TGFβ Inhibits CD1d Expression on Dendritic Cells

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TGFβ Inhibits CD1d Expression on Dendritic Cells S. Ronger-Savle, J. Valladeau, A. Claudy, D. Schmitt, J. Peguet-Navarro, C. Dezutter-Dambuyant, L. Thomas, D. Jullien Journal of Investigative Dermatology Volume 124, Issue 1, Pages 116-118 (January 2005) DOI: 10.1111/j.0022-202X.2004.23315.x Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 CD1d expression on Langerhans cells and dendritic cells generated in vitro from monocytes: TGFβ decrease CD1d expression. (A) Detection of CD1d mRNA in DC by RT-PCR: specimens were normalized to yield equivalent GAPDH PCR products. The CD1d specific signal was observed in all populations, but it was stronger in DC without TGFβ in culture. Marker molecular are shown (M). (B) Surface expression of CD1d molecules using flow cytometric analysis: the open histograms show staining with anti-CD1d mAb (42.1). The solid histograms show staining with an isotype-matched non-binding control mAb. CD1d protein was not expressed on DC derived monocytes with TGFβ (Langerhans cells like) but there was a signal on DC derived monocytes that have not TGFβ on cultures, that are DDC. Journal of Investigative Dermatology 2005 124, 116-118DOI: (10.1111/j.0022-202X.2004.23315.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 CD1d expression along differentiation of CD34+ hematopoietic progenitor cells toward Langerhans cells. LC were generated from CD34+ HPC in the presence of GM-CSF, TNFα and TGFβ. FACS analysis performed at various stage of differentiation show that as opposed to CD1a, CD1d molecule expression at the cell surface decrease during the differentiation process. When TGFβ was omitted CD1d membrane expression remained constant. The open histograms show staining with anti-CD1d mAb (CD1d 42.1) or anti-CD1a mAb (NA1/34). The solid histograms show staining with isotype-matched non-binding control mAbs. Comparison of CD1d mRNA at various stages of differentiation: the cDNA derived from cells harvested at day 5, 7 and at day 14 (i.e., LC) were normalized to yield equivalent GAPDH PCR products. When LC were generated from CD34+ HPC in the presence of GM-CSF, TNFα and TGFβ1, CD1d mRNA expression decreased progressively and very low levels were obseved at day 14 in differentiated LC. When TGFβ was omitted, CD1d mRNA expression levels remained constant. Journal of Investigative Dermatology 2005 124, 116-118DOI: (10.1111/j.0022-202X.2004.23315.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Ex vivo, LC do not express CD1d. Fresh Langerhans cells were obtained from fresh normal skin donors. Staining on FACS analysis with CD1d 42.1 was negative, and CD1a was overexpressed. Journal of Investigative Dermatology 2005 124, 116-118DOI: (10.1111/j.0022-202X.2004.23315.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions