il virus dell’epatite b -...
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![Page 1: Il virus dell’epatite B - fad-connecta.ecm33.itfad-connecta.ecm33.it/cm/pdf/fad_connecta_modulo_1.pdfIl virus dell’epatite B Giovanni Raimondo Epatologia Clinica e Biomolecolare](https://reader034.vdocuments.us/reader034/viewer/2022050717/5e1514286325d5597f546bf2/html5/thumbnails/1.jpg)
Il virus dell’epatite BGiovanni RaimondoEpatologia Clinica e BiomolecolarePoliclinico Universitario di Messina
La patogenesi dell’epatite BCarlo FerrariUnità Operativa di Malattie Infettive ed EpatologiaAzienda Ospedaliero-Universitaria di Parma
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Il virus dell’epatite BGiovanni RaimondoEpatologia Clinica e BiomolecolarePoliclinico Universitario di Messina
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VIROLOGY OF HEPATITIS B VIRUS (HBV)
Modulo 1
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Virology of Hepatitis B Virus (HBV)Family: Hepadnaviridae
Genus: HepadnavirusOrthohepadnavirus (Mammalian)Human Hepatitis B-Virus (HBV)Chimpanzee Hepatitis B Virus (ChHBV)Gibbon Hepatitis B-Virus (GiHBV)Orang Utan Hepatitis B Virus (OuHBV)Gorilla Hepatitis B-Virus (GoHBV)Woolly Monkey Hepatitis B-Virus (WMHBV)Woodchuck Hepatitis-Virus (WHV)Ground squirrel Hepatitis-Virus (GSHV)Arctic Ground squirrel Hepatitis-Virus (AGSHV)
AvihepadnavirusDuck Hepatitis B-Virus (DHBV)Heron Hepatitis-Virus (HHV)Ross Goose Hepatitis BVirus (RGHBV)Snow Goose Hepatitis BVirus (SGHBV)Maned Duck Hepatitis BVirus (MDHBV)Grey Teal Hepatitis B Virus (GTHBV)Stork Hepatitis B-Virus (STHBV)
• DNA virus• 3.2 kilobases• partial-double-stranded• enveloped
Mature virus (Dane particle) 42 nm diameter
Target cells: hepatocytesReceptor: unknownLigand: preS1-protein
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HEPATITIS B VIRUS
HBV GENOMES gene: pre-S1 (large S), pre-S2 (middle S), S (small S)
X gene: X
Core gene: core, ePol gene: P
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C
A
B
HEPATITIS B VIRUS
C
A
B
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HEPATITIS B VIRUS Genome Organization
• 3.2 kb pdDNA• 3 promoters = Core, S-, and X• 4 ORF = preC/Core, pre-S1/pre-S2/S,
Polymerase, X-gene
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HBsAg and Reverse Transcriptase (RT) Protein Synthesis
Met- ……… Ala - Arg - Phe - Ser - TrpTrp - Leu - Ser - Leu - Leu
Met- ………… Pro - Phe - Leu - Leu - AlaAla Gln - Phe - Thr
GENOMIC REGIONS
ATG..... AGC CCG TTT CTC CTG GCT CAG TTT ACT
ATG ..... GCC CGT TTC TCC TGG CTC AGT TTA CTA
Transcription
RT
HBsAg
mRNA … … … … … … … … … … … … … … … … …
mRNA … … … … … … … … … … … … … … … … … …
Translation
RT
S
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HBsAg and RT Protein Syntheses in HBV Strains with A181V aa Change in RT
GENOMIC REGIONS
Transcription
Translation
Met- ……… Ala - Arg - Phe - Ser - STOPSTOP
ATG..... AGC CCG TTT CTC CTG ACT CAG TTT ACT
ATG ..... GCC CGT TTC TCC TGA CTC AGT TTA CTA
Met- ………… Pro - Phe - Leu - Leu - ThrThr Gln - Phe - Thr
… … … … … … … … … … … … … … … … …
… … … … … … … … … … … … … … … … … …
RT
S
RT
HBsAg
mRNA
mRNA
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HBV Life Cycle
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HBV Life Cycle
RECYCLING
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HBV GENETIC HETEROGENEITY
Modulo 1
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Geographic Distribution of HBV Genotypes
Genotype A Worldwide
“ “ B Asia
“ “ C Asia
“ “ D South Europe, Americas, Australia
“ “ E Africa
“ “ F Native Americans and Polynesians
“ “ G Europe, USA
“ “ H Native Americans
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HBV Genomic Variants…
• Naturally occurring
• Therapeutically induced
…When Are They Clinically Relevant?
