Frank O. Nestle Journal of Investigative Dermatology

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Human Dermal Dendritic Cells Process and Present Soluble Protein Antigens Frank O. Nestle Journal of Investigative Dermatology Volume 110, Issue 5, Pages 762-766 (May 1998) DOI: 10.1046/j.1523-1747.1998.00189.x Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Ultrastructural analysis of TT uptake and processing in epidermal Langerhans cells. (A) Overview of Langerhans cell with typical dendritic membrane processes. (B) No membrane binding of TT was found but occasionally colloidal gold (4 nm) labeled TT (arrowhead) is found in an invagination of the surface membrane at the beginning of macropinocytosis. (C) TT-gold complex (arrowhead) in an endocytotic vesicle near to a Birbeck granule (arrow). (D) TT-gold complexes (arrowhead) in a tubular (one arrowhead) and in a multivesicular (two arrowheads) vesicle in the area of the early lysosomal compartment near to Birbeck granules (arrow). Journal of Investigative Dermatology 1998 110, 762-766DOI: (10.1046/j.1523-1747.1998.00189.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Ultrastructural analysis of TT uptake and processing in DDC. (A) Overview of a DDC with typical dendritic membrane protrusions. (B) Colloidal gold (4 nm) labeled TT (arrowhead) bound to the surface membrane of a DDC. (C) TT-gold complexes (arrowhead) are visualized bound to the membrane of endocytotic vesicles near to the cell surface of a DDC. (D) TT-gold complexes (arrowhead) are visualized bound to membrane structures in lamellar vesicles in the area of the early lysosomal compartment. Journal of Investigative Dermatology 1998 110, 762-766DOI: (10.1046/j.1523-1747.1998.00189.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Induction of TT specific T cell proliferation by increasing numbers of stimulator cells. Langerhans cells, DDC, and monocytes (Mono) were pulsed with TT (10 μg per ml) for 2 h and cultured at 50, 500, and 5000 cells per well in triplicates with 5 × 104 autologous resting CD4 positive T cells, representing stimulator:responder ratios of 1:1000, 1:100, and 1:10, respectively. Proliferative responses in these cultures were measured after 3 d. T cells alone or DDC alone or in the presence of TT incorporated less than 500 cpm. Note the strong stimulatory capacity of Langerhans cells and DDC relative to monocytes. Journal of Investigative Dermatology 1998 110, 762-766DOI: (10.1046/j.1523-1747.1998.00189.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Effects of chloroquine on antigen processing. Langerhans cells and DDC were used as APC and pulsed with TT (TTprot; 10 μg per ml) or a TT peptide p30 (TTpept at 10 ng per ml) in the presence or absence of 300 μM chloroquine (Chloro). Five × 102 APC were incubated with 5 × 104 resting CD4 positive T cells (stimulator:responder ratio of 1:100) and proliferative responses were measured after 3 d. In the presence of chloroquine there was a 68% decrease (16744 ± 2210 vs 5511 ± 466) in the ability of DDC and a 64% decrease (14493 ± 1031 vs 5445 ± 447) in the ability of Langerhans cells to induce antigen specific T cell proliferation. T cell proliferation in the absence of antigen was 2122 ± 332 for Langerhans cells and 1880 ± 322 for DDC. Journal of Investigative Dermatology 1998 110, 762-766DOI: (10.1046/j.1523-1747.1998.00189.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 TT specific T cell proliferation induced by subpopulations of DDC. Cell populations were differentially sorted using immunofluorescence staining for CD1a. CD1a negative and positive, as well as unsorted DDC (50, 500, or 5000 per well in triplicates), were pulsed with TT (10 μg per ml) for 2 h and incubated with 5 × 104 autologous resting CD4 positive T cells, representing stimulator:responder ratios of 1:1000, 1:100, and 1:10, respectively. Proliferative responses in these cultures were measured after 3 d. T cells alone or DDC alone or in the presence of TT incorporated less than 500 cpm. Note that CD1a positive and negative, as well as unsorted DDC, have similar capacities in inducing TT specific T cell proliferation. Journal of Investigative Dermatology 1998 110, 762-766DOI: (10.1046/j.1523-1747.1998.00189.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions