Towards personalized p immunotherapy of cancer Hans-Georg - PowerPoint PPT Presentation

NCT Conference 2013 New Cancer Targets Towards personalized p immunotherapy of cancer Hans-Georg Rammensee Interfakultäres Institut für Zellbiologie Abteilung Immunologie Prof. Dr. Hans ‐ Georg Rammensee

Towards patient-specific tumor antigen selection for vaccination. i ti Rammensee HG, Weinschenk T, Gouttefangeas C, Stevanovi ć S. I Immunol Rev 188:164-76, 2002 l R 188 164 76 2002

1. MHC, peptides, and cancer 2. Our strategy to identify cancer associated peptides 3 Cli i 3. Clinical studies l t di 4. Why mutated antigens should be the better targets y g g 5. Problems

this can be recognized by T cells protein fragment This is true for all p This is true for all g proteins inside a = peptid cell, also for tumor Antigen presentation Antigen presentation associated changes, by HLA molecule including mutations Antigen processing Antigen processing

Cancer cells differ from normal cells from normal cells eg., in gene expression This can be sensed by T cells - no matter which cellular compartment is affected by the change a ected by t e c a ge

3 classes of tumor antigens

Limited to few cancer We know plenty of such antigens; This is our vision entities immune responses tend to be weak but stronger immune responses may lead but stronger immune responses may lead to autoimmunity

I I I I so la tio n o f Na tura lly Pre se nte d HL so la tio n o f Na tura lly Pre se nte d HL so la tio n o f Na tura lly Pre se nte d HL so la tio n o f Na tura lly Pre se nte d HL A- A L A A -L L L ig a nds ig a nds ig a nds ig a nds L a b Me e ting Da nie l K o wa le wski 07.07.2011

Peptide preparation Andy Weinzierl General survey y • Protein A affinity chromatography using W6/32-antibody i W6/32 tib d • elution with citrate buffer, pH 3

Technological progress

HLA ligandome analysis, next generation Daniel Kowalewski Daniel Kowalewski D D

Technological advance: ultraHPLC lt HPLC

Comparative HLA ligandome analysis

Off ‐ the ‐ shelf vaccines: Allele ‐ specific target identification (A*02) p g ( ) 6000+ peptides exclusively identified in A*02+ tumor ligandomes 362 peptides identified in ≥ 50 % of A*02+ tumor ligandomes tumor ligandomes

Oncology Department of Hematology Immunology Immunology Rheumatology Ligandomics in leukemia Juliane Stickel

HLA ligand isolation g ells ignant ce Affinity Purification Chromatography Mali Cell lysate HLA antibody Leukemia Patient PBMCs s Acid elution id l i Ultrafiltration 10 kDa Isolated peptides LC-MS/MS od_70521_28 1 (0.019) Sm (Mn, 2x3.00) TOF MSMS 497.93ES+ od_70521_28 1 (0.019) Sm (Mn, 2x3.00) TO F MSMS 497.93ES+ 183.17 9.45 183.17 9.45 100 100 685.73 685.73 193.16 193.16 572.60 572.60 402.40 402.40 211.19 211.19 253.25 367.33 686.69 253.25 367.33 686.69 % % 515.53 515.53 292.27 310.28 292.27 310.28 270.31 555.54 270.31 555.54 385.38 385.38 440.48 440.48 Healthy donor Healthy donor 573.62 573.62 86.15 516.55 86.15 516.55 171.17 458.53 628.69 171.17 458.53 628.69 671.69 671.69 112 14 112.14 498.54 112.14 112 14 498.54 687.77 687.77 212.18 784.79 212.18 784.79 654.65 654.65 165.13 403.44 165.13 403.44 574.63 574.63 157.17 157.17 349.29 688.76 349.29 688.76 72.14 625.65 786.84 72.14 625.65 786.84 749.76 749.76 0 m/z 0 m /z 50 100 150 200 250 300 350 400 450 500 550 600 650 700 750 800 850 900 950 50 100 150 200 250 300 350 400 450 500 550 600 650 700 750 800 850 900 950 Peptide sequences & Source protein annotation Overall comparison HLA Ligandome