Immunomodulatory fusion proteins and uses thereof

Abstract

The present disclosure relates to immunomodulatory fusion proteins containing an extracellular binding domain and an intracellular signaling domain, wherein binding of a target can generate a modulatory signal in a host cell, such as a T cell. The present disclosure also relates to uses of immune cells expressing such immunomodulatory fusion proteins to treat certain diseases, such as cancer or infectious disease.

What is claimed is: 
     
         1 . A fusion protein, comprising (a) an extracellular component comprised of a binding domain that specifically binds a target, (b) an intracellular component comprised of an intracellular signaling domain, and (c) a hydrophobic component connecting the extracellular and intracellular components,
 wherein the extracellular portion of a complex formed by specific binding of the fusion protein to the target (fusion protein::target complex) is of a size, or spans a distance, of (i) up to about a distance between two cell membranes of an immunological synapse, (ii) up to about or substantially the same as a distance spanned by the extracellular portion of a complex between a T cell receptor (TCR) and an MHC-peptide complex specifically bound by the TCR, (iii) up to about or substantially the same as a distance spanned by the extracellular portion of a complex between a natural molecule comprising the binding domain and its cognate binding partner; (iii) less than or up to about 40 nm, 25 nm, 20 nm, 15 nm, or 14 nm; or (iv) any combination thereof.   
     
     
         2 . The fusion protein according to  claim 1 , wherein the fusion protein::target complex localizes to a supramolecular activation cluster (SMAC). 
     
     
         3 . The fusion protein according to  claim 1  or  2 , wherein the fusion protein::target complex localizes to a central region supramolecular activation cluster (cSMAC). 
     
     
         4 . The fusion protein according to  claim 2  or  claim 3 , wherein the SMAC has a width of a T cell receptor (TCR) associated with an antigen::human leukocyte antigen (HLA) complex. 
     
     
         5 . The fusion protein according to any one of  claims 1 - 4 , wherein the extracellular component is associated with a negative signal and the intracellular component is associated with a positive signal. 
     
     
         6 . A fusion protein, comprising (a) an extracellular component comprised of a binding domain that specifically binds a target, (b) an intracellular component comprised of an intracellular signaling domain, and (c) a hydrophobic component connecting the extracellular and intracellular components,
 wherein the binding domain is, or has at least 95% identity to, an inhibitory molecule binding domain and the intracellular signaling domain is, or contains at least 95% identity to, a costimulatory or stimulatory molecule binding domain, and   wherein the inhibitory molecule is not a B7-CD28 superfamily member, is not CTLA4, is not PD1, does not bind to B7, is a glycoprotein, or any combination thereof.   
     
     
         7 . The fusion protein according to any of  claims 1 - 6 , wherein the expression of the fusion protein in a T cell comprising a TCR or chimeric antigen receptor specific for an antigen results in at least about a 1.5-fold, 2-fold, or 3-fold increase in survival, expansion, cytotoxicity, cytokine secretion, and/or response to multiple rounds of stimulation, by the T cell, in response to binding of the antigen and/or following administration to a subject, and/or results in at least about a 1.5-fold, 2-fold, or 3-fold increase in time of survival, disease-free survival, or amelioration of one or more disease symptom, of a subject to which the cell is administered, as compared to a cell substantially the same as the T cell but not containing the fusion protein. 
     
     
         8 . The fusion protein according to any of  claims 1 - 7 , wherein the fusion protein is capable, when expressed in a T cell, of co-localizing with a TCR or CAR expressed by the T cell. 
     
     
         9 . The fusion protein according to any of  claims 6 - 8 , wherein the binding domain and/or the inhibitory molecule specifically binds to a CD200 or a CD47. 
     
     
         10 . The fusion protein according to any of  claims 6 - 9 , wherein the binding domain is derived from, and/or the inhibitory molecule is, a CD200R, a SIRPα, a TIM3, a CD2, a CD95 (Fas), a CD223 (LAG3), an A2aR, a KIR, TIM3, a CD300, or a LPA5. 
     
