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PLoS One. 2014 Apr 16;9(4):e89648. doi: 10.1371/journal.pone.0089648. eCollection 2014.

Whole CMV proteome pattern recognition analysis after HSCT identifies unique epitope targets associated with the CMV status.

Author information

1
Department of Medicine Huddinge, Karolinska Institutet; Dept. of Hematology, Karolinska University Hospital, Stockholm, Sweden; CAST (Center for allogeneic stem cell transplantation), Karolinska Hospital.
2
CAST (Center for allogeneic stem cell transplantation), Karolinska Hospital.
3
Microbiology, Tumor and Cell Biology (MTC), Stockholm, Sweden.
4
The Swedish Institute for Infectious Disease Control (SMI), Stockholm, Sweden.
5
Department of Enzymology, Institute for Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, Halle, Germany.
6
Division of Therapeutic Immunology (TIM), LabMed Karolinska Institutet, Stockholm, Sweden.
7
Department of Genome Biology, John Curtin School of Medical Research, The Australian National University, Canberra, Australia.
8
Department of Medicine Huddinge, Karolinska Institutet; Dept. of Hematology, Karolinska University Hospital, Stockholm, Sweden.
9
CAST (Center for allogeneic stem cell transplantation), Karolinska Hospital; Division of Therapeutic Immunology (TIM), LabMed Karolinska Institutet, Stockholm, Sweden.

Abstract

Cytomegalovirus (CMV) infection represents a vital complication after Hematopoietic Stem Cell Transplantation (HSCT). We screened the entire CMV proteome to visualize the humoral target epitope-focus profile in serum after HSCT. IgG profiling from four patient groups (donor and/or recipient +/- for CMV) was performed at 6, 12 and 24 months after HSCT using microarray slides containing 17174 of 15mer-peptides overlapping by 4 aa covering 214 proteins from CMV. Data were analyzed using maSigPro, PAM and the 'exclusive recognition analysis (ERA)' to identify unique CMV epitope responses for each patient group. The 'exclusive recognition analysis' of serum epitope patterns segregated best 12 months after HSCT for the D+/R+ group (versus D-/R-). Epitopes were derived from UL123 (IE1), UL99 (pp28), UL32 (pp150), this changed at 24 months to 2 strongly recognized peptides provided from UL123 and UL100. Strongly (IgG) recognized CMV targets elicited also robust cytokine production in T-cells from patients after HSCT defined by intracellular cytokine staining (IL-2, TNF, IFN and IL-17). High-content peptide microarrays allow epitope profiling of entire viral proteomes; this approach can be useful to map relevant targets for diagnostics and therapy in patients with well defined clinical endpoints. Peptide microarray analysis visualizes the breadth of B-cell immune reconstitution after HSCT and provides a useful tool to gauge immune reconstitution.

PMID:
24740411
PMCID:
PMC3989190
DOI:
10.1371/journal.pone.0089648
[Indexed for MEDLINE]
Free PMC Article

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