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Relative Replication Yield (RC) of HBV Mutants
0 1 2 3 4 5 6 7 8 9 10L180M+A181V+N236T
A181V+N236TL180M
L180M+T184G+S202I+M204V*A181V
M250V*M204I
L180M+A181V+M204VI169T+V173L+L180M+M204V+M250V*
N236TL180M+M204V*
WTA181T+N236TL180M+A181V
A181TI169T+V173L+L180M+M204V*
V173L+L180M+M204V*
Modified from Edwards R, et al. Global Antiviral Journal 2005;1(Suppl2):77
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Secondary Mutations at the RT Level in HBV Isolates from Naïve Patients
aa substitution No. of cases
N94R 1
V173M 1
A181D * 1
T207I 1
V214A/E 2
Q215S/H/P 9
R217L 1
S219A 3
F221Y 3
I233V 3
P237T 2
N238H 4
*Previously unreported aa substitution in a position critical for primary resistance to Lamivudine and Adefovir.
aa substitutions No. of Cases
Q215S/H/P +V173M 1
F221Y 1
P237T 2
N238H 1
L217RF221Y +
N238T
3
2
V214EN238H +
N94R
1
1
S219A + S185I 1
F221Y + I233V + N238D 1
Q215S + R217L + F221Y 1
S202T +L217R + S219A + F221Y 1
Pollicino T, et al. Antivir Ther 2009;14:649-654
Single Mutation Multiple Mutations
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MOST COMMON, NATURALLY OCCURING HBV GENETIC VARIANTS
Modulo 1
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PreC/C-gene
np 1814-6ATG
TGG
HBcAg
HBeAgPredicted precore protein
np 1896
np 1901-3ATG
np = nucleotide position
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PreC/C-gene
np 1814ATG
TGG TAG stop codon
HBcAg
HBeAgPredicted precore protein
np 1896
np 1901ATG
np = nucleotide position
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Basal-Core Promoter Mutations
Pre-C C P Enhancer II/basal core promoter
np 1762 1764
X
A T G A
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Hepatitis B Virus Genome Organization
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PreS/S Gene Transcription and Envelope Proteins (ORFs: Pre-S1/Pre-S2/S)
Pre-S1 RNA (2.4kb)
S
S
Pre-S2/S RNA (2.1kb)
ATG
p r e-S1 p r o t e i n
p r e-S2 p r o t e i n
Pre-S2/S RNA (2.1kb)
Pre-S2Pre-S1ATG
ATG
ATG
S
ATG
ATG SPre-S2
S p r o t e i n
LHBs
MHBs
SHBs
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Envelope Gene and Proteins
S-gene
Large protein (390-399 aa)
Middle protein (281 aa)
Small protein (226 aa)
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Natural Occurring Pre-S1 HBV Variant
PRE-S2 S
183-bp deletion
PRE-S1ATG
ATG
ATG
Gerken G, et al. Virology 1991;183:555-565
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Pre-S2 Minus HBV Variants
SPRE-S1ATG
ATG
AGG
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HBV Variants Carrying Deletion in Pre-S2 Region
SPRE-S1ATG
ATG
ATG
PRE-S2
In framedeletion
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PreS2-Defective HBV Variants: Evidence of Clinical Impact
- Fulminant hepatitis
- Fibrosing cholestatic hepatitis
- Cirrhosis and HCC
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Distribution of pre-S2-defective variants in HBsAg+ patients (pts) with different clinical picturesRaimondo G, et al. J Hepatol 2004;40:515-519
°p=0.002*p=0.02
All pts infected with genotype D.
Prevalence of pre-S mutants in patients infected with genotype B, C, or mixed genotypes Chen BF, et al. Gastroenterology 2006;130:1153-1168
cc : chronic (inactive) carriersCH : chronic hepatitisLC : liver cirrhosis
DeletionWild-type + deletionWild-type
Diagnosis No. pts No. pS2 variants
Inactive carriers° 15 2
Chronic hepatitis°* 50 25
Cirrhosis°* 26 13
HCC°* 19 16
Infection with HBV PreS2-Defective Variants is associated with HCC
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HBV Surface Antigen “a” Determinant
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Clinically Relevant Mutations in the “a” Determinant of HBsAg
124137
139 147
Cys
Ile
Cys Ser
Lys
Thr
Gly
Cys Cys
Tyr
8 amino acid insertion)
Asn, Thr
Tyr
Thr
IIe,Glu Met
Arg
145
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Hepatits B Virus
…leads to an infection that lasts indefinitely and is referred to as “occult infection” when it is mantained without HBsAg
…replication process takes place through a reverse transcription phase like retroviruses, and like retroviruses it may be integrated into the host genome
…has a compact genomic structure, with partial overlapping of its different genomic regions
…cannot eliminate the viral mutants that develop during each replicative cycle, so that they may accumulate leading to the formation of a complex quasispecies, often influencing the clinical evolution of the liver disease
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La patogenesi dell’epatite BCarlo FerrariUnità Operativa di Malattie Infettive ed EpatologiaAzienda Ospedaliero-Universitaria di Parma
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INNATE IMMUNE RESPONSES IN HBV INFECTION
Modulo 1
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Role of Innate Immunity in Acute Viral Infections
Time (months) after infection1 2 3 4 5 6
Viral load
Innate response T cell response
Antibody response
Early viral containment
Priming and maturation of adaptive immune responses
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Kinetics of Virus Replication and Liver Damage in HBV InfectionMajority of viral clearance prior to peak of liver inflammation
non-cytolytic mechanisms likely implicatedWebster GR, et al. Hepatology 2000;32:1395-1406
Thimme R, et al. J Virol 2003;77:68-76Guidotti L, et al. Science 1999;284:825
ALT (U
/L)
0
100
200
300
400
500
ALTLiverHBV DNA
Live
r HB
V-D
NA
(%)
100
10
1
0
2 4 6 8 10 12 14 16 18
1
10
100
HBV
1,000
2,000
ALT
Weeks after infection
242080 4 6 10 12 18 2214 162
Chimpanzee infectionHuman infection
0
Weeks after infection
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Intrahepatic CTL Frequency, HBV Replication, and Liver Damage in Acute Hepatitis B
0
100
200
300
400
0 2 4 6 8 10 12 14 16 18 20 22 24
20
40
60
80
100
sAL
T (U
/L)
HBV-DNA ALT %
HB
V-D
NA
liver
Intrahepatic HBV-specific CD8 T cells in chimps
00
1010
55
HBV-specific
CD8 cells
Thimme R, et al. J Virol 2003;77:68
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In Vivo Chimpanzee DataLack of IFN-inducible genes in chimpanzee model of HBV infection
HBV seems unable to induce innate responses: a ‘stealth virus’
Genes associated with entry and expansionof the virus, reflecting activation and effector
function of the innate immune responseNo gene
Genes associated with viral clearance(brought into the liver by cells of the adaptive
immune response)110 genes
Wieland S, et al. PNAS 2004;101:6669-6674
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HBV Is a Good Inducer of Innate Responses: Proofs in Favor
• Intrahepatic expression of innate immune response genes immediately afterinfection in woodchucks infected with WHV (Guy CS. J Virol 2008;82:8579-8591)
• Induction of IFN-I in HepaRG hepatocytes infected with baculovirus carrying HBVgenome results in non-cytolytic control of HBV (Lucifora J. Hepatology 2009;51:63-72)
• Primary human hepatocytes infected with HBV stimulate Kupffer cells to produceIL-6 with anti-HBV activity (Hösel J. Hepatology 2009;50:1773-1782)
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Flaws of These Studies
• In woodchuck infection, activation of innate responses is transient and it isstimulated by inocula with very high viral concentrations; moreover, this animalmodel may only partially reproduce human infection
• IFN-I production by HepaRG cells is stimulated by high intracellular HBVreplication levels which likely do not reproduce the early kinetics of HBVreplication in natural infection
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HBV Is a Poor Inducer of Innate Responses: Proofs in Favor
Cytokine production in acute HBV infection
is significantly more modest and delayed compared with
acute HIV infection (Stacey AR, et al. J Virol
2009;83:3719-3733)
Low production of type 1 IFN, IL-15 and IFN-1, associated with high serum IL-10 levels,
at the early stages of HBV infection
(Dunn C, et al. Gastroenterology 2009;137:1289-1300)
IFN-α IL-15 IFN-1A B C
-10 -5 0 5 10 15 20 -10 -5 0 5 10 15 20
Time (days since T0) Time (days since T0)Fold
cha
nges
in
grou
p m
ean
cyto
kine
leve
l
HIV infection HBV infection
Healthy DNA
high
Resolution phase
Acute HAV
n.27 n.11 n.17 n.13
p=0.05
p=0.02
pg/m
l
100
200
300
400
Healthy DNA high
Resolution phase
Acute HAV
n.10 n.11 n.16 n.13pg
/ml
2
4
6
8
p=0.03p=0.05
Healthy DNA high
Resolution phase
Acute HAV
n.25 n.11 n.18 n.13
p=0.0001p=0.0002
p=0.0001
pg/m
l
200
400
400
800
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Can the lack of IFN-I detection in acute HBV infection be attributable to active suppression rather than a complete
failure of induction?
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Regulation of Toll-like Expression by HBV
• HBeAg can suppress TLR-2 expression in chronic hepatitis B (Visvanathan K, et al.Hepatology 2007;45:102-110)
• Low levels of TLR-1, 2, 4, and 6 messenger RNA transcripts in PBMC of CAHpatients (Chen Z, et al. Clin Immunol 2008;128:400-408)
• Hepatitis B virus suppresses Toll-like receptor mediated innate immuneresponses in murine parenchimal and nonparenchimal liver cells (Wu J, et al.Hepatology 2009;49:1132-1140)
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VirionsHBsAgHBeAg ISGF3
IFN receptor
Interferon loop
TYK2 JAK1
SOCS1
SOCS3
IRF-9
STA
T1
STA
T2
IFN- IFN-
IFN receptor
IFN
-
PKR, ADAR1, 2-5OAS, IFIT1, Viperin, ISG6-16,
MxA
ISG expression
TLR2 TLR4 TLR3
Extracellular sensing (TLR) Intracellular sensing (RIG-1)
NF-kB
TRIF
IRF-3PP
RIG-1
PP
IRF-3IRF-7
PRDNF-kB target genes
IFN-
Intracellular Host Innate Immune Defences
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What Role Do NK Cells Play in Acute HBV Infection?
Hepatocytes express very low levels of MHC Class I, such that any up-
regulation of cellular stress ligands able to engage NK activatory
receptors should be able to induce local NK cell effector function
NK cells are extremely abundant in the liver, constituting 30-40% of
intrahepatic lymphocytes
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Kinetics of NK Cell Responses
FUNCTION
HBV-DNA HBV-DNA copies/mL
IFN
Normalized51Cr release
050,000
100,000150,000200,000250,000
0 2 5 7 10
Weeks from HBsAg detection
% IF
N
0 13102 5 70
1234
0
5
10
15
51C
r rel
ease
NK cellsCD56+CD3-
PHENOTYPE%
CD
69+
0
12.5
25
0 2 5 7 100
35
70
NK
G2D
MFI
% CD69
NKG2D MFINK cells
CD56+CD3-
NK IFN
CD8 IFNCD4 IFN
% C
D56
+CD
3-IF
Ng+
00.51
1.52
2.533.5
0 5 7 10 1300.10.20.30.40.50.6
% C
D8/
4+IF
Ng+
Weeks from HBsAg detection
Early induction after infection(Fisicaro P, et al. Gut 2009;58:974-982)
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Kinetics of NK Cell ResponsesDelayed induction after infection
(Dunn C, et al. Gastroenterology 2009;137:1289-1300)
0
510
15
20
25
Perc
ent C
D69
+ N
K c
ells
0
5
10
15
Healthy Resolution phase
n.15 n.6 n.10
DNA highPe
rcen
t CD
69+
NK
cel
ls
0
5
10
15
Healthy Resolution phase
n.27 n.11 n.17
DNA high
P=0.03
0.009
0.003
0.02
Weeks 0 5 10 5 20Pe
rcen
t IFN
+N
K c
ells
510
15
20
25
NK cells
HBV-DNA
NK cells
HBV-DNA
Perc
ent I
FN+
NK
cel
ls
Weeks 0 5 10 5 20
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ADAPTIVE IMMUNE RESPONSES IN HBV INFECTION
Modulo 1
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Kinetics of Virus Replication and HBV-specific Responses During Acute Infection
Webster GR, et al. Hepatology 2000;32:1395-1406
CD4+ T cells
1
10
100
Weeks after infection2 4 6 8 10 12 14 16 18 20
20
40
0
HBV-DNA% tet.+
CD8+ T cells
nd1
10
100
0.4
0.8
HBsAg - - - - + + + + + + + +
HBV-DNA
nd
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CD8 Cells Are the Main Effectors of Viral Clearance and Disease in Acute Hepatitis B(Thimme R, et al. J Virol 2003;77:68-76)
No depletion
sALT (U/L)
0
100
200
300
400
500
sALTLiver
HBV-DNA
Live
r HB
V-D
NA
(%) 100
10
1
0
Control Ab
CD4 Ab
CD8 Ab
00
sALT (U/L)
100
200
300
400
500
sALTLiver
HBV-DNA
Live
r HB
V-D
NA
(%) 100
10
1
CD4 depletion
Weeks after infection242080 4 6 10 12 18 2214 162
sALT (U/L)
0
100
200
300
400
500
sALT
LiverHBV-DNA
Live
r HB
V-D
NA
(%) 100
10
1
0
CD8 depletion
Weeks after infection242080 4 6 10 12 18 2214 162
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Role of HBV-specific CTL in Liver Damage and Virus ControlHBV is not directly cythopatic; HBV-specific CD8 cells not only protect, but also damage
CD8 CELLCD8 CELL
CLEARANCE BY CYTOKINESIFNγ/TNFα
CYTOPATHIC NON-CYTOPATHIC
CLEARANCE BY LYSIS OF INFECTED CELLS
CD8 CELLCD8 CELL
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HBV Pathogenesis
Cyt
okin
es
IFN/TNFα
FasL
Perforins
Liver cell killing
Chemokines
HBV-specific CTL
Infected liver cell
Cured liver cells
Amplification of liver damage
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HBV InfectionSelf-limited
Vigorous, T1-oriented, multispecific acute phase
responses
Long-lasting protective responses
%Lysis
0
10
20
30
0
10
20
30
40
POLIMERASE X CORE ENVELOPE
ACUTE PHASE
RECOVERY PHASE(20 years from recovery)
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CD4-mediated Proliferative Response to HBV Core in Acute HBV Infection
Urbani S, et al. Hepatology 2005;41:826-831
50
Evolution of infection
