PMID- 26599878
OWN - NLM
STAT- MEDLINE
DCOM- 20160620
LR  - 20181113
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 10
IP  - 11
DP  - 2015
TI  - A Simple Proteomics-Based Approach to Identification of Immunodominant Antigens 
      from a Complex Pathogen: Application to the CD4 T Cell Response against Human 
      Herpesvirus 6B.
PG  - e0142871
LID - 10.1371/journal.pone.0142871 [doi]
LID - e0142871
AB  - Most of humanity is chronically infected with human herpesvirus 6 (HHV-6), with 
      viral replication controlled at least in part by a poorly characterized CD4 T 
      cell response. Identification of viral epitopes recognized by CD4 T cells is 
      complicated by the large size of the herpesvirus genome and a low frequency of 
      circulating T cells responding to the virus. Here, we present an alternative to 
      classical epitope mapping approaches used to identify major targets of the T cell 
      response to a complex pathogen like HHV-6B. In the approach presented here, 
      extracellular virus preparations or virus-infected cells are fractionated by 
      SDS-PAGE, and eluted fractions are used as source of antigens to study cytokine 
      responses in direct ex vivo T cell activation studies. Fractions inducing 
      significant cytokine responses are analyzed by mass spectrometry to identify 
      viral proteins, and a subset of peptides from these proteins corresponding to 
      predicted HLA-DR binders is tested for IFN-gamma production in seropositive donors 
      with diverse HLA haplotypes. Ten HHV-6B viral proteins were identified as 
      immunodominant antigens. The epitope-specific response to HHV-6B virus was 
      complex and variable between individuals. We identified 107 peptides, each 
      recognized by at least one donor, with each donor having a distinctive footprint. 
      Fourteen peptides showed responses in the majority of donors. Responses to these 
      epitopes were validated using in vitro expanded cells and naturally expressed 
      viral proteins. Predicted peptide binding affinities for the eight HLA-DRB1 
      alleles investigated here correlated only modestly with the observed CD4 T cell 
      responses. Overall, the response to the virus was dominated by peptides from the 
      major capsid protein U57 and major antigenic protein U11, but responses to other 
      proteins including glycoprotein H (U48) and tegument proteins U54 and U14 also 
      were observed. These results provide a means to follow and potentially modulate 
      the CD4 T-cell immune response to HHV-6B.
FAU - Becerra-Artiles, Aniuska
AU  - Becerra-Artiles A
AD  - Department of Pathology, University of Massachusetts Medical School, Worcester, 
      MA, United States of America.
FAU - Dominguez-Amorocho, Omar
AU  - Dominguez-Amorocho O
AD  - Department of Pathology, University of Massachusetts Medical School, Worcester, 
      MA, United States of America.
FAU - Stern, Lawrence J
AU  - Stern LJ
AD  - Department of Pathology, University of Massachusetts Medical School, Worcester, 
      MA, United States of America.
AD  - Department of Biochemistry and Molecular Pharmacology, University of 
      Massachusetts Medical School, Worcester, MA, United States of America.
FAU - Calvo-Calle, J Mauricio
AU  - Calvo-Calle JM
AD  - Department of Pathology, University of Massachusetts Medical School, Worcester, 
      MA, United States of America.
LA  - eng
GR  - U19 AI057319/AI/NIAID NIH HHS/United States
GR  - HHSN27220140046C/PHS HHS/United States
GR  - U19 AI109858/AI/NIAID NIH HHS/United States
GR  - U19-AI057319/AI/NIAID NIH HHS/United States
GR  - U19-AI109858/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20151123
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Antigens, Viral)
RN  - 0 (HLA-DRB1 Chains)
RN  - 0 (Immunodominant Epitopes)
RN  - 0 (Peptides)
RN  - 0 (Viral Proteins)
RN  - 82115-62-6 (Interferon-gamma)
SB  - IM
MH  - Algorithms
MH  - Alleles
MH  - Amino Acid Sequence
MH  - Antigens, Viral/*immunology
MH  - CD4-Positive T-Lymphocytes/*immunology
MH  - Cell Separation
MH  - Chromatography, Liquid
MH  - HLA-DRB1 Chains/immunology
MH  - Haplotypes/genetics
MH  - Herpesvirus 6, Human/*immunology
MH  - Humans
MH  - Immunodominant Epitopes/*immunology
MH  - Interferon-gamma/metabolism
MH  - Mass Spectrometry
MH  - Molecular Sequence Data
MH  - Peptides/chemistry
MH  - Protein Binding
MH  - Proteomics/*methods
MH  - Reproducibility of Results
MH  - Tissue Donors
MH  - Viral Proteins/immunology
PMC - PMC4658110
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2015/11/26 06:00
MHDA- 2016/06/21 06:00
PMCR- 2015/11/23
CRDT- 2015/11/25 06:00
PHST- 2015/09/21 00:00 [received]
PHST- 2015/10/27 00:00 [accepted]
PHST- 2015/11/25 06:00 [entrez]
PHST- 2015/11/26 06:00 [pubmed]
PHST- 2016/06/21 06:00 [medline]
PHST- 2015/11/23 00:00 [pmc-release]
AID - PONE-D-15-41761 [pii]
AID - 10.1371/journal.pone.0142871 [doi]
PST - epublish
SO  - PLoS One. 2015 Nov 23;10(11):e0142871. doi: 10.1371/journal.pone.0142871. 
      eCollection 2015.