PD-1, SOCS-1, Tim-3 in HCV infection
This presentation is the property of its rightful owner.
Sponsored Links
1 / 43

PD-1, SOCS-1, Tim-3 in HCV infection -WHY WE CARE? PowerPoint PPT Presentation


  • 62 Views
  • Uploaded on
  • Presentation posted in: General

PD-1, SOCS-1, Tim-3 in HCV infection -WHY WE CARE?. Yao, Z. Q. M.D. Ph.D. Director, Hepatitis (HCV/HIV) Program, JHQ-VAMC Associate Professor, Division of Infectious Diseases Department of Internal Medicine, Quillen COM ETSU. Disclosures.

Download Presentation

PD-1, SOCS-1, Tim-3 in HCV infection -WHY WE CARE?

An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -

Presentation Transcript


Pd 1 socs 1 tim 3 in hcv infection why we care

PD-1, SOCS-1, Tim-3 in HCV infection

-WHY WE CARE?

Yao, Z. Q. M.D. Ph.D.

Director, Hepatitis (HCV/HIV) Program, JHQ-VAMC

Associate Professor, Division of Infectious Diseases

Department of Internal Medicine, Quillen COM ETSU


Disclosures

Disclosures

  • Grant funding from NIH NIAID, NIDDK, and ETSU/WFU

  • No other financial interests involved in this presentation


Pd 1 socs 1 tim 3 in hcv infection why we care

In this Presentation

We’ll talk:

  • 1. Clinical features and immunodysregulations of HCV infection

  • Negative signaling molecules such as PD-1, SOCS-1, and Tim-3

  • in control of human innate to adaptive immune responses

  • We expect to know:

  • A) how HCV employ negative signaling molecules to establish

  • chronic infection;

  • B) why we care about this – its application in the HCV

  • pathogenesis, treatment, and vaccine development


Pd 1 socs 1 tim 3 in hcv infection why we care

200 M WW

4 M U.S.

Clinical features of HCV infection

15%

PD-1, SOCS-1, Tim-3

HIV

Why the majority of infected individuals become chronic?


Pd 1 socs 1 tim 3 in hcv infection why we care

B cell hyperactivation

mixed cryoglobulinemia

T cell dysfunction

and exhaustion

non-Hodgkins lymphoma

Immunodysregulation in chronic HCV infection

Impaired Monocyte maturation into DC

Decreased IL-12

Viral Persistence

B cell clonal expansion

Th17 cell and Foxp3+ Treg cell expansions

What is the underlying mechanism leading to these immunodysregulations?


Pd 1 socs 1 tim 3 in hcv infection why we care

Mechanism leading to these immunodysregulations

The primary site of HCV infection is within the liver, where hepatic sinusoids lack basal membrane with a very low velocity of blood flow

PD-1

SOCS-1

HCV+ Huh-7 HCV- Huh-7

PD-1

Tim-3

So HCV-infected hepatocytes has ample opportunity to contact circulating or infiltrating immune cells

Tim-3+ CD14+ M/MØ

6 h 12 h 24 h 48 h


What is pd 1

  • Programmed Death-1, first identified on apoptotic cells

  • Inducible expressed receptor on immune cells upon activation

  • Provides a negative signaling to TCR positive signaling pathway

  • A powerful negative feedback mechanism to balance the +/- signal

  • Blocking PD-1 signaling will reverse T cell dysfunction

What is PD-1


Pd 1 socs 1 tim 3 in hcv infection why we care

PD-1 and T cell function / exhaustion


Pd 1 socs 1 tim 3 in hcv infection why we care

Tim-3 : a molecule different from PD-1

A new negative molecule first identified on Th1, but not Th2,

and now also found on other cell types: M/MФ, NK cells


Pd 1 socs 1 tim 3 in hcv infection why we care

Suppressor of cytokine signaling (SOCS)

– a family of negative inhibitors of cell signaling

Cytokine


Pd 1 socs 1 tim 3 in hcv infection why we care

Why we care about this?

