Modification of the human allergic immune response by allergen-DNA–transfected dendritic cells in vitro Bettina Klostermann, MSc, Iris Bellinghausen,

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Presentation transcript:

Modification of the human allergic immune response by allergen-DNA–transfected dendritic cells in vitro Bettina Klostermann, MSc, Iris Bellinghausen, PhD, Ingo Böttcher, MSc, Arnd Petersen, PhD, Wolf-Meinhard Becker, PhD, Jürgen Knop, MD, Joachim Saloga, MD Journal of Allergy and Clinical Immunology Volume 113, Issue 2, Pages 327-333 (February 2004) DOI: 10.1016/j.jaci.2003.10.067

FIG 1 Efficiency of the adenoviral transfection of human mature DCs. DCs were generated as described and transfected at day 6 with adenoviral vectors encoding green fluorescent protein at different MOIs. One representative example of 8 independent experiments is shown. Journal of Allergy and Clinical Immunology 2004 113, 327-333DOI: (10.1016/j.jaci.2003.10.067)

FIG 2 Expression of the protein Phl p 1 by transfected DCs. The cells were lysed 48 hours after transfection. Phl p 1 protein (0.5 ng) was used as a positive control, and as a negative control, we used untransfected mature DCs. A representative example of 3 separate experiments is shown. Journal of Allergy and Clinical Immunology 2004 113, 327-333DOI: (10.1016/j.jaci.2003.10.067)

FIG 3 Surface marker expression of DCs after adenoviral transfection or exposure to allergen protein. DCs were generated as described and transfected with Ad-Phl p 1 at day 6. After 48 hours, cells were stained with the markers indicated. A representative example of 3 separate experiments is shown. Journal of Allergy and Clinical Immunology 2004 113, 327-333DOI: (10.1016/j.jaci.2003.10.067)

FIG 4 Proliferation of autologous human CD4+ (A) and CD8+ (B) T cells activated by transfected DCs. T cells were stimulated with allergen protein–pulsed, transfected, or control DCs. After 5 days, cells were pulsed with 1 μCi of tritiated thymidine. Results are expressed as the means ± SD from 8 atopic donors. ∗P ≤ .05 compared with medium or Psi 5. Journal of Allergy and Clinical Immunology 2004 113, 327-333DOI: (10.1016/j.jaci.2003.10.067)

FIG 5 Cytokine production by CD4+ (A) and CD8+ (B) T cells after stimulation with autologous DCs. T cells were stimulated twice with allergen protein–pulsed, transfected, or control DCs, and cytokines were measured by means of ELISA. Results are expressed as means ± SD from 6 donors. ∗P ≤ .05 compared with medium or Psi 5. †P ≤ .03 comparing allergen-DNA conjugate– transfected and allergen protein–pulsed DCs. Journal of Allergy and Clinical Immunology 2004 113, 327-333DOI: (10.1016/j.jaci.2003.10.067)