David Wraith is founder and CSO of Apitope International NV
Immune system depends on T cell directed mechanisms for effective immunological tolerance ď‚Ą Ig genes undergo somatic hypermutation in response to foreign antigen - risk of autoantibodies ď‚Ą Immune responses must be governed by T cells, since TCR genes do not undergo somatic hypermutation, through controlled T cell help for B cells and regulatory mechanisms ď‚Ą
Thymic: positive and negative selection Thymic: generation of FoxP3+ natural regulatory T (nTreg) cells Peripheral: sequestration, privileged site, homeostasis and anergy Peripheral: generation of FoxP3+ inducible regulatory T (iTreg) cells & negative feedback mechanisms
Strength Of Signal
Death by TCR Signalling ‘Negative Selection’
Selection of FoxP3 Treg Pop’n Survival ‘Positive Selection’
Death by neglect MHC + Peptide Antigen
Signal strength is the product of TCR & +ve/-ve costimulatory signals Liu et al. Immunity (1995) 3, 407-415 Verhagen, J., et al. Proc. Natl. Acad. Sci. (USA) (2009) 106, 3306-3311
Spleen CD4 cells
Induced Regulators Soluble Peptide 2x Per week
92-95% CD25-ve 5-8% CD25&FoxP3+ve
TCR (Va4,Vb8.2) specific for N-terminal epitope of MBP Ac1-9
Burkhart et al. Int. Immunol. (1999) 11, 1625 Sunstedt et al. J. Immunol. (2003) 170, 1240
â€˘ injection of antigen in CFA induces EAE â€˘ 5-10 doses of soluble peptide antigen induces tolerance and prevents induction of EAE
Anergy Suppression of inflammatory cytokines Induction of anti-inflammatory cytokine (IL10) Bystander suppression Upregulation of negative costimulatory molecules
Cytokines act on innate and/or adaptive immune cells to control the effector response
CD4+ naïve T cell
Th1 cell T-bet
T-bet IFN-g & IL-10
Intracellular Pathogens Autoimmunity
Th2 cell GATA-3
Regulatory T cell FoxP3
IL-4, IL-5, IL-13, IL-10 Extracellular Pathogens Allergy
None, TGF-b, IL-10 Immune Regulation
Th17 cell ROR-gT
IL-17 IL-10 Bacterial Pathogens Autoimmunity
Gabryšová et al: J. Exp. Med. (2009) 206: 1755-1767
IFN-g IFN-g IL-10
TCR signal strength
Is this approach suitable for treatment of allergic and autoimmune diseases? Gabryšová & Wraith Eur. J. Immunol. (2010) 40: 1386-1395
In 1911, Drs John Freeman and Leonard Noon published an account of a novel treatment for hay fever. Their method of desensitisation consisted of injecting increasing doses of an extract of pollen subcutaneously until the hypersensitivity reaction was diminished or abolished “there seems little doubt that there has been a distinct amelioration of symptoms. This improvement took several forms; a greater freedom from attack, the attack not so bad as in former years, and the attack sooner over, the constitutional disturbance not so great”
Noon, L. Prophylactic innoculation against hay fever. Lancet i, 1572-1573. 1911
Intact antigen can activate mast cells and basophils by cross-linking IgE Intact antigen can stimulate antibody secretion (MOG in EAE) Intact antigen given orally can stimulate cytotoxic T lymphocytes (Insulin in diabetes) Use CD4 T cell epitopes*
*Harrison, L. C. and D. Hafler (2000). "Antigen-specific therapy for autoimmune disease." Current Opinion in Immunology 12: 704-711. Larche, M. and Wraith, D.C. Peptide Vaccines for Allergic and Autoimmune Diseases Nature Medicine (2005) 11: s69-76.
Nature Immunology 3, 175.2002
Not all T cell epitopes induce tolerance Peptides must be designed to mimic naturally processed epitopes Such peptides are defined as antigen processing independent epitopes or apitopes
Journal of Experimental Medicine 178, 1783.
Peptide epitopes induce linked suppression
Dose/animal & route
100 mg i.n.
Metzler et al 1993 Burkhart et al 1999
100 mg i.p.
Liu et al 1995
100 mg i.n.
Anderton et al 1998
5 x 120 mg i.n
Liu, et al 1998
Collagen II 245-270
3 x 100 mg i.n.
Chu et al 1999
3 x 100 mg i.n. or s.c.
Prakken et al 1997
4 GAD peptides
200 mg i.n.
Tian et al 1996
100 mg i.n. or s.c.
Daniel et al 1996
50 mg i.p.
Elias et al 1991
Band 3 861-874
100 mg i.n.