-Negative signaling molecules in HCV pathogenesis

PD-1 & Tim-3 in Monocyte IL-12 regulation


Pd 1 socs 1 tim 3 in hcv infection why we care

Immunodysregulation in chronic HCV infection

Impaired Monocyte maturation into DC

Decreased IL-12

Viral Persistence


Pd 1 socs 1 tim 3 in hcv infection why we care

Monocyte IL-12 expression is significantly suppressed in chronic HCV infection

Healthy HCV

15.2%

Gating strategy

CD14

IL-12

60%

**

__________________________

50%

*

____________

**

____________________________

40%

PD-1+ CD14+ cells

60%

*

_______________

30%

50%

20%

40%

10%

30%

IL-12+ CD14+ cells

0%

20%

HCV-InfectedHCV-Resolved Healthy

10%

0%

HCV-InfectedHCV-Resolved Healthy

Ma et al. Immunology 2010; Zhang et al. J Immunol 2011


Pd 1 socs 1 tim 3 in hcv infection why we care

PD-1 is inversely associated with IL-12 production by monocytes

B)

Pearson Correlation = -0.464*

A)

LPS/R848

PD-1

PD-1+ CD14+ cells

IL-12

IL-12 production

C)

D)

50%

50%

P = 0.003

P = 0.034

40%

40%

PD-1+ CD14+ Cells

IL-12+ CD14+ Cells

30%

30%

20%

20%

10%

10%

0

0

before after

IFN/RBV IFN/RBV

before after

IFN/RBV IFN/RBV


Pd 1 socs 1 tim 3 in hcv infection why we care

Tim-3 is a negative molecule expressed on resting monocytes

to control IL-12 expression

Positive cells (%)

isoun-stimulatedTLR-stimulated

isoun-stimulatedTLR-stimulated

2.7%

0.2%

1.8%

Tim-3

55.3%

0.4%

2.2%

Tim-3

1.2%

3.0%

45.8%

0.1%

0.1%

38.9%

Time of TLR stimulation

IL-12

CD14

CD14

0.3%

0.2%

27.2%

IL-12

Zhang et al. JLB 2011

Tim-3 functions as a break, and TLR as the driving force for IL-12 expression


Pd 1 socs 1 tim 3 in hcv infection why we care

Tim-3/IL-12 expression in resting and activated monocytes in HCV patients

A

Healthy Subject HCV Patient

Isotype control Naïve Activated Naïve Activated

2.8

4.1

4.4

54.6

3.4

8.7

7.4

38.3

Tim-3

47.6

18.8

74.4

45.5

22.3

13.5

27.3

27.0

CD14

40.9

33.9

54.0

15.3

71.3

0.2

67.7

0.4

CD14

2.5

1.0

0.1

22.7

0.2

29.6

28.5

31.8

IL-12

**

***

B

C

NS

**

***

***

***

***

% of IL12+ CD14+ M/MØ

% of Tim-3+ CD14+ M/MØ

Healthy Subjects Chronic HCV Patients

Naïve Activated Naïve Activated

Naïve Activated Naïve Activated

Zhang et al. PLoS One 2011


Pd 1 socs 1 tim 3 in hcv infection why we care

It’s not because of TLR expression, but due to defect of intracellular signaling

A)

Healthy Subjects HCV patients

B)

Healthy Subjects HCV patients

99.5%

10.3%

40.7%

99.7%

*

TLR7+ CD14+ Cells

TLR4+ CD14+ Cells

Healthy Subjects HCV patients

Healthy Subjects HCV patients

C)

Healthy subject HCV-infected HCV-resolved

STAT-1+ CD14+ Cells

21.6%

9.6%

23.7%

*

Healthy HCV-infected HCV-resolved

D)

IgG anti-Tim-3

+ + Core

Phospho Stat1

Total Stat1


Pd 1 socs 1 tim 3 in hcv infection why we care

gC1qR

PD-1

Isotype 1.2%

LPS R848 11.7%

LPS R848+core+IgG 3.2%

LPS R848+core+a-PDL-1 8.9%

Control IgG a-PDL-1

+ + Core

SOCS-1

β-actin

HCV core

TLR

Count

* *

M/MФ IL-12 production

SOCS-1

% IL-12+CD14+ cells

* *

IL-12


Pd 1 socs 1 tim 3 in hcv infection why we care

Silencing SOCS-1 inhibits PD-1 expression and improve IL-12 production

A)

