Fusion proteins comprising an anti-cd40 antibody and hiv antigenic peptides

Abstract

The present invention includes compositions and methods for the expression, secretion and use of novel compositions for use as, e.g., vaccines and antigen delivery vectors, to delivery antigens to antigen presenting cells. In one embodiment, the vector is an anti-CD40 antibody, or fragments thereof, and one or more antigenic peptides linked to the anti-CD40 antibody or fragments thereof, including humanized antibodies.

1 - 56 . (canceled) 
     
     
         57 . An antibody-antigen fusion protein comprising:
 a dendritic cell (DC)-targeting antibody or antigen binding fragment thereof;   one or more viral antigens; and   one or more flexible linkers between the DC-targeting antibody or antigen binding fragment thereof and the one or more antigens; wherein the viral antigen is not a tumor associated antigen.   
     
     
         58 . The fusion protein of  claim 57 , wherein the one or more flexible linkers comprises a polypeptide with an amino acid sequence corresponding to one or more of: SEQ ID NO:11-14 and 145. 
     
     
         59 . The fusion protein of  claim 57 , wherein the one or more antigens comprise one or more of gag, pol, nef, and env HIV antigens. 
     
     
         60 . The fusion protein of  claim 59 , wherein the one or more antigens comprise a HIV polypeptide with an amino acid sequence corresponding to one or more of SEQ ID NOS:1-5. 
     
     
         61 . The fusion protein of  claim 57 , wherein the DC targeting antibody or antigen binding fragment is selected from an antibody or antigen binding fragment that specifically binds to MHC class I, MHC class II, CD1, CD2, CD3, CD4, CD8, CD11b, CD14, CD15, CD16, CD19, CD20, CD29, CD31, CD40, CD43, CD44, CD45, CD54, CD56, CD57, CD58, CD83, CD86, CMRF-44, CMRF-56, DCIR, CLEC-6, CD40, BDCA-2, MARCO, DEC-205, mannose receptor, Langerin, DECTIN-1, B7-1, B7-2, IFN-γ receptor and IL-2 receptor, ICAM-1, Fcγ receptor, LOX-1, or ASGPR. 
     
     
         62 . The fusion protein of  claim 61 , wherein the DC targeting antibody or antigen binding fragment specifically binds to CD40. 
     
     
         63 . The fusion protein of  claim 62 , wherein the CD40 antibody comprises:
 a heavy chain variable domain (VH) which comprises in sequence hypervariable regions CDR1H, CDR2H and CDR3H, the CDR1H having the amino acid sequence GFTFSDYYMY (SEQ ID NO.:45), the CDR2H having the amino acid sequence YINSGGGSTYYPDTVKG (SEQ ID NO.:46), and the CDR3H having the amino acid sequence RGLPFHAMDY (SEQ ID NO.:47); and   a light chain variable domain (VL) which comprises in sequence hypervariable regions CDR1L, CDR2L and CDR3L, the CDR1L having the amino acid sequence SASQGISNYLN (SEQ ID NO.:41), the CDR2L having the amino acid sequence YTSILHS (SEQ ID NO.:42), and the CDR3L having the amino acid sequence QQFNKLPPT (SEQ ID NO.:43).   
     
     
         64 . The fusion protein of  claim 62 , wherein the CD40 antibody comprises anti-CD40_12B4.2C10 (ATCC Submission No. HS446, Accession No. PTA-10653) or a humanized version thereof. 
     
     
         65 . The fusion protein of  claim 62 , wherein the CD40 antibody comprises anti-CD40_11B6.1C3 (ATCC Submission No. HS440, Accession No. PTA-10652) or a humanized version thereof. 
     
     
         66 . The fusion protein of  claim 57 , wherein the (DC)-targeting antibody is a human antibody or a humanized antibody. 
     
     
         67 . A vaccine comprising the fusion protein of  claim 57 . 
     
     
         68 . A nucleic acid expression vector encoding the fusion protein of  claim 57 . 
     
     
         69 . A method for treating or preventing HIV in a patient, the method comprising administering a vaccine comprising the fusion protein of  claim 59 . 
     
     
         70 . A method of making HIV peptide-specific T cells comprising: immunizing a subject with a fusion protein of  claim 59 . 
     
     
         71 . A method of enhancing T cell responses comprising: immunizing a subject in need of vaccination with an effective amount of a vaccine of  claim 67 . 
     
     
         72 . The method of  claim 69 , wherein the vaccine is able to elicit an HIV-specific T cell immune response. 
     
     
         73 . The method of  claim 69 , wherein the one or more antigens elicit at least one of a humoral or a cellular immune response in a host. 
     
     
         74 . A method of generating antigen-loaded, activated dendritic cells comprising: obtaining previously isolated patient dendritic cells and exposing the dendritic cells to activating amounts of the fusion protein of  claim 57 . 
     
     
         75 . A method of producing a vaccine of  claim 67 , the method comprising introducing a flexible linker between a DC-targeting antibody or antigen binding fragment thereof and an antigen. 
     
     
         76 . A method of making a DC-targeting antibody-antigen fusion protein, the method comprising expressing the fusion protein of  claim 57  in a host cell and isolating the protein from the host cell.