The role of scavenger receptor BI in hepatitis - eTheses Repository ...

vi
List of Figures.
Figure 1-1 Phylogentic analysis of HCV........................................................... 4
Figure 1-2 A cartoon of a Hepatitis C virion. .................................................. 15
Figure 1-3 HCV genome and polyprotein....................................................... 17
Figure 1-4 Cellular receptors for HCV............................................................ 34
Figure 1-5 The basic structure of a lipoprotein............................................... 44
Figure 1-6 SR-BI-lipoprotein interactions within a hepatocyte. ...................... 51
Figure 1-7 Lipo-viro-particle assembly........................................................... 59
Figure 2-1 In vitro transcribed HCV RNA....................................................... 75
Figure 2-2 Electroporated Huh-7.5 cells. ....................................................... 77
Figure 2-3 Basic plasmid map of pJZ189....................................................... 82
Figure 2-4 Restriction Map............................................................................. 83
Figure 2-5 SR-BII restriction digest................................................................ 84
Figure 2-6 Basic plasmid map of PCDNA3.1 H E2....................................... 86
Figure 2-7 PCDNA3.1 H E2 digest................................................................. 86
Figure 2-8 sE2 coding region PCR. ............................................................... 88
Figure 2-9 sE2 sequencing. ........................................................................... 89
Figure 2-10 Expression of sE2....................................................................... 90
Figure 2-11 Titration of sE2............................................................................ 92
Figure 3-1 Expression of SR-BI on CHO cells. .............................................. 94
Figure 3-2 Activity of rat anti-E2 monoclonal antibodies................................ 96
Figure 3-3 Ability of anti-E2 mAb to recognise SR-BI bound E2.................... 97
Figure 3-4 Soluble HCV E2 binding to cell expressed SR-BI......................... 98
Figure 3-5 Position of SR-BI mutations........................................................ 101
Figure 3-6 Expression of SR-BI mutants in CHO cells................................. 102
Figure 3-7 Antigenic characterisation of SR-BI mutants. ............................. 104
Figure 3-8 Expression of SR-BII in CHO cells. ............................................ 106
Figure 4-1 Over expression of human SR-BI/II in Huh-7.5 cells. ................. 112
Figure 4-2 Over expression of SR-BI and SR-BII in Huh-7.5 cells enhances
HCVcc infection............................................................................................ 114
Figure 4-3 Over expression of SR-BI and SR-BII in Huh-7.5 cells increases
infected cell focus size. ................................................................................ 116
Figure 4-4 Over expression of SR-BI and SR-BII in Huh-7.5 cells enhances
HCVpp-JFH infection. .................................................................................. 118
Figure 4-5 Expression of murine SR-BI in Huh-7.5 cells does not enhance
JFH-1 infection............................................................................................. 120
Figure 4-6 Over expression of SR-BI and SR-BII in Huh-7.5 cells enhances
the infectivity of cell culture- and plasma derived J6/JFH. ........................... 123
Figure 4-7 Anti-SR-BI serum inhibits cell culture- and plasma-derived J6/JFH
infectivity. ..................................................................................................... 125
Figure 4-8 Infection of Huh-7.5 over expression SR-BI by cell culture adapted
JFH-1 HCVcc. .............................................................................................. 127
Figure 5-1 JFH-1 G451R has an altered dependence on SR-BI. ................ 133
Figure 5-2 CD81 dependence of JFH-1 wt and G451R infection................. 135
Figure 5-3 JFH-1 wt and G451R soluble E2 interaction with CHO cells
expressing SR-BI and CD81........................................................................ 138
Figure 5-4 JFH-1 wt and G451R soluble E2 interaction with recombinant
CD81............................................................................................................ 139