Efficacy of Combinatorial anti-cancerchemotherapy and immunotherapy:

A central role of dendritic cellsMohamed Labib Salem, PhD

Prof. of ImmunologyZoology Department, Faculty of Science

Director, Competitive Project UnitTanta University, Egypt

Mohamed Labib Salem, PhDProf. of Immunology

Zoology Department, Faculty of ScienceDirector, Competitive Project Unit

Tanta University, Egypt

Key Lab. Infection and ImmunityCenter for Infection and Immunity

Institute of BiophysicsChinese Academy of Sciences

February 9, 2012

The Beginning of Cancerous Growth:

A process that mimics the cancer grows in Egyptbefore January 25th, 2011

Underlying tissue

EGYPT

Cancer TherapyIn clinic

• Surgery:Effective but doesn’t cure tumormetastasis or prevent tumorrecurrence

• Radiotherapy/Chemotherapy:Effective but doesn’t preventtumor recurrence besides itsserous side effects

• Surgery:Effective but doesn’t cure tumormetastasis or prevent tumorrecurrence

• Radiotherapy/Chemotherapy:Effective but doesn’t preventtumor recurrence besides itsserous side effects

Cancer TherapyIn clinical trials

• Immunotherapy:can cure local and distanttumors and prevent tumorrecurrence through developingtumor-specific memory T cellresponses

• Immunotherapy:can cure local and distanttumors and prevent tumorrecurrence through developingtumor-specific memory T cellresponses

Cancer ImmunotherapyCancer Immunotherapy AntibodiesAntibodies

Cytokines (ILCytokines (IL--22, IFN, IFN--a)a) Cytokines (ILCytokines (IL--22, IFN, IFN--a)a)

Adoptive T Cell TherapyAdoptive T Cell Therapy

VaccinesVaccines

Activation of DCs is essential to prime T cells

?

? ?

??

?

Cancer cells originate from self cells (i.e. not foreign)• Poorly immunogenic• Induce very weak (undetectable) immune response

Microbes are foreign• Immunogenic• Induce strong immune response

Cancer cells can be immunogenic if a danger signalinduced by microbial products

Challenge to cancer Immunotherapy:1- Tumor antigens are self

Cancer cells originate from self cells (i.e. not foreign)• Poorly immunogenic• Induce very weak (undetectable) immune response

Microbes are foreign• Immunogenic• Induce strong immune response

Cancer cells can be immunogenic if a danger signalinduced by microbial products

Down regulate the receptors required to interactwith immune cells

Upregulate receptors that suppress immune cellsupon interaction

Produce factors that suppress immune cellfunctions

Favors expansion of suppressor immune cells

Challenge to cancer Immunotherapy:2- Tumor cells escape from immune cells

Down regulate the receptors required to interactwith immune cells

Upregulate receptors that suppress immune cellsupon interaction

Produce factors that suppress immune cellfunctions

Favors expansion of suppressor immune cells

1:10,000 T cells 1:50 T cells 1:2 T cells

Dis

ease

Bur

den

Challenge to cancer Immunotherapy:3- High tumor cells/T cell ratio

ClinicallyUn-detectable

LocalizedDisease

AdvancedDisease

Dis

ease

Bur

den

Adoptive T cell transfer therapy

2- Grow T cells

With IL-2

3- Test T cellfunction

5- Infusionof T cells+ vaccine

4- Clonalexpansion of Tcells with IL-2

& CD3 Ab

6- Chemoor IrradiationBefore T cell

transfer

5- Infusionof T cells+ vaccine

1- Tumor excision

Chemotherapy and Immunotherapy““Removal of weeds, and planting and watering good seeds”Removal of weeds, and planting and watering good seeds”

Generation ofT cells in vitro

Strategies to Improving Adoptive T Cell Therapy

Vaccineadjuvant

ChemotherapyIrradiation

In vitro In vivoIn vivo

Our Model of ChemotherapyOur Model of Chemotherapy

Systemic injection ofSystemic injection ofcyclophosphamidecyclophosphamide (CTX(CTX))200200 mg/kg (mg/kg (44 mg/mouse)mg/mouse)

Systemic injection ofSystemic injection ofcyclophosphamidecyclophosphamide (CTX(CTX))200200 mg/kg (mg/kg (44 mg/mouse)mg/mouse)

Our models for adoptive T cell transfer

(Donor)Transgenic

Ly 5.1 mouse

(Recipient)Naive

Ly 5.2 mouse

CD8+ T cellsCD8 T cells are engineered torecognize tumor antigens

1 x 106

I.V. injection

Donor CD8+ T cells(0.2 %) are clearlyvisualized &quantified

CD49low

CD44low

CD69low

CD62Lhigh

Chemotherapy and T cell TherapyChemotherapy and T cell TherapyProtocolProtocol

PBSCTX

CD8+ T cell1 x 106

Antigenpriming

Antigenboosting

Readout(Number of Donor Cells)

d0 d1 d2 d3 d5 d7 d70

CTX increased Ag-specific expansion of transferredCD8 cells during effector and memory responses

0

5

10

15

20

25

3 6 7 8 14 21

%O

T-1

cells

in P

BL

PBSCTX 1 mgCTX 4 mg

Of priming

00.5

11.5

22.5

33.5

4

PBS CTX

%O

T-1

cells

in P

BL

Day 70

0

2

4

6

8

10

12

PBL Spleen

% O

T-1

cells

Re-vaccination

0

2

4

6

8

10

12

PBL Spleen

%O

T-1

cells

in P

BL PBS

CTX

Day 7 Day 14

3 6 7 8 14 21Days post vaccination

Salem et al. J Immunother 30(1): 40-53 (2007)

How chemotherapy alters hostHow chemotherapy alters hostmicroenvironmentmicroenvironment

0

2000

4000

6000

8000

10000#

PBL

(106 /L

)Blood

00 3 6 9 12 15 18

0

20

40

60

80

100

0 3 6 9 12 15 18

Days post CTX treatment

# Le

ucoc

ytes

(106 )/o

rgan

SpleenBM

Spleen and Bone marrow

Before vaccination

0.2

0.4

0.6

0.8

1

1.2

%O

T-1

cells

in P

BL

Tota

l num

ber (

106 )

per

sple

en

1020304050607080

**

**

Mechanisms of action of CTX:1- Induces homeostatic expansion of transferred cells

0

0.2

PBS CTX -6 hr CTX -24 hr CTX -48hr%

OT-

1ce

lls in

PB

L

0

10

20

30

40

50

60

70

PBS CTX -6 hr CTX -24 hr CTX -48hr

%O

T-1

cells

in P

BL

on d

ay7

After vaccination

Tota

l num

ber (

106 )

per

sple

en

010

0 6 24 72Hr after CTX

Salem et al. J Immunother 30(1): 40-53 (2007)

1820

%O

T-1

cells

in P

BL

on d

ay3

Mechanisms of action of CTX:2- A space “niche” is not a critical factor

d0 d1

ACTi.v.

Vacs.c.

d-2

CTXi.p.

Measuring Ag-specific expansion of adoptivelytransferred CD8+ T cells

Filling the space

02468

1012141618

PBS CTX%O

T-1

cells

in P

BL

on d

ay3

Salem et al. J Immunother 30(1): 40-53 (2007)

CTX+50m

CTX+100m

CTX+150m

Mechanisms of action of CTX:3- Decreases the numbers of regulatory T cells

Mechanisms of action of CTX:4- Type I IFNs are critical factors

1 4 8 24 48 72 96

IFN

-(p

g/m

l)

0102030405060708090

100

PBS

CTX

1 4 8 24 48 72 96Hours post CTX (4 mg) treatment

Days post vaccination

% C

D8

T ce

lls in

PB

L

0

2

4

6

8

10

12

3 5 7 45

PBS

CTX/Wild Type

CTX/IFNaKO

Salem et al. J Immunother 30(1): 40-53 (2007)

Mechanisms of action of CTX:5- Inducing activation of dendritic cells

Salem et al. J Immunother 30(1): 40-53 (2007)

Induction ofactivation of

dendritic cells

Eliminationof

suppressivecells

Creation of a space “niche”

How Does Chemotherapy BenefitImmunotherapy?

