Recombinant
The present disclosure relates generally to therapeutic compositions comprising recombinant bacteria. Further, the disclosure elaborates upon methods of utilizing the taught therapeutic compositions to treat autoimmune and inflammatory disease. The present teachings also relate to the disclosed recombinant bacteria and methods of producing the recombinant bacteria utilized in the compositions and methods. Further taught herein are dietary supplements and food additive compositions comprising the taught recombinant bacteria.
1. A composition for the treatment of an autoimmune or inflammatory disease, the composition comprising:
a recombinant Lactococcus lactis bacterial cell comprising a nucleotide sequence coding for enterotoxigenic Escherichia coli colonization factor antigen I fimbriae genes cfaA, cfaB, cfaC, and cfaE, wherein the bacterial cell expresses E. coli colonization factor antigen 1 fimbriae genes cfaA, cfaB, cfaC, and cfaE; and wherein the cfaA gene comprises SEQ ID NO: 3, the cfaB gene comprises SEQ ID NO: 2, the cfaC gene comprises SEQ ID NO: 4, and the cfaE gene comprises SEQ ID NO: 5, and an acceptable carrier.
2. The composition of claim 1 , wherein the composition induces an anti-inflammatory response in a subject treated with the composition.
3. The composition of claim 1 , wherein said E. coli colonization factor antigen I fimbriae genes are oriented in a nucleotide sequence operon in the following non-native order: cfaB, cfaA, cfaC, and cfaE.
4. The composition of claim 1 , wherein the nucleotide sequence is operably linked to a composite promoter.
5. The composition of claim 4 , wherein the composite promoter comprises an inducible promoter.
6. The composition of claim 1 , wherein the nucleotide sequence is operably linked to a composite promoter comprising at least one sequence selected from the group consisting of: SEQ ID NO:6, SEQ ID NO:7, and SEQ ID NO:8.
7. The composition of claim 1 , wherein the nucleotide sequence is operably linked to a composite promoter comprising SEQ ID NO:6, SEQ ID NO:7, and SEQ ID NO:8.
8. A method comprising administering to a subject the composition of claim 1 .
9. The method of claim 8 , wherein the composition administered increases the level of a regulatory cytokine selected from IL-10 or TGF-β in the subject as compared to the level of the regulatory cytokine IL-10 or TGF-β present in the subject before said administering.
10. The method of claim 8 , wherein the composition administered decreases the level of at least one cytokine selected from the group consisting of IFN-γ, TNF-α, and IL-17 as compared to the level of at least one of the cytokines selected from the group consisting of IFN-γ, TNF-α, and IL-17 present in the subject before said administering.
11. A composition for the treatment of an autoimmune or inflammatory disease, the composition comprising:
a recombinant Lactococcus lactis bacterial cell comprising a nucleotide sequence coding for enterotoxigenic Escherichia coli colonization factor antigen I fimbriae genes cfaA, cfaB, cfaC, and cfaE, wherein the bacterial cell expresses E. coli colonization factor antigen 1 fimbriae genes cfaA, cfaB, cfaC, and cfaE; and wherein the nucleotide sequence comprises a polynucleotide sequence sharing at least 95% sequence identity with SEQ ID NO: 1, and an acceptable carrier.
12. The composition of claim 11 , wherein the polynucleotide sequence shares 100% sequence identity with SEQ ID NO:1.
13. A recombinant Lactococcus lactis bacterial cell comprising a nucleotide sequence coding for enterotoxigenic Escherichia coli colonization factor antigen I fimbriae genes cfaA, cfaB, cfaC, and cfaE, wherein the bacterial cell expresses E. coli colonization factor antigen I fimbriae genes comprising cfaA, cfaB, cfaC, and cfaE; and wherein the cfaA gene comprises SEQ ID NO: 3, the cfaB gene comprises SEQ ID NO: 2, the cfaC gene comprises SEQ ID NO: 4, and the cfaE gene comprises SEQ ID NO: 5.
14. The recombinant Lactococcus lactis bacterial cell of claim 13 , wherein said E. coli colonization factor antigen I fimbriae genes are oriented in a nucleotide sequence operon in the following non-native order: cfaB, cfaA, cfaC, and cfaE.
15. The recombinant Lactococcus lactis bacterial cell of claim 13 , wherein the nucleotide sequence is operably linked to a composite promoter.
16. The recombinant Lactococcus lactis bacterial cell of claim 15 , wherein the composite promoter comprises an inducible promoter.
17. The recombinant Lactococcus lactis bacterial cell of claim 13 , wherein the nucleotide sequence is operably linked to a composite promoter comprising at least one sequence selected from the group consisting of: SEQ ID NO:6, SEQ ID NO:7, and SEQ ID NO:8.
18. The recombinant Lactococcus lactis bacterial cell of claim 13 , wherein the nucleotide sequence is operably linked to a composite promoter comprising SEQ ID NO:6, SEQ ID NO:7, and SEQ ID NO:8.
19. A recombinant Lactococcus lactis bacterial cell comprising a nucleotide sequence coding for enterotoxigenic Escherichia coli colonization factor antigen I fimbriae genes cfaA, cfaB, cfaC, and cfaE, wherein the bacterial cell expresses E. coli colonization factor antigen I fimbriae genes comprising cfaA, cfaB, cfaC, and cfaE; wherein the nucleotide sequence comprises a polynucleotide sequence sharing at least 95% sequence identity with SEQ ID NO:1.
20. The recombinant Lactococcus lactis bacterial cell of claim 19 , wherein the polynucleotide sequence shares 100% sequence identity with SEQ ID NO:1.
21. A method for producing a composition for the treatment of an autoimmune or inflammatory disease, the method comprising:
(a) introducing a nucleotide sequence coding for enterotoxigenic Escherichia coli colonization factor antigen I fimbriae genes into a recipient Lactococcus lactis bacterial cell, wherein the antigen I fimbriae genes comprise cfaA, cfaB, cfaC, and cfaE; and wherein the nucleotide sequence comprises a polynucleotide sequence sharing at least 95% sequence identity with SEQ ID NO:1.
22. The method of claim 21 , wherein the polynucleotide sequence shares 100% sequence identity with SEQ ID NO:1.
23. The method of claim 21 , further comprising:
(b) culturing the bacterial cell under conditions which allow for expression of the enterotoxigenic Escherichia coli colonization factor.