Multi-epitopic construct

Abstract

The invention relates to multiple epitope constructs, immunogenic and vaccine compositions comprising recombinant molecules presenting inserted multiple and different epitopes from a variety of antigens. The antigenic determinants being associated with different pathways leading to atherosclerosis. In particular, the invention relates to such compositions for eliciting an immune response against antigens and pathogens involved in the development of atherosclerosis. The invention includes inter alia methods of treating and/or preventing the disease and recombinant protein products.

What is claimed is: 
     
         1 . A recombinant construct comprising:
 (i) a scaffold portion;   (ii) a first species of epitope capable of eliciting an anti-arteriosclerotic vascular disease response via a first pathway, wherein the first species of epitope is a C5a receptor (C5aR) polypeptide; and   (iii) a second species of epitope capable of eliciting an anti-arteriosclerotic vascular disease via a second pathway that is independent from the said first pathway, wherein the second species of epitope is a  chlamydia pneumonia  epitope.   
     
     
         2 . The construct according to  claim 1  further comprising a third species epitope capable of eliciting an anti-arteriosclerotic vascular disease response via a third pathway that is independent from the first pathway wherein the third species epitope is a heat-shock protein (HSP) epitope. 
     
     
         3 . The construct according to  claim 2  comprising a plurality of first, second and/or third species of epitopes. 
     
     
         4 . The construct according to  claim 1 , wherein the C5aR epitope is a polypeptide comprising an amino acid sequence from 5 to 45 contiguous amino acid residues from SEQ ID NO:6. 
     
     
         5 . The construct according to  claim 1 , wherein C5aR epitope is either a C or N terminus sequence and optionally wherein the epitope is a polypeptide comprising, or consisting of an amino acid sequence selected from the group comprising MNSFNYTTPDYGHYDDKDTLD (SEQ ID NO:7), TLDLNTPVDKTSN (SEQ ID NO:8) and MNSFNYTTPDYGHYDDKDTLDLNTPVDKTSN (SEQ ID NO:9) or a functional fragment thereof that has antigenic activity. 
     
     
         6 . The construct according to  claim 2 , wherein the HSP is either a HSP 60 or a HSP 65 and optionally wherein in the instance that the HSP is an HSP 60 it is a human HSP 60 or a  Mycobacterium bovis  HSP. 
     
     
         7 . The construct according to  claim 6 , wherein the HSP is a HSP60 epitope that comprises an amino acid sequence selected from the group comprising, or consisting of peptide 1 (AA) 153-160: AELKKQSK; (SEQ ID NO:10), peptide 1 (AA) 153-163: AELKKQSKPVT; (SEQ ID NO:11), peptide 1 (AA) 303-312: PGFGDNRKNQ (SEQ ID NO:12), peptide 2: AA 277-286 PGFGDNRKNQ (SEQ ID NO:13), peptide (AA) 516-528: KGIIDPTKWRTA (SEQ ID NO:14), and  mycobacterium  (AA) 253-268: EGEALSTLVVNKIRGT (SEQ ID NO:15) or a functional fragment thereof that has antigenic activity. 
     
     
         8 . The construct according to  claim 7 , wherein the construct comprises the HSP60 epitope of peptide 1 (AA) 153-163: AELKKQSKPVT (SEQ ID NO: 10); and peptide 1 (AA) 303-312: PGFGDNRKNQ (SEQ ID NO:12), optionally wherein the constructs is RHHC. 
     
     
         9 . The construct according to  claim 1 , wherein the  chlamydia pneumonia  epitope is Cpn1 or Cpn2 and optionally is a polypeptide comprising, or consisting of an amino acid sequence selected from the group comprising the major outer membrane protein (MOMP) (amino acid sequence (AA) 67-74: GDYVFDRI (SEQ ID NO:16), and putative outer membrane protein (Pomp) 5 of Cpn (amino acid sequence (AA) 283-291: QAVANGGAI SEQ ID NO:17) or a functional fragment thereof that has antigenic activity. 
     
     
         10 . The construct according to  claim 1 , wherein the scaffold portion is a dendroaspin scaffold protein as depicted in SEQ ID NO:1 or a fragment or variant thereof. 
     
     
         11 . The construct according to  claim 10 , wherein the first and/or second species of epitope is incorporated into any one or more of the following positions: (a) loop I and/or loop II; (b) loop I and/or loop III; (c) loop II and/or loop III; (d) loop I, loop II and loop III; (e) an Nor C terminus of the dendroaspin scaffold protein. 
     
     
         12 . An expression vector comprising nucleic acids encoding the epitopes incorporated into the construct according to  claim 1 . 
     
     
         13 . An antigenic composition comprising the construct according to  claim 1  optionally wherein the composition comprises an antigenic hydrophobic complex comprising: (i) an isolated microsome optionally wherein it as an inverted microsome or (ii) an MHC protein or (iii) an inverted micelle or (iv) a synthetic product. 
     
     
         14 . A pharmaceutical composition comprising the recombinant construct according to  claim 1  formulated as an injectable or oral product, optionally wherein the pharmaceutical composition further includes a suitable adjuvant, excipient, diluent and/or carrier. 
     
     
         15 . A pharmaceutical composition comprising the construct according to  claim 1 . 
     
     
         16 . A vaccine comprising the construct according to  claim 1 . 
     
     
         17 . A method of eliciting an anti-atherosclerosis response in a mammal comprising administering to an individual a recombinant construct of  claim 1 . 
     
     
         18 . A method of treating, preventing or reducing atherosclerosis comprising administering to an individual a recombinant construct of  claim 1 . 
     
     
         19 . A method of treating an individual with early stage atherosclerosis or an individual identified as at risk of developing atherosclerosis comprising administering to an individual a recombinant construct of  claim 1 . 
     
     
         20 . A method of eliciting an immune response against epitopes associated with two independent pathways associated with atherosclerosis formation, the method comprising:
 1. constructing and expressing a dendroaspin scaffold protein comprising at least one first and at least one second epitope according to  claim 1 ;   2. incubating eukaryotic cells with said dendroaspin scaffold protein;   3. using said eukaryotic cells to prepare microsomes;   4. incorporating said microsomes and dendroaspin scaffold protein with one or more pharmaceutically acceptable constituents to produce an orally or injectable administratable preparation; and   5. administering said preparation to a mammal or human.