     
         11 . The fusion protein according to any of  claims 6 - 9 , wherein the inhibitory molecule is or comprises a CD200R. 
     
     
         12 . The fusion protein according to any of  claims 6 - 9 , wherein the inhibitory molecule is or comprises a SIRPα. 
     
     
         13 . The fusion protein according to any one of  claims 1 - 12 , wherein the binding domain is an antibody binding fragment, a receptor ectodomain, a cytokine, or a ligand. 
     
     
         14 . The fusion protein according to any one of  claims 1 - 13 , wherein the extracellular component comprises an extracellular portion of a CD200R, SIRPα, CD279 (PD-1), CD2, CD95 (Fas), CD152 (CTLA4), CD223 (LAG3), CD272 (BTLA), A2aR, KIR, TIM3, CD300, or LPA5. 
     
     
         15 . The fusion protein according to any of  claims 1 - 14 , wherein the extracellular component further comprises an additional extracellular portion, wherein the additional extracellular portion optionally is from or shares identity with an extracellular portion of a molecule that is distinct from the binding domain source molecule or does not contain the binding domain. 
     
     
         16 . The fusion protein according to any one of  claims 1 - 15 , wherein the extracellular component further comprises an extracellular portion from a the hydrophobic component, or that contains the hydrophobic component or a portion thereof. 
     
     
         17 . The fusion protein according to  claim 15  or  16 , wherein the additional extracellular portion comprises a multimerization domain and/or a spacer. 
     
     
         18 . The fusion protein according to any one of  claims 1 - 17 , wherein the extracellular component comprises an extracellular portion of a CD200R or wherein the binding domain comprises a CD200-binding portion of a CD200R or binding variant thereof. 
     
     
         19 . The fusion protein according to any one of  claims 1 - 17 , wherein the extracellular component comprises an extracellular portion of a SIRPα or wherein the binding domain comprises a CD47 binding portion of a SIRPα or binding variant thereof. 
     
     
         20 . The fusion protein according to any one of  claims 1 - 19 , wherein the extracellular component or the additional extracellular portion comprises a multimerization domain. 
     
     
         21 . The fusion protein according to  claim 20 , wherein the multimerization domain comprises a cysteine residue. 
     
     
         22 . The fusion protein according to  claim 20 , wherein the multimerization domain comprises an extracellular component modified to contain a cysteine residue within about 2 to about 15 amino acids from the hydrophobic component. 
     
     
         23 . The fusion protein according to any one of  claims 1 - 22 , wherein the hydrophobic component comprises a transmembrane domain of a CD2, CD3ε, CD3δ, CD3ζ, CD25, CD27, CD28, CD40, CD79A, CD79B, CD80, CD86, CD95 (Fas), CD134 (OX40), CD137 (4-1BB), CD150 (SLAMF1), CD152 (CTLA4), CD200R, CD223 (LAG3), CD270 (HVEM), CD272 (BTLA), CD273 (PD-L2), CD274 (PD-L1), CD278 (ICOS), CD279 (PD-1), CD300, CD357 (GITR), A2aR, DAP10, FcRα, FcRβ, FcRγ, Fyn, GALS, KIR, Lck, LAT, LRP, NKG2D, NOTCH1, NOTCH2, NOTCH3, NOTCH4, PTCH2, ROR2, Ryk, Slp76, SIRPα, pTα, TCRα, TCRβ, TIM3, TRIM, LPA5, or Zap70. 
     
     
         24 . The fusion protein according to any one of  claims 1 - 23 , wherein the hydrophobic component comprises a transmembrane domain of a CD28. 
     
     
         25 . The fusion protein according to any one of  claims 1 - 23 , wherein the hydrophobic component comprises a transmembrane domain of a 4-1BB. 
     
     
         26 . The fusion protein according to any one of  claims 1 - 25 , wherein the intracellular signaling domain comprises an intracellular signaling domain of a costimulatory molecule. 
     
     
         27 . The fusion protein of  claim 26 , wherein the costimulatory molecule comprises CD28, CD137 (4-1BB), or ICOS. 
     
