Figure 1

Proportions of dendritic cells and intracellular expression of cytokines by them in healthy controls (n = 20) and in untreated patients with HIV infection: recent (n = 10) and chronic (n = 10). (A) Myeloid dendritic cells –mDCs- (percentage of leukocytes). mDCs were identified by their reactivity with anti-CD1c (BDCA-1, clone AD5-8E7) (Miltenyi Biotec GMBH, Bergisch Gladbach, Germany) after exclusion of monocytes, B lymphocytes and dead cells using a cocktail of anti-CD14/anti-CD19/dead cells (Cocktail Lin clones CD3-SK3, CD14-Mφ9, CD16-3G8, CD19-SJ25L1, CD20-L27, CD56-NCAM16.2; BD Biosciences, Franklin Lakes, NJ, USA). (B) Intracellular expression of interleukin 12 (percentage of myeloid dendritic cells expressing interleukin 12), determined using specific antibodies (anti-IL12p40/70, clone C8.6, Miltenyi Biotec GMBH), after stimulation with LPS (1 μg/ml) for 5 hours. (C) Plasmocytoid dendritic cells –pDCs- (percentage of leukocytes). pDCs were identified by their reactivity with anti-CD303 (BDCA-2, clone AC-144) (Miltenyi Biotec GMBH, Bergisch Gladbach, Germany) after exclusion of monocytes, B lymphocytes and dead cells using a cocktail of anti-CD14/anti-CD19/dead cells (Cocktail Lin clones CD3-SK3, CD14-Mφ9, CD16-3G8, CD19-SJ25L1, CD20-L27, CD56-NCAM16.2; BD Biosciences, Franklin Lakes, NJ, USA). (D) Intracellular expression of interferon alpha (percentage of plasmacytoid dendritic cells expressing interferon alpha), determined using specific antibodies (clone LT 27:295, Miltenyi Biotec GMBH), after stimulation with CpG-A (3 μM) for 20 hours. Above each graphic, flow cytometry data for representative cases of a healthy control and a patient with untreated chronic HIV infection are shown. A grey line in (B,C) figures represents isotype controls, used to confirm specificity of staining and to discriminate background staining. Data are provided as median, interquartile values and range.