noi principii in vaccinare c leclerc 2014

Nouvelles perspectives en

vaccinologie

AAEIP, Université Paris Sud, 31 Mars 2014

Claude Leclerc

DEVELOPPEMENT of HUMAN VACCINES

Live attenuated vaccines

Genetically engieneered

Purified protein or polysaccharide Killed vaccines

Smallpox, 1798 Rabies, 1885 BCG, 1927 Yellow fever, 1935 Polio (oral) Measles Mumps Rubella Adenovirus Typhoid (Ty21a) Varicella Rotavirus

Diphteria, 1923 Tetanus, 1927 Pneumococcus Meningococcus Haemophilus influenzae PRP Hepatitis B (plasma derived) Tick-birne encephalitis H. influenzae PRP conjugate Typhoid (Vi) Acellular pertussis

Typhoid 1896 Cholera, 1896 Plague, 1897

Pertussis, 1926 (killed bacteria) Influenza, 1936 Rickettsia, 1938

Polio (injected) Rabies (new) Japanese Encephalitis Hepatitis A

Hepatitis B (recombinant) Human Papilloma virus Rotavirus

18th Century

19th Century

Early 20th Century

After World War II (cellular culture)

Vaccines have been made for 36 of >400 human pathogens

Immunological Bioinformatics, The MIT press.

+HPV & Rotavirus

The different types of vaccines

Attenuated Vaccines

Killed Vaccines

Acellular sub-unit vaccines

Pertussis Diphteria Hepatitis B Tetanus

Cholera Pertussis Hepatitis A Polio

Polio Yellow fever BCG

New and improved technologies and resulting vaccines

R Rappuoli, CW. Mandl, S Black & E De Gregorio Nature Reviews Immunology Published online 4 November 2011

Dengue epidemiology

Nature Reviews Microbiology 2010

Dengue vaccines under development

Dengue vaccines under development

Sanofi Pasteur dengue vaccine enters phase III clinical study in October 2010

The yellow fever 17D virus as a platform for new live attenuated vaccines

Worldwide map of phase II/III dengue clinical trials, and major results obtained so far in humans

Guy et al, Vaccine, 2011, 7229-7241

Lancet, Published Online, September 11, 2012

Serotype-specific and overall efficacy of CYD tetravalent dengue vaccine against virologically confirmed dengue disease

Reverse cumulative distribution of serotype-specific PRNT 50 antibody titres curves for DENV serotypes 1–4 by baseline FV-serostatus, pre-vaccination and after two and three doses of CYD-TDV (Full Analysis Set).

Vaccine, Volume 31, 2013, 5814 - 5821

sReuters, March 25, 2014

Schematic representation of the CSP and the RTS,S vaccine

P D. Crompton, SK. Pierce, L H. Miller J Clin Invest. 2010

Malaria cuts risk in half in late-stage trial

H Waters Nature Medicine Nov 2011

N Eng J Med 2012

Malaria cuts risk in half in late-stage trial

H Waters Nature Medicine Nov 2011

31%

How to discover protective

antigens?

Identification of new target antigens: impact of genomics

Whole genome sequences of most bacterial pathogens and parasites completed

E. coli K-12 B. burgdorferi B. subtilis M. tuberculosis R. prowazekii H. influenzae C. pneumoniae C. trachomatis N. gonorrhoeae S. aureus H. pylori P. horikoshü E. faecalis N. meningitidis S. epidermitis M. genitalium S. pneumoniae L. pneumophila P. falciparum S. pyogenes M. pneumoniae T. pallidum L. major P. aeruginosa T. cruzi

M. leprae P. aerophilum V. cholerae

Genomic-based vaccine development

Whole genomic sequence

Computer prediction

Expression of recombinant proteins

DNA preparation

In silico vaccine candidates

Immunogenicity testing in animal models

Vaccine development

600 potential vaccine candidates identified

350 proteins successfully expressed in E.coli

344 proteins purified and used to immunize mice

355 sera tested 91 novel surface-exposed

proteins identified 28 novel proteins have bactericidal

activity

Meningoccocal B Vaccine: A Genomic Approach

5 vaccine candidates Rappuoli et al, 2002 Clinical trials

2000 2013

Front Immunol 2014

Structural vaccinology

A new computational method to design epitope-focused vaccines, illustrated with a neutralization epitope from RSV

Nature 507, 201–206 (13 March 2014)

Nature 507, 201–206 (13 March 2014)