     
         28 . The fusion protein of any of  claims 1 - 27 , wherein the intracellular signaling domain comprises an intracellular signaling domain of a CD3ε, CD3δ, CD3ζ, CD25, CD27, CD28, CD40, CD47, CD79A, CD79B, CD134 (OX40), CD137 (4-1BB), CD150 (SLAMF1), CD278 (ICOS), CD357 (GITR), CARD11, DAP10, DAP12, FcRα, FcRβ, FcRγ, Fyn, Lck, LAT, LRP, NKG2D, NOTCH1, NOTCH2, NOTCH3, NOTCH4, ROR2, Ryk, Slp76, pTα, TCRα, TCRβ, TRIM, Zap70, PTCH2, or any combination thereof. 
     
     
         29 . The fusion protein according to any one of  claims 1 - 28 , wherein the intracellular signaling domain comprises a costimulatory domain of a CD137 (4-1BB), CD27, CD28, ICOS, OX40 (CD134), or any combination thereof. 
     
     
         30 . The fusion protein according to any one of  claims 1 - 29 , wherein the intracellular signaling domain comprises a costimulatory domain of a CD137 (4-1BB) or CD28, or any combination thereof. 
     
     
         31 . The fusion protein according to any one of  claims 1 - 30 , wherein the intracellular signaling domain comprises a costimulatory domain of a CD28. 
     
     
         32 . The fusion protein according to any one of  claims 1 - 31 , wherein the intracellular signaling domain comprises a costimulatory domain of a CD137 (4-1BB). 
     
     
         33 . The fusion protein according to any one of  claims 1 - 32 , wherein the target is an immunosupressive ligand. 
     
     
         34 . The fusion protein according to any one of  claims 1 - 32 , wherein the target is selected from a CD47, a CD58, a CD95L (FasL), a CD200, a CD270 (HVEM), a CD274 (PD-L1), and a GAL9. 
     
     
         35 . The fusion protein according to any of  claims 1 - 34 , wherein (a) the extracellular component comprises an extracellular portion of a CD200R, (b) the hydrophobic component comprises a transmembrane domain of a CD28, and (c) the intracellular component comprises an intracellular signaling domain of a CD28 or a 41BB. 
     
     
         36 . The fusion protein according to  claim 35 , wherein the extracellular portion of the CD200R comprises the entire extracellular domain of CD200R. 
     
     
         37 . The fusion protein according to  claim 35 , wherein the extracellular portion of the CD200R comprises at least 200 amino acids from the N-terminus of CD200R. 
     
     
         38 . The fusion protein according to  claim 35 , wherein the extracellular portion of the CD200R comprises at least about 225 amino acids to at least about 235 amino acids from the N-terminus of CD200R. 
     
     
         39 . The fusion protein according to  claim 35 , wherein the extracellular portion of the CD200R comprises at least about 234 amino acids from the N-terminus of CD200R. 
     
     
         40 . The fusion protein according to any of  claims 35 - 39 , wherein the CD200R is a human CD200R. 
     
     
         41 . The fusion protein according to any one of  claims 35 - 40 , wherein the intracellular component comprises a second intracellular signaling domain. 
     
     
         42 . The fusion protein according to  claim 41 , wherein the second intracellular signaling domain comprises an intracellular signaling domain of a CD137 (4-1BB). 
     
     
         43 . The fusion protein according to any one of  claims 35 - 42 , wherein the extracellular component comprises an extracellular portion of a CD28 extending from the CD28 transmembrane domain. 
     
     
         44 . The fusion protein according to  claim 43 , wherein the extracellular portion of the CD28 comprises a cysteine residue. 
     
     
         45 . The fusion protein according to any of  claims 1 - 44 , wherein the intracellular signaling component does not contain a CD3ζ signaling domain and/or does not contain a signaling domain capable of delivering a primary signal to a T cell and/or does not contain a signaling domain from a molecule naturally capable of delivering a primary signal to a T cell. 
     
     
         46 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:2, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:3, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         47 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:2, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         48 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:8, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises the amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         49 . The fusion protein according to  claim 41 , wherein the extracellular component further comprises a multimerization domain having an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9. 
     
