1. Vaccination against allergy Ulla Seppälä, PhD Vaccine Research

2. What is Allergy ? Non-IgE mediated hypersensitivity IgE- antibodies X Allergic response(s) Immediate-type Delayed- type sensitization

3. What is causing the symptoms ? - Sensitisation > < Clinical allergy - T-cell B-cell HLA - class II IgM/D T-cell APC B IgE Mediator release Mast cell IgE 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.

4. Most common sources of inhalant allergens Betula verrucosa Felis domesticus Phleum pratense Dermatophagoides pteronyssinus

5. Vespid & Bee venom allergens Vespula vulgaris Apis mellifera

6. Allergens and Allergen Diagnostics Allergen Diagnostics: Clinical diagnosis In vitro: specific IgE In vivo: SPT / allergen extracts

7. What is allergy vaccination? Tolerance Reaction - Allergy vaccination - Allergen  Allergens are administered: in different doses at different sites } ”the immune response is changed”

8. What is allergy vaccination ? Subcutaneous Immunotherpy /SCIT

9. What is allergy vaccination ? Sublingual Immunotherapy / SLIT

10. Allergen Specific Immunotherapy I. Anergy = clonal silence or actual clonal deletion of allergen-reactive T-cells II. Immunoregulation-induced immune deviation, leading into different 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.

11. 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. T regs / CD4 + CD25 +

12. Targeting the therapeutic tools ? Cell mediated immunity IgG- production DTH IFN-  Th0 Th1 Th2 Mast cells and Basophils IL-4 PGE 2 IL-12 APC IFN-  CD8 + T-cells IL-10 - + IFN-  -  + + + + + IL-4 -4 - - TNF-  + T-helper-cell differentiation APC B-cell Mast Cell Allergen IL-4 (IL-13) IL-5 Eosinophil IgE Late Phase Reaction SensitisationAllergic Reaction Immediate Reaction

13. natural allergen extracts anti-allergy drugs e.g. antihistamines What are the tools to cure allergy? CURRENT METHODS

14. 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. FUTURE METHODS

15. Therapeutical recombinant proteins / rDNAs rIgs Fc  -Fel d 1 allergen + CpG/ISS Daocheng Z et al. A chimeric human-cat fusion protein blocks cat-induced allergy. Nature Medicine 2005;11(4):446-49.

16. Formulation of allergen vaccines by use of various adjuvants 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. ”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.”antigenicimmune systemvaccine

17. 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.

18. How to monitor allergen immunotherapy ? A prequisite for a vaccine is the knowledge of how to induce T reg 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

19. ” 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.

20. 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.

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 Schweitzer B et al. Multiplexed protein profiling on microarrays by rolling-circle amplification. Nature Biotechnology, 2002;20:359-65.

22. 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/T reg – profiles – up / down regulation of signal transduction pathways, marker molecules plasma / serum proteomics characterization cellular responses against natural and/or modified rDNA Chromy BA et al. Proteomic Analysis of Human Serum by Two-Dimentional Differential Gel Electrophoresis after Depletion of High-Abundant Proteins. J Proteome Res. 2004;3:1120-1127.

23. 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.

24. Identification of the disease genes or biomarkers 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.

25. 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 ! HormonesCytokines 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.

26. Conclusions ABCD “New vaccines”