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The association between human cytomegalovirus (hCMV) reactivation and thedevelopment of graft-versus-host-disease (GVHD) has been observed in stem celltransplantation (SCT). Seventy seven SCT donor-recipient pairs (DRP) (HLAmatched unrelated donor (MUD), n=50; matched related donor (MRD), n=27)underwent whole exome sequencing to identify single nucleotide polymorphisms(SNPs) generating alloreactive peptide libraries for each DRP (9-merpeptide-HLA complexes); Human CMV CROSS (Cross-Reactive Open Source Sequence)Database was compiled from NCBI; HLA class I binding affinity for each DRPs HLAwas calculated by NetMHCpan 2.8 and hCMV- derived 9-mers algorithmicallycompared to the alloreactive peptide-HLA complex libraries. Short consecutive(6 or greater) amino acid (AA) sequence homology matching hCMV to recipientpeptides was considered for HLA-bound-peptide (IC50<500 nM) cross reactivity.Of the 70,686 hCMV 9-mers contained within the hCMV CROSS database, 29,658.8+/- 9038.5 were found to match MRD DRP alloreactive peptides and 52,910.2 +/-16121.8 matched MUD DRP peptides (Student's T-test, p<0.001). In silicoanalysis revealed multiple high affinity, immunogenic CMV-Human peptide matches(IC50<500 nM) expressed in GVHD-affected tissue-specific manner (proteinsexpressed at 10 RPKM or greater). hCMV+GVHD was found in 18 patients, 13developing hCMV viremia before GVHD onset with a subset analysis of 7 instancesof hCMV viremia prior to acute GVHD onset (n=3), chronic GVHD (n=2) and acute +chronic GVHD (n=2) indicating cross reactive peptide expression within affectedorgans. We propose that based on our analysis and preliminary clinicalcorrelations that hCMV immune cross-reactivity may cause antigenic mimicry ofhuman alloreactive peptides triggering GVHD.

Cytomegalovirus Antigenic Mimicry of Human Alloreactive Peptides: A Potential Trigger for Graft versus Host Disease