Figure 1
Figure 1. TH cells are juxtaposed with monocytes in inflamed skin and drive monocyte differentiation into DCs. (A) Representative skin lesions from psoriasis and atopic dermatitis patients were stained with antibodies against CD4 (brown) and CD14 (blue). Images are shown at 400× magnification and were acquired using the equipment described in “Immunohistochemistry.” Red arrows indicate potential monocyte/T cell interactions. Scale bar equals 20 μm. (B-F) CD14+ monocytes were cultured with IL-2 (Mono), autologous TH cells and IL-2 (DCTh), or GM-CSF and IL-4 (DCGM) for 6 days. (B) Hoffman modulation contrast microscopy shown at 400× magnification. Images were acquired using the equipment described in “Monocyte/T-cell cocultures.” Data are representative of > 10 donors. (C) Median fluorescence intensities (MFIs) of DC associated molecules are shown after excluding TH cells. Dashed lines indicate the average MFIs of isotype control Abs used for staining from all culture conditions. Data are from 4 independent experiments and > 10 donors (mean and SEM); *P < .05, **P < .01, ***P < .001 compared with DCGM. (D-F) On day 6, Mono, DCTh and DCGM were cultured for 36 hours in the presence (+ LPS) or absence (Unstim) of LPS. (D) MFIs of CD80 and CD86 are shown after excluding TH cells. Dashed lines indicate the average MFIs of isotype control Ab used for staining from all culture conditions. Data are from 2 independent experiments and 6 donors (mean and SEM). (E) Unstimulated (black histograms) and LPS-stimulated (red histograms) DCTh and DCGM were incubated with DQ-OVA for 30 minutes on ice (open histograms) or at 37°C (shaded histograms). DQ-OVA expression is shown after excluding TH cells. Data are representative of 2 independent experiments and 2 donors. (F) Allogeneic naive CD4+ T cells were cultured alone (T Alone) or with irradiated Mono, DCTh or DCGM with and without LPS stimulation at a 1:5 APC to T cell ratio in an MLR. After 6 days, 3H-thymidine was added for an additional 20 hours and T cell proliferation was assessed. Data are from 2 independent experiments and 5 donors (mean and SEM).

TH cells are juxtaposed with monocytes in inflamed skin and drive monocyte differentiation into DCs. (A) Representative skin lesions from psoriasis and atopic dermatitis patients were stained with antibodies against CD4 (brown) and CD14 (blue). Images are shown at 400× magnification and were acquired using the equipment described in “Immunohistochemistry.” Red arrows indicate potential monocyte/T cell interactions. Scale bar equals 20 μm. (B-F) CD14+ monocytes were cultured with IL-2 (Mono), autologous TH cells and IL-2 (DCTh), or GM-CSF and IL-4 (DCGM) for 6 days. (B) Hoffman modulation contrast microscopy shown at 400× magnification. Images were acquired using the equipment described in “Monocyte/T-cell cocultures.” Data are representative of > 10 donors. (C) Median fluorescence intensities (MFIs) of DC associated molecules are shown after excluding TH cells. Dashed lines indicate the average MFIs of isotype control Abs used for staining from all culture conditions. Data are from 4 independent experiments and > 10 donors (mean and SEM); *P < .05, **P < .01, ***P < .001 compared with DCGM. (D-F) On day 6, Mono, DCTh and DCGM were cultured for 36 hours in the presence (+ LPS) or absence (Unstim) of LPS. (D) MFIs of CD80 and CD86 are shown after excluding TH cells. Dashed lines indicate the average MFIs of isotype control Ab used for staining from all culture conditions. Data are from 2 independent experiments and 6 donors (mean and SEM). (E) Unstimulated (black histograms) and LPS-stimulated (red histograms) DCTh and DCGM were incubated with DQ-OVA for 30 minutes on ice (open histograms) or at 37°C (shaded histograms). DQ-OVA expression is shown after excluding TH cells. Data are representative of 2 independent experiments and 2 donors. (F) Allogeneic naive CD4+ T cells were cultured alone (T Alone) or with irradiated Mono, DCTh or DCGM with and without LPS stimulation at a 1:5 APC to T cell ratio in an MLR. After 6 days, 3H-thymidine was added for an additional 20 hours and T cell proliferation was assessed. Data are from 2 independent experiments and 5 donors (mean and SEM).

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal