In vivo cell surface engineering

Abstract

The present invention provides methods and compositions for the in vivo engineering of cell surfaces, such as tumor cell surfaces, with one or more immune co-stimulatory polypeptides. The methods, compositions and engineered cells are useful, for example, to stimulate an immune response against the cells. When the engineered cell surfaces are tumor cell surfaces, the methods, compositions and engineered cells are useful for improving a patient's immune response against the cancer and for reducing tumor size and inhibiting tumor growth.

1 . A method of engineering the surface of a cell to display one or more immune co-stimulatory polypeptides, comprising administering to an individual containing said cell:
 a) a bifunctional molecule comprising a first member of a binding pair and a molecule that binds to the surface of a cell;   b) a first immune co-stimulatory moiety comprising a first immune co-stimulatory polypeptide and a second member of said binding pair; and, optionally,   c) a second immune co-stimulatory moiety comprising a second immune co-stimulatory polypeptide and a second member of said binding pair,   wherein said first and optional second immune co-stimulatory polypeptides are displayed on said cell surface via binding between said first member of said binding pair and said second member of said binding pair, and   wherein at least one of said first and optional second immune co-stimulatory polypeptides is selected from the group consisting of CD86, ICOSL, PD-L1, PD-L2, B7-H3, B7-H4, OX40L, 4-1BBL, CD27L, CD30L, LIGHT, BAFF, and APRIL.   
     
     
         2 . The method of  claim 1 , wherein said cell surface is a tumor cell surface. 
     
     
         3 . The method of  claim 2 , wherein said bifunctional molecule comprising a first member of said binding pair is administered via intratumoral injection. 
     
     
         4 . The method of  claim 2 , wherein at least one of said first and optional second immune co-stimulatory moieties are administered via intratumoral injection. 
     
     
         5 . The method of  claim 1 , wherein said optional second immune co-stimulatory moiety is administered, and said first and second immune co-stimulatory moieties are administered substantially simultaneously. 
     
     
         6 . The method of  claim 1 , wherein said optional second immune co-stimulatory moiety is administered, and wherein said first and second immune co-stimulatory moieties are administered sequentially on different days. 
     
     
         7 . The method of  claim 1 , wherein at least about 5% of cells initially displaying said first or second immune co-stimulatory moiety continue to display said first or second immune co-stimulatory moiety at about 2 days post-injection. 
     
     
         8 . The method of  claim 1 , wherein said first member of said binding pair comprises biotin and said second member of said binding pair comprises core streptavidin. 
     
     
         9 . The method of  claim 1 , wherein said optional second immune co-stimulatory moiety is administered, and wherein said first and second immune co-stimulatory polypeptides are selected from the group consisting of CD86, ICOSL, PD-L1, PD-L2, B7-H3, B7-H4, OX40L, 4-1BBL, CD27L, CD30L, LIGHT, BAFF, APRIL, CD80 and CD40L. 
     
     
         10 . The method of  claim 1 , further comprising:
 administering a third immune co-stimulatory moiety comprising a third immune co-stimulatory polypeptide and a second member of said binding pair.   
     
     
         11 . The method of  claim 10 , wherein at least two of said first, second and third immune co-stimulatory polypeptides are selected from the group consisting of CD86, ICOSL, PD-L1, PD-L2, B7-H3, B7-H4, OX40L, 4-1BBL, CD27L, CD30L, LIGHT, BAFF, APRIL, CD80 and CD40L. 
     
     
         12 . The method of  claim 11 , wherein said first, second and third immune co-stimulatory polypeptides are LIGHT, CD80, and 4-1BBL. 
     
     
         13 . The method of  claim 10 , wherein at least one of said first, second and third immune co-stimulatory moieties is administered by a route different from at least one other of said first, second and third immune co-stimulatory moieties. 
     
     
         14 . The method of  claim 10 , wherein each of said first, second and third immune co-stimulatory moieties are administered by the same route. 
     
     
         15 . The method of  claim 14 , wherein said first, second and third immune co-stimulatory moieties are administered as components of a single composition. 
     
     
         16 . A composition comprising:
 a first fusion protein comprising a first immune co-stimulatory polypeptide and a member of a binding pair and   a second fusion protein comprising a second immune co-stimulatory polypeptide and the same member of a binding pair,   wherein at least one of said immune co-stimulatory polypeptides is selected from the group consisting of CD86, ICOSL, PD-L1, PD-L2, B7-H3, B7-H4, OX40L, 4-1BBL, CD27L, CD30L, LIGHT, BAFF, and APRIL.   
     
     
         17 . The composition of  claim 16 , wherein said first and second immune co-stimulatory polypeptides are selected from the group consisting of CD86, ICOSL, PD-L1, PD-L2, B7-H3, B7-H4, OX40L, 4-1BBL, CD27L, CD30L, LIGHT, BAFF, APRIL, CD80 and CD40L. 
     
     
         18 . The composition of  claim 16 , wherein said member of a binding pair comprises core streptavidin. 
     
     
         19 . The composition of  claim 16 , wherein at least one of said fusion proteins comprises a sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, and SEQ ID NO:9. 
     