     
         50 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:11, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         51 . The fusion protein according to  claim 50 , wherein the extracellular component further comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9. 
     
     
         52 . The fusion protein according to  claim 51 , wherein the intracellular component further comprises a second intracellular signaling domain having an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:13. 
     
     
         53 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an extracellular portion of a CD200R, (b) the hydrophobic component comprises a transmembrane domain of a CD28, and (c) the intracellular component comprises an intracellular signaling domain of a CD137 (4-1BB). 
     
     
         54 . The fusion protein according to  claim 53 , wherein the extracellular portion of the CD200R comprises the entire extracellular domain of CD200R. 
     
     
         55 . The fusion protein according to  claim 53 , wherein the extracellular portion of the CD200R comprises at least 200 amino acids from the N-terminus of CD200R. 
     
     
         56 . The fusion protein according to  claim 53 , wherein the extracellular portion of the CD200R comprises at least about 225 amino acids to at least about 235 amino acids from the N-terminus of CD200R. 
     
     
         57 . The fusion protein according to any one of  claims 53 - 56 , wherein the extracellular component comprises an extracellular portion of a CD28 extending from the CD28 transmembrane domain. 
     
     
         58 . The fusion protein according to  claim 57 , wherein the extracellular portion of the CD28 comprises a cysteine residue. 
     
     
         59 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an extracellular portion of a CD200R, (b) the hydrophobic component comprises a transmembrane domain of a CD137 (4-1BB), and (c) the intracellular component comprises an intracellular signaling domain of a CD137 (4-1BB). 
     
     
         60 . The fusion protein according to  claim 59 , wherein the extracellular portion of the CD200R comprises the entire extracellular domain of CD200R. 
     
     
         61 . The fusion protein according to  claim 59 , wherein the extracellular portion of the CD200R comprises at least a portion of the extracellular domain at least 230 amino acids from the N-terminus of CD200R. 
     
     
         62 . The fusion protein according to any one of  claims 59 - 61 , wherein the extracellular component comprises an extracellular portion of a CD137 (4-1BB) extending from the CD137 (4-1BB) transmembrane domain. 
     
     
         63 . The fusion protein according to  claim 62 , wherein the extracellular portion of the CD137 (4-1BB) comprises a cysteine residue. 
     
     
         64 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises a binding domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:8 and a multimerization domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:13. 
     
     
         65 . The fusion protein according to  claim 64 , wherein the intracellular component further comprises a second intracellular signaling domain having an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         66 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises a binding domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:11 and a multimerization domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:13. 
     
     
         67 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an extracellular portion of a SIRPα, (b) the hydrophobic component comprises a transmembrane domain of a CD28, and (c) the intracellular component comprises an intracellular signaling domain of a CD28. 
     
     
         68 . The fusion protein according to  claim 67 , wherein the extracellular portion of the SIRPα comprises the entire extracellular domain of SIRPα. 
     
     
         69 . The fusion protein according to  claim 67 , wherein the extracellular portion of the SIRPα comprises at least 361 amino acids from the N-terminus of SIRPα. 
     
     
         70 . The fusion protein according to any one of  claims 67 - 69 , wherein the intracellular component comprises a second intracellular signaling domain. 
     
     
         71 . The fusion protein according to  claim 70 , wherein the second intracellular signaling domain comprises an intracellular signaling domain of a CD137 (4-1BB). 
     
     
         72 . The fusion protein according to any one of  claims 67 - 71 , wherein the extracellular component comprises an extracellular portion of a CD28 extending from the CD28 transmembrane domain. 
     
     
         73 . The fusion protein according to  claim 72 , wherein the extracellular portion of the CD28 comprises a cysteine residue. 
     
     
         74 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:17, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:18, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         75 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:17, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         76 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises a binding domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:21 and a multimerization domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9, (b) the hydrophobic component comprises the amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises the amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         77 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises a binding domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:21 and a multimerization domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9, (b) the hydrophobic component comprises the amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises the amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:13. 
     