Induction of neutralizating antibodies against RSV

Overview of the problems and methodologies of systems vaccinology

Seminars in Immunology, 2013, 209 - 218

Alum adjuvants are non-cystalline gels based on aluminum oxyhydroxide (referred to as Aluminum hydroxide gel), aluminum hydroxyphosphate (referred to as aluminum phosphate gel) or various proprietary salts such as aluminum hydroxy-sulfate) Alum is used in several licensed vaccines including:

•  diphtheria-pertusis-tetanus •  diphtheria-tetanus (DT) •  DT combined with Hepatitis B (HBV) •  Haemophilus influenza B •  inactivated polio virus •  Hepatitis A (HAV) •  Streptoccucus pneumonia •  Menngococccal •  Human papilloma virus (HPV)

Vaccines containing Alum Adjuvant

Dendritic cells initiate antigen-specific immune responses

•  most efficient of all APCs •  high MHC class I, II & costimulators •  efficient cross presentation •  stimulate naïve T cells (CD4, CD8)

All immunization strategies must target DCs

Initiate Ag-specific immune responses

Multiple inducers of DC maturation

Immature DC Mature DC

various T cell responses

Microbial products / TLR ligands Viral products Inflammatory cytokines Signaling receptors

Antigen-presenting cells serve as the bridge between innate and antigen-specific responses

2003, 2, 727-735

Rappuoli, CW. Mandl, S Black & E De Gregorio Nature Reviews Immunology Nov 2011

Vaccine adjuvants

Innate immune responses

Innate Lymphoid Cells (ILC)

T cell differentiation pathways

Coomes S M et al. Open Biol. 2013;3:120157 ©2013 by The Royal Society

Therapeutic vaccines

for chronic infections or cancers

Cancer, a worldwide burden

  1st cause of mortality in France

  In Europ, in 2012: - 1.75 million deaths from cancer -  3.45 million new cases of cancer

  In the world, in 2012: - 8.2 millions deaths - 14 million new cases diagnosed

Cancer, a cell disease

uncontrolled proliferation

Tumor

Surgery

Chimiotherapy Radiotherapy

Anti-angiogenic drugs

Immune responses can control the growth of tumor cells

The immunosurveillance theory

“It is by no means inconceivable that small accumulations of tumour cells may develop

and because of their possession of new antigenic potentialities provoke an effective immunological reaction, with regression of

the tumor and no clinical hint of its existence”

British Med Journal, April 1957

Burnet

Tumor specific/associated antigens

Overexpressed self antigens

Differentiation antigens

Mutated self antigens

Non self oncoviral antigens

Altered self antigens: Abnormal post-

translational/transcriptional modification:

underglycosylation

The concept of therapeutic anti-cancer vaccines

  Induction of specific immune responses against tumor specific/associated antigens to kill tumor cells or prevent their growth without affecting normal cells

Tumor vaccines - Whole tumor cells: + BCG or DETOX, e.g. Melacine vaccine (cell lysates), CancerVax

( irradiated melanoma cell lines), M-Vax (hapten-treated autologous cells) and gene-modified, irradiated tumor cells (GM-CSF)

- Tumor antigens: MAGE-1, MAGE-3, MART-1/Melan-A, tyrosinase, gp100, MUC-1,

CEA, etc. - Peptide vaccines: mutated ras, mutated p53, Her-2/neu, MART -1, gp100, MUC-1 - Heat shock proteins - DNA vaccines - Dendritic cell vaccines

Response rate = 3. 8%

Current human cancer vaccines show very low objective clinical response rate

Rosenberg, Yang & Restifo Nature Med 10:909 (2004)

Response rate = 3. 8%

Current human cancer vaccines show very low objective clinical response rate

Rosenberg, Yang & Restifo Nature Med (2004)

Benefit of passive immunotherapy (antibodies)

in cancer patients Lack of efficacy of most

current therapeutic cancer vaccines

Problems

Tumor derived antigens are weakly immunogenic

Need for better adjuvants or immunisation strategies

Dendritic cells initiate antigen-specific immune responses

•  most efficient of all antigen-presenting cells

•  stimulate naïve T cells (CD4, CD8)

All immunization strategies must target DCs

An Approach to Initiating Immunity to Cancer: Dendritic Cells Loaded with Tumor Antigens ex vivo

DC precursors expanded immature DCs

add disease- related antigens

maturing DCs presenting antigen(s)

