Vaccination against allergy

Ulla Seppälä, PhD

Vaccine Research

What is allergy?

An immunological over-reaction against specific molecules in our environment.

Allergens are specific molecules / proteins against which the sensitized indivuduals are responding by producing IgE antibodies.

Phleum pratense

What is causing the symptoms ?What is causing the symptoms ?- Sensitisation > < Clinical allergy -- Sensitisation > < Clinical allergy -

T-cell

B-cell

HLA-class II

IgM/D

T-cellAPC

BIgE

Mediator release

Mast cellIgE

CD23

Allergens

“other components”

Andersson A-C, Seppälä U, Rudin A. Activation of human neonatal monocyte-derived dendritic cells by lipopolysaccharide: Down-regulation of birch allergen-induced Th2 responses. Eur J Immunol. 2004;34(12) :3516-24.

Most common sources of inhalant allergens

Betula verrucosa

Felis domesticus

Phleum pratense

Dermatophagoidespteronyssinus

Vespid & Bee venom allergens

Vespula vulgarisApis mellifera

Allergens and Allergen Diagnostics

Allergen Diagnostics:

Clinical diagnosisIn vitro: specific IgEIn vivo: SPT / allergen extracts

What is allergy vaccination?What is allergy vaccination?

Tolerance

Reaction

- Allergy vaccination

- Allergen

Allergens are administered: in different doses at different sites } ”the immune response is changed”

Allergy vaccination at its best ;

Decrease in symptoms

Decrease in use of medication

Long-term effectiveness

Acting on basic immunological mechanisms

Anti-inflammatory treatment

Causal treatment

Preventive treatment Immunotherapy is the only treatmentthat influence the basic course of theallergic diseases.WHO Position Paper 1998

What is allergy vaccination ?

Subcutaneous Immunotherpy /SCIT

What is allergy vaccination ?

Sublingual Immunotherapy / SLIT“Targeting the sublingual mucosa”, using single-dose droplets or tablets.

Allergen Specific Immunotherapy

I.I. Anergy Anergy = clonal silence or actual clonal deletion of = clonal silence or actual clonal deletion of allergen-reactive T-cellsallergen-reactive T-cells

II.II. Immunoregulation-induced Immunoregulation-induced immune deviationimmune deviation, leading into , leading into different cytokine milieu in a target tissuedifferent cytokine milieu in a target tissue

Induction a state of tolerance by utilizing T - lymphocytes as targets

Rossenwasser LJ and Gelfand EW. Immunotherapy with antigens and epitopes.Am. J Repir.Cell. Mol. Biol. 1999:21;4-6.

Th2/Th1 Th2/Th1

SIT

IL-10/TGF-

Proposed mechanisms in immunotherapy

+ IgG4

Till SJ et al. Mechanisms of immunotherapy. J Allergy Clin Immunol 2004;113:1025-34.

Tregs/CD4+CD25+

Shift from TH2 to TH1-like responsefollowing SIT for grass pollen allergy

0

10

20

30

40

50

60

70

Before SIT After 3 months After 12 months

TH2 TH1 TH0

Ebner et al, Clin. Exp. Allergy, 1997, 27, 1007-1015

Targeting the therapeutic tools ?

Cell mediated immunityIgG- productionDTH

IFN-

Th0

Th1

Th2

Mast cells and Basophils

IL-4PGE2

IL-12

APC

APC

IFN-

CD8+ T-cells

IL-10 -

+ IFN-

-

++

++

+ IL-4IL-4

-

-

TNF-+

T-helper-cell differentiation

APC

B-cell

Mast Cell

Allergen

Allergen

IL-4

(IL-13)

IL-5

Eosinophil

IgE

Late Phase Reaction

Sensitisation Allergic Reaction

Immediate Reaction

• natural allergen extracts• anti-allergy drugs e.g. antihistamines

What are the tools to cure allergy?

CURRENT METHODS

What are the tools to cure allergy?

DNA - vaccines rDNA - vaccines

Reichert JM and Paquette. Therapeutic recombinant proteins: Trends in US approvals 1982 – 2002. Curr Opin Mol Ther. 2003:5;139-47.

Donnelly JJ,Wahren B, Liu MA. DNA vaccines: progress and challenges. J Immunol. 2005;15;175:633-9.

FUTURE METHODS

Therapeutical recombinant proteins / rDNAs

rIgs Fc-Fel d 1

allergen + CpG/ISS/other adjuvants

Daocheng Z et al. A chimeric human-cat fusion protein blocks cat-induced allergy. Nature Medicine 2005;11(4):446-49.

Formulation of allergen vaccines by use of various adjuvants

Aluminium hydroxide Aluminium hydroxide Bacterial origin mucosal adjuvants Bacterial origin mucosal adjuvants – CpG ODN, CpG ODN, – Monophosphoryl lipid A (MPL)Monophosphoryl lipid A (MPL)– Cholera toxin /CT (Cholera toxin /CT (Vibrio choleraVibrio cholera)),, entero toxin /LT entero toxin /LT

((Escherichia coliEscherichia coli))Carbohydrates / (CBPs)Carbohydrates / (CBPs)Microencapsulated allergen vaccinesMicroencapsulated allergen vaccines

Francis JN, Durham SR. Adjuvants for allergen immunotherapy: experimental results and clinical perspectives. Curr Opin Allergy Clin Immunol. 2004 Dec;4(6):543-8.

Freytag LC, Clements JD. Mucosal adjuvants. Vaccine. 2005 Mar 7;23(15):1804-13.

”An adjuvant is an agent which, while not having any specific antigenic effect in itself, may stimulate the immune system, increasing the response to a vaccine.”

How to monitor allergen immunotherapy ?

– improved clinical phenotype

– serum IgE / IgG4 –levels IgE / IgG4

– decrease in number of mast cells and eosinophils after allergen provocation

– modified and/or reduced production of cytokines

– BIOMARKERS ? !

Walker C and Zuany-Amorim. New trends in immunotherapy to prevent atopic diseases.TRENDS Pharm Sci. 2001;22(2):84-90.

Allergen specific IgG is induced by SIT

IgE =

IgG4

Gabrielsson S et al, Allergy 56:293, 2001.

IgE

IL-4

IL-5 Allergicresponse

Eosinophils

CD4

APC

CD80/86

Th2

T cell

Allergen

CD3HLA

CD28

B-cell

+

+

Tregs

IL-10

TGF-

--

Th1

IgG

IFN-

B-cell

+

IT

IT

-

DCs

Immunotherapy: Immune deviation

How to monitor allergen immunotherapy ?

A prequisite for a vaccine is the knowledge of how to induce Treg cells in vivo

Sauer S et al. Miniaturization in functional genomics and proteomics. Nature Reviews Genetics. 2005;6:465-76.

DNA / protein arrays+

”omics” technologies

”Genomics and proteomics of allergic disease”

Identification of the disease genes; for design of new classes of anti-inflammatory compounds

Identification of expression and function of proteins; to obtain increased knowledge of mechanisms underlying allergic disease

Identifiction of novel biomarkers

TODA M and ONO SJ. Genomics and proteomics of allergic disease. Immunol. 2002;106:1-10.

Microarray technology

DNA – microarrays:

• High – throughput analysis and expression of multiple genes or single nucleotide polymorphisms (SNIPs).

Karp CL et al. Identification of complement factor 5 as a susceptibility locus for experimental allergic asthma. Nat Immunol 2000;1:181-7.

Miklos GL, Maleszka R. Microarray reality checks in the context of a complex disease. Nat Biotechnol. 2004 May;22(5):615-21.

Microarray technology

Protein –microarrays:

• examine the time course of cytokine secretion pattern by cell cultures, T- cells, DCs, Mast Cells / Basophils

• examine expression profiles of cells expressing recombinant allergens

Harwanegg C & Hiller R. Protein microarrays for the diagnosis of allergic diseases: state-of-the-art and future development. Clin Chem Lab Med 2005;43:1321-6.

Proteomics technology

• investigation of the influence of SNIPs in gene expression and function of the proteins = elucidation of disease gene expression

transcriptome = proteome

• follow-up of the Th1/Th2/Treg – profiles – up / down regulation of signal transduction pathways, marker molecules

• plasma / serum proteomics / other fluids

• characterization cellular responses against natural and/or modified rDNA

Ghafouri et al. Comparative proteomics of nasal fluid in seasonal allergic rhinitis. J Proteome Res. 2006;5:330-8.

Proteomics in Allergy ResearchProteomics in Allergy Research

Fehninger T.E. et al. Exploring the context of lung proteome within the airway mucosa following allergen challenge. J Proteome Res. 2004;3:307-20.

Identification of the disease genes or Identification of the disease genes or biomarkersbiomarkers

Identification of the proteins / peptides by Mass Spectrometry

DATABASE SEARCH

MW

pI

Weingarten P et al. Application of proteomics and protein analysis for biomarker and target finding for immunotherapy. Methods Mol Med. 2005;109:155-74.

Biomarker discovery:Biomarker discovery: PEPTIDOMICS PEPTIDOMICS

Peptides are ideal candidates for biomarkersPeptides are ideal candidates for biomarkers

Isolation/ measurement of biomarkers from blood - clinical applicationIsolation/ measurement of biomarkers from blood - clinical application

Proteases liberate biomarkers – processing and specific degradation Proteases liberate biomarkers – processing and specific degradation products – discovery tool !products – discovery tool !

HormonesHormonesCytokinesCytokinesGrowth factorsGrowth factorsetc.etc.

Schulte I. et al. Peptides in body fluids and tissues as markers of disease. Expert Rev Mol Diagn. 2005 Mar;5(2):145-57.

Crameri R. The potential of proteomics and peptidomics for allergy and asthma research. Allergy 2005 Oct;60(10):1227-37.

ConclusionsConclusions

A B C D

“New vaccines”

Optimal treatment of the allergic patient

Allergy

Disease

Specific allergy vaccination Medication

Patient education

Allergen avoidance

SLIT Preventive medication

Jacobsen,L. Preventive aspects of immunotherapy: prevention for children at risk of developing asthma. Ann.Allergy Asthma Immunol. 2001; 87(1 Suppl 1):43-46.