     
         78 . The fusion protein according to  claim 70 , wherein the intracellular component further comprises the amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ. ID. NO:5. 
     
     
         79 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an extracellular portion of a CD279 (PD-1), (b) the hydrophobic component comprises a transmembrane domain of a CD28, and (c) the intracellular component comprises an intracellular signaling domain of a CD28. 
     
     
         80 . The fusion protein according to  claim 79 , wherein the extracellular portion of the CD279 (PD-1) comprises the entire extracellular domain of CD279 (PD-1). 
     
     
         81 . The fusion protein according to  claim 79 , wherein the extracellular portion of the CD279 (PD-1) comprises a portion of the extracellular domain at least 100 amino acids from the N-terminus of CD279 (PD-1). 
     
     
         82 . The fusion protein according to  claim 79 , wherein the extracellular portion of the CD279 (PD-1) comprises a portion of the extracellular domain at least 149 amino acids from the N-terminus of CD279 (PD-1). 
     
     
         83 . The fusion protein according to any one of  claims 79 - 82 , wherein the extracellular component comprises an extracellular portion of a CD28 extending from the CD28 transmembrane domain. 
     
     
         84 . The fusion protein according to  claim 83 , wherein the extracellular portion of the CD28 comprises a cysteine residue. 
     
     
         85 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:89 and a multimerization domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         86 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:91 and a multimerization domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         87 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:95 and a multimerization domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         88 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an extracellular portion of a CD95 (Fas), (b) the hydrophobic component comprises a transmembrane domain of a CD28, and (c) the intracellular component comprises an intracellular signaling domain of a CD28. 
     
     
         89 . The fusion protein according to  claim 88 , wherein the extracellular portion of the CD95 (Fas) comprises the entire extracellular domain of CD95 (Fas). 
     
     
         90 . The fusion protein according to  claim 88 , wherein the extracellular portion of the CD95 (Fas) comprises at least a portion of the extracellular domain of at least 175 amino acids from the N-terminus of CD95 (Fas). 
     
     
         91 . The fusion protein according to  claim 88 , wherein the extracellular portion of the CD95 (Fas) comprises at least a portion of the extracellular domain of at least 173 amino acids from the N-terminus of CD95 (Fas). 
     
     
         92 . The fusion protein according to  claim 88 , wherein the extracellular portion of the CD95 (Fas) comprises at least a portion of the extracellular domain of at least 166 amino acids from the N-terminus of CD95 (Fas). 
     
     
         93 . The fusion protein according to  claim 88 , wherein the extracellular portion of the CD95 (Fas) comprises at least a portion of the extracellular domain of at least 161 amino acids from the N-terminus of CD95 (Fas). 
     
     
         94 . The fusion protein according to any one of  claims 88 - 93 , wherein the extracellular component comprises an extracellular portion of a CD28 extending from the CD28 transmembrane domain. 
     
     
         95 . The fusion protein according to  claim 94 , wherein the extracellular portion of the CD28 comprises a cysteine residue. 
     
     
         96 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:71 and a multimerization domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         97 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:73 and a multimerization domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         98 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:75 and a multimerization domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         99 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an extracellular portion of a CD2, (b) the hydrophobic component comprises a transmembrane domain of a CD28, and (c) the intracellular component comprises an intracellular signaling domain of a CD28. 
     
     
         100 . The fusion protein according to  claim 99 , wherein the extracellular portion of the CD2 comprises the entire extracellular domain of CD2. 
     
     
         101 . The fusion protein according to  claim 99 , wherein the extracellular portion of the CD2 comprises at least a portion of the extracellular domain of at least 175 amino acids from the N-terminus of CD2. 
     
     
         102 . The fusion protein according to any one of  claims 99 - 101 , wherein the extracellular component comprises an extracellular portion of a CD28 extending from the CD28 transmembrane domain. 
     
     
         103 . The fusion protein according to  claim 102 , wherein the extracellular portion of the CD28 comprises a cysteine residue. 
     
