Patent ID: 11912751

Abstract:
This invention discloses a bifunctional TGF-B/immune checkpoint fusion gene and protein with anti-inflammatory activity that represents a new class of therapy for Immune disorders, immune dysregulation, and autoimmune diseases. The bifunctional TGF-B/immune checkpoint fusion gene and protein include: (i) a TGF-B domain consists of TGF-B1 (ii) an immune checkpoint consisting of PD-L1, (iii) a flexible peptide linker that links two TGF-B domains resulting in a dimeric TGF-B construct wherein the dimeric form of the TGF-B ligand is important for its binding and functional activity, and (iv) a rigid peptide linker, wherein the dimeric TGF-B ligand is linked to the immune checkpoint ligand.A unique feature of this invention is the engineering of a dimeric TGF-B1 domain in the TGF-B1/PD-L1 fusion protein which was experimentally shown to be important in its binding to its TFGBR1 receptor and its functional activity. The functional domains of the fusion protein act in concert to maintain tolerance for self-antigens by stimulating regulatory cells and to regulate the activities of effector cells, respectively. The immune regulatory activity of the bifunctional TGF-B/immune checkpoint fusion gene and protein was demonstrated using a TGF-B1/PD-L1 fusion pair, showing that the domains functionally bind their respective receptors and inhibit NF-kB activation in a reporter assay, downregulate NF-kB-responsive genes, suppress the growth of PHA/IFN-γ-activated effector-like Jurkat T cells with subsequent upregulation of Foxp3 and downregulation of NF-kB1 and IL-6. The fusion protein described in this invention elicits a high frequency of cell-to-cell contact that induced upregulation of Treg-associated markers (Foxp3 and IL2RA). The bifunctional fusion protein described herein modulates immune reactions and represents a new class of therapy for Immune disorders, immune dysregulation, and autoimmune diseases.