Adiponectin modulates NK-cell function
Article first published online: 1 MAR 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
European Journal of Immunology
Volume 43, Issue 4, pages 1024–1033, April 2013
How to Cite
Wilk, S., Jenke, A., Stehr, J., Yang, C.-A., Bauer, S., Göldner, K., Kotsch, K., Volk, H.-D., Poller, W., Schultheiss, H.-P., Skurk, C. and Scheibenbogen, C. (2013), Adiponectin modulates NK-cell function. Eur. J. Immunol., 43: 1024–1033. doi: 10.1002/eji.201242382
- Issue published online: 16 APR 2013
- Article first published online: 1 MAR 2013
- Accepted manuscript online: 11 FEB 2013 04:08AM EST
- Manuscript Accepted: 1 FEB 2013
- Manuscript Revised: 19 DEC 2012
- Manuscript Received: 11 JAN 2012
- Sonderforschungsbereich (SFB) Transregio 19 of the Deutsche Forschungsgemeinschaft (DFG)
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Figure 1A, 1B and 1 C. For the expression analysis of AdipoR1 and R2 „doublets “and dead cells were excluded. Human PBMCs were gated on CD3-CD56+ NK cells and for CD3+CD4+ T cells (Fig. 1A). Further CD56+ cells were subdivided into CD56high and CD56dim cells. Surface and intracellular expression of AdipoR1/R2 was analyzed.
Figure 2A, B, C, D and E. For identification of IFN-γ producing and CD107a+ degranulating cells, CD3-CD56+ cells were used. Dead cells and doublets were excluded from alalysis. INF-γ and CD107a was analyzed on CD56+ NK cells.For identification of sorted IFN-γ producing NK cells, dead cells, doublets and remained CD3+ cells were excluded. IFN-γ secretion was analyzed (Fig. 2B, D).
Figure 2F. For the analysis of cytotoxicity, K562 were labeled with a fluorescence dye and incubated with sorted NK cells. PI was used to gate on dead cells.
Figure 3A. For the identification of of DX5+Nkp46+ cells, ex vivo splenocytes of uninfected and infected WT and APN-KO mice were used. Doublets and CD3+ cells were excluded.
Figure 3B and C. For the identification of NK cell frequencies and numbers, ex vivo splenocytes of uninfected and infected WT and APN-KO mice were used. Doublets and CD3+ cells were excluded.
Figure 4. Intra- and extracellular AdipoR1 and 2 expression was analyzed on CD3e+ and CD3e-Nkp46+ splenocytes. Doublets were excluded from analysis.
Figure 5. Expression of the degranulation marker CD107a, NKG2D and CD94low/high was analyzed on CD3- Nkp46+ splenoycytes. In the flow cytometric analysis for CD94 expression we included the maturation marker CD11b. NK cell subsets were analyzed by CD11b and CD27 on Nkp46+ cells.
Figure 6. Expression of IFN-γ was analyzed ex vivo on CD3e-Nkp46+ splenocytes.
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