title: Lipidated promiscuous peptide augments the expression of MHC-II molecules on dendritic cells and activates T cells.
abstract: OBJECTIVE
  In spite of the fact that BCG is the most widely used vaccine, tuberculosis (TB) continues to be a major killer disease in TB-endemic regions. Recently, many emerging evidences from the published literature indicate the role of environmental mycobacteria in blocking the processing and presentation of BCG antigens and thereby impairing with suboptimal generation of protective T cells. To surmount this problem associated with BCG, we constructed a novel lipopeptide (L91) by conjugating a promiscuous peptide consisting of CD4 + T-helper epitope of sequence of 91-110 of 16 kDa antigen of Mycobacterium tuberculosis to Pam2Cys, an agonist of Toll-like receptor-2.
  
  
  METHODS
  Mice were immunized subcutaneously with 20 nmol of L91, followed by a booster with 10 nmol, after an interval of 21 days of primary immunization. Animals were sacrificed after seven days of post-booster immunization. L91 induced immune response was characterized by the expression of MHC-II and CD74 on the surface of dendritic cells (DCs) by flowcytometry. Cytokines (IL-4, IL-10, IFN-γ) secretion and anti-peptide antibodies were measured by ELISA.
  
  
  RESULTS
  Self-adjuvanting lipopeptide vaccine (L91) was directly bound to MHC-II molecules and without requiring extensive processing for its presentation to T cells. It stimulated and activated dendritic cells and augmented the expression of MHC-II molecules. Further, it activated effector CD4 T cells to mainly secrete interferon (IFN)-γ but not interleukin (IL)-4 and IL-10. L91 did not elicit anti-peptide antibodies.
  
  
  CONCLUSIONS
  The findings suggest that L91 evokes maturation and upregulation of MHC class II molecules and promotes better antigen presentation and, therefore, optimum activation of T cells. L91 mainly induces effector Th1 cells, as evidenced by predominant release of IFN-γ, consequently can mount favourable immune response against M. tuberculosis . As L91 does not provoke the generation of anti-peptide antibodies, there is no fear of the efficacy of the vaccine being neutralized by pre-existing anti-mycobacterial antibodies in TB-endemic population. In conclusion, L91 may be considered as a future potential candidate vaccine against TB.
authors.forename: Uthaman
authors.initials: U
authors.lastname: Gowthaman
authors.forename: Pradeep K
authors.initials: PK
authors.lastname: Rai
authors.forename: Weiguang
authors.initials: W
authors.lastname: Zeng
authors.forename: David C
authors.initials: DC
authors.lastname: Jackson
authors.affiliation: CSIR-Institute of Microbial Technology, Chandigarh, India.
authors.forename: Javed N
authors.initials: JN
authors.lastname: Agrewala
mesh_terms: D000818:Animals; D017951:Antigen Presentation / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology*; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology; D005802:Genes, MHC Class II / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D015847:Interleukin-4 / Q000276:immunology / Q000378:metabolism; D008055:Lipids / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D009163:Mycobacterium bovis / Q000276:immunology; D009169:Mycobacterium tuberculosis / Q000276:immunology / Q000378:metabolism; D010455:Peptides / Q000276:immunology* / Q000494:pharmacology; D014376:Tuberculosis / Q000235:genetics / Q000276:immunology* / Q000382:microbiology
chemical_list: D008055:Lipids; D010455:Peptides; D015847:Interleukin-4; D007371:Interferon-gamma