Induction ofactivation of

dendritic cells

Eliminationof

suppressivecells

Induction ofsurvival

cytokines

Generation ofT cells in vitro

Strategies to Improving Adoptive T Cell Therapy

Vaccineadjuvant

ChemotherapyIrradiation

In vitro In vivoIn vivo

Focusing on Dendritic CellsAfter Chemotherapy

Focusing on Dendritic CellsAfter Chemotherapy

Dendritic Cells are A Critical player in Anti-Tumor Immunity

No response

TLRLs

No response

Mechanisms of action of CTX:Inducing expansion of DCs during restoration phase

CD

11c

CD11b

5.6± 1.1 3.5± 0.9 6.4± 0.8 18.7± 4.125.2± 1.1

Lymphopenic phase Restoration phaseDay 0 Day 2 Day 6 Day 9 Day 12

CD11b

0200400600800

100012001400

0 2 6 10 14Days post CTX treatment

# C

D11

c+ CD

11b+ c

ells

(106 /L

)

PBS CTX

CTX treatment induces an expansion of DCs

Post CTX expanded DCs express the phenotype of immature myeloid cells

d4 d14d0 d2 d10

CD80

4.5% 25% 35% 2.1%7.2%

Lymphopenic phase Restoration phase

35

Myeloid DCs (mDCs): CD11chighCD11bhighLy6GlowB220low

Plasmacytoid DCs (pDCs): CD11chighCD11blowLy6GhighB220high

Day 12 post treatment

0

5

10

15

20

25

30

35

mDCs pDCs

% C

D11

c+ce

lls

PBSCTX

Is dose-dependentInduces proliferation of progenitors of

dendritic cells in bone marrowInduces production of stem cell

mobilizing factors Flt3 and CCR2 signaling is critical

How CTX Induces Dendritic cellexpansion?

Is dose-dependentInduces proliferation of progenitors of

dendritic cells in bone marrowInduces production of stem cell

mobilizing factors Flt3 and CCR2 signaling is critical

Salem et al. 2009 J ImmunologySalem et al., 2010 J ImmunologySalem et al., 2010 Cell Immunology

Phases post CTX treatment relative toPhases post CTX treatment relative toadoptive T cell therapyadoptive T cell therapy (Salem CII(Salem CII 20102010))

• Creation of a space niche due tothe induced lymphopenia

• Homeostatic expansion of T cells• Elimination of regulatory cells• Elimination of cytokine

competition• Microbial translocation• Activation of dendritic cells

• Cellular recovery from lymphopenia• Less of lymphopenia• Less homeostatic proliferation of T cells• Expansion of immature dendritic cells

Num

ber o

f den

driti

c ce

lls

Lymphopenic phase Recovery phase

DonorT cells

3 6 9 12 15 18Days after CTX treatment

Num

ber o

f den

driti

c ce

lls

0-1

ACT Antigen priming+ TLR agonists

Antigen boosting+ TLR agonists

20

DCs

DonorT cells

Can post CTX expanded DCs be matured andbenefit anti-tumor CD8+ T cell responses?

Can post CTX expanded DCs be matured andbenefit anti-tumor CD8+ T cell responses?

How to Mature Dendritic CellsHow to Mature Dendritic Cells

TollToll--Like Receptor (TLR) ligandsLike Receptor (TLR) ligandsDendritic cells express TLRsDendritic cells express TLRsCytokinesCytokines

TollToll--Like Receptor (TLR) ligandsLike Receptor (TLR) ligandsDendritic cells express TLRsDendritic cells express TLRsCytokinesCytokines

Maturation of dendritic cells with TLRLs

Bachmann et al. Nature Reviews Immunology 6: 159–164 (2006)

TLRagonists

TLRs Alert Hosts From Invaders

NK cellsMacrophagesDendritic cells

TLRs and their specific TLR ligands

Microbial products

Dendritic cells

TL3 agonist (Poly (I:C))• Poly (I:C) is a synthetic double stranded RNA

which mimics viral products

• It binds specifically to TLR3 expressed in DCs.

• TLR3/TLRL triggers inflammatory mediatorsand activate DCs.

• We and others have established that poly(I:C) isa potent adjuvant for CD8+ T cell responses indifferent vaccination settings.

Salem et al. J Immunother 28:220-228 (2005)Salem et al. Vaccine 24:5119-5132 (2006)

• Poly (I:C) is a synthetic double stranded RNAwhich mimics viral products

• It binds specifically to TLR3 expressed in DCs.

• TLR3/TLRL triggers inflammatory mediatorsand activate DCs.

• We and others have established that poly(I:C) isa potent adjuvant for CD8+ T cell responses indifferent vaccination settings.

Can Poly(I:C)-activated dendritic cellsbenefit T cell responses to vaccination with a

self/tumor antigen?

Can Poly(I:C)-activated dendritic cellsbenefit T cell responses to vaccination with a

self/tumor antigen?

Can Poly(I:C) Activate Dendritic CellsExpanded after CTX Treatment?

2.36 1.35 36.3

40.7 28.2 83.3

None PolyPBS

LymphNodes

CTXNone PolyPBS

CTX

1.8 1.6 4.8

49.534.7

Blood

85%

CCR7CD80

TLR3 agonist activates DCs and drives theirhoming from blood to lymph nodes

15

20

25

30

15

20

25

30%

CD

11c+

CD

11b+

DC

s

0

5

10

15

0

5

10

15

Blood Lymph nodes

%C

D11

c+C

D11

b+ D

Cs

CTXPBS

CTXPoly

CTXPBS

CTXPoly

PBSCTX

T cells106

Melanomaantigen± poly(I:C)

Melanomaantigen± poly(I:C)

B16s.c.

Testing prime-boost vaccination strategy

Prime Boost

d0 d1 d2 d12d-12 d15

Readout

PmelPmel--11 adoptive transfer mouse modeladoptive transfer mouse model

Naïve spleenCD8+ T cells

(500,000)

Transgenicmice

Tumorcells

Vaccine+Poly(I:C)

Vaccine+Poly(I:C)

CD8 T cells react against gp100 melanoma peptide

Naïve spleenCD8+ T cells

(500,000)Tumorcells

D-2CTX

Vaccine+Poly(I:C)

D -1 D 0 D 12D -10

DCs are expandedT cells can grow

Vaccine+Poly(I:C)

Stimulation of post CTX expanded DCs is essential foraugmenting CD8+ T cell responses

0

0.05

0.1

0.15

0.2

0.25

0

500

1000

1500

2000

2500

#pm

el-1

in D

LNs

(106 )

0.02% 3.1% 0.6%

39.4%

0.04%0.05%

0.1% 0.2%4.5%

2.1%0.1%

12.5%#

pmel

-1in

PB

L (1

06 /L)

+-+---Poly I:C++++--gp100++--+-CTX++++--B16

+-+---++++--++--+-++++--

# D

Cs i

n PB

L (1

06 /L)

# D

Cs i

n D

LNs

(106 )

0

0.03

0.06

0.09

0.12

0.15

0.18

0

250

500

750

1000

1250

1500

6.0%14.9%

7.6%

18.2%

14.6%

6.5%

1.9% 1.7%3.6%

4.6%

9.1%

2.2%

Boosting at the peak of post CTX DC expansion is essential toestablish therapeutic anti-tumor immunity

050

100150200250300350400450

9 12 15 18 21 24 27 30 33Days post tumor inoculation

Tum

or ar

ea (m

m2 )

