Strategic internal covalent cross-linking of TNF produces a stable TNF trimer with improved TNFR2 signaling.
BACKGROUND
Soluble TNF superfamily (TNFSF) ligands are less stable and less active than their transmembrane (tm) analogues. This is a problem for the therapeutic use of recombinant TNFSF ligands in diverse diseases including cancer and autoimmunity. Creating TNFSF ligand analogues with improved targeting of their respective receptors is important for research and therapeutic purposes.


RESULTS
Covalent internal cross-linking of TNF monomers by double mutations, S95C/G148C, results in stable trimers with improved TNFR2 function. The resulting mutein induced the selective death of autoreactive CD8 T cells in type-1 diabetic patients and demonstrates targeted proliferation and expansion of human CD4 Tregs.


CONCLUSIONS
Stable TNF trimers, created by internal covalent cross-linking, show improved signaling. The high structural homology within the TNF superfamily provides an opportunity to extend internal cross-linking to other TNF superfamily proteins to produce active trimers with improved stability and receptor signaling, and with potential applications for cancer, autoimmunity, infections, and transplantation.
Immunobiology Laboratory, Harvard Medical School and Massachusetts General Hospital, Boston, MA USA.
Liqin
L
Ban
Immunobiology Laboratory, Harvard Medical School and Massachusetts General Hospital, Boston, MA USA.
Willem
W
Kuhtreiber
Immunobiology Laboratory, Harvard Medical School and Massachusetts General Hospital, Boston, MA USA.
John
J
Butterworth
Immunobiology Laboratory, Harvard Medical School and Massachusetts General Hospital, Boston, MA USA.
Yoshiaki
Y
Okubo
Immunobiology Laboratory, Harvard Medical School and Massachusetts General Hospital, Boston, MA USA.
Éva S
ÉS
Vanamee
Immunobiology Laboratory, Harvard Medical School and Massachusetts General Hospital, Boston, MA USA.
Denise L
DL
Faustman
Cell signaling; TNF; TNFR2; Transmembrane TNF; Trimer; Tumor necrosis factor; Tumor necrosis factor receptor