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1
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2
Hepatitis C specific prevention
BASIC PARAMETERS OF INFECTION INDUCED BY
ISOLATED HCV VARIANT IN PRIMARY HUMAN ENDOTELIAL
CELLS
OF CELLS
lg TCD50
HCV RNA
HCV RNA
IN MONOLAYER
8
100
7
80
6
5
60
4
40
3
2
20
1
1
2
3
4
5
9
8
6
7
TIME AFTER CELL INFECTION BY HCV (DAYS)
3
Hepatitis C specific prevention
HCV SPECIFIC ANTIGENS IN HCV INFECTED CHICK
EMBRYO FIBROBLASTS DETECTED BY INDIRECT IMMUNE
FLUORESCENCE METHOD USING MONOCLONAL ANTIBODIES
TO RECOMBINANT HCV CORE PROTEIN.
4
Hepatitis C specific prevention

HCV REPLICATION IN VITRO IS INHIBITED BY
HUMAN ANTI-HCV NEUTRALIZING ANTIBODIES a)
RT PCR

Before
After neutralization
neutralization
tenfold virus dilutions used
10-1 10-2 10-3 10-4 10-5 10-1 10-2 10-3 10-4 10-5 10-1 10-2 10-3 10-4 10-5 10-1 10-2 10-3 10-4 10-5 10-1 10-2 10-3 10-4 10-5 10-1 10-2 10-3 10-4 10-5 10-1 10-2 10-3 10-4 10-5 10-1 10-2 10-3 10-4 10-5 10-1 10-2 10-3 10-4 10-5 10-1 10-2 10-3 10-4 10-5 10-1 10-2 10-3 10-4 10-5
0 0 0 0 0 0
b) HCV CYTOPATHOGENIC EFFECT
5
Hepatitis C specific prevention
a- INTERFERON ANTIVIRAL ACTIVITY STUDY IN SW-13
CELL CULTURES INFECTED WITH HEPATITIS C VIRUS
lg ?CD 50
Contol HCV infected cells
Interferon ffter virus absorbtion
Interferon 24 h p.i.
Interferon 24 h and 48 h p.i.
6th day p.i.
5th day p.i.
4th day post infection
Time after HCV infection of SW-13 cell cultures
6
Hepatitis C specific prevention
INFECTION OF RABBITS INDUCED BY CYTOPATHOGENIC
HCV VARIANT (STUDY OF BLOOD SERA
SAMPLES COLLECTED ON 24-th DAY POSTINFECTION
)
HCV INFECTED RABBITS lg TCD50 RT PCR HCV HA-activity Anti-HCV- antibodies Detected by Anti-HCV- antibodies Detected by Anti-HCV- antibodies Detected by
HCV INFECTED RABBITS lg TCD50 RT PCR HCV HA-activity HIT NT (index lg TCD50) EIA
No 1 10 () 1320 1320 1,5 1160
No 2 7,5 (-) 1160 180 5 180
7
Hepatitis C specific prevention
HCV infected mouse liver (56 days after virus
administration by intravenously way)
a) liver of uninfected mouse
b) liver of HCV infected mouse
8
Hepatitis C specific prevention
Liver of uninfected mouse. Liver has normal
structure. A central vena can not iced. Normal
hepatocytes. Zoom x 100
Liver of HCV infected mouse. (56 day after the
infection) Large periportal cell infiltration.
Zoom x 400
9
Hepatitis C specific prevention
HCV SPECIFIC ANTIBODIES TO DIFFERENT HCV
SPECIFIC PROTEINS DETECTED BY WESTERN BLOT
(RecomBlot HCV IgG) IN MOUSE BLOOD SERUM SAMPLES
r Core (aa 1- aa 115)
r NS 3 (aa 1007 aa 1534)
r NS 4 (aa 1616 aa 1862)
r NS 4 1 (aa 1680 aa 1796)
10
Hepatitis C specific prevention
Identification of cytopathogenic HCV isolated
from infected mice
11

Hepatitis C specific prevention
AN ABILYTY OF INACTIVATED HCV CONTAINING MATERIAL
TO INDUCE HCV SPECIFIC ANTIBODIES IN MICE
Dilutions of inactivated HCV HCV-specific antibodies induced in mice Methods of detection HCV-specific antibodies induced in mice Methods of detection HCV-specific antibodies induced in mice Methods of detection
NT (index of neutralization) HIT ELISA
0 3.0 140 20
12 2.0 1160 80
110 1.0 140 0