Species Specific Differences of CD1d Oligomer Loading In Vitro
Please always quote using this URN: urn:nbn:de:bvb:20-opus-124879
- 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, weCD1d 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.…
Author: | Daniel Paletta, Alina Suzann Fichtner, Lisa Starick, Steven A. Porcelli, Paul B. Savage, Thomas Herrmann |
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URN: | urn:nbn:de:bvb:20-opus-124879 |
Document Type: | Journal article |
Faculties: | Medizinische Fakultät / Institut für Virologie und Immunbiologie |
Language: | English |
Parent Title (English): | PLoS One |
Year of Completion: | 2015 |
Volume: | 10 |
Issue: | 11 |
Pagenumber: | e0143449 |
Source: | PLoS ONE 10(11): e0143449. doi:10.1371/journal. pone.0143449 |
DOI: | https://doi.org/10.1371/journal.pone.0143449 |
Dewey Decimal Classification: | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
Tag: | T cells; binding analysis; cell staining; glycolipids; lipids; major histocompatibility complex; oligomers; surfactants |
Release Date: | 2016/01/26 |
Collections: | Open-Access-Publikationsfonds / Förderzeitraum 2015 |
Licence (German): | CC BY: Creative-Commons-Lizenz: Namensnennung |