Two vaccinations

Single vaccination

0 10 20 30 400

20

40

60

80

100 PBS (No pmel-1)CTX (No pmel-1)PBS/gp100 + poly I:C d1

CTX/gp100 + poly I:C d1CTX/gp100 + poly I:C d1 and d12

PBS/gp100 + poly I:C d1 and d12

Days post tumor inoculation

% Su

rviva

l

050

100150200250300350400450

9 12 15 18 21 24 27 30 33Days post tumor inoculation

Tum

or ar

ea (m

m2 )

Boosting with antigen/poly(I:C) at the peak of post CTXDC expansion is effective against advanced tumor

250

300

350

400

450

Tum

or a

rea

(mm

2 )

PBS (No T cells)CTX(No T cells)

CTX/Vac + IL-2

PBS/Vac + poly(I:C)

0

50

100

150

200250

12 14 17 20 22 24 26 28 32

Days post tumor inoculation

Tum

or a

rea

(mm

2 )

CTX/Vac+ poly(I:C)

Tripartite regimen generates memory T cells with robustTripartite regimen generates memory T cells with robustantianti--tumor immunitytumor immunity

250

300

350

400

450

Tum

or a

rea

(mm

2 ) PBS (no 2nd ACT) CTX (no 2nd ACT)

CTX/hgp100 (2nd ACT)

0

50

100

150

200

Days post tumor inoculation

Tum

or a

rea

(mm

2 )

11 14 16 18 22 26 28

CTX/hgp100 (2nd ACT)

CTX/hgp100+poly(I:C)(2nd ACT)

Do DCs play a critical role in mediation of the beneficialeffects of CTX?

Do DCs play a critical role in mediation of the beneficialeffects of CTX?

Depletion of post CTX expanded DCs abrogates CTX effects

0

1

2

3

4

5

PBS CTX PBS CTX0

0.2

0.4

0.6

0.8

PBS CTX PBS CTX

#pm

el-1

in sp

leen

(106 )

#pm

el-1

in D

LNs (

106 )

1.5%

11.6%

4.5%2.2%

25.2%

Spleen LNs

0

1

2

3

4

5

PBS CTX PBS CTX0

0.2

0.4

0.6

0.8

PBS CTX PBS CTX

#pm

el-1

in sp

leen

(106 )

#pm

el-1

in D

LNs (

106 )

WT DTR Tg WT DTR Tg

0.8%

2.2%

0.03% 2.4%

Poly(I:C) induces DCs to produce inflammatorycytokines

0

200400600800

100012001400

16001800

TNFα

(pg/

ml s

erum

)

14000

16000

MC

P1(p

g/m

l ser

um)

0

2000400060008000

10000

1200014000

1600018000

IL-6

(pg/

ml s

erum

)

300

350

IFN

-γ(p

g/m

l ser

um)

0

2000

4000

6000

8000

10000

12000

14000

MC

P1(p

g/m

l ser

um)

PBS

Poly

CTX

/pol

y

DTR

/CTX

+ po

ly

0

50

100

150

200

250

300

IFN

-γ(p

g/m

l ser

um)

PBS

Poly

CTX

/pol

y

DTR

/CTX

+po

ly

Treatment with G-CSF post CTX therapy increasesthe numbers of DCs

1.5

2

2.5

3

3.5

#D

endr

itic

cells

(10^

6/m

l)

CTX

PBS

CTX + G-CSF

Day 7 Day 9 Day12

Days post CTX treatment

0

0.5

1

1.5

#D

endr

itic

cells

(10^

6/m

l)

DCs number increased in breast cancer patientsDCs number increased in breast cancer patientstreated with CTXtreated with CTX--based chemotherapybased chemotherapy