     
         104 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:61 and a multimerization domain with an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:9, (b) the hydrophobic component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:4, and (c) the intracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:5. 
     
     
         105 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an extracellular portion of a TIM3, (b) the hydrophobic component comprises a transmembrane domain of a CD28, and (c) the intracellular component comprises an intracellular signaling domain of a CD28. 
     
     
         106 . The fusion protein according to  claim 105 , wherein the extracellular portion of the TIM3 comprises the entire extracellular domain of TIM3. 
     
     
         107 . The fusion protein according to  claim 105 , wherein the extracellular portion of the TIM3 comprises at least a portion of the extracellular domain of at least 180 amino acids from the N-terminus of TIM3. 
     
     
         108 . The fusion protein according to any one of  claims 105 - 107 , wherein the extracellular component comprises an extracellular portion of a CD28 extending from the CD28 transmembrane domain. 
     
     
         109 . The fusion protein according to  claim 108 , wherein the extracellular portion of the CD28 comprises a cysteine residue. 
     
     
         110 . The fusion protein according to  claim 1 , wherein (a) the extracellular component comprises an extracellular portion of a LAG3, (b) the hydrophobic component comprises a transmembrane domain of a CD28, and (c) the intracellular component comprises an intracellular signaling domain of a CD28. 
     
     
         111 . The fusion protein according to  claim 110 , wherein the extracellular portion of the LAG3 comprises the entire extracellular domain of LAG3. 
     
     
         112 . The fusion protein according to  claim 110 , wherein the extracellular portion of the LAG3 comprises at least a portion of the extracellular domain of at least 410 amino acids from the N-terminus of LAG3. 
     
     
         113 . The fusion protein according to any one of  claims 110 - 112 , wherein the extracellular component comprises an extracellular portion of a LAG3 extending from the CD28 transmembrane domain. 
     
     
         114 . The fusion protein according to  claim 113 , wherein the extracellular portion of the CD28 comprises a cysteine residue. 
     
     
         115 . A fusion protein according to  claim 5 , wherein the extracellular component comprises a binding domain with an amino acid sequence as set forth in SEQ ID NO.:31 and a multimerization domain with an amino acid sequence as set forth in SEQ ID NO.:32, (b) the hydrophobic component comprises an amino acid sequence as set forth in SEQ ID NO.:27, and (c) the intracellular component comprises an amino acid as set forth in SEQ ID NO.:36. 
     
     
         116 . The fusion protein according to  claim 115 , wherein the intracellular component further comprises a second intracellular signaling domain having an amino acid sequence as set forth in SEQ ID NO.:28. 
     
     
         117 . The fusion protein according to  claim 5 , wherein (a) the extracellular component comprises a binding domain with an amino acid sequence as set forth in SEQ ID NO.:34 and a multimerization domain with an amino acid sequence as set forth in SEQ ID NO.:32, (b) the hydrophobic component comprises an amino acid sequence as set forth in SEQ ID NO.:27, and (c) the intracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:36. 
     
     
         118 . The fusion protein according to  claim 5 , wherein (a) the extracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:40, (b) the hydrophobic component comprises an amino acid sequence as set forth in SEQ ID NO.:41, and (c) the intracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:28. 
     
     
         119 . The fusion protein according to  claim 5 , wherein (a) the extracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:40, (b) the hydrophobic component comprises an amino acid sequence as set forth in SEQ ID NO.:27, and (c) the intracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:28. 
     
     
         120 . The fusion protein according to  claim 5 , wherein (a) the extracellular component comprises a binding domain with an amino acid sequence as set forth in SEQ ID NO.:44 and a multimerization domain with an amino acid sequence as set forth in SEQ ID NO.:32, (b) the hydrophobic component comprises the amino acid sequence as set forth in SEQ ID NO.:27, and (c) the intracellular component comprises the amino acid sequence as set forth in SEQ ID NO.:28. 
     
