Tumor necrosis factor (TNF)-functionalized nanostructured particles for the stimulation of membrane TNF-specific cell responses.
Most members of the tumor necrosis factor (TNF) ligand family occur in both a membrane-bound and a soluble form, which can possess differential bioactivities. The aim of this work was the construction of a synthetic-biological hybrid system consisting of chemically nanostructured core-shell particles with a diameter of 100 nm, 1 microm, or 10 microm and the cytokine TNF to obtain a tool that mimics the bioactivity of naturally occurring membrane-bound TNF. Synthetic core-shell nanoparticles consisting of an inorganic silica core and an ultrathin organic shell bearing a maleimide group at the shell surface which allowed for a covalent and site-directed coupling of CysHisTNF mutants were prepared. The TNF mutants were modified at the N-terminus by PCR cloning by introducing a His-Tag for purification and a free cysteine group for reaction with the particle-attached maleimide group. The resulting nanostructured hybrid particles initiated strong TNF receptor type 2 specific responses, otherwise only seen for the membrane-bound form of TNF, but not the soluble cytokine, thus clearly demonstrating new and membrane TNF-like properties of the bioconjugated soluble TNF.
Bryde
Susanne
S
Institute of Cell Biology and Immunology, University of Stuttgart, Allmandring 31, D-70569 Stuttgart, Germany.
Grunwald
Ingo
I
Hammer
Angela
A
Krippner-Heidenreich
Anja
A
Schiestel
Thomas
T
Brunner
Herwig
H
Tovar
Günter E M
GE
Pfizenmaier
Klaus
K
Scheurich
Peter
P
D000818:Animals; D045744:Cell Line, Tumor; D015195:Drug Design; D006801:Humans; D008024:Ligands; D008301:Maleimides; D008565:Membrane Proteins / Q000737:chemistry; D051379:Mice; D018716:Molecular Mimicry; D009154:Mutation; D049329:Nanostructures / Q000737:chemistry*; D010316:Particle Size; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000378:metabolism*; D012822:Silicon Dioxide; D012995:Solubility; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry* / Q000235:genetics
D008024:Ligands; D008301:Maleimides; D008565:Membrane Proteins; D047889:Receptors, Tumor Necrosis Factor, Type II; D014409:Tumor Necrosis Factor-alpha; C043592:maleimide; D012822:Silicon Dioxide