Patient P14 P17 P26 P29 P36 P40 P45 P47 P54 P56 P66 P74

Baseline 175.1 264.6 696.6 810.3 591.6 660.9 883.8 141.0 489.3 253.3 395.6 252.0

AC 216.5 701.5 872.9 1750.7 865.8 921.0 854.8 144.6 592.8 881.4 938.9 561.4

% Change 20 160 20 115 50 40 -3 2.5 21.1 240 137 123

Blood were collected from newly diagnosed breast cancer patients (n=12; stage I-IV) who hadreceived four courses of standard ddAC: doxorubicin 60 mg/m2 plus CT 600 mg/m2 on day 1,followed by paclitaxel 175 mg/m2 (ddT) on day 1 for four cycles. The numbers of HLA-DR+ cells wereanalyzed by flow cytometry and expressed as the changes relative to the healthy donor.

Chemotherapy with CTX induces markedChemotherapy with CTX induces markedincreases in the numbers of DCs inincreases in the numbers of DCs in 1212 days.days.

GG--CSF corrects the lymphopenia induced byCSF corrects the lymphopenia induced byCTX but increases the expansion of DCs.CTX but increases the expansion of DCs.

CTXCTX--expanded DCs express immatureexpanded DCs express immaturephenotypephenotype

Poly(I:C) can mature CTXPoly(I:C) can mature CTX--expanded DCs inexpanded DCs invivovivo

Poly(I:C)Poly(I:C)--matured DCs induce therapeuticmatured DCs induce therapeuticCDCD88+ mediated anti+ mediated anti--tumor immunitytumor immunity

ConclusionsConclusions Chemotherapy with CTX induces markedChemotherapy with CTX induces marked

increases in the numbers of DCs inincreases in the numbers of DCs in 1212 days.days. GG--CSF corrects the lymphopenia induced byCSF corrects the lymphopenia induced by

CTX but increases the expansion of DCs.CTX but increases the expansion of DCs. CTXCTX--expanded DCs express immatureexpanded DCs express immature

phenotypephenotype Poly(I:C) can mature CTXPoly(I:C) can mature CTX--expanded DCs inexpanded DCs in

vivovivo Poly(I:C)Poly(I:C)--matured DCs induce therapeuticmatured DCs induce therapeutic

CDCD88+ mediated anti+ mediated anti--tumor immunitytumor immunity

Strategies to Improving Adoptive T Cell Therapy

Vaccineadjuvant

Generation of Tcells in vitro

ChemotherapyIrradiation

In vivoIn vitro In vivo

CD8+ T cells arespecific to the MHC-1gp10025-33 melanoma

peptide

DonorTg Pmel

Culture T cells and stimulate with +gp10025-33 peptide ± IL12 for 3~5 days

Pept + IL12(pmelIL12 )

Pept alone(pmelsham )

Improving T cell functions in vitro beforetheir adoptive transfer in vivo

CD8+ T cells arespecific to the MHC-1gp10025-33 melanoma

peptide

CD44

CD

62L

Pept + IL12(pmelIL12 )

Pept alone(pmelsham )

PBS

CTX

PBS

CTX

Vaccination+ Poly (I:C)

Conditioning of pmelConditioning of pmel--11 cells with ILcells with IL--1212induces activation phenotypeinduces activation phenotype

Conditioning pmelConditioning pmel--11 cells with ILcells with IL--1212 enhancesenhancestheir function in vitro and expansion in vivotheir function in vitro and expansion in vivo

In press

30405060708090

% P

mel

cel

ls in

PB

L

Conditioning pmelConditioning pmel--11 cells with ILcells with IL--1212enhances their systemic expansion in vivoenhances their systemic expansion in vivo

0102030

% P

mel

cel

ls in

PB

L

CD8sham CD8IL12

PBS CTX PBS CTX

Improving T cell functions in vitro before their adoptivetransfer in vivo increases anti-tumor responses

Days post pmel-1 cell transfer

0

50

100

150

200

250

300

350

9 12 15 18 21 24 27

Tum

or a

rea

(mm

2 )

Pmelsham (5 x 106)PmelIL12 (5 x 106)

