Evaluation of the Capacity of Dendritic Cells Derived from Cord Blood CD34+ Precursors to Present Haptens to Unsensitized Autologous T Cells In Vitro

Abstract

Langerhans cells play a key role in contact hypersensitivity reactions. The application of haptens on the skin leads to many modifications of these cells, including the increase of major histocompatibility complex II expression, allogeneic stimulation potency, and migration towards lymph nodes to activate T cells. Moreover, it has been shown that Langerhans cells cultured in vitro are able to prime naive T cells in response to hapten contact. From CD34+ progenitors present in cord blood, we generated dendritic cells of which some presented the phenotypic markers of Langerhans cells. We show that these cells are able to sensitize syngeneic naive (CD45RA+) T cells to haptens such as trinitrophenyl conjugate of trinitrobenzene sulfonic acid (TNP) and fluoroscein isothiocyanate. The response to TNP is higher than to fluoroscein isothiocyanate, whereas sodium dodecyl sulfate, an irritant molecule used as a control, never caused this effect. Phenotypic analysis of cellular suspensions and experiments of cell sorting lead to the conclusion that only CD1a+ cells are able to induce a primary response of syngeneic T cells to TNP or fluoroscein isothiocyanate. Furthermore, we have shown a close relationship between the differentiation state of dendritic cells and their ability to prime T lymphocytes. Dendritic cells are able to present haptens in an efficient manner between day 10 and 14 of culturing CD34+ progenitors, whereas they were efficient in presenting alloantigens from day 6 until after day 20. This dissociation suggests the need of an active metabolic process for hapten presentation in the direct treatment of dendritic cells with haptens. This model of hapten presentation was used for a panel of fragrance molecules and other molecules considered as weaker haptens than TNP and fluoroscein isothiocyanate.

Nickel and DNCB Induce CCR7 Expression on Human Dendritic Cells Through Different Signalling Pathways: Role of TNF-α and MAPK

“…Contact sensitizers have been shown to induce DC maturation in vitro (Aiba et al, 1997;Degwert et al, 1997;Rougier et al, 1998;Coutant et al, 1999;Arrighi et al, