Species specific differences of CD1d oligomer loading in vitro

Daniel Paletta, Alina Suzann Fichtner, Lisa Starick, Steven A. Porcelli, Paul B. Savage, Thomas Herrmann

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

CD1d molecules are MHC class I-like molecules that present glycolipids to iNKT cells. The highly conserved interaction between CD1d:α-Galactosylceramide (αGC) complexes and the iNKT TCR not only defines this population of αβ T cells but can also be used for its direct identification. Therefore, CD1d oligomers are a widely used tool for iNKT cell related investigations. To this end, the lipid chains of the antigen have to be inserted into the hydrophobic pockets of the CD1d binding cleft, often with help of surfactants. In this study, we investigated the influence of different surfactants (Triton X-100, Tween 20, Tyloxapol) on in vitro loading of CD1d molecules derived from four different species (human, mouse, rat and cotton rat) with αGC and derivatives carrying modifications of the acyl-chain (DB01-1, PBS44) and a 6-acetamido-6-deoxy-addition at the galactosyl head group (PBS57). We also compared rat CD1d dimers with tetramers and staining of an iNKT TCR transductant was used as readout for loading efficacy. The results underlined the importance of CD1d loading efficacy for proper analysis of iNKT TCR binding and demonstrated the necessity to adjust loading conditions for each oligomer/glycolipid combination. The efficient usage of surfactants as a tool for CD1d loading was revealed to be species-specific and depending on the origin of the CD1d producing cells. Additional variation of surfactant-dependent loading efficacy between tested glycolipids was influenced by the acyl-chain length and the modification of the galactosyl head group with PBS57 showing the least dependence on surfactants and the lowest degree of species-dependent differences.

Original languageEnglish (US)
Article numbere0143449
JournalPLoS One
Volume10
Issue number11
DOIs
StatePublished - Nov 1 2015

Fingerprint

Oligomers
Surface-Active Agents
surfactants
CD1d Antigen
glycolipids
Glycolipids
Galactosylceramides
Rats
Natural Killer T-Cells
Sigmodon
Sigmodontinae
T-cells
Polysorbates
rats
Octoxynol
cells
Chain length
Dimers
Cotton
T-lymphocytes

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Paletta, D., Fichtner, A. S., Starick, L., Porcelli, S. A., Savage, P. B., & Herrmann, T. (2015). Species specific differences of CD1d oligomer loading in vitro. PLoS One, 10(11), [e0143449]. https://doi.org/10.1371/journal.pone.0143449

Species specific differences of CD1d oligomer loading in vitro. / Paletta, Daniel; Fichtner, Alina Suzann; Starick, Lisa; Porcelli, Steven A.; Savage, Paul B.; Herrmann, Thomas.

In: PLoS One, Vol. 10, No. 11, e0143449, 01.11.2015.

Research output: Contribution to journalArticle

Paletta, D, Fichtner, AS, Starick, L, Porcelli, SA, Savage, PB & Herrmann, T 2015, 'Species specific differences of CD1d oligomer loading in vitro', PLoS One, vol. 10, no. 11, e0143449. https://doi.org/10.1371/journal.pone.0143449
Paletta D, Fichtner AS, Starick L, Porcelli SA, Savage PB, Herrmann T. Species specific differences of CD1d oligomer loading in vitro. PLoS One. 2015 Nov 1;10(11). e0143449. https://doi.org/10.1371/journal.pone.0143449
Paletta, Daniel ; Fichtner, Alina Suzann ; Starick, Lisa ; Porcelli, Steven A. ; Savage, Paul B. ; Herrmann, Thomas. / Species specific differences of CD1d oligomer loading in vitro. In: PLoS One. 2015 ; Vol. 10, No. 11.
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