PmelIL12 (1 x 106)

NonePmelsham (1 x 106)

Days post pmel cell transfer

0

50

100

150

200

250

300

350

400

9 12 15 18 21 24

Tum

or a

rea

(mm

2 )

NonePmelsham

Pmelsham+gp100PmelIL12

PmelIL12+gp100

Days post pmel-1 cell transfer

0 5 10 15 20 25 30 35 400

20

40

60

80

100

Days post pmel cell adoptive transfer

Per

cent

sur

viva

l

Days post pmel-1 cell transfer

Tum

or a

rea

(mm

2)

050

100150200250300350400450

4 8 12 16 20 24 28 32 36 40 44

No CTX, no pmelCTXPmelsham

Pmelsham+gp100PmelIL12

PmelIL12+gp100

Days post pmel cell transfer

ILIL--1212 induces pmelinduces pmel--11 cells to acquirecells to acquirestem cell phenotypestem cell phenotype

Current Preclinical Protocols developed atNational Cancer Institute, USA

1.Total body irradiation2. Adoptive transfer of tumor-specific T cells3. Vaccination with Viral vector encoding tumor

antigen or dendritic cells loaded with antigen4. Treatment with high dose of IL-2 for 5 days

1.Total body irradiation2. Adoptive transfer of tumor-specific T cells3. Vaccination with Viral vector encoding tumor

antigen or dendritic cells loaded with antigen4. Treatment with high dose of IL-2 for 5 days

This strategy is significantly effective in melanomasetting but very toxic

Our treatment strategy

1.Treatment with cyclophosphamide (CTX)2. Adoptive transfer of tumor-specific T cells3. Priming with tumor antigen during the

lymphopenic phase post CTX therapy +Poly(I:C)

4. Boosting with tumor antigen at the peak ofdendritic cell expansion + Poly(I:C)

1.Treatment with cyclophosphamide (CTX)2. Adoptive transfer of tumor-specific T cells3. Priming with tumor antigen during the

lymphopenic phase post CTX therapy +Poly(I:C)

4. Boosting with tumor antigen at the peak ofdendritic cell expansion + Poly(I:C)

This strategy is significantly effective in melanomasetting but non toxic and less expensive

Replacement of ex vivoReplacement of ex vivo--based DCbased DCvaccination with post chemotherapyvaccination with post chemotherapyexpanded DCsexpanded DCs

Replacement of highly toxic ILReplacement of highly toxic IL--22 withwitheffective and noneffective and non--toxic poly(I:C)toxic poly(I:C)

NonNon--viral delivery of antigen forviral delivery of antigen forvaccinationvaccination

Adoptive transfer of ILAdoptive transfer of IL--1212 activated Tactivated Tcells can further augments the anticells can further augments the anti--tumor immunity.tumor immunity.

Significance of our FindingsSignificance of our Findings

Replacement of ex vivoReplacement of ex vivo--based DCbased DCvaccination with post chemotherapyvaccination with post chemotherapyexpanded DCsexpanded DCs

Replacement of highly toxic ILReplacement of highly toxic IL--22 withwitheffective and noneffective and non--toxic poly(I:C)toxic poly(I:C)

NonNon--viral delivery of antigen forviral delivery of antigen forvaccinationvaccination

Adoptive transfer of ILAdoptive transfer of IL--1212 activated Tactivated Tcells can further augments the anticells can further augments the anti--tumor immunity.tumor immunity.

Cancer immunotherapyCombination, Combination, Combination …..

Nature 446, 964-966 (26 April 2007)

Future StudiesFuture Studies

Does expansion of DCs occur in cancerDoes expansion of DCs occur in cancerpatients?patients?

Can poly(I:C)Can poly(I:C) mature DCs in cancermature DCs in cancerpatinetspatinets??

Is vaccination/poly(I:C)Is vaccination/poly(I:C) of cancer patientsof cancer patientssufficient to generate antisufficient to generate anti--tumor responses?tumor responses?