Shen et al. 2003
600 mg i.v./month
Riemekasten et al 2004
3 AChR peptides
50 mg i.n.
Karachunski et al 1997
10 x 6 mg i.n.
Zou et al 1999
Peptides therapy in MS MOG
R. O. Weller
Epitopes in MBP
MS 1 30-44 DQ 6
MS 7 83-99 DR 15
MS 4 MS 6 131-145 140-154 DQ 6 DR 15
Mice express HLA-DR15 and human TCR specific for MBP
Treatment with ATX-MS1467 prevents disease onset and suppresses ongoing disease
Treatment suppresses secretion of inflammatory cytokines (IL-1a, IL-2, IL-6, IL-17, IFN-g, TNF-a, GMCSF)
Secretion of antiinflammatory cytokine (IL10) sustained
Subjects – Patients (6) with secondary progressive multiple sclerosis (SPMS)
Design: – Open-label – Dose-escalation of five doses, plus repeat of highest dose
Posology – Dose frequency: 7 to 14 days apart – Dose Escalation: 25, 50, 100, 400, 800, 800 mg i.d.
Primary objective – Assess safety and tolerability of ATX-MS-1467
Secondary objective – Monitor immunological parameters in response to ATX-MS-1467 – Monitor disease status in the CNS using MRI
HLA DR Type
Improvement in vision Right eye: 6/24 (V1) to 6/9 (V9) Left eye : 6/9 (V1) to 6/6 (V9)
Improvement in Gd-enhanced MRI on Visit 8 (1 month follow-up)
3H.thymidine Incorporation (ccpm)
Visit 8 P=0.3438
Visit 1: prior to treatment Visit 8: one month after last dose
Visit 8 P=0.0313
3H.thymidine Incorporation (ccpm)
Immune responses 15000
Visit 1: prior to treatment Visit 8: one month after last dose Visit 9: three months after last dose
Repeated treatment with ATX-MS1467 will be required to maintain suppression of the immune response to myelin antigen
Patterns of MS â€˘ A further trial comparing ID and SC routes of ATX-MS1467 is in progress â€˘ This second trial is treating patients with relapsing multiple sclerosis Years
ClinicalTrials.gov Identifier: NCT01097668 Dr Jeremy Chataway
J. Clin. Invest. 1998. 102, 98-106
PLA 45-62, 82-92, 113-124
J. Allergy Clin Immunol. 1998. 101, 747-754
â€œOur results provide, for the first time, in vivo evidence of linked epitope suppression and IL-10 induction in both human allergic disease and a mouse model designed to closely mimic peptide therapy in humansâ€?
Chronic stimulation through infection or repeated dose antigen-specific therapy
Th1 Th1 IFN-g
T-bet STAT-4 Na誰ve CD4+ T cell
IL-4 GATA-3 STAT-6
Th2 IL-4/5/13 IL-10
IL-12 (ERK/STAT-4) IL-27 (STATs-1/3) c-MAF/SMAD-4
c-MAF/ERK/JUN GATA-3 STAT-6
Th1 IFN-g IL-10
Th2 IL-4/5/13 IL-10
IL-6 TGF-b, IL-1b STAT3 Rorgt, Rora
IL-21 (STAT-3/c-MAF) IL-6 (STAT-3/c-MAF) IL-27 (STATs-1/3/c-MAF) ERK
Th17 IL-17 IL-10
Sabatos-Peyton, C., Verhagen, J. & Wraith, D.C. Antigen-specific immunotherapy of autoimmune and allergic diseases. Current Opinion in Immunology (2010) 22:609-615
Bristol Collaborators Graham Britton Stephan Strobel Bronwen Burton Immanuel Luescher Leona GabryĹĄovĂĄ Raphael Genolet Elaine Hill Arlene Sharpe Graziella Mazza Graham Anderson Kirsty Nicolson Julian Dyson Srihari Pillai Catherine Sabatos-Peyton Neil Scolding Ella Shepard Heather Streeter Johan Verhagen
Our laboratory is supported by the Wellcome Trust and the Multiple Sclerosis Society of Great Britain and Northern Ireland. The phase I/IIA clinical trial in MS was funded by a Wellcome Trust UTA to Apitope
SIGNAL 1 & 2
SIGNAL 1, 2 & 3
TCR activation in the absence of costimulation
TCR activation with costimulation (TLR)
TCR activation with CD40 ligation, cytokine secretion and co-stimulation
T cell anergy T cell depletion
T cell activation and peripheral tolerance: Expansion of FoxP3+ and FoxP3- regulatory T cells
Activation and polarization of effector cells