TY - JOUR
T1 - The second and third amino acids of Pam2 lipopeptides are key for the proliferation of cytotoxic T cells
AU - Takeda, Yohei
AU - Azuma, Masahiro
AU - Hatsugai, Ryoko
AU - Fujimoto, Yukari
AU - Hashimoto, Masahito
AU - Fukase, Koichi
AU - Matsumoto, Misako
AU - Seya, Tsukasa
N1 - Funding Information:
This work was in part supported by the Uehara Memorial Foundation and Smoking Research Foundation. We are grateful to Drs K. Funami, T. Akazawa, H. Shime, J. Kasamatsu, M. Tatematsu and A. Maruyama for their thoughtful discussions. We are also grateful to Drs M. Kasahara (Hokkaido University), M. Fukushima and H. Nishimura (Translational Research Informatics Center, Kobe) for their support of our study. We had non-profit support from AID Co. Ltd (Kobe, Japan) through the university-company contract, which we acknowledge gratefully.
Publisher Copyright:
© The Author(s) 2018.
PY - 2018/7/1
Y1 - 2018/7/1
N2 - 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.
AB - 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.
KW - CTL proliferation
KW - MyD88-mediated dendritic cell priming
KW - Pam2 lipopeptide
KW - TLR2 agonist
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U2 - 10.1177/1753425918777598
DO - 10.1177/1753425918777598
M3 - Article
C2 - 29848176
AN - SCOPUS:85047962771
SN - 1753-4259
VL - 24
SP - 323
EP - 331
JO - Journal of Endotoxin Research
JF - Journal of Endotoxin Research
IS - 5
ER -
