CD8a and t cell receptor variants and methods of using same in modulating immune cell responses
Novel costimulatory fusion proteins and DNA sequences that enhance T cell responses to weakly immunogenic and/or lowly expressed antigens and that confer T cell resistance against MDSC-mediated suppression are disclosed. The fusion proteins comprise portions of CD4, CD8α or the T cell receptor linked to a specific region of MyD88 or other signaling molecules. These fusion proteins and sequence variants thereof improve T cell activation and responsiveness. Also disclosed is the use of these molecules in host cells as a means to enhance and costimulate responses of immune cells including cytotoxic CD8 + T cells and the use of these cells to treat cancer, infectious agents and other diseases.
1. A fusion protein comprising an amino-terminal domain linked to a region of MyD88 lacking the TIR domain,
wherein the amino-terminal domain is selected from the group consisting of:
(a) extracellular and transmembrane regions of CD8α,
(b) a transmembrane region of CD8α,
(c) extracellular and transmembrane regions of CD4, and
(d) a T cell receptor; and
wherein the region of MyD88 lacking the TIR domain corresponds to amino acids 1-155 of human MyD88 (SEQ ID NO:24) plus or minus up to 10 amino acids from either end or from both ends.
2. The fusion protein of claim 1 , wherein (a) the extracellular and transmembrane regions of CD8α correspond to amino acids 1-217 of mouse CD8α (SEQ ID NO:16) or amino acids 1-203 of human CD8α (SEQ ID NO:12) and (b) the region of MyD88 lacking the TIR domain corresponds to amino acids 1-155 of human MyD88 (SEQ ID NO:24).
3. The fusion protein of claim 1 , wherein the fusion protein comprises mCD8α-hMyD88 as set forth in SEQ ID NO:17 or hCD8α-hMyD88 as set forth in SEQ ID NO:14.
4. The fusion protein of claim 1 , wherein the transmembrane region of CD8α corresponds to amino acids 1-83 of SEQ ID NO:18 and the region of MyD88 lacking the TIR domain corresponds to amino acids 1-155 of human MyD88 (SEQ ID NO:24).
5. The fusion protein of claim 1 , wherein the fusion protein comprises hCD8αTM-hMyD88 as set forth in SEQ ID NO:18.
6. The fusion protein of claim 1 , wherein (a) the extracellular and transmembrane regions of CD4 correspond to amino acids 1-417 of mouse CD4 (amino acids 1-417 of SEQ ID NO:21) or human CD4 (amino acids 1-418 of SEQ ID NO:20) and (b) the region of MyD88 lacking the TIR domain corresponds to amino acids 1-155 of human MyD88 (SEQ ID NO:24).
7. The fusion protein of claim 1 , wherein the fusion protein comprises mCD4-hMyD88 as set forth in SEQ ID NO:21 or hCD4-hMyD88 as set forth in SEQ ID NO:20.
8. The fusion protein of claim 1 , wherein the TCR is the DMFS TCR having the amino acid sequence of residues 1-603 of SEQ ID NO:22 and the region of MyD88 lacking the TIR domain corresponds to amino acids 1-155 of human MyD88 (SEQ ID NO:24).
9. The fusion protein of claim 1 , wherein the fusion protein comprises hTCR-hMyD88 as set forth in SEQ ID NO:22.
10. An isolated population of cells expressing at least one fusion protein of claim 1 .
11. The isolated population of cells of claim 10 , wherein the isolated population of cells expresses at least one fusion protein selected from the group consisting of mCD8α-hMyD88 (SEQ ID NO:17), hCD8α-hMyD88 (SEQ ID NO:14), hCD8αTM-hMyD88 (SEQ ID NO:18), mCD4-hMyD88 (SEQ ID NO:21), hCD4-hMyD88 (SEQ ID NO:20) and hTCR-hMyD88 (SEQ ID NO:22).
12. A method of treating a subject having cancer or an infectious disease, comprising administering to a subject having cancer or an infectious disease a therapeutically-effective amount of at least one population of cells as defined in claim 10 .
13. A method of treating a subject having cancer or an infectious disease, comprising administering to a subject having cancer or an infectious disease a therapeutically-effective amount of at least one population of cells as defined in claim 11 .
14. A method of conferring T cell resistance against myeloid derived suppressor cells (MDSC)-mediated suppression, comprising expressing at least one fusion protein of claim 1 in a T cell.
15. The method of claim 14 , wherein the fusion protein is selected from the group consisting of mCD8α-hMyD88 (SEQ ID NO:17), hCD8α-hMyD88 (SEQ ID NO:14), hCD8αTM-hMyD88 (SEQ ID NO:18), mCD4-hMyD88 (SEQ ID NO:21), hCD4-hMyD88 (SEQ ID NO:20) and hTCR-hMyD88 (SEQ ID NO:22).
16. A method of enhancing immune cell recognition of an antigen, comprising expressing at least one fusion protein of claim 1 in an immune cell.
17. The method of claim 16 , wherein the fusion protein is selected from the group consisting of mCD8α-hMyD88 (SEQ ID NO:17), hCD8α-hMyD88 (SEQ ID NO:14), hCD8αTM-hMyD88 (SEQ ID NO:18), mCD4-hMyD88 (SEQ ID NO:21), hCD4-hMyD88 (SEQ ID NO:20) and hTCR-hMyD88 (SEQ ID NO:22).
18. The method of claim 16 , wherein the antigen is present at a low concentration in vitro or in vivo.
19. The method of claim 16 , wherein the antigen is a weakly antigenic antigen.