Transcript Allergen

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 ?
- Sensitisation > < Clinical allergy CD23
Allergens
IgE
APC
B
T-cell
HLA- class II
IgE
Mast cell
T-cell
“other components”
B-cellIgM/D
Mediator release
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
Phleum pratense
Betula verrucosa
Dermatophagoides
pteronyssinus
Felis domesticus
Vespid & Bee venom allergens
Apis mellifera
Vespula vulgaris
Allergens and Allergen Diagnostics
Allergen Diagnostics:
Clinical diagnosis
In vitro: specific IgE
In vivo: SPT / allergen
extracts
What is allergy vaccination?
 Allergens are administered:
in different doses
at different sites
}
”the immune response is changed”
- Allergy vaccination
Tolerance
Reaction
- Allergen
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
Immunotherapy is the only treatment
Preventive treatment
that influence the basic course of the
allergic 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
Induction a state of tolerance by utilizing T - lymphocytes as targets
I.
II.
Anergy = clonal silence or actual clonal deletion of
allergen-reactive T-cells
Immunoregulation-induced immune deviation, leading into
different cytokine milieu in a target tissue
Rossenwasser LJ and Gelfand EW. Immunotherapy with antigens and epitopes.
Am. J Repir.Cell. Mol. Biol. 1999:21;4-6.
Proposed mechanisms in immunotherapy
Tregs/
CD4+CD25+
IL-10/TGF-b
Th2/Th1
Th2/Th1
SIT
+ IgG4
Till SJ et al. Mechanisms of immunotherapy. J Allergy Clin Immunol 2004;113:1025-34.
Shift from TH2 to TH1-like response
following SIT for grass pollen allergy
70
60
50
40
30
20
10
0
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 ?
Sensitisation
Allergic Reaction
T-helper-cell differentiation
Late Phase Reaction
CD8+ T-cells
Allergen
APC
IL-10
- IL-12
TNF-a
APC
+
+
+
+
Th1
-g
-
+
+
+
Eosinophil
IFN-g
Th0
IL-4
IL-4
IL-5
Immediate Reaction
Th2
PGE2
APC
Cell mediated immunity
IFN-g IgG- production
DTH
IFN-g
IL-4
Mast cells and Basophils
IgE
IL-4
(IL-13)
Mast Cell
B-cell
Allergen
What are the tools to cure allergy?
CURRENT METHODS
• natural allergen extracts
• anti-allergy drugs e.g. antihistamines
What are the tools to cure allergy?
FUTURE METHODS
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.
Therapeutical recombinant proteins / rDNAs
allergen + CpG/ISS/
other adjuvants
rIgs
Fcg-Fel d 1
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
”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.”
Aluminium hydroxide
Bacterial origin mucosal adjuvants
– CpG ODN,
– Monophosphoryl lipid A (MPL)
– Cholera toxin /CT (Vibrio cholera), entero toxin /LT (Escherichia
coli)
Carbohydrates / (CBPs)
Microencapsulated 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.
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.
Immunotherapy:
Immune deviation
IgE
IL-4
B-cell
Allergen
APC
DCs
CD80/86
CD28
IL-5
HLA
CD3
-
IT
T cell
CD4
Eosinophils
Th2
IT
+
Allergic
+ response
-
TGF-b
Tregs
IL-10
IFN-g
Th1
+
B-cell
IgG
How to monitor allergen immunotherapy
?
A prequisite for a vaccine is the knowledge of how to induce Treg cells in vivo
DNA / protein arrays
+
”omics” technologies
Sauer S et al. Miniaturization in functional genomics and proteomics. Nature Reviews
Genetics. 2005;6:465-76.
”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 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
biomarkers
pI
MW
Identification of the proteins / peptides by Mass Spectrometry
DATABASE SEARCH
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:
PEPTIDOMICS
Peptides are ideal candidates for biomarkers
Isolation/ measurement of biomarkers from blood - clinical application
Proteases liberate biomarkers – processing and specific degradation
products – discovery tool !
Hormones
Cytokines
Growth factors
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.
Conclusions
A
B
C
“New vaccines”
D
Optimal treatment of the allergic patient
Allergen
avoidance
Patient
education
Allergy
SLIT
Specific allergy
vaccination
Preventive medication
Disease
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.