Skip to main content
. Author manuscript; available in PMC: 2010 Jul 31.
Published in final edited form as: J Immunol Methods. 2009 May 14;346(1-2):38–44. doi: 10.1016/j.jim.2009.05.003

Fig. 1.

Fig. 1

Schematic diagram of CD1d-based aAPC. (A) In this system, CD1d-Ig is used to provide the cognate antigen specific signal through the TCR (signal 1) and anti-CD28 Abs or other costimulatory molecules provide signal 2. These molecules can be easily loaded with lipid antigens, such as α-GalCer, simply by incubating them with an excess of the lipid of interest. (B) aAPC were analyzed for surface expression of CD1d-Ig and anti-CD28 by flow cytometry. Open histograms indicate isotype control; filled histograms represent the indicated antibodies. (C) DN32.D3 (5×104) NKT cell hybridomas were cocultured with media, soluble α-GalCer (100 ng/ml), or aAPC (2.5 × 105) in 96-well U- bottom plates for 20–24hr. Culture supernatants were harvested and standard sandwich ELISA was used to measure IL-2 production. (D) human CD1d-Ig Expressing aAPC can Stimulate IL-2 Production by NKT cells. Unloaded or α–GalCer loaded human CD1d-based aAPC were cocultured with the indicated Vαl4+ NKT cell hybridomas. Culture supernatants were harvested and standard sandwich ELISA was used to measure IL-2 production.