Synthetic immunogen useful for generating long lasting immunity and protection against pathogens
The present invention relates to a synthetic immunogen represented by the general formula 1, useful for generating long lasting protective immunity against various intracellular pathogens which are the causative agents of tuberculosis, leishmaniasis, AIDS, trypanosomiasis, malaria and also allergy, cancer and a process for the preparation thereof. The developed immunogen is able to circumvent HLA restriction in humans and livestock. The invention further relates to a vaccine comprising the said immunogen for generating enduring protective immunity against various diseases. The said vaccine is targeted against intracellular pathogens, more particularly the pathogen M. tuberculosis in this case. In the present invention, promiscuous peptides of M. tuberculosis are conjugated to TLR ligands especially; Pam2Cys to target them mainly to dendritic cells and therefore elicit long-lasting protective immunity. wherein, X 1 =a promiscuous CD4 T helper epitope selected from SEQ ID No. 1 to 98 OR nil; X 2 =a promiscuous CD8 T cytotoxic epitope selected from SEQ ID No. 99 to 103 OR nil; when X1=nil; X2=SEQ ID No. 99 to 103 and when X2=nil; X1=SEQ ID No. 1 to 98; Y=Lysine; and S=Serine.
1. A synthetic immunogen of general formula 1
wherein,
X 1 =a promiscuous CD4 T helper epitope having binding ability to HLA class II peptide, said HLA selected from the group consisting of HLA-DR, HLA-DP and HLA-DQ, wherein said CD4 T helper epitope is SEQ ID No. 1;
X 2 =a promiscuous CD8 T cytotoxic epitope having binding ability to HLA class I peptide, said HLA selected from the group consisting of HLA-A, HLA-B and HLA-C, wherein said CD8 T cell epitope is selected from SEQ ID No. 99 to 103;
Y=Lysine; and
S=Serine;
wherein the promiscuous epitopes represented by SEQ ID No. 1 and SEQ ID NO: 99 to 103 are from Mycobacterium tuberculosis.
2. A synthetic immunogen represented by the formula:
wherein X 1 =a promiscuous CD4 T helper epitope having binding ability to HLA class II peptide, said HLA selected from the group consisting of HLA-DR, HLA-DP and HLA-DQ, wherein said CD4 T helper epitope is SEQ ID No. 1; and
Y=Lysine; and
S=Serine,
wherein the promiscuous epitopes represented by SEQ ID No. 1 are from Mycobacterium tuberculosis.
3. An immunogen as claimed in claim 1 , represented by the formula:
wherein, Y=Lysine and S=Serine.
4. An immunogen as claimed in claim 1 , wherein the TLR ligand is selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, TLR11, TLR12 and TLR13 ligands; or wherein the TLR ligand is selected from the group consisting of diacyl lipopeptides, triacyl lipopeptides, lipoarabinomanan and lipopolysaccharides.
5. An immunogen as claimed in claim 1 , wherein the TLR ligand is S-[2,3-bis(palmitoyloxy)propyl]cysteine.
6. An immunogen as claimed in claim 1 , wherein the promiscuous CD4 T helper epitope from M. tuberculosis is identified based on T cell proliferation and secretion of IFN-γ and IL-4; or wherein the promiscuous CD4 T helper epitope from M. tuberculosis is identified based on the secretion of cytokines IL-2, IL-4, IL-12 and IFN-γ.
7. An immunogen as claimed in claim 1 , wherein the promiscuous CD4 T helper epitope from M. tuberculosis enhances MHC/HLA expression; or wherein the promiscuous CD4 T helper epitope from M. tuberculosis enhances the expression of co-stimulatory molecules selected from CD80, CD86 and CD40.
8. An immunogen as claimed in claim 1 , wherein the promiscuous epitopes from M. tuberculosis enhance the proliferation of CD4 + and CD8 + T cells by upregulating the expression of CD69 and CD44; or wherein the promiscuous epitopes enhances CD4 + and CD8 + T cell memory, including both central and effector T cell memory.
9. An immunogen as claimed in claim 1 , wherein the promiscuous epitopes from M. tuberculosis modulate the secretion of cytokines IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IFN-γ and TNF-α; wherein the promiscuous epitopes modulate the expression of CD44, CD62L and CD127 on memory CD4 + and CD8 + T cells.
10. An immunogen as claimed in claim 1 , wherein the promiscuous epitopes from M. tuberculosis exploit[s] TLR ligands as adjuvants and hence extra adjuvants are not required.
11. An immunogen as claimed in claim 1 , wherein it is coated to/encapsulated in nanoparticles.
12. An immunogen as claimed in claim 1 , wherein it is covalently coupled to/entrapped in mannosylated liposomes or liposomes which are tagged with anti-DEC-205 antibody.
13. A pharmaceutical injectable composition comprising the immunogen as claimed in claim 1 with at least one of a pharmaceutically acceptable carrier, diluent and excipient.
14. A method of inducing an immune response against tuberculosis in a subject, comprising administering to the subject a therapeutically effective amount of the immunogen as claimed in claim 1 optionally along with pharmaceutically acceptable carriers, diluents or excipients.