S78. Proffered paper: High-affinity CD20-specific TCRs suitable for adoptive immunotherapy can be...
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INVITED SPEAKER PRESENTATION Open Access S78. Proffered paper: High-affinity CD20-specific TCRs suitable for adoptive immunotherapy can be readily isolated from the allo-repertoire using reverse immunology L Jahn 1* , P Hombrink 2 , C Hassan 3 , MGD Kester 1 , MP Schoonakker 1 , JHF Falkenburg 1 , P van Veelen 3 , MHM Heemskerk 1 From 1st Immunotherapy of Cancer Conference (ITOC1) Munich, Germany. 12-14 March 2014 Studies using T-cell receptor (TCR) or chimeric antigen receptor (CAR) transduced T-cells have shown the effec- tiveness of adoptive immunotherapy to treat different malignancies. The efficacy and safety of such interventions greatly depends on good target selection to prevent on- target toxicity. Furthermore, the broad application of TCR-based adoptive immunotherapy is hampered by a lack of an effective immune response against self-antigens. Through self-tolerance, T-cells carrying high-affinity TCRs reactive to self-antigens are deleted during thymic selec- tion. An attractive strategy is to exploit the immunogeni- city of foreign human leukocyte antigen (HLA) molecules to generate an effective immune response against these antigens. Here, we describe a protocol to efficiently isolate high-avidity alloHLA-restricted T-cells targeting the B-cell compartment. From a B-cell peptide elution library 15 peptides derived from genes exhibiting B-cell restricted expression patterns were identified and peptide-MHC multimers (pMHC) of HLA-A*0201 were generated. Via MACSort- ing and FACSorting a plethora of pMHC-multimer bind- ing T-cell clones from HLA-A*0201-negative individuals were isolated. Generated T-cell clones were selected based on peptide-specificity and avidity for further characterization. We successfully isolated two distinct T-cell clones carry- ing high-affinity TCRs specific for a CD20 peptide pre- sented in HLA-A*0201. CD20 dependent recognition could be demonstrated by genetically engineering CD20- negative K562-A2 cells to express CD20. The isolated T-cell clones efficiently recognized CD20-expressing HLA-A*0201 primary chronic lymphocytic leukaemia (CLL), acute lymphoblastic leukaemia (ALL) and mantle cell lymphoma (MCL), while recognition of CD20-negative hematopoietic and non-hematopoietic cell-subsets was absent. In addition, the CD20-specific T-cell clones were able to more efficiently recognise ALL cell-lines than CD20 specific antibodies. We demonstrated that on ALL cell lines with only very low CD20 surface expression, the CD20-specific T cell clones could still efficiently recognise endogenously processed CD20-derived peptides in the context of HLA-A*0201. In conclusion, we developed a platform for the rapid identification of high-affinity TCRs of therapeutic rele- vance targeted to self-antigens by combining gene expres- sion data with valuable information on peptide processing from peptide elution studies and exploiting the immuno- genicity of foreign HLA. Using this platform we success- fully isolated CD20-specific TCRs which could broaden the application of immunotherapies targeted to CD20 in cases were CD20-cell surface expression is low. Based on its general principle the developed platform could easily be adapted to target other malignancies. Authors’ details 1 Leiden University Medical Center, Hematology, Leiden, the Netherlands. 2 Sanquin Research, Hematopoiesis, Amsterdam, the Netherlands. 3 Leiden University Medical Center, Immunohematology and Blood Transfusion, Leiden, the Netherlands. 1 Leiden University Medical Center, Hematology, Leiden, the Netherlands Full list of author information is available at the end of the article Jahn et al. Journal for ImmunoTherapy of Cancer 2014, 2(Suppl 2):I16 http://www.immunotherapyofcancer.org/content/2/S2/I16 © 2014 Jahn et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Transcript of S78. Proffered paper: High-affinity CD20-specific TCRs suitable for adoptive immunotherapy can be...