Tumor- specific T cells

responding to

dendritic cells

2010: FDA panel passes first cancer vaccine

Original Article Sipuleucel-T Immunotherapy for Castration-

Resistant Prostate Cancer Philip W. Kantoff, M.D., Celestia S. Higano, M.D., Neal D. Shore, M.D., E. Roy

Berger, M.D., Eric J. Small, M.D., David F. Penson, M.D., Charles H. Redfern, M.D., Anna C. Ferrari, M.D., Robert Dreicer, M.D., Robert B. Sims, M.D., Yi Xu, Ph.D., Mark

W. Frohlich, M.D., Paul F. Schellhammer, M.D., for the IMPACT Study Investigators

N Engl J Med Volume 363(5):411-422

July 29, 2010

Source: www.provenge.com/

Provenge clinical trials : prostate cancer

Cancer vaccine pipeline

Problems

Tumor derived antigens are weakly immunogenic

Need for better adjuvants or immunisation strategies

CD8+ T cell

CD4+ T cell

Dendritic cell

Induction of optimized T cell responses by in vivo dendritic cells targeting

Antigen targeting

Maturation signals 2

1 Adjuvant

CyaA: a new proteinic vector targeted to dendritic cells

Bordetella pertussis

Dermonecrotic Toxin

BrkA FHA

TCF FIM

TCT

Pertussis Toxin

cAMP

Pertactin

Adenylate cyclase Toxin

DendriticCell

CD11b/CD18

AC domain

RTX domain

1 400 1706

InternalizationEndosomes

Cytosol

CyaA binds to CD11b allowing efficient targeting

to dendritic cells

Guermonprez et al, J. Exp. Med, 2001

Adenylate cyclase (CyaA)

Recombinant CyaA

+

Activation of CD8+ Cytotoxic T lymphocytes

Dendritic Cell ϕ

CD11b/CD18

Antigen

Th CD4+ MHC-II

endosomes

lysosomes

ϕ ϕ

MHC class II presentation

Activation of CD4+ Helper T lymphocytes

MHC class I presentation

Endoplasmic Reticulum

CTL CD8+ MHC-I/β2

ϕ Translocation Endocytosis

Antigens grafted in CyaA are delivered to both MHC class I & MHC class II presentation pathways

Immunisation in mice and non-human primates by

recombinant CyaA carrying a variety of antigens (such as

from M. tuberculosis or HIV) stimulates strong CTL and Th1

responses, even in the absence of adjuvant.

Préville et al, Cancer Res, 2005, Mascarell et al, J. Virol 2005, Majlessi et al, Inf Immun, 2005, Hervas-Stubb et al, Inf Immun, 2006, Mascarell et al, Vaccine 2006, Berraondo et al,

Cancer Res, 2007, Fayolle et al, Vaccine 2010.

CyaA: a new proteinic vector targeted to dendritic cells

80

HPV infection life cycle Few months to few years Up to 20 years

Goodman A., Wilbur D. C.

Human papilloma virus

�� - The HPV E6 and E7 oncoproteins are expressed throughout the replicative cycle of the virus and are necessary for the onset and maintenance of malignant transformation.�� - HPV E6 and E7 antigens are potential targets for specific

immunotherapy.

Rational

MHGDTPTLHEYMLDLQPETTDLYCYEQLN

CyaAE5

GQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKP

CyaA-E7∆30-42

Catalytic domain

224 235 319 320

1 400 1706

LQ

LQ

Recombinant CyaA carrying E7 from HPV16

GMP batch produced in recombinant E. coli

Therapeutic vaccination with recombinant��HPV16-E7 CyaAs eradicates established tumors

No treatment CyaA-OVA

0 500

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0 10

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0/10

20 30 40 50 60 70 80 90 0 500

1000 1500 2000

CyaA-HPV16E7∆30-42

-  Graft of TC-1 cells at day 0

- At day 10, one injection of:

-  50 µg of CyaA-E7 or of control CyaA-OVA

Preville et al, Cancer Res. 2005; Berraondo, K. et al. Cancer Res. 2007

A therapeutic vaccine candidate against HPV chronic infection and/or cervical cancer

Clinical trials started in 2010

0 102030405060708090

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0 102030405060708090100

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0 5 10 15 20 25 30 35

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0.2

10

20

daysdays days

CyaAE5 HPV16E7∆30-42 CyaAE5 CysOVA

Controls

One injection at Day 10

CyaAE5 HPV16E7∆30-42

Préville et al, Cancer Res, 2005

http://www.genticel.com

ProCervix - Phase 2 Clinical Trial

Merci de votre attention !

[email protected]

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