Vaccine adjuvants: the good, the bad and the

ugly

Professor Nikolai PetrovskyDirector of Endocrinology, Flinders Medical Centre

Research Director, Vaxine Pty LtdEditor-in-Chief, Journal of Vaccines and Vaccination

nikolai.petrovsky@flinders.edu.au

What are adjuvants?

•Any substance that increases the memory immune response to a co-administered antigen

•Discovered by trial and error, e.g. alum, oil emulsions, saponins

•Common feature – induction of inflammation – encapsulated in danger-hypothesis

•Postulated to work via innate immune cell activation, inflammatory cytokine production and APC activation leading to co-stimulation of memory T- and B-cell responses

Why do we need adjuvants

•Enhance antibody titers

•Generate CTL responses

•Improve cross-protective responses

•Antigen-sparing

•Speed kinetics of protection

•Decrease no. vaccine doses

•Extend immune memory

•Improve protection in neonates and the elderly

“After 2 doses of an H7N9 formulation containing 15 µg of hemagglutinin given without adjuvant, with AS03 adjuvant, or with MF59 adjuvant, the proportion achieving an HIA titer of 40 or higher was 2% without adjuvant, 84% with AS03 adjuvant, and 57% with MF59 adjuvant”

Potential adverse effects

LOCALpain bruisingswellingtenderness granuloma abscessulcer

SYSTEMICfever headachemyalgia/arthralgia fatigueLymphadenopathyeosinophilia allergy/anaphylaxisautoimmunity

Potential to trigger or exacerbate autoimmunity

Can we avoid adjuvant toxicity?

LOCAL•pain •bruising•swelling•tenderness •abscess

SYSTEMIC•fever •headaches•myalgia/arthralgia •fatigue•autoimmunity

REACTOGENICITY/TOXICITY

INFLAMMATION

Traditional adjuvants work via induction of inflammation

Delta inulin (Advax™) adjuvant breaks the

danger-signal paradigm

Soluble form –kidney testing

Particulate form –

Vaccine adjuvant

β-D-(2→1) polyfructofuranosyl α-D-glucose (inulin)

Advax enhances H5N1 influenza vaccine protection

in ferret model

Vaccine. 2011 Aug 26;29(37):6242-51.

Advax enhances JEV vaccine heterologous protection against

West Nile virus

Advax enhances neonatal influenza vaccine

protection

Advax provides influenza protection in adult B-cell deficient mice (uMT mice)

Nonsegmented negative-stranded RNA genome containing 7 structural and regulatory genes. )

Family: FiloviridaeFilamentous form with diameter ~80 nm and

variable length. 

DoD VLP vaccine based on VP40 and glycoprotein (GP) expressed in

HEK293 cells

Ebola vaccine project with USAMRIID

VLP+Advax2 CD8 T-cell responses 2x PolyICLC

Preclinical models confirming efficacy of Advax delta inulin

adjuvant

Human clinical trials confirming efficacy of Advax

delta inulin adjuvant

• Trivalent inactivated seasonal Influenza

vaccine

• Recombinant HA pandemic influenza

(H1N1/2009pdm) vaccine

• T cell fluropeptide-based influenza vaccine

• Hepatitis B vaccine

• Insect venom allergy immunotherapy vaccine

Next-generation pandemic influenza vaccine project with Protein Sciences Corporation (Meriden, CT)

21 April 2009 - new H1N1/2009 virus isolated

and sequenced

Haemagglutinin gene cloned into baculovirus

Insect cellexpression

GMP cell culture and protein purification

Formulation with Advax adjuvant

17

Human trial commenced

Adelaide – 18 July 2009

Animal study

confirms need for adjuvant

Vaccine 2012 Aug 3;30(36):5407-16

Advax enhances human T-cell IFNresponses to influenza

peptide vaccine

Confidential slide

Human trials of Advax

HBVvaccine

Seasonal inactivated influenza vaccine

Bee sting anaphylaxisImmuno-therapy

rH1N1/2009pdm vaccine

Peptide-based universal influenza

Totalsubjects

>30 >1400 >20 271 80

Proportionreceiving Advax

2:1 2:1 2:1 1:1 1:1

Number of immunizations

3 1-4 >30 2 2

Why is Advax different to other

adjuvants?

Delta inulin is non-reactogenic

and targets mDC

Holden Maecker, Stanford Uni

Advax provides single-dose anthrax vaccine

protection

Clin Vaccine Immunol. 2014 Apr;21(4):580-6.

Advax induces minimal reactogenicity

Clin Vaccine Immunol. 2014 Apr;21(4):580-6.

Adjuvant reactogenicity comparison study in camels

Enhanced immunogenicity Enhanced immunogenicity versus reactogenicityversus reactogenicity

•  Positive-stranded RNA virus• Spike protein mediates target cell

entry via ACE2• Alum-adjuvanted vaccine induces

lung eosinophilic pathology

SARS coronavirus vaccine

J. Virol. 2014 Dec 17.

Alzheimer’s vaccine• Abeta and p-tau proteins play

pathogenic role• Monoclonal Ab. can work but

expensive & inconvenient

1

10

100

1000

10000

Montanide ISA51

Montanide ISA720

MPLA Alhydrogel®

Quil-A AdvaxMT

Abeta Multitep vaccine

Con

c. o

f an

ti-A

Ab

s (µ

g/m

l)β

Abeta

P-tau

Abeta pTau

How does Advax enhance humoral

and cellular immunity without reactogenicity?

Traditional adjuvants

Advax is effective in innate immune knockout models

•MYD88/TRIF•TLR4, TLR9 •NALP3•IL1 receptor•Complement C3, Factor B •IL12, IL33•IFNR1•NSAID treatment•NFkB inhibition

Advax is effective in MYD88/TRIF double knockouts

Advax is the first non-inflammatory

adjuvant

CD4T cell

CD8T cell

Advax

Advax

Chemokines/Chemokine receptors

(MCP1, IL-8 CCR1, CCR2)

Antigen presentation (MHC1, MHC2,

CD40, CD40L, CD80, CD86)

Inflammatory cytokines

(TNFa, IL1, IL6)

TLR agonist

MyD88/TRIF

TRAF6

NFkB

MHC 1MHC 2

Monocyte/mDC differentiation

(CD11c, CD71)

Advax mechanisms of action

Advax

Advax

CD71

CD11c

CD86

B cellB cellMCP1

IL-8MIP1b

TLRsMF59QS21Alum

Opsonisation

Endosomalregulation

Chemotaxis

DC differentiation

Co-stimulation

Adaptive adjuvant

Inflammation

Innate adjuvants

Antigen presentation NALP3

A new way to think about adjuvants?

NIH Adjuvant Discovery Program

TLR9 ECD

Advax combined adjuvants for mTB

vaccines

Triccas, unpublished data

Alternative immunization routes

Lung immunization with Advax

Enhanced neonatal immunization

Y. Honda-Okuba Vaccine, in press

Adjuvant evolutionAdjuvant evolution

1st Generation (CFA, oil emulsions, bacterial extracts) • Emulsions/bacterial extracts generate danger signals by

inducing tissue damage– high reactogenicity• Induce autoimmunity in animal models

2nd Generation (alum, MPL, CpG, squalene, QS21) • Better characterised – stimulate TLR or inflammasome• Induce inflammatory cytokines (IL-1, TNF-a)• Can cause pyrexia• Remaining potential for autoimmunity

Next generation adjuvants (Advax)• Non-inflammatory • Don’t induce autoimmunity• Don’t induce pyrexia• Specific enhancement adaptive immunity

AcknowledgementsVaxinePeter CooperSu HeinzelBruce LyonsY. Honda-OkubaFadi SaadeSamay TrecLei LiHarinda R.

Mario LobigsDavid GordonDimitar Sajkov

Manon Cox, Protein Sciences

John Morrey and Dale Barnard, Utah State University

Fred Coster, Lovelace Respiratory Research Institute

Tod Merkel, CBER, FDA

Kitasato Institute, Japan

Dr Uli KinniCVRL Dubai

NIH ContractsU01AI061142 HHSN272200800039CHHSN272201400053C