+ + + + Core

Isotype 0.73%

LPS R848 Core control siRNA 39.1%

Isotype 2.88%

48 h after transfection72 h after transfection

Control siRNASOCS-1 siRNAControl siRNASOCS-1 siRNA

LPS R848 Core SOCS-1 siRNA 2.9%

LPS R848 Core control siRNA 10.1%

LPS R848 Core SOCS-1 siRNA 19.4%

B)

SOCS-1

Count

β-Actin

PD-1

C)

Count

IL-12


Pd 1 socs 1 tim 3 in hcv infection why we care

Crosstalk between PD-1 and SOCS-1 to inhibit STAT-1/5 phosphorylations

A)

B)

Control IgG a-PDL-1

Control siRNA SOCS-1 siRNA

+ + Core

- + + + Core

pSTAT-1

pSTAT-1

Total STAT-1

Total STAT-1

D)

Control IgG Anti-PDL-1

Control IgG Anti-PDL-1

STAT-1

STAT-5

CD14

CD14

**

*

%STAT-5+CD14+ cells

%STAT-1+CD14+ cells


Pd 1 socs 1 tim 3 in hcv infection why we care

gC1qR

Our Model

LPS/R848

Gal-9

HCV core

PD-L1

Tim-3

PD-1

TLRs

Signaling pathways for

IL-12 expression (Jak/STAT)

SOCS-1

Th1/Tc1 dysregulation

Viral persistence

Viral clearance


Pd 1 socs 1 tim 3 in hcv infection why we care

B cell hyperactivation

mixed cryoglobulinemia

T cell dysfunction

/ exhaustion

non-Hodgkins lymphoma

Viral Persistence

Immunodysregulation in chronic HCV infection

Impaired Monocyte maturation into DC

Decreased IL-12

B cell clonal expansion

What is the underlying mechanism leading to these immunodysregulations?


Pd 1 socs 1 tim 3 in hcv infection why we care

Why we care about this?

-Negative signaling molecules in HCV pathogenesis

HCV infection lead to a differential effect on T/B lymphocytes - what is the underlying mechanism ?


Pd 1 socs 1 tim 3 in hcv infection why we care

CD20

CD20

CD20

CD4

CD8

19.4%

45.3%

Cell Immunology & Biology 2011

5.8%

10.4%

96.2%

83.3%

IgM

IgG

TALL-1

HCV-NHL

CD69

HCV-NHL

HCV-Tetramer

19.1%

4.2%

72.3%

48.8%

HS

HS

70.0%

6.9%

HCV-NHL

HCV-NHL

PD-1

PD-1

PD-1

47.3%

37.2%

53.5%

13.7%

HCV

HS

CD4

Tetramer CD8

CD20


Pd 1 socs 1 tim 3 in hcv infection why we care

T cells B cells

+ - + - Core

SOCS-1

-Actin

Differential regulation of T/B lymphocyte activation in patients with HCV-NHL

Differential regulation of T / B lymphocyte signaling by HCV core protein

T cells B cells

T cells B cells

T cells B cells

____________

____________

____________

____________

+ - + - Core

HCV-NHL HS HCV-NHL HS

HCV-NHL HS HCV-NHL HS

SOCS-1

SOCS-1

pSTAT1

SOCS-1

-Actin

hβ2M

Conclusion: HCV induces a differential regulation of PD-1/SOCS-1 expression, which translate into

a differential regulation of T/B lymphocyte functions through Jak/STAT pathway


Pd 1 socs 1 tim 3 in hcv infection why we care

Blocking PD-1 signaling restores T cell activation and proliferation

A)

B)

Anti-PD-L1

Control Ab

Anti-PD-L1

Control Ab

29.8%

17.1%

80%

10%

9%

1%

46%

13%

36%

5%

CD69

HS

T cell Counts

CD4

22%

36%

38%

4%

1%

9%

77%

13%

13.7%

21.3%

HCV-NHL

HCV-Tetramer

CFSE

CD8


Pd 1 socs 1 tim 3 in hcv infection why we care

Why we care about this?

-Negative signaling molecules in HCV pathogenesis

Differential regulation of IL-12/IL-23 expressions by M/MФ leads to TH17 cell and Foxp3+ Treg development


Pd 1 socs 1 tim 3 in hcv infection why we care

Differential regulation of IL-12/IL-23 expressions by M/MФ

leads to TH17 cell development during HCV infection

HCV

HS

**

IL-17A

HCV HS

CD4

IL-12 p35

IL-23 p19

**

*

Pearson r=0.465

p<0.05

IL-23/IL-12 by CD14+ cells


Pd 1 socs 1 tim 3 in hcv infection why we care

Hepatocyte

Hepatocyte

Hepatocyte

TLR

TLR

monocyte

monocyte

monocyte

monocyte

 Tim-3

STAT-3

STAT-1

 IL-23

 IL-12

HCV

 Foxp3+ Tregs

Tim-3

 TH17

Gal-9


Pd 1 socs 1 tim 3 in hcv infection why we care

Differential regulation of IL-12/IL-23 expressions by M/MФ leads to

TH17 cell and Foxp3+ Treg development during HCV infection

HS

HCV

A)

**

11.94

20.85

8.91

13.54

11.93

74.75

53.68

4.40

B)

HS

HCV

***

*

5.68

9.41

1.54

2.44

39.95

25.45

52.82

62.70


Pd 1 socs 1 tim 3 in hcv infection why we care

A)

B)

HS

HCV

HS

HCV

15.78

7.42

4.89

25.86

20.06

15.94

7.26

14.53

29.85

27.44

16.7

28.50

45.08

62.47

19.03

59.18

NS

NS

*

**

Pearson Correlation = 0.75

Sig. (1-tailed)=0.0002; (2-tailed)=0.0004

CD4+

CD4+

CD25+

CD4+

CD25+

Foxp3+

CD4+

CD25+

Foxp3-


Pd 1 socs 1 tim 3 in hcv infection why we care

Differential regulation of IL-12/IL-23 expressions by monocytes/macrophages

leads to TH17 cell and CD4+CD25+Foxp3+development during HCV infection

HCV-infected hepatocytes

produce Gal-9 and TGF-β

CD4+ T cell

 IL-23/IL-12

CD25+ FoxP3- T eff

 IL-17

CD25+ FoxP3+ T reg

Tim-3 is up-regulated more

on Foxp3+ Tregs than on Teffs

TGF-β/IL-10

monocyte

IL-2

 proliferation

 apoptosis

 apoptosis

 proliferation

Tim-3/Gal-9 interactions shift the balance of Tregs/Teffs by regulating

T cell proliferation and apoptosis

Teff

Treg

Treg

Treg

α-Tim-3

α-Tim-3 may correct the imbalance

of Tregs/Teffs ratio induced by HCV

Tim-3

/Gal-9

Moorman JP et al J Immunol 2012


Pd 1 socs 1 tim 3 in hcv infection why we care

Immunodysregulation during chronic HCV infection

HCV-infected Hepatocytes

Increased PD-1, SOCS-1, Tim-3

Decreased IL-12

Impaired CD14+ M/MΦ maturation into DC

Increased IL-23

Increased IL-17

HCV chronic infection

Accumulated TH17 & Foxp3+ Treg cells

Increased IL-10

Increased TGF-β

Autoimmune disorders

Decreased IL-2

Diminished CD4+/ CD8+ T cells

Decreased TNF-α

Decreased IFN-γ

Increased IgG

Aberrant CD19+ B cell activation

Increased IgM


Pd 1 socs 1 tim 3 in hcv infection why we care

Why we care about this?

-Negative signaling molecules in Vaccine response

HBV vaccine response and HCV vaccine development


Pd 1 socs 1 tim 3 in hcv infection why we care

Tim-3 on HBV vaccine failure in HCV-infected individuals

HBV Vaccine response: 90% in Healthy Subjects; 50% in HCV-infected patients

A)

***

HBV-R

**

HBV-NR

Isotype

*

B)

% Tim-3/CD14+ cells

% IL-23p19/CD14+ cells

% IL-12p35/CD14+ cells


Pd 1 socs 1 tim 3 in hcv infection why we care

PD-1/SOCS-1 on HBV vaccine failure in HCV-infected individuals

9.4% vs 4.9%, P=0.007

12.1% vs 7.0%, P=0.002

5.6% vs 4.5%, P>0.05

Pearson Corr. = - 0.374**

Sig.(2-tailed) = 0.001

PD-1 expression on CD4+ T cells

CD69+ CD4+ T cells

PD-1+ CD4+ T cells

HCV patients

HBV-NR (n=29)