What cancer patients and what cancerWhat cancer patients and what cancerpopulation can benefit from post CTXpopulation can benefit from post CTXexpansion of DCs?expansion of DCs?

Does expansion of DCs occur in cancerDoes expansion of DCs occur in cancerpatients?patients?

Can poly(I:C)Can poly(I:C) mature DCs in cancermature DCs in cancerpatinetspatinets??

Is vaccination/poly(I:C)Is vaccination/poly(I:C) of cancer patientsof cancer patientssufficient to generate antisufficient to generate anti--tumor responses?tumor responses?

What cancer patients and what cancerWhat cancer patients and what cancerpopulation can benefit from post CTXpopulation can benefit from post CTXexpansion of DCs?expansion of DCs?

ACKNOLWEDGEMENTSMedical UniversityMedical University University of Miamiof South Carolinaof South Carolina Alberto Montero, MDAlberto Montero, MDDavid J.David J. Cole, MDCole, MD Diaz Montero, MDDiaz Montero, MDMike Nishimura,Mike Nishimura, PhDPhDMarkMark RubsteinRubstein, PhD, PhD OncovirOncovir, Inc, Washington, Inc, WashingtonAmir AAmir A AlkhamiAlkhami,, PhDPhD Andreas Salazar, MDAndreas Salazar, MDMarcela Montero,Marcela Montero, PhDPhDAndreAndre KadimaKadima, MD, MD Tanta University, EgyptTanta University, EgyptYianYian Chen,Chen, PhDPhD SabrySabry ELEL--NaggarNaggar, PhD, PhDOsama Naga,Osama Naga, DDSDDS RandaRanda AlAl--NaggarNaggar,, PhdPhdElizabeth Little,Elizabeth Little, BScBSc WaelWael Y.Y. AttiaAttia, PhD, PhDRickRick PepplerPeppler,, MSMSNarenderNarender NathNath,, PhDPhD

Suez Canal University, EgyptSuez Canal University, EgyptAhmedAhmed KhafagyKhafagy,, PhDPhD

Medical UniversityMedical University University of Miamiof South Carolinaof South Carolina Alberto Montero, MDAlberto Montero, MDDavid J.David J. Cole, MDCole, MD Diaz Montero, MDDiaz Montero, MDMike Nishimura,Mike Nishimura, PhDPhDMarkMark RubsteinRubstein, PhD, PhD OncovirOncovir, Inc, Washington, Inc, WashingtonAmir AAmir A AlkhamiAlkhami,, PhDPhD Andreas Salazar, MDAndreas Salazar, MDMarcela Montero,Marcela Montero, PhDPhDAndreAndre KadimaKadima, MD, MD Tanta University, EgyptTanta University, EgyptYianYian Chen,Chen, PhDPhD SabrySabry ELEL--NaggarNaggar, PhD, PhDOsama Naga,Osama Naga, DDSDDS RandaRanda AlAl--NaggarNaggar,, PhdPhdElizabeth Little,Elizabeth Little, BScBSc WaelWael Y.Y. AttiaAttia, PhD, PhDRickRick PepplerPeppler,, MSMSNarenderNarender NathNath,, PhDPhD

Suez Canal University, EgyptSuez Canal University, EgyptAhmedAhmed KhafagyKhafagy,, PhDPhD

• TWAS, The Academy of Sciences forThe Developing World (Third WorldAcademy of Sciences)

• CAS, Chinese Academy of Sciences• Prof. Shengedian Wang, MD/PhD

Director, Key Lab. Infection and Immunity,Institute of Biophysics, Chinese Academyof Sciences, Beijing

SPECIAL THANKS• TWAS, The Academy of Sciences for

The Developing World (Third WorldAcademy of Sciences)

• CAS, Chinese Academy of Sciences• Prof. Shengedian Wang, MD/PhD

Director, Key Lab. Infection and Immunity,Institute of Biophysics, Chinese Academyof Sciences, Beijing

THANKYOUTHANKYOU