.丙型肝炎病毒的发现

丙型肝炎病毒的发现
High titer NANBH chimpanzee plasma
Ultracentrifugation at 104,000g for 5 hours
Extract nuclei acid, random primed cDNA into gt11 phage
Immunoscreened with NANBH patient serum
292: 767; Gastro. 69:1278; JID 139:511; Vox Sang. 44:231)
• non-A, non-B hepatitis (NANBH)
– Might be a small, enveloped virus transmissible to chimpanzees (Gastro 88:773; JID 156:636) – Standard immunological methods failed to identify viral antibodies or antigens – Chiron group
HCV Replication----病毒的入侵需要细胞膜表面的受体
HCV Replication
Assembly and Release • Capsid protein interacts with 5’ NCR • Homotypic capsid protein interaction • E1 and E2 on ER membrane:
Screen with HCV Patient serum
Western blot
Clone 5-1-1 expressed as SOD fusion protein called PS5
NANBH patient serum
Experimentally infected chimp serum
– Nucleocapsid precursors bud into ER to aquire envelope – Pass through host secretory pathway, released
Nature Review Microbiology
-HCV core protein drives assembly at the lipid droplet （LD） -LD is bound by core, then NS5A and other NS proteins
HCV病原学
+ssRNA
核衣壳
包膜 糖蛋白 包膜
• 黄病毒科（Flaviviridae）丙型肝炎病毒属
Baidu Nhomakorabea
Genomic RNA is infectious
HCV Life Cycle
Typical virus life cycle: Binding, internalization, uncoating, protein expression/replication, assembly, release
HCV Replication
Huh-7 cells infected
by hepatitis C virus. The expression of core protein and actin filaments
丙型肝炎病毒的发现
丙型肝炎病毒的发现
• 1970’s: Hepatitis A and B diagnostics
– Most transfusion associated hepatitis not caused by HAV and HBV or other known viral agents (NEJM
Protein from clone 5-1-1 closely associated with NANBH infections
黑猩猩易感
HCV was the first virus to be isolated and characterized solely by molecular methods, ie without culture methods
• Thought that there was not enough viral antigen in NANBH • So they decided to try and concentrate the virus
丙型肝炎病毒的发现
1.传染性肝炎(HA) 血清型肝炎(HB) 2.动物模型（黑猩猩） 3.基因文库 4.放射免疫标记 5.全基因测序
Clone 5-1-1
Larger overlapping Clone 81
丙型肝炎病毒的发现
Phage with portions of HCV genome
Infect E.coli
Phage kills E.coli and leaves plaques
Library Screening