Direct stimulation of cellular immune response via TLR2 signaling triggered by contact with hybrid glyco-biointerfaces composed of chitohexaose and cellohexaose

Recognition of carbohydrates by cell receptors activates various cell signaling processes. In this report, hybrid self-assembled monolayers (SAMs) of chitohexaose (βGlcNAc6) and cellohexaose (βGlc6) are prepared to induce a highly sensitive, glyco-density-dependent inflammatory response by HEK293 cells expressing toll-like receptor 2 (TLR2), which recognizes oligo-βGlcNAc moieties. Thiosemicarbazide-labeled βGlcNAc6 and βGlc6, as a bio-signal inducer and a spacer molecule, respectively, were immobilized on a gold substrate via spontaneous chemisorption to control the βGlcNAc6 density on the SAM surface. The βGlcNAc6 density ranged from 0 to 0.65 chains nm ^–2 . The inflammatory response of HEK293 cells varied as a function of the surface glyco-density, and the hybrid SAM with 0.146 chains nm ^–2 of βGlcNAc6 induced a stronger response than pure βGlcNAc6-SAM with 0.654 chains nm ^–2 . Thus, direct activation of TLR2-mediated cellular stimulation triggered by as-designed glyco-biointerfaces was possible through a mutual interaction between the fittingly βGlcNAc6 moieties assembled on hybrid glyco-SAMs and TLR2 on the surface of HEK293 cells.