Ii-Key/antigenic epitope hybrid peptide vaccines
Disclosed is a nucleic acid molecule comprising a first expressible sequence encoding a protein of interest or polypeptide of interest which contains an MHC Class II-presented epitope. In addition, the nucleic acid molecule comprises a second expressible nucleic acid sequence encoding an antigen presentation enhancing hybrid polypeptide. The antigen presentation enhancing hybrid polypeptide includes the following elements: i) an N-terminal element consisting essentially of 4–16 residues of the mammalian Ii-Key peptide LRMKLPKPPKPVSKMR (SEQ ID NO: 1) and non-N-terminal deletion modifications thereof that retain antigen presentation enhancing activity; ii) a C-terminal element comprising an MHC Class II-presented epitope in the form of a polypeptide or peptidomimetic structure which binds to the antigenic peptide binding site of an MHC class II molecule, the MHC Class II-presented epitope being contained in the protein of interest of step a); and iii) an intervening peptidyl structure linking the N-terminal and C-terminal elements of the hybrid, the peptidyl structure having a length of about 20 amino acids or less.
1. An isolated nucleic acid molecule comprising:
a) a first expressible nucleic acid sequence encoding a protein of interest or polypeptide of interest which contains a first MHC Class II-presented epitope; and
b) a second expressible nucleic acid sequence encoding an antigen presentation enhancing hybrid polypeptide comprising:
i) an N-terminal element consisting essentially of the residues LRMK (amino acids 1–4) of SEQ ID NO.: 1 and 0–12 additional sequential residues of SEQ ID NO.: 1, and modifications of SEQ ID No: 1 that retain antigen presentation enhancing activity;
ii) a C-terminal element comprising a second an MHC Class II-presented epitope, said second MHC Class II-presented epitope being in the form of a polypeptide or peptidomimetic structure which binds to the antigenic peptide binding site of an MHC class II molecule, the second MHC Class II-presented epitope being from the same protein or polypeptide of interest encoded by the nucleic acid sequence a); and
iii) an intervening peptidyl structure linking the N-terminal and C-terminal elements of the hybrid, the peptidyl structure having a length of about 20 amino acids or less.
2. The isolated nucleic acid of claim 1 wherein the modifications of element b) i) are selected from the group consisting of:
a) deletion of amino acids from the C-terminus of SEQ ID No: 1;
b) N-terminal extensions of SEQ ID No: 1;
c) amino acid substitutions of SEQ ID No: 1 excluding said residues LRMK.
3. The isolated nucleic acid of claim 1 wherein the C-terminal element further comprises an MHC Class I-presented epitope, or a portion thereof, the amino acid residues comprising the MHC Class I-presented epitope or portion thereof being constituent residues of the MHC Class II-presented epitope.
4. The isolated nucleic acid of claim 1 wherein the C-terminal element further comprises an antibody-recognized determinant, or a portion thereof, the amino acid residues comprising the antibody-recognized determinant or a portion thereof being constituent residues of the MHC Class II-presented epitope.
5. The isolated nucleic acid sequence of claim 1 wherein the protein or polypeptide of interest corresponds to a protein or polypeptide encoded by of an infectious pathogen.
6. The isolated nucleic acid sequence of claim 5 wherein the infectious pathogen is selected from the group consisting of anthrax, EBOLA, HIV and influenza.
7. The isolated nucleic acid sequence of claim 5 wherein the infectious pathogen is vaccinia virus.
8. The isolated nucleic acid sequence of claim 1 wherein the protein of interest is gp42.