     
         121 . The fusion protein according to  claim 5 , wherein (a) the extracellular component comprises a binding domain with an amino acid sequence as set forth in SEQ ID NO.:44 and a multimerization domain with an amino acid sequence as set forth in SEQ ID NO.:32, (b) the hydrophobic component comprises the amino acid sequence as set forth in SEQ ID NO.:27, and (c) the intracellular component comprises the amino acid sequence as set forth in SEQ ID NO.:36. 
     
     
         122 . The fusion protein according to  claim 121 , wherein the intracellular component further comprises the amino acid sequence as set forth in SEQ. ID. NO:28. 
     
     
         123 . The fusion protein according to  claim 5 , wherein (a) the extracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:90 and a multimerization domain with an amino acid sequence as set forth in SEQ ID NO.:32, (b) the hydrophobic component comprises an amino acid sequence as set forth in SEQ ID NO.:27, and (c) the intracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:28. 
     
     
         124 . The fusion protein according to  claim 5 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:92 and a multimerization domain with an amino acid sequence as set forth in SEQ ID NO.:32, (b) the hydrophobic component comprises an amino acid sequence as set forth in SEQ ID NO.:27, and (c) the intracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:28. 
     
     
         125 . The fusion protein according to  claim 5 , wherein (a) the extracellular component comprises an amino acid sequence encoded by a nucleic acid molecule as set forth in SEQ ID NO.:96 and a multimerization domain with an amino acid sequence as set forth in SEQ ID NO.:32, (b) the hydrophobic component comprises an amino acid sequence as set forth in SEQ ID NO.:27, and (c) the intracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:28. 
     
     
         126 . The fusion protein according to  claim 5 , wherein (a) the extracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:72 and a multimerization domain with an amino acid sequence as set forth in SEQ ID NO.:32, (b) the hydrophobic component comprises an amino acid sequence as set forth in SEQ ID NO.:27, and (c) the intracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:28. 
     
     
         127 . The fusion protein according to  claim 5 , wherein (a) the extracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:74 and a multimerization domain with an amino acid sequence as set forth in SEQ ID NO.:32, (b) the hydrophobic component comprises an amino acid sequence as set forth in SEQ ID NO.:27, and (c) the intracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:28. 
     
     
         128 . The fusion protein according to  claim 5 , wherein (a) the extracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:76 and a multimerization domain with an amino acid sequence as set forth in SEQ ID NO.:32, (b) the hydrophobic component comprises an amino acid sequence as set forth in SEQ ID NO.:27, and (c) the intracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:28. 
     
     
         129 . The fusion protein according to  claim 5 , wherein (a) the extracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:62 and a multimerization domain with an amino acid sequence as set forth in SEQ ID NO.:32, (b) the hydrophobic component comprises an amino acid sequence as set forth in SEQ ID NO.:27, and (c) the intracellular component comprises an amino acid sequence as set forth in SEQ ID NO.:28. 
     
     
         130 . A nucleic acid molecule encoding a fusion protein according to any one of  claims 1 - 129 . 
     
     
         131 . A vector comprising a nucleic acid molecule according to  claim 130 . 
     
     
         132 . The vector according to  claim 131 , wherein the vector is a viral vector. 
     
     
         133 . The vector according to  claim 132 , wherein the viral vector is a lentiviral or retroviral vector. 
     
     
         134 . The vector according to  claim 133 , wherein the viral vector is a lentiviral vector. 
     
     
         135 . The vector according to any one of  claims 131 - 134 , further encoding an antigen-specific TCR. 
     
     
         136 . The vector according to  claim 135 , wherein the TCR is exogenous to a host cell. 
     
     
         137 . The vector according to  claim 135  or  claim 136 , wherein the TCR is specific to a HLA class I restricted antigen. 
     
     
         138 . The vector according to any one of  claims 135 - 137 , wherein the antigen is a cancer-specific antigen. 
     
     
         139 . The vector according to  claim 138 , wherein the cancer-specific antigen comprises WT-1, mesothelin, or cyclin-A1. 
     
     
         140 . The vector according to any one of  claims 131 - 139 , further encoding a ligand. 
     
     
         141 . The vector according to  claim 140 , wherein the ligand is CD200, CD47, PD-L1, or CD58. 
     
     
         142 . The vector according to any one of  claims 131 - 141 , further encoding an siRNA for reducing the expression of an endogenous receptor. 
     
     
         143 . The vector according to  claim 142 , wherein the endogenous receptor comprises CD200R, SIRPα, CD279 (PD-1), CD95 (Fas), or CD2 or is a TCR or portion thereof. 
     
     
         144 . A host cell, comprising a fusion protein according to any one of  claims 1 - 129 . 
     
     
         145 . A host cell, comprising at least two different fusion proteins according to any one of  claims 1 - 129 . 
     
     
         146 . A host cell according to  claim 145 , comprising:
 a fusion protein according to  claim 79 ; and   a fusion protein according to  claim 105 .   
     
     
         147 . A host cell, comprising a nucleic acid molecule according to  claim 130 . 
     
     
         148 . A host cell, comprising a vector according to any one of  claims 131 - 143 . 
     
     
         149 . The host cell according to any one of  claims 144 - 148 , wherein the host cell is an immune system cell. 
     
     
         150 . The host cell according to  claim 149 , wherein the immune system cell is a T cell. 
     
     
         151 . The host cell according to  claim 150 , wherein the T cell is a CD4+ T cell. 
     
     
         152 . The host cell according to  claim 150 , wherein the T cell is a CD8+ T cell. 
     
     
         153 . The host cell according to any one of  claims 144 - 152 , further comprising an antigen receptor, which optionally is an antigen-specific TCR. 
     
     
         154 . The host cell according to any one of  claim 153 , wherein the antigen-specific TCR is exogenous to the cell or the host. 
     
     
         155 . The host cell according to  claim 153  or  claim 154 , wherein the TCR binds to an antigen::HLA complex with high affinity. 
     
     
         156 . The host cell according to  claim 155 , wherein the high affinity binding has a K a  equal to or greater than 10 7  M −1 . 
     
     
         157 . The host cell according to any one of  claims 153 - 156 , wherein the TCR is specific to a HLA class I restricted antigen. 
     
     
         158 . The host cell according to any one of  claims 153 - 156 , wherein the antigen is a cancer-specific antigen. 
     
     
         159 . The host cell according to  claim 158 , wherein the cancer-specific antigen comprises WT-1, mesothelin, or cyclin-A1. 
     
     
         160 . The host cell according to any one of  claims 153 - 156 , wherein the antigen is a viral antigen. 
     
     
         161 . The host cell according to  claim 153 , wherein the antigen receptor is a chimeric antigen receptor. 
     
     
         162 . The host cell according to  claim 161 , wherein the chimeric antigen receptor comprises an extracellular antigen binding domain and an intracellular signaling domain capable of delivering a primary signal to a T cell and optionally a costimulatory domain. 
     
     
         163 . The host cell according to any one of  claims 144 - 162 , further encoding a ligand. 
     
     
         164 . The host cell according to  claim 163 , wherein the ligand is CD200, CD47, PD-L1, or CD58. 
     
     
         165 . The host cell according to any one of  claims 144 - 164 , further encoding an siRNA for reducing the expression of an endogenous receptor. 
     
     
         166 . The host cell according to  claim 165 , wherein the endogenous receptor comprises CD200R, SIRPα, CD279 (PD-1), CD95 (Fas) or CD2. 
     
     
         167 . A method of treating a disease in a subject comprising administering a fusion protein according to any one of  claims 1 - 129  to the subject. 
     
     
         168 . A method of treating a disease in a subject comprising administering a vector according to any one of  claims 131 - 143  to the subject. 
     
     
         169 . A method of treating a disease in a subject comprising administering a host cell according to any one of  claims 144 - 166  to the subject. 
     
     
         170 . The method according to any one of  claims 167 - 169 , wherein the disease is selected from the group consisting of viral infection, bacterial infection, cancer, and autoimmune disease. 
     
     
         171 . The method according to any one of  claims 167 - 170 , wherein the subject is human. 
     
     
         172 . A fusion protein according to any one of  claims 1 - 129 , for use in treating cancer.