0
5
10
15
20
25
30
35
40
45
1
Chronic
1 2 3 4 5 6
Self-limited
Stim
ulat
ion
Inde
x
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HBV InfectionChronic evolution
Inefficient acute phase immune responses
Progressive impairment of protective responses
CD4 RESPONSES
CD8 RESPONSES
HBVPOL
HBVX
HBVCORE
HBVENV
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2 4 6 8 10 12 14 16 18
1
10
100
HBV
1,000
2,000
ALT
0
Poor induction of early
intracellular innate responses
1Efficient and
timely induction of adaptive responses
3
Early HBV-DNA clearance by non-
cytolytic mechanisms
4
Liver damage and elimination of infected
cells by cythopatic mechanisms
5
NK cells activation
2
0
Summary of the Early Immune Events in HBV Infection
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Helper T cellsExpansion of
immune response
B cellsAb production
Blocking cell to cell spreadNeutralization of circulating virus
ANTIBODY PRODUCTION IN ACUTE HBV INFECTIONAcute HBV infection
Th1Th2
BB
Antigen excess Partial neutralization
Anti-HBs negative
EARLY
Th1Th2
BB
Antibody excess Complete neutralization
Anti-HBs positive
LATE
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Hierarchy of HBV Control
Partial controlChronic inactive infection
Inefficient controlChronic active infection
Efficient controlOccult infection
0
5
10
1 2 3 4 5 6 7 8 9 10 111213 14 1516mea
n %
IFN
/tot
0
5
10
1 2 3 4 5 6 7 8 9 10 111213 14 1516mea
n %
IFN
/tot
02468
10
1 2 3 4 5 6 7 8 9 1011 1213141516mea
n %
IFN+
/tot
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HBV-specific T Cell Responses are Stronger in Acute than in Chronic Hepatitis B
Boni C, et al. J Virol 2007;81:4215-4225
050100
12 34 5 67 8910111213141516
Pt. C1Pt. C1
Pt. C2Pt. C2
Pt. C3Pt. C3
050100
1 2345 678910111213141516
050100
1 2345 678910111213141516
050100
1 234 5 67 8 910111213141516
050100
12 34 5 67 8910111213141516
050100
1 234 5 67 8910111213141516
Pt. C4Pt. C4
Pt. C5Pt. C5
050100
1234 5 678910111213141516050100
1 23 456 78 910111213141516
Pt. C6Pt. C6
Pt. C7Pt. C7
050100
123 4567 8 910111213141516
050100
123 4567 8 910111213141516
050100
1234 5 678910111213141516
050100
12 345 67 8910111213141516
050100
1234 5 67 8 910111213141516
050100
1 23 45678 910111213141516
100
12345 67 8 910111213141516050
050100
12 34 56 78 910111213141516
050100
12 345 678 910111213141516
050100
12 34 56 78 910111213141516
100
1234 56 78910111213141516050
050100
12 34 56 78 910111213141516050100
1234 5 67 8910111213141516
xx corecore envenv polpol xx corecore envenv polpol xx corecore envenv polpol
SFU
/2 x
105
PBM
C
Chronic patients
Pt. A3
Pt. A2
Pt. A1
100
Pt. A4
Pt. A6050
250
1 2 3 4 5 6 7 8 9 1011121314 1516
050
100
1 2 3 4 5 6 7 8 9 10111213141516
050
1 2 3 4 5 6 7 8 9 10111213141516
050
100
1 2 3 4 5 6 7 8 9 10111213141516
Pt. A5
050350
1 2 3 4 5 6 7 8 9 10111213141516
050100
1 2 3 4 5 6 7 8 9 10111213141516
x core env pol
Acute patients
SFU
/2 x
105
PBM
C
Ex Vivo Elispot Analysis
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Effect of Antigen Stimulation on HBV-specific T Cell Responses and Correlation with Viral Load
0
0.2
0.4
0.6
0.8
1
1.2
1.4
17-01-2003 25-03-2003 12-05-20030
1,000,000
2,000,000
3,000,000
4,000,000
5,000,000
6,000,000
50
100
150
200
250
ALT
HB
V-D
NA
% IF
Nγ
+ ce
lls/C
D4
Pt. C1
Pt. C2
Pt. C3
Pt. C4
Pt. C5
Pt. C6
Pt. C7
050100
050100
050100
050100
050100
050100
050100
x core env pol x core env pol x core env pol
SFU
/ 2 x
105
PBM
C
Time point 1 Time point 2 Time point 3
