GM-CSF and IL-4 Conjugates, Compositions, and Methods Related Thereto

Abstract

In certain embodiments, this disclosure relates to conjugates comprising GM-CSF and IL-4 and uses related thereto, e.g., enhancing the immune system. Typically, the GM-CSF and IL-4 are connected by a linker. In certain embodiments, the disclosure relates to isolated nucleic acids encoding these polypeptide conjugates, vectors comprising nucleic acid encoding polypeptide conjugates, and protein expression systems comprising these vectors such as infectious viral particles and host cells comprising such a nucleic acid.

1 . A method of treating cancer comprising administering an effective amount of a conjugate comprising a granulocyte macrophage colony stimulating factor (GM-CSF) polypeptide and an interleukin 4 (IL-4) polypeptide, wherein said GM-CSF polypeptide and said IL-4 polypeptide are connected by a peptide linker consisting of amino acids selected from glycine, serine, threonine, asparagine, alanine and proline, wherein the conjugate is at least 70% identical to SEQ ID NO: 8, wherein said conjugate induces proliferation of B-cells, to a subject in need thereof. 
     
     
         2 . The method of  claim 1 , wherein the cancer is a hematological malignancy. 
     
     
         3 . The method of  claim 2 , wherein the hematological malignancy is selected from leukemia, lymphoma, acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), chronic myelogenous leukemia, acute monocytic leukemia (AMOL), Hodgkin's lymphomas, and non-Hodgkin's lymphomas such as Burkitt lymphoma, B-cell lymphoma and multiple myeloma. 
     
     
         4 . The method of  claim 1 , wherein the cancer is melanoma. 
     
     
         5 . The method of  claim 1 , wherein said GM-CSF polypeptide is located closer to the N-terminus of said conjugate relative to said IL-4 polypeptide. 
     
     
         6 . The method of  claim 1 , wherein said GM-CSF polypeptide comprises the amino acid sequence of SEQ ID NO: 6 and said IL-4 polypeptide comprises the amino acid sequence of SEQ ID NO: 3. 
     
     
         7 . The method of  claim 1 , wherein said GM-CSF polypeptide and said IL-4 polypeptide are connected by a peptide linker consisting of amino acids selected from glycine and serine. 
     
     
         8 . The method of  claim 1 , wherein the linker comprises repeats of serine or repeats of glycine. 
     
     
         9 . The method of  claim 1 , wherein the linker is 1 to 5 amino acids. 
     
     
         10 . The method of  claim 1 , wherein the linker comprises GGGGS (SEQ ID NO:5). 
     
     
         11 . A method of treating cancer comprising administering an effective amount of a nucleic acid encoding a conjugate comprising a granulocyte macrophage colony stimulating factor (GM-CSF) polypeptide and an interleukin 4 (IL-4) polypeptide, wherein said GM-CSF polypeptide and said IL-4 polypeptide are connected by a peptide linker consisting of amino acids selected from glycine, serine, threonine, asparagine, alanine and proline, wherein the conjugate is at least 70% identical to SEQ ID NO: 8, to a subject in need thereof. 
     
     
         12 . The method of  claim 12 , wherein the nucleic acid is a recombinant vector comprising the nucleic acid. 
     
     
         13 . The method of  claim 11 , wherein the cancer is a hematological malignancy. 
     
     
         14 . The method of  claim 13 , wherein the hematological malignancy is selected from leukemia, lymphoma, acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), chronic myelogenous leukemia, acute monocytic leukemia (AMOL), Hodgkin's lymphomas, and non-Hodgkin's lymphomas such as Burkitt lymphoma, B-cell lymphoma and multiple myeloma. 
     
     
         15 . The method of  claim 11 , wherein the cancer is melanoma. 
     
     
         16 . The method of  claim 11 , wherein said GM-CSF polypeptide comprises the amino acid sequence of SEQ ID NO: 6 and said IL-4 polypeptide comprises the amino acid sequence of SEQ ID NO: 3. 
     
     
         17 . The method of  claim 11 , wherein said GM-CSF polypeptide and said IL-4 polypeptide are connected by a peptide linker consisting of amino acids selected from glycine and serine. 
     
     
         18 . The method of  claim 11 , wherein the linker comprises repeats of serine or repeats of glycine. 
     
     
         19 . The method of  claim 11 , wherein the linker is 1 to 5 amino acids. 
     
     
         20 . The method of  claim 1 , wherein the linker comprises GGGGS (SEQ ID NO:5).