HCV patients

HBV-R (n=32)

HCV resolved

individuals(n=6)

Healthy

Subjects(n=10)

HBsAg stimulation a-CD3/28 stimulation

*

HBV-R HBV-NR HBV-R HBV-NR

*

SOCS-1/actin

SOCS-1

β-actin

HBV-R HBV-NR HBV-R HBV-NR

HBsAg stimulation a-CD3/28 stimulation


Pd 1 socs 1 tim 3 in hcv infection why we care

Control Ab a-PD-L1 Ab

A)

IgG

*

18.1%

29.9%

a-PDL-1

HBsAg stimulation

*

% CD69+ in CD4+ T cells

31.6%

57.8%

HBsAg stimulation a-CD3/28 stimulation

a-CD3/28 stimulation

IgG

Control Ab a-PD-L1 Ab

B)

a-PDL-1

0% 0.3% 7% 92%

1.5% 8% 18% 73%

CFSE / HBsAg

*

HBsAg stimulation

*

M1 M2 M3 M4

IgG

1% 21% 58% 20%

0% 1.5% 19% 79%

*

a-PDL-1

a-CD3/28 stimulation

*

*

CFSE / a-CD3/28

M1 M2 M3 M4


Pd 1 socs 1 tim 3 in hcv infection why we care

Why do we care - Listeriamonocytogenes (Lm)-based DC-targeting HCV vaccine Development

HCV vaccine development: HCV-quasispecies; HCV-delivery; HCV-models; HCV-exhaustion

Listeria

monocytogenes

HCV

antigens

Virulence

determinants

Lm-NS5B

∆actA/∆inlB

Lm vector

NS5B

Lm-infected DC


Pd 1 socs 1 tim 3 in hcv infection why we care

Why do we care?

Improve Lm-based DC-targeting HCV vaccine by blocking Tim-3 signaling

M/MФ

BSA-FITC Uptake (ΔMFI)

iDC

%Tim-3+ cells

%IL-12 + cells

mDC

Un-infected HCV HCV+IgG HCV+a-Tim-3


Pd 1 socs 1 tim 3 in hcv infection why we care

Why do we care?

Improve Lm-based DC-targeting HCV vaccine by blocking Tim-3 signaling

%HCV-Tet+/CD3+CD8+

HCV-Tetramer

%Tim-3+/HCV-Tet+CD3+CD8+

Lm-control

Lm-NS5B

SSC

HCV-Tetramer

α-Tim-3

IgG

IgG

α-Tim-3

A)

Gating strategy

CD3+CD8+

Tim-3

CD3+CD8+

B)

Iso

HCV-resolved

HCV-infected


Pd 1 socs 1 tim 3 in hcv infection why we care

Why do we care?

Listeria monocytogenes (Lm)-based DC-targeting HCV vaccine

A)

%IFN-r+/HCV-Tet+CD3+CD8+

IFN-γ

B)

%GranzymeB+/HCV-Tet+CD3+CD8+

Granzyme-B


Pd 1 socs 1 tim 3 in hcv infection why we care

Why do we care?- novel therapeutics

HCV Core

MHC/peptide/B7

PD-L1

LPS

Gal-9

gC1qR

TLR

PD-1

CD3

CD28

Tim-3

STAT

Monocyte IL-12

α-gC1qR

α-HCV core

SOCS

T cell activation

α-PD-1

α-Tim-3

Negative T cell regulators

Improve HBV vaccine response

in HCV/HIV-infected individuals

T cell dysfunction

viral clearance

Improve HCV - DC

therapeutic Vaccine

HCV persistence


Pd 1 socs 1 tim 3 in hcv infection why we care

Acknowledgements

Dr. T. Niki: President of GalPharm, Japan; Dr. T.J. Liang, Chief Liver Dis, NIH NIDDK

Dr. T Wakita, Director Virology Lab, NIH, Japan; Dr. D. Brockstedt, VP of Aduro BioTech, CA


  • Login