Jashodeep Datta, Shuwen Xu, Cinthia Rosemblit, Jenessa B. Smith, Jessica A. Cintolo, Daniel J. Powell, Brian J. Czerniecki

Vaccination strategies incorporating the immunodominant HLA-A2-restricted HER2/neu-derived peptide 369-377 (HER2369-377) are increasingly utilized in HER2/neu-expressing cancer patients. The failure of post-vaccination HER2369-377-specific CD8(+) T-cells to recognize HLA-A2(pos) HER2/neu-expressing cells in vitro, however, has been attributed to impaired MHC class I/HLA-A2 presentation observed in HER2/neu-overexpressing tumors. We reconcile this controversy by demonstrating that HER2369-377 is directly recognized by high functional-avidity HER2369-377-specific CD8(+) T-cells-either genetically modified to express a novel HER2369-377-TCR or sensitized using HER2369-377-pulsed type 1-polarized dendritic cells (DC1)-on class I-abundant HER2(low), but not class I-deficient HER2(high), cancer cells. Importantly, a critical cooperation between CD4(+) T-helper type-1 (Th1) cytokines IFN-γ/TNF-α and HER2/neu-targeted antibody trastuzumab is necessary to restore class I expression in HER2(high) cancers, thereby facilitating recognition and lysis of these cells by HER2369-377-specific CD8(+) T-cells. Concomitant induction of PD-L1 on HER2/neu-expressing cells by IFN-γ/TNF and trastuzumab, however, has minimal impact on DC1-sensitized HER2369-377-CD8+ T-cell-mediated cytotoxicity. Although activation of EGFR and HER3 signaling significantly abrogates IFN-γ/TNF-α and trastuzumab-induced class I restoration, EGFR/HER3 receptor blockade rescues class I expression and ensuing HER2369-377-CD8(+) cytotoxicity of HER2/neu-expressing cells. Thus, combinations of CD4(+) Th1 immune interventions and multivalent targeting of HER family members may be required for optimal anti-HER2/neu CD8(+) T-cell-directed immunotherapy.