Major histocompatibility complex (MHC) class I molecules present peptides that are derived from endogenous proteins1. These antigens can also be transferred to professional antigen-presenting cells in a process called cross-presentation, which precedes initiation of a proper T-cell response2,3; but exactly how they do this is unclear. We tested whether peptides can be transferred directly from the cytoplasm of one cell into the cytoplasm of its neighbour through gap junctions. Here we show that peptides with a relative molecular mass of up to ∼1,800 diffuse intercellularly through gap junctions unless a three-dimensional structure is imposed. This intercellular peptide transfer causes cytotoxic T-cell recognition of adjacent, innocent bystander cells as well as activated monocytes. Gap-junction-mediated peptide transfer is restricted to a few coupling cells owing to the high cytosolic peptidase activity. We present a mechanism of antigen acquisition for cross-presentation that couples the antigen presentation system of two adjacent cells and is lost in most tumours: gap-junction-mediated intercellular peptide coupling for presentation by bystander MHC class I molecules and transfer to professional antigen presenting cells for cross-priming.
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We thank R. van Beem and E. Sellink for human monocyte and HUVEC isolations, W. Moolenaar, B. Giepmans and L. van Zeijl for A431/Cx43 cells and Cx43 reagents, R. Luiten and H. Spits for T-cell clone (InfA13TGA), W. E. Benckhuijsen for peptide synthesis, E. Mesman and M. Tjin-A-Koeng for immunohistochemistry, K. Jalink for experimental support, and H. Pickersgill, A. Griekspoor and M. Wolkers for critical reading. This work was supported by grants from the Dutch Cancer Society KWF.
Authors' contributions. J.N. and C.H. performed most experiments with support from E.R. Constructs were made by L.J. and peptides by J.W.D. Supervision by J.N.
The authors declare that they have no competing financial interests.
The fluorescence in the bottom-right cell was photo-inactivated (bleached) and recovered over time to the cost of fluorescence in the micro-injected cell. (AVI 3568 kb)
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