Ex vivo
050100
050100
050100
050100
050100
050100
050100
050100
050100
050100
050100
050100
050100
050100
Pt. C1
Pt. C2
Pt. C3
Pt. C4
Pt. C5
Pt. C6
Pt. C7
x coreenv pol x coreenv pol x core env pol
Time point 1 Time point 2 Time point 3
In vitro
250500
250500
0
0250500
0250500
0250500
0250500
0
0250500
250500
250500
0
0250500
0250500
0250500
0250500
0
0250500
250500
250500
0
0250500
0250500
0250500
0250500
0
0250500
SFU
/ 2 x
105
PBM
C
Boni C, et al. J Virol 2007;81:4215-4225
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Frequency of Intrahepatic and Circulating HBV-specific CD8 Cells in Chronic HBV Infection
Fisicaro P, et al. Gastroenterology 2010;138:682-93, 693.e1-4
HBV-DNA TITER IU/mL
Mea
n %
TET
+/C
D8+
in
trah
epat
ic c
ells
0
1
2
3
4
5
6
0-104 104-105 >105
p=0.006
p=0.03
% H
BV-
TET+
/CD
8+0
0.020.040.060.080.1
12468
10LIVER BLOOD
p=0.0002
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HBV-specific T Cell Dysfunction
Possible causes
Dendritic cellimpairment
T regsuppression
Defective innate responses
DysfunctionalHBV-T cells
Hyper-production of regulatory cytokines
Persistent exposure to high Ag doses
(HBeAg-HBsAg)
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HBV-specific T Cell DysfunctionPossible causes
Persistent exposure to high Ag dosesPersistent inflammation
Amplification of negative costimulatory pathways
Impairment of TCR signaling by -chain
down-regulation
Enhanced T cell apoptosis caused by Bim up-
regulation
DysfunctionalHBV-specific T cells
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APC
T CELL
B7 FAMILY TNF SUPERFAMILY
B7.1 or B7.2
PD-L1PD-L2
PD-1 CTLA-4 CD28
ICOSL
ICOS
INHIBITION
OX40L 4-1BBL
CD40CD70
OX40 CD27 CD40L 4-1BB
COSTIMULATORY SIGNALS
Costimulatory Pathways in T Cell Activation
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PD-1/PD-L1 Pathway
Barber DL, et al. Nature 2006;439:682-687
PD-1PD-L1
Dendritic cellsLiver cells T lymphocyte
Up-regulation in viral infections
Up-regulation - activated T cells
- high antigen doses
T cell inactivation and functional exhaustion
Anti-PD-L1
T cell functional restoration
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Effect of Anti-PD-L1 Treatment on HBV-specific CD8 T Cell Function
Boni C, et al. J Virol 2007;81:4215-4225
Anti-PD-L1
T cell functional restorationBarber DL, et al. Nature 2006;439:682-687
EXPANSION IL-2 PRODUCTION
0
0.5
1
1.5
0 1 2 105
0
10
20
30
40
0 1 2 100
5
10
15
20
0 2 5 10
IFN PRODUCTION
g/mL anti-PD-L1
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CONCLUSIONS
HBV-DNA negative HBV-DNA <104 HBV-DNA >104
Antigen load
T cell efficiency+++ ++ +
Immune subjects
Asymptomatic carriers
Chronic patients
Chronic HBV infection comprises a wide spectrum of different virological and immunologicalsituations which are the expression of a complex natural history, where levels of virusreplication and levels of T cell reactivity appear to be inversely correlated and where HBVpersistence is likely caused by the synergistic effect of different mechanisms, includingexhaustion by high antigen concentrations.
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FUTURE PERSPECTIVES OF ANTI-HBV IMMUNE THERAPY(Ferrari C. Gastroenterology 2008;134:1601-1604)
T CELL DYSFUNCTION
Antiviral treatmentDecline of antigen load
Inhibition of negative costimulatory pathways
RECOVERY OF T CELL RESPONSIVENESS
VACCINATION
CONTROL OF INFECTION
Blocking antibodies
DC CD8
Viru
s/an
tigen
load