INVITED SPEAKER PRESENTATION Open Access
S78. Proffered paper: High-affinity CD20-specificTCRs suitable for adoptive immunotherapy canbe readily isolated from the allo-repertoire usingreverse immunologyL Jahn1*, P Hombrink2, C Hassan3, MGD Kester1, MP Schoonakker1, JHF Falkenburg1, P van Veelen3,MHM Heemskerk1
From 1st Immunotherapy of Cancer Conference (ITOC1)Munich, Germany. 12-14 March 2014
Studies using T-cell receptor (TCR) or chimeric antigenreceptor (CAR) transduced T-cells have shown the effec-tiveness of adoptive immunotherapy to treat differentmalignancies. The efficacy and safety of such interventionsgreatly depends on good target selection to prevent on-target toxicity. Furthermore, the broad application ofTCR-based adoptive immunotherapy is hampered by alack of an effective immune response against self-antigens.Through self-tolerance, T-cells carrying high-affinity TCRsreactive to self-antigens are deleted during thymic selec-tion. An attractive strategy is to exploit the immunogeni-city of foreign human leukocyte antigen (HLA) moleculesto generate an effective immune response against theseantigens. Here, we describe a protocol to efficiently isolatehigh-avidity alloHLA-restricted T-cells targeting the B-cellcompartment.From a B-cell peptide elution library 15 peptides
derived from genes exhibiting B-cell restricted expressionpatterns were identified and peptide-MHC multimers(pMHC) of HLA-A*0201 were generated. Via MACSort-ing and FACSorting a plethora of pMHC-multimer bind-ing T-cell clones from HLA-A*0201-negative individualswere isolated. Generated T-cell clones were selectedbased on peptide-specificity and avidity for furthercharacterization.We successfully isolated two distinct T-cell clones carry-
ing high-affinity TCRs specific for a CD20 peptide pre-sented in HLA-A*0201. CD20 dependent recognition
could be demonstrated by genetically engineering CD20-negative K562-A2 cells to express CD20. The isolatedT-cell clones efficiently recognized CD20-expressingHLA-A*0201 primary chronic lymphocytic leukaemia(CLL), acute lymphoblastic leukaemia (ALL) and mantlecell lymphoma (MCL), while recognition of CD20-negativehematopoietic and non-hematopoietic cell-subsets wasabsent. In addition, the CD20-specific T-cell clones wereable to more efficiently recognise ALL cell-lines thanCD20 specific antibodies. We demonstrated that on ALLcell lines with only very low CD20 surface expression, theCD20-specific T cell clones could still efficiently recogniseendogenously processed CD20-derived peptides in thecontext of HLA-A*0201.In conclusion, we developed a platform for the rapid
identification of high-affinity TCRs of therapeutic rele-vance targeted to self-antigens by combining gene expres-sion data with valuable information on peptide processingfrom peptide elution studies and exploiting the immuno-genicity of foreign HLA. Using this platform we success-fully isolated CD20-specific TCRs which could broadenthe application of immunotherapies targeted to CD20 incases were CD20-cell surface expression is low. Based onits general principle the developed platform could easilybe adapted to target other malignancies.
Authors’ details1Leiden University Medical Center, Hematology, Leiden, the Netherlands.2Sanquin Research, Hematopoiesis, Amsterdam, the Netherlands. 3LeidenUniversity Medical Center, Immunohematology and Blood Transfusion,Leiden, the Netherlands.
1Leiden University Medical Center, Hematology, Leiden, the NetherlandsFull list of author information is available at the end of the article
Jahn et al. Journal for ImmunoTherapy of Cancer 2014, 2(Suppl 2):I16http://www.immunotherapyofcancer.org/content/2/S2/I16
© 2014 Jahn et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative CommonsAttribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction inany medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Published: 12 March 2014
doi:10.1186/2051-1426-2-S2-I16Cite this article as: Jahn et al.: S78. Proffered paper: High-affinityCD20-specific TCRs suitable for adoptive immunotherapy can be readilyisolated from the allo-repertoire using reverse immunology. Journal forImmunoTherapy of Cancer 2014 2(Suppl 2):I16.
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Jahn et al. Journal for ImmunoTherapy of Cancer 2014, 2(Suppl 2):I16http://www.immunotherapyofcancer.org/content/2/S2/I16
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