長的核心蛋
白HCc1-190及HCc1-98兩個蛋白質與C型肝炎病患血清之間有作用. 而經B
型肝炎病毒核心抗原重組後的蛋白質,同時具有B型肝炎病毒核心與C型核
心的抗原性. HBcPreS1PreS2HCc1-98和HCc1-98兩個蛋白質在兔子 及小老
鼠(BABL/c)體內,均能提引出抗C型肝炎病毒核心蛋白的抗體 ,而前者亦可
提引出抗B型肝炎病毒核心抗原的抗體,且較後者提引出較高的抗C型肝炎
病毒核心蛋白的抗體. 一套由這兩個蛋白質當固定相的抗原 ,來一起偵測
抗C型肝炎核心蛋白抗體和抗B型肝炎病毒核心蛋白抗體,或單獨測抗C型肝
炎病毒核心但捱抗體的免疫酵素聯結方法已經研發出來.這個結果顯示,B
型肝炎病毒核心抗原上攜帶有C型肝炎病毒核心蛋白的重組蛋白,能在捐血
者中偵測出抗B型核心和 C型核心蛋白的抗體存在.
Haptitis C virus (HCV) is a positive single-stranded RNA virus
which frequently causes persistent infection leading to chronic
liver disease and hepatocellular carcinoma. The nucleocapsid
(core) protein of HCV consists of 191 amino acids, among which,
the major immunodomint epitopes within the core protein
identified mostly lie in the amino-terminal half. In the study,
two sequences encoding the full-length and truncated core
protein (HCc) of HCV, encompassing amino acids 1 to 190 and 1
to 98,respectively, were fused to E.coli expression vector
,pRSET-B. The plasmid, pHBcPreS1PreS2HCc1-98, which directs the
expression of the hybrid hepatitis B virus core(HBcAg)
particles carrying HBcAg (3-144), preS1(1-20) and preS2(1-26)
segment from HBV as well as the core protei(1-98) from HCV, was
also constructed for expression in E.coli. The full-length and
truncated HCV-infected individuals in the Western blot
analysis. The hybrid HBcAg protein exhibited both HBV core and
HCV core antigenicities. HBcPreS1PreS2HCc1-98 and HCc1-98
induced antibodies against HBcAg or/and the HCV core protein in
rabbits and in BABL/c mice. Moreover, the former is more
immunogenic than the latter in terms of anti-HCc induction. An
ELISA using these two proteins as solid-phase antigens to
detect anti-HCc or/and anti-HBc antibodies was developed. The
resluts also indicate the potential value of the hybrid HBcAg
protein carrying the HCV core protein for simultaneous
detection of antibodies to HBV core and HCV in donated blood.
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