Mixed alkanethiol monolayers on submicrometric gold patterns: a controlled platform for studying cell-ligand interactions.
Nanoscale organization of surface ligands often has a critical effect on cell-surface interactions. We have developed an experimental system that allows a high degree of control over the 2-D spatial distribution of ligands. As a proof of concept, we used the developed system to study how T-cell activation is independently affected by antigen density and antigen amount per cell. Arrays of submicrometer gold islands at varying surface coverage were defined on silicon by electron beam lithography (EBL). The gold islands were functionalized with alkanethiol self-assembled monolayers (SAMs) containing a small antigen, 2,4,6-trinotrophenyl (TNP), at various densities. Genetically engineered T-cell hybridomas expressing TNP-specific chimeric T-cell antigen receptor (CAR) were cultured on the SAMs, and their activation was assessed by IL-2 secretion and CD69 expression. It was found that, at constant antigen density, activation increased monotonically with the amount of antigen, while at constant antigen amount activation was maximal at an intermediate antigen density, whose value was independent of the amount of antigen.
Department of Physics, Technion-Israel Institute of Technology, Haifa 32000, Israel.
Rami
R
Fishler
Arbel
A
Artzy-Schnirman
Elad
E
Peer
Ron
R
Wolchinsky
Reuven
R
Brener
Tova
T
Waks
Zelig
Z
Eshhar
Yoram
Y
Reiter
Uri
U
Sivan
D000473:Alkanes / Q000737:chemistry*; D006046:Gold / Q000737:chemistry*; D007118:Immunoassay / Q000379:methods*; D008422:Materials Testing; D054802:Molecular Imprinting / Q000379:methods; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D040081:Protein Array Analysis / Q000379:methods*; D025941:Protein Interaction Mapping / Q000379:methods*; D013438:Sulfhydryl Compounds / Q000737:chemistry*
D000473:Alkanes; D013438:Sulfhydryl Compounds; D006046:Gold