Fine Tuning and Efficient T Cell Activation with Stimulatory aCD3 Nanoarrays

Abstract

Anti-CD3 (aCD3) nanoarrays fabricated by self-assembled nanopatterning combined with site-directed protein immobilization techniques represent a novel T cell stimulatory platform that allows tight control over ligand orientation and surface density. Here, we show that activation of primary human CD4+ T cells, defined by CD69 upregulation, IL-2 production and cell proliferation, correlates with aCD3 density on nanoarrays. Immobilization of aCD3 through nanopatterning had two effects: cell activation was significantly higher on these surfaces than on aCD3-coated plastics and allowed unprecedented fine-tuning of T cell response.

Full control of ligand positioning reveals spatial thresholds for T cell receptor triggering

“…S8a), and it also increased with decreasing inter-NP spacing (Fig. 2b), in line with previous observations 10, 11, 20 . Similar spatial effects were observed for cell spreading area and the number of adherent cells, which was also generally increased by including the mobile ICAM1 layer (Supplementary Fig.…”

“…Nanofabricated surfaces have previously been used to study the effect of ligand spacing on TCR triggering 9, 10, 11 . Those studies used extended arrays (meaning much larger than the cell) of gold nanoparticle (NP)-bound TCR ligands surrounded by a non-adhesive background 9, 10 .…”

“…Those studies used extended arrays (meaning much larger than the cell) of gold nanoparticle (NP)-bound TCR ligands surrounded by a non-adhesive background 9, 10 . A graded increase in TCR triggering with decreasing inter-ligand spacing between 200 nm and 40 nm was observed, but there was no evidence of a spacing threshold 10, 11 . Those arrays, created by self-assembly, could not decouple ligand spacing from density.…”

Size-Tunable Organic Nanodot Arrays: A Versatile Platform for Manipulating and Imaging Cells

“…The second variation is STONA supported on an elastomeric substrate, which better mimics soft cells and also lend themselves to eventual use as a tool for force measurements. 43,44 NAV-STONA were transferred by reverse-contact-printing 13 to silicone rubber coated on glass, with a measured elasticity of 300 kPa (Fig. 1g, 4g, Fig.…”

Plasmonic Surfaces for Cell Growth and Retrieval Triggered by Near‐Infrared Light

“…[10] Au nanoparticles were deposited on ag lass substrate by micellar block copolymer nanolithography (BCML), in which inverse micelles of the block copolymer polystyrene-b-poly-2-vinylpyridine (PS-b-P2VP) containing Au ions in the hydrophilic core,a re deposited by dip-coating onto as ubstrate, forming at wo-dimensional ordered hexagonal array. [11] The monolayer of inverse micelles is then subjected to as hort treatment of oxygen plasma for four minutes,l eading to formation of Au nanoparticles with an average diameter of 15 nm. Subsequent chemical growth was induced by immersion in asolution of AuCl 4 À ions,ascorbic acid, and hexadecyltrimethylammonium bromide (CTAB), which yields anisotropic Au nanostructures with elongated branches pointing out in random orientations.…”