These authors contributed equally to this work.
Complement opsonization of HIV-1 results in a different intracellular processing pattern and enhanced MHC class I presentation by dendritic cells
Article first published online: 24 APR 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
European Journal of Immunology
Volume 43, Issue 6, pages 1470–1483, June 2013
How to Cite
Tjomsland, V., Ellegård, R., Burgener, A., Mogk, K., Che, K. F., Westmacott, G., Hinkula, J., Lifson, J. D. and Larsson, M. (2013), Complement opsonization of HIV-1 results in a different intracellular processing pattern and enhanced MHC class I presentation by dendritic cells. Eur. J. Immunol., 43: 1470–1483. doi: 10.1002/eji.201242935
- Issue published online: 21 JUN 2013
- Article first published online: 24 APR 2013
- Accepted manuscript online: 25 MAR 2013 03:20AM EST
- Manuscript Accepted: 19 MAR 2013
- Manuscript Revised: 20 FEB 2013
- Manuscript Received: 22 AUG 2012
- VINNMER. Grant Number: AI52731
- The Swedish Research Council
- University Hospital Research Fund
- National Cancer Institute, National Institutes of Health. Grant Number: HHSN261200800001E
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Table S1. Primer sequences.
Figure S1: Complement opsonization of HIV-1 enhanced MHCI presentation by IDCs and MDCs (A-D) IDCs and MDCs (0.15 x106) were incubated over night with mock, free HIV-1BaL (F-HIV), complement opsonized HIV-1BaL (C-HIV), IgG opsonized HIV-1BaL (IgG-HIV), or complement and IgG opsonized HIV-1BaL (C-IgG-HIV) (75ng p24CA equivalent/group). After the incubation the different groups of DCs were washed and cocultured with a HIV-1 gag p17 SL9 (SLYNTVATL) specific CD8+ T-cell clone to assess MHCI presentation (A-B) or HIV-1 p24 LI13 (LNKIVRMYSPTS) specific CD4+ T-cell clone to assess MHCII presentation (C-D) for 12h. The T-cell activation was assessed by IFN-g ELISPOT assay and the amount of spot forming cells (SFC) was measured. Data are shown as mean ±SEM and one representative experiment, with triplicate values, out of 6-28 experiments performed.
Figure S2: Complement opsonization of HIV did not affect DC expression of costimulatory molecules and their ability to prime naïve T cell responses. (A-D) DCs were exposed to mock, F-HIV or C-HIV for 48h and the level of expression of CD40, CD80, CD86, and HLA DR was assessed by staining with PE-conjugated antibodies and analyzing by flow cytometry. Poly I:C (25 ug/mL) was used as a positive control for maturation. Data are shown as mean ±SEM of three experiments performed (E) DC were pulsed with mock, F-HIV or C-HIV overnight, washed twice, and cocultured with naïve bulk T cells at a ratio of 1:10. Priming cultures were restimulated with 10 000 DC/well after 7 days of coculture and T-cell proliferation measured on day 8 by 3H-Thymidine incorporation. Data are shown as mean ±SEM of six experiments performed.
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