PMID- 29603667
OWN - NLM
STAT- MEDLINE
DCOM- 20190710
LR  - 20240426
IS  - 1615-9861 (Electronic)
IS  - 1615-9853 (Print)
IS  - 1615-9853 (Linking)
VI  - 18
IP  - 11
DP  - 2018 Jun
TI  - A Targeted LC-MS Strategy for Low-Abundant HLA Class-I-Presented Peptide 
      Detection Identifies Novel Human Papillomavirus T-Cell Epitopes.
PG  - e1700390
LID - 10.1002/pmic.201700390 [doi]
LID - 1700390
AB  - For rational design of therapeutic vaccines, detailed knowledge about target 
      epitopes that are endogenously processed and truly presented on infected or 
      transformed cells is essential. Many potential target epitopes (viral or 
      mutation-derived), are presented at low abundance. Therefore, direct detection of 
      these peptides remains a challenge. This study presents a method for the 
      isolation and LC-MS(3) -based targeted detection of low-abundant human leukocyte 
      antigen (HLA) class-I-presented peptides from transformed cells. Human 
      papillomavirus (HPV) was used as a model system, as the HPV oncoproteins E6 and 
      E7 are attractive therapeutic vaccination targets and expressed in all 
      transformed cells, but present at low abundance due to viral immune evasion 
      mechanisms. The presented approach included preselection of target 
      antigen-derived peptides by in silico predictions and in vitro binding assays. 
      The peptide purification process was tailored to minimize contaminants after 
      immunoprecipitation of HLA-peptide complexes, while keeping high isolation yields 
      of low-abundant target peptides. The subsequent targeted LC-MS(3) detection 
      allowed for increased sensitivity, which resulted in successful detection of the 
      known HLA-A2-restricted epitope E7(11-19) and ten additional E7-derived peptides 
      on the surface of HPV16-transformed cells. T-cell reactivity was shown for all 
      the 11 detected peptides in ELISpot assays, which shows that detection by our 
      approach has high predictive value for immunogenicity. The presented strategy is 
      suitable for validating even low-abundant candidate epitopes to be true 
      immunotherapy targets.
CI  - (c) 2018 The Authors. Proteomics Published by Wiley-VCH Verlag GmbH & Co. KGaA.
FAU - Blatnik, Renata
AU  - Blatnik R
AD  - Immunotherapy and Immunoprevention, German Cancer Research Center (DKFZ), Im 
      Neuenheimer Feld 280, 69120 Heidelberg, Germany.
AD  - Molecular Vaccine Design, German Center for Infection Research (DZIF), Partner 
      Site Heidelberg, Heidelberg, Germany.
FAU - Mohan, Nitya
AU  - Mohan N
AUID- ORCID: 0000-0002-8189-1399
AD  - Immunotherapy and Immunoprevention, German Cancer Research Center (DKFZ), Im 
      Neuenheimer Feld 280, 69120 Heidelberg, Germany.
FAU - Bonsack, Maria
AU  - Bonsack M
AUID- ORCID: 0000-0003-2087-7451
AD  - Immunotherapy and Immunoprevention, German Cancer Research Center (DKFZ), Im 
      Neuenheimer Feld 280, 69120 Heidelberg, Germany.
AD  - Molecular Vaccine Design, German Center for Infection Research (DZIF), Partner 
      Site Heidelberg, Heidelberg, Germany.
FAU - Falkenby, Lasse G
AU  - Falkenby LG
AD  - Department of Biochemistry and Molecular Biology, University of Southern Denmark, 
      Odense M, Denmark.
FAU - Hoppe, Stephanie
AU  - Hoppe S
AD  - Immunotherapy and Immunoprevention, German Cancer Research Center (DKFZ), Im 
      Neuenheimer Feld 280, 69120 Heidelberg, Germany.
AD  - Molecular Vaccine Design, German Center for Infection Research (DZIF), Partner 
      Site Heidelberg, Heidelberg, Germany.
FAU - Josef, Kathrin
AU  - Josef K
AUID- ORCID: 0000-0001-9026-7048
AD  - Immunotherapy and Immunoprevention, German Cancer Research Center (DKFZ), Im 
      Neuenheimer Feld 280, 69120 Heidelberg, Germany.
