The second and third amino acids of Pam2 lipopeptides are key for the proliferation of cytotoxic T cells

Innate Immun. 2018 Jul;24(5):323-331. doi: 10.1177/1753425918777598. Epub 2018 May 31.

Abstract

The TLR2 agonist, dipalmitoyl lipopeptide (Pam2LP), has been used as an immune adjuvant without much success. Pam2LP is recognised by TLR2/6 receptors in humans and in mice. This study examined the proliferative activity of cytotoxic T lymphocytes (CTL) using mouse Ag-presenting dendritic cells (DCs) and OT-I assay system, where a library of synthetic Pam2LP was utilised from the Staphylococcus aureus database. Ag-specific CTL expansion and IFN-γ levels largely depended on the Pam2LP peptide sequence. The first Aa is cysteine (Cys), which has an active SH residue to bridge fatty acids, and the second and third Aa are hydrophilic or non-polar. The sequence structurally adapted to the residual constitution of the reported TLR2/6 pocket. The inactive sequence contained proline or leucine/isoleucine after the first Cys. Notably, no direct activation of OT-I cells was detected without DCs by stimulation with the active Pam2LP having the Cys-Ser sequence. MyD88, but not TICAM-1 or IFN pathways, in DCs participates in DC maturation characterised by upregulation of CD40, CD80 and CD86. Hence, the active Pam2LPs appear suitable for dimeric TLR2/6 on DCs, resulting in induction of DC maturation.

Keywords: CTL proliferation; MyD88-mediated dendritic cell priming; Pam2 lipopeptide; TLR2 agonist.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adjuvants, Immunologic
  • Animals
  • Antigen Presentation
  • Cell Differentiation
  • Cell Proliferation
  • Cells, Cultured
  • Cysteine / metabolism*
  • Dendritic Cells / immunology*
  • Interferon-gamma / metabolism
  • Lipopeptides / genetics
  • Lipopeptides / metabolism*
  • Lymphocyte Activation
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Myeloid Differentiation Factor 88 / genetics
  • Myeloid Differentiation Factor 88 / metabolism
  • Peptide Library
  • Protein Conformation
  • Serine / metabolism*
  • Staphylococcus aureus / metabolism
  • Staphylococcus aureus / physiology*
  • T-Lymphocytes, Cytotoxic / immunology*
  • Toll-Like Receptor 2 / agonists

Substances

  • Adjuvants, Immunologic
  • Lipopeptides
  • Myeloid Differentiation Factor 88
  • Pam2CSK4 lipopeptide
  • Peptide Library
  • Tlr2 protein, mouse
  • Toll-Like Receptor 2
  • Serine
  • Interferon-gamma
  • Cysteine