CD1d protein structure determines species-selective antigenicity of isoglobotrihexosylceramide (iGb3) to invariant NKT cells
Version of Record online: 31 JAN 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
European Journal of Immunology
Volume 43, Issue 3, pages 815–825, March 2013
How to Cite
Sanderson, J. P., Brennan, P. J., Mansour, S., Matulis, G., Patel, O., Lissin, N., Godfrey, D. I., Kawahara, K., Zähringer, U., Rossjohn, J., Brenner, M. B. and Gadola, S. D. (2013), CD1d protein structure determines species-selective antigenicity of isoglobotrihexosylceramide (iGb3) to invariant NKT cells. Eur. J. Immunol., 43: 815–825. doi: 10.1002/eji.201242952
- Issue online: 11 MAR 2013
- Version of Record online: 31 JAN 2013
- Accepted manuscript online: 26 DEC 2012 08:47AM EST
- Manuscript Accepted: 17 DEC 2012
- Manuscript Revised: 2 DEC 2012
- Manuscript Received: 28 AUG 2012
- Higher Education Funding Council for England (HEFCE)
- National Health and Medical Research Council of Australia (NHMRC)
- Senior Principal Research Fellowship
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Figure S1. Alignment of α2-helix forming amino acids of published CD1d sequences, showing conservation of W155 in catarrhines (apes and Old World monkeys) and of G153 in nearly all other species. 1. Pseudogene. 2. CD1 protein, but not orthologous to any specific eutherian isoform.
Figure S2. Gating strategy for enumeration of iNKT populations in lipid-treated PBMC cultures (as used for generating data shown in figure 1a). Shown is an example of a KRN7000 stimulated in vitro culture: Forward/side scatter characteristics were used to define region 1 (R1) containing lymphocytes (left density plot). Propidium iodide exclusion and staining with FITC-conjugated anti-CD3 antibody were used to define region 2 (R2), and from this live T lymphocytes were identified by gating cells on (R1+R2)(middle density plot), and live iNKT cells were identified as αGC-CD1d tetramer positive cells in gate(R1+R2) (right density plot). All flow cytometry data were acquired on a BD FACSCalibur, and data were analysed using CellQuest Pro software.
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