Invariant natural killer T cells are not affected by lysosomal storage in patients with Niemann-Pick disease type C
Article first published online: 14 JUN 2012
© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
European Journal of Immunology
Volume 42, Issue 7, pages 1886–1892, July 2012
How to Cite
Speak, A. O., Platt, N., Salio, M., te Vruchte, D., Smith, D. A., Shepherd, D., Veerapen, N., Besra, G. S., Yanjanin, N. M., Simmons, L., Imrie, J., Wraith, J. E., Lachmann, R. H., Hartung, R., Runz, H., Mengel, E., Beck, M., Hendriksz, C. J., Porter, F. D., Cerundolo, V. and Platt, F. M. (2012), Invariant natural killer T cells are not affected by lysosomal storage in patients with Niemann-Pick disease type C. Eur. J. Immunol., 42: 1886–1892. doi: 10.1002/eji.201141821
- Issue published online: 17 JUL 2012
- Article first published online: 14 JUN 2012
- Accepted manuscript online: 14 MAY 2012 07:41AM EST
- Manuscript Accepted: 22 MAR 2012
- Manuscript Revised: 17 FEB 2012
- Manuscript Received: 1 JUN 2011
- MRC. Grant Number: G0700851
- MRC. Grant Number: G0800158
- Action Medical Research. Grant Number: SP4023
- Cancer Research UK. Grant Number: C399/A2291
Disclaimer: Supplementary materials have been peer-reviewed but not copyedited.
Figure S1. Gating Doublets were excluded by FSC-H versus FSC-A and lymphocytes identified by size and granularity (FSC-A versus SSC-A). Viable lymphocytes were selected on the basis of exclusion of live/dead aqua stain. Total T cells were identified as CD3+ viable lymphocytes and iNKT cells as either 6B11+CD3+ or tetramer+CD3+ cells. Fluorescence minus one controls with isotype control antibodies were used to establish gating for iNKT cells and CD161, CD4 and CD8.
Figure S2. Gating strategy for identification of B cells and monocytes. Doublets were excluded by FSC-H versus FSC-A and viable cells were selected on the basis of exclusion of live/dead aqua. B cells were identified as CD19+SSClow and monocytes as CD14+SSCmid. CD1d expression was determined by MFI of the PE channel and specificity determined by fluorescence minus one with isotype control antibody controls.
Figure S3. Example of human CD1d expression on EBV-B cells after transfection. EBV-B cells were identified on the basis of size (FSC vs SSC) and dead cell excluded with the use of a viability dye, human CD1d was detected at the cell surface with a fluorescent antibody.
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