Materials and Reagents

1. Mouse-anti-human CD107a PE (clone H4A3) (BD Biosciences, catalog number: 555801 )
   
2. Mouse-anti-human CD3 PE-Cy7 (clone UCHT1) (Biolegend, catalog number: 300420 )
   
3. Mouse-anti-human CD8 Alexa-Fluor-700 (clone 3B5) (Life Technologies, Invitrogen™, catalog number: MCHD0824 )
   Note: The above antibodies have been tested by the author and may be substituted with the antibodies desired by users.
   
4. CFSE (Molecular Probes Europe BV)
   
5. Iscove’s Modified Dulbecco’s Medium (IMDM) (Life Technologies, Invitrogen™)
   
6. Heat-inactivated fetal calf serum (FCS) (Integro)
   
7. Human serum (HS) (Sanquin Blood Bank)
   
8. Recombinant human IL-2 (Proleukin, Chiron)
   
9. Recombinant human IL-15 (Immunotools, catalog number: 11340155 )
   
10. Antigen-specific peptide (For example MiHA or CMV-peptide, Thinkpeptides)
    
11. APC-conjugated antigen-specific tetramer (Kindly provided by Prof. dr. J. H. F. Falkenburg, Leiden University Medical Centre, the Netherlands)
    
12. Sytox blue (Life Technologies, Invitrogen™, catalog number: S34857 )
    
13. Mature monocyte-derived DC [cultured following the protocol described in Hobo et al. (2010)]
    
14. FACS buffer (see Recipes)
    

Equipment

1. Beckman Coulter Navios flow cytometer
   
2. 24-wells plate
   
3. 37 °C incubator
   

Procedure