ABSTRACT HLA‐E and ‐G class Ib molecules were considered unrelated to viral antigen presentation. HLA‐E binds nonamers from the leader sequences of other HLA‐I molecules and the human cytomegalovirus (HCMV) UL40 protein, interacting with CD94/NKG2 NK cell receptors. Yet, evidence that HLA‐E may present some pathogen‐derived peptides to CD8+ T lymphocytes has been reported. By contrast, HLA‐G binds a broad spectrum of endogenous sequences but its role in antigen presentation is unknown. An experimental approach was set up to search for HCMV antigens displayed by HLA‐G in infected cells. Among the analysed peptidome, 22 sequences corresponding to 16 HCMV molecules were identified; 17 peptides were confirmed to interact in vitro with HLA‐G of which 10 displayed characteristic anchor residues. As compared to the response in short‐term (6 h) assays to immunodominant IE‐1 and pp65 antigens, none of the HLA‐G‐binding peptides stimulated cytokine production by CD8+ T cells from HCMV‐seropositive blood donors ( n = 15). Following a 14‐day peptide stimulation of PBMC and expansion with IL‐2, CD8+ T cells specifically responding to a subset of these viral antigens were detected in some individuals, yet were not restricted by HLA‐G in functional assays. A subset of viral peptides did bind to both HLA‐G and ‐E but were not recognised by CD94/NKG2 NK cell receptors. Our results provide the first evidence that HLA‐G may display potentially immunogenic viral peptides in HCMV‐infected cells, yet do not support their ability to promote HLA‐G‐restricted CD8+ T cell responses nor to modulate NK cell functions.