AD  - Molecular Vaccine Design, German Center for Infection Research (DZIF), Partner 
      Site Heidelberg, Heidelberg, Germany.
FAU - Steinbach, Alina
AU  - Steinbach A
AD  - Immunotherapy and Immunoprevention, German Cancer Research Center (DKFZ), Im 
      Neuenheimer Feld 280, 69120 Heidelberg, Germany.
AD  - Molecular Vaccine Design, German Center for Infection Research (DZIF), Partner 
      Site Heidelberg, Heidelberg, Germany.
FAU - Becker, Sara
AU  - Becker S
AUID- ORCID: 0000-0001-5182-2360
AD  - Immunotherapy and Immunoprevention, German Cancer Research Center (DKFZ), Im 
      Neuenheimer Feld 280, 69120 Heidelberg, Germany.
FAU - Nadler, Wiebke M
AU  - Nadler WM
AUID- ORCID: 0000-0002-7673-1147
AD  - Division of Stem Cells and Cancer, German Cancer Research Center (DKFZ) and 
      Heidelberg Institute for Stem Cell Technology and Experimental Medicine 
      (HI-STEM), Heidelberg, Germany.
FAU - Rucevic, Marijana
AU  - Rucevic M
AUID- ORCID: 0000-0003-1003-2666
AD  - Massachusetts General Hospital, Center for Cancer Research, Charlestown, MA, USA.
FAU - Larsen, Martin R
AU  - Larsen MR
AUID- ORCID: 0000-0001-6203-0123
AD  - Department of Biochemistry and Molecular Biology, University of Southern Denmark, 
      Odense M, Denmark.
FAU - Salek, Mogjiborahman
AU  - Salek M
AUID- ORCID: 0000-0002-6664-4752
AD  - Immunotherapy and Immunoprevention, German Cancer Research Center (DKFZ), Im 
      Neuenheimer Feld 280, 69120 Heidelberg, Germany.
AD  - Molecular Vaccine Design, German Center for Infection Research (DZIF), Partner 
      Site Heidelberg, Heidelberg, Germany.
FAU - Riemer, Angelika B
AU  - Riemer AB
AUID- ORCID: 0000-0002-5865-0714
AD  - Immunotherapy and Immunoprevention, German Cancer Research Center (DKFZ), Im 
      Neuenheimer Feld 280, 69120 Heidelberg, Germany.
AD  - Molecular Vaccine Design, German Center for Infection Research (DZIF), Partner 
      Site Heidelberg, Heidelberg, Germany.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20180502
PL  - Germany
TA  - Proteomics
JT  - Proteomics
JID - 101092707
RN  - 0 (Epitopes, T-Lymphocyte)
RN  - 0 (Histocompatibility Antigens Class I)
RN  - 0 (Peptide Fragments)
SB  - IM
MH  - Chromatography, Liquid/*methods
MH  - Epitopes, T-Lymphocyte/immunology/*metabolism
MH  - Female
MH  - Histocompatibility Antigens Class I/immunology/*metabolism
MH  - Humans
MH  - Papillomaviridae/*immunology
MH  - Papillomavirus Infections/immunology/metabolism/virology
MH  - Peptide Fragments/*analysis
MH  - Tandem Mass Spectrometry/*methods
MH  - Tumor Cells, Cultured
MH  - Uterine Cervical Neoplasms/immunology/*metabolism/virology
PMC - PMC6033010
OTO - NOTNLM
OT  - human papillomavirus (HPV)
OT  - immunopeptidomics
OT  - immunotherapy
OT  - neoepitopes
OT  - targeted mass spectrometry (MS)
EDAT- 2018/04/01 06:00
MHDA- 2019/07/11 06:00
PMCR- 2018/07/05
CRDT- 2018/04/01 06:00
PHST- 2017/10/21 00:00 [received]
PHST- 2018/03/16 00:00 [revised]
PHST- 2018/04/01 06:00 [pubmed]
PHST- 2019/07/11 06:00 [medline]
PHST- 2018/04/01 06:00 [entrez]
PHST- 2018/07/05 00:00 [pmc-release]
AID - PMIC12864 [pii]
AID - 10.1002/pmic.201700390 [doi]
PST - ppublish
SO  - Proteomics. 2018 Jun;18(11):e1700390. doi: 10.1002/pmic.201700390. Epub 2018 May 
      2.