Glycoconjugate Journal

, Volume 36, Issue 1, pp 69–78 | Cite as

The effects of trehalose glycolipid presentation on cytokine production by GM-CSF macrophages

  • Bridget L. StockerEmail author
  • Kristel Kodar
  • Kanu Wahi
  • Amy J. Foster
  • Jacquie L. Harper
  • Daiki Mori
  • Sho Yamasaki
  • Mattie S. M. TimmerEmail author
Original Article


Trehalose glycolipids (TGLs) are promising vaccine adjuvants, however effects of glycolipid presentation in the in vitro evaluation, and ultimate selection, of lead vaccine adjuvants are often overlooked. To this end, we synthesised a variety of TGLs and determined how the physicochemical presentation of these lipids influenced the cytokine response by bone marrow derived macrophages (BMMs). The TGLs were presented to wild-type and Mincle−/− BMMs as micellar solutions, coated on plates, coated on beads or surfactant solubilised. Medium to long-chain TGLs, either coated on plates or surfactant solubilised, resulted in the highest BMM activation. Stimulation of BMMs with TGLs coated on beads led to a decreased cytokine response, as compared to TGLs alone. All the TGL responses were Mincle dependent, however the mode of presentation did not have the same effect for each individual TGL. This was most apparent for the C22 trehalose monoester, which showed reduced activity compared to its diester counterpart when presented on a plate, but similar activity to the diester when presented as micelles or on beads. Taken together, our findings support the use of several in vitro assays for selecting lead vaccine adjuvants, particularly if structural differences between the adjuvants are pronounced.

Graphical abstract

The mode of glycolipid presentation, such as micellar solutions, coated on plates, coated on beads or surfactant solubilised, influences the immune response to trehalose glycolipids


Trehalose glycolipid Mincle Macrophage Adjuvant 



The authors would like to thank the Health Research Council (Hercus Fellowship, BLS, 2013/33) and the Cancer Society of New Zealand (2013/33) for financial support.

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflicts of interest.

Ethical approval

All experimental procedures were approved by the Victoria University Animal Ethics Committee in accordance with their guidelines for the care of animals (protocol nr 22,371).

Supplementary material

10719_2018_9857_MOESM1_ESM.pdf (729 kb)
ESM 1 (PDF 729 kb)


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Authors and Affiliations

  1. 1.School of Chemical and Physical SciencesVictoria University of WellingtonWellingtonNew Zealand
  2. 2.Centre for BiodiscoveryVictoria University of WellingtonWellingtonNew Zealand
  3. 3.Division of Molecular Immunology, Medical Institute of BioregulationKyushu UniversityFukuokaJapan
  4. 4.Department of Molecular Immunology, Research Institute for Microbial DiseasesOsaka UniversityOsakaJapan
  5. 5.Laboratory of Molecular Immunology, Immunology Frontier Research CenterOsaka UniversitySuitaJapan
  6. 6.Division of Molecular Immunology, Medical Mycology Research CenterChiba UniversityChibaJapan

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