T-cell modulatory multimeric polypeptides and methods of use thereof
The present disclosure provides variant immunomodulatory polypeptides, and fusion polypeptides comprising the variant immunomodulatory peptides. The present disclosure provides T-cell modulatory multimeric polypeptides, and compositions comprising same, where the T-cell modulatory multimeric polypeptides comprise a variant immunomodulatory polypeptide of the present disclosure. The present disclosure provides nucleic acids comprising nucleotide sequences encoding the T-cell modulatory multimeric polypeptides, and host cells comprising the nucleic acids. The present disclosure provides methods of modulating the activity of a T cell; the methods comprise contacting the T cell with a T-cell modulatory multimeric polypeptide of the present disclosure.
1. A nucleic acid comprising:
a) a first nucleotide sequence encoding a first polypeptide comprising:
i) a human papillomavirus epitope; and
ii) first class I major histocompatibility complex (MHC) polypeptide, wherein the first MHC polypeptide is a β2-microglobulin (β2M) polypeptide;
b) a second nucleotide sequence encoding a second polypeptide comprising:
i) a second class I MHC polypeptide, wherein the second MHC polypeptide is an MHC Class I HLA-A heavy chain polypeptide;
ii) at least one immunomodulatory polypeptide, wherein the at least one immunomodulatory polypeptide is an IL-2 variant polypeptide that comprises an amino acid sequence having at least 98% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:44, wherein amino acid 16 is other than histidine and amino acid 42 is other than phenylalanine, wherein the percent sequence identity is determinable by a sequence alignment performed using BLAST; and
iii) an immunoglobulin (Ig) Fc polypeptide.
2. A nucleic acid of claim 1 , wherein the β2M polypeptide comprises an amino acid sequence having at least 95% percent amino acid sequence identity to amino acids 21 to 119 of SEQ ID NO:95.
3. A nucleic acid of claim 1 , wherein the β2M polypeptide comprises an amino acid sequence as set forth in SEQ ID NO:17.
4. A nucleic acid of claim 1 , wherein the Ig Fc polypeptide comprises an amino acid sequence having at least about 95% percent amino acid sequence identity to SEQ ID NO:57, wherein the percent sequence identity is determinable by a sequence alignment performed using BLAST.
5. A nucleic acid of claim 1 , wherein the multimeric polypeptide comprises two copies of the IL-2 variant polypeptide, and wherein each copy comprises the amino acid sequence of SEQ ID NO:84.
6. A nucleic acid of claim 5 ,
wherein the Ig Fc polypeptide comprises an amino acid sequence having at least about 95% percent amino acid sequence identity to SEQ ID NO:57,
wherein the β2M polypeptide comprises an amino acid sequence having at least 95% percent amino acid sequence identity to amino acids 21 to 119 of SEQ ID NO:95,
wherein the percent sequence identity is determinable by a sequence alignment performed using BLAST,
wherein the first polypeptide comprises a peptide linker between the epitope and the β2M polypeptide, and
wherein the second polypeptide comprises a peptide linker between one or more of:
a) a first copy of the IL-2 polypeptide and a second copy of the IL-2 polypeptide;
b) one of the two copies of the IL-2 polypeptide and the MHC class I heavy chain polypeptide; and
c) the MHC class I heavy chain polypeptide and the Ig Fc polypeptide.
7. A plurality of cells genetically modified with the nucleic acid of claim 1 .
8. A plurality of cells genetically modified with the nucleic acid of claim 6 .
9. A method comprising culturing a plurality of genetically modified cells according to claim 7 in a culture medium under conditions such that the cell synthesizes the first and second polypeptides.
10. A method comprising culturing a plurality of genetically modified cells according to claim 8 in a culture medium under conditions such that the cell synthesizes the first and second polypeptides.
11. First and second nucleic acids, wherein
a) the first nucleic acid comprises a nucleotide sequence encoding a first polypeptide comprising:
i) a human papillomavirus epitope; and
ii) first class I major histocompatibility complex (MHC) polypeptide, wherein the first MHC polypeptide is a β2-microglobulin (β2M) polypeptide;
b) the second nucleic acid comprises a nucleotide sequence encoding a second polypeptide comprising:
i) a second class I MHC polypeptide, wherein the second MHC polypeptide is an MHC Class I HLA-A heavy chain polypeptide;
ii) at least one immunomodulatory polypeptide, wherein the at least one immunomodulatory polypeptide is an IL-2 variant polypeptide that comprises an amino acid sequence having at least 98% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:44, wherein amino acid 16 is other than histidine and amino acid 42 is other than phenylalanine, wherein the percent sequence identity is determinable by a sequence alignment performed using BLAST; and
iii) an immunoglobulin (Ig) Fc polypeptide.
12. The nucleic acids of claim 11 , wherein the β2M polypeptide comprises an amino acid sequence having at least 95% percent amino acid sequence identity to amino acids 21 to 119 of SEQ ID NO:95.
13. The nucleic acids of claim 12 , wherein the β2M polypeptide comprises an amino acid sequence as set forth in SEQ ID NO:17.
14. The nucleic acids of claim 11 , wherein the Ig Fc polypeptide comprises an amino acid sequence having at least about 95% percent amino acid sequence identity to SEQ ID NO:57, wherein the percent sequence identity is determinable by a sequence alignment performed using BLAST.
15. The nucleic acids of claim 11 , wherein the multimeric polypeptide comprises two copies of the IL-2 variant polypeptide, and wherein each copy comprises the amino acid sequence of SEQ ID NO:84.
16. The nucleic acids of claim 15 ,
wherein the Ig Fc polypeptide comprises an amino acid sequence having at least about 95% percent amino acid sequence identity to SEQ ID NO:57,
wherein the β2M polypeptide comprises an amino acid sequence having at least 95% percent amino acid sequence identity to amino acids 21 to 119 of SEQ ID NO:95,
wherein the percent sequence identity is determinable by a sequence alignment performed using BLAST,
wherein the first polypeptide comprises a peptide linker between the epitope and the β2M polypeptide, and
wherein the second polypeptide comprises a peptide linker between one or more of:
a) a first copy of the IL-2 polypeptide and a second copy of the IL-2 polypeptide;
b) one of the two copies of the IL-2 polypeptide and the MHC class I heavy chain polypeptide; and
c) the MHC class I heavy chain polypeptide and the Ig Fc polypeptide.
17. A plurality of cells genetically modified with the nucleic acids of claim 11 .
18. A plurality of cells genetically modified with the nucleic acids of claim 16 .
19. A method comprising culturing a plurality of genetically modified cells according to claim 17 in a culture medium under conditions such that the cell synthesizes the first and second polypeptides.
20. A method comprising culturing a plurality of genetically modified cells according to claim 18 in a culture medium under conditions such that the cell synthesizes the first and second polypeptides.