Combinatorial platform for the display of surface adjuvants and antigens
Engineered bacteria are provided that produce modified lipid A and a polypeptide or polysaccharide antigens. In some aspects, immunogenic compositions are provided comprising a modified a lipid A and a polypeptide or polysaccharide antigen.
1. An isolated engineered bacteria strain comprising
(a) at least one expression vector comprising at least one polynucleotide encoding a lipid A modification enzyme, wherein the gene encoding a lipid A modification enzyme is selected from the group consisting of lpxE, lpxF, lpxO, lpxR, pagL, and pagP; and
(b) at least one expression vector comprising at least one polynucleotide encoding a polysaccharide antigen biosynthesis protein,
wherein the engineered bacteria strain produces the polysaccharide antigen covalently conjugated to a modified lipid A; and
wherein the engineered bacteria strain:
(i) comprises a deletion of the Eck1673 gene;
(ii) comprises at least lpxE, lpxO, pagL and pagP;
(iii) produces the Vibrio cholerae O antigen, Salmonella typhimurium O antigen, or Shigella spcs O antigen antigen covalently conjugated to a modified lipid A; or
(iv) produces the Streptococcus pneumonia capsule, Staphylococcus aureus capsule, or Neisseria meningitidis capsule antigen covalently conjugated to a modified lipid A.
2. The isolated engineered strain of claim 1 , further comprising a deletion of the Eck1673 gene.
3. The isolated engineered strain of claim 1 , further comprising a deletion of the rfbD gene.
4. The isolated engineered strain of claim 1 , further comprising a deletion of the rfbB-wbbL genetic region.
5. The isolated engineered strain of claim 1 , comprising at least lpxE, pagL and pagP.
6. The isolated engineered strain of claim 1 , comprising at least lpxE, lpxO, pagL and pagP.
7. The isolated engineered strain of claim 1 , wherein the polysaccharide antigen is Vibrio cholerae O antigen, Salmonella typhimurium O antigen, or Shigella spcs O antigen.
8. The isolated engineered strain of claim 7 , wherein the at least one polynucleotide encoding a polysaccharide antigen biosynthesis protein is selected from the group consisting of a gmhD gene, a manC gene, a manB gene, a gmd gene, a wbeE gene, a wbeG gene, a wzm, a wzt gene, a wbeK gene, a wbeL gene, a wbeM gene, a wbeN gene, a wbeO gene, a wbeP gene, a wbeT gene, a wbeU gene, a wbeV gene, a gale gene, and a wbeW gene, and a wbf region gene.
9. The isolated engineered strain of claim 1 , wherein the polysaccharide antigen is Streptococcus pneumonia capsule, Staphylococcus aureus capsule, or Neisseria meningitidis capsule.
10. The isolated engineered strain of claims 1 , wherein the bacteria is an E. coli bacteria.
11. The isolated engineered strain of claim 9 , wherein the at least one polynucleotide encoding a polysaccharide antigen biosynthesis protein is selected from the group consisting of a wzm gene, a wzt gene, a wzx gene, and a wzy gene.
12. A composition isolated from the engineered bacteria strain of claim 1 , wherein the composition comprises the polysaccharide antigen covalently conjugated to a modified lipid A.