     
         20 . The composition according to  claim 16 , comprising a soluble fusion protein comprising LIGHT and said member of a binding pair, a soluble fusion protein comprising CD80 and said member of a binding pair, and a soluble fusion protein comprising 4-1BBL and said member of a binding pair. 
     
     
         21 . The composition of  claim 20 , wherein said member of a binding pair comprises core streptavidin. 
     
     
         22 . A combination comprising:
 a first composition comprising a first fusion protein comprising a first immune co-stimulatory polypeptide and a member of a binding pair and   a second composition comprising a second fusion protein comprising a second immune co-stimulatory polypeptide and the same member of a binding pair,   wherein at least one of said immune co-stimulatory polypeptides is selected from the group consisting of CD86, ICOSL, PD-L1, PD-L2, B7-H3, B7-H4, OX40L, 4-1BBL, CD27L, CD30L, LIGHT, BAFF, and APRIL.   
     
     
         23 . The combination of  claim 19 , wherein said first and second immune co-stimulatory polypeptides are selected from the group consisting of CD86, ICOSL, PD-L1, PD-L2, B7-H3, B7-H4, OX40L, 4-1BBL, CD27L, CD30L, LIGHT, BAFF, APRIL, CD80 and CD40L. 
     
     
         24 . The combination of  claim 22 , wherein said member of a binding pair comprises streptavidin. 
     
     
         25 . The combination of  claim 22 , wherein at least one of said fusion proteins comprises a sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, and SEQ ID NO:9. 
     
     
         26 . The combination according to  claim 22 , comprising a composition comprising a soluble fusion protein comprising LIGHT and said member of a binding pair, a composition comprising a soluble fusion protein comprising CD80 and said member of a binding pair, and a composition comprising a soluble fusion protein comprising 4-1BBL and said member of a binding pair. 
     
     
         27 . A method of reducing the size of a tumor or inhibiting tumor growth, comprising administering to an individual containing said tumor:
 a) a bifunctional molecule comprising a first member of a binding pair and a molecule that binds to the surface of a tumor cell of said tumor;   b) a first immune co-stimulatory moiety comprising a first immune co-stimulatory polypeptide and a second member of said binding pair; and, optionally,   c) a second immune co-stimulatory moiety comprising a second immune co-stimulatory polypeptide and a second member of said binding pair,   wherein said first and optional second immune co-stimulatory polypeptides are displayed on said tumor cell surface via binding between said first member of said binding pair and said second member of said binding pair, and   wherein at least one of said first and optional second immune co-stimulatory polypeptides is selected from the group consisting of CD86, ICOSL, PD-L1, PD-L2, B7-H3, B7-H4, OX40L, 4-1BBL, CD27L, CD30L, LIGHT, BAFF, and APRIL.   
     
     
         28 . A method of engineering the surface of a cell to display an immune co-stimulatory polypeptide, comprising contacting a cell surface with:
 a) a bifunctional molecule comprising a first member of a binding pair and a molecule that binds to the surface of said cell; and   b) an immune co-stimulatory moiety comprising an immune co-stimulatory molecule and a second member of said binding pair,   wherein said immune co-stimulatory polypeptide is selected from the group consisting of CD86, ICOSL, PD-L1, PD-L2, B7-H3, B7-H4, OX40L, 4-1 BBL, CD27L, CD30L, LIGHT, BAFF, and APRIL, and   wherein said immune co-stimulatory polypeptide is displayed on said cell surface via binding between said first member of said binding pair and said second member of said binding pair.   
     
     
         29 . The method of  claim 28 , wherein said method is effected in vitro. 
     
     
         30 . The method of  claim 28 , wherein said method is effected in vivo. 
     
     
         31 . The method of  claim 28 , wherein said first member of a binding pair comprises biotin and said second member of a binding pair is selected from the groups consisting of avidin and streptavidin. 
     
     
         32 . The method of  claim 31 , wherein said streptavidin is core streptavidin. 
     
     
         33 . A chimeric protein comprising an immune co-stimulatory polypeptide and a member of a binding pair, wherein said immune co-stimulatory polypeptide is selected from the group consisting of CD86, ICOSL, PD-L1, PD-L2, B7-H3, B7-H4, OX40L, 4-1BBL, CD27L, CD30L, LIGHT, BAFF, and APRIL. 
     
     
         34 . The chimeric protein of  claim 33 , wherein said member of a binding pair is selected from the group consisting of avidin and streptavidin. 
     
     
         35 . The chimeric protein of  claim 34 , wherein said streptavidin is core streptavidin. 
     
     
         36 . A pharmaceutical composition comprising
 a chimeric protein comprising an immune co-stimulatory polypeptide and a member of a binding pair, wherein said immune co-stimulatory polypeptide is selected from the group consisting of CD86, ICOSL, PD-L1, PD-L2, B7-H3, B7-H4, OX40L, 4-1BBL, CD27L, CD30L, LIGHT, BAFF, and APRIL, and   a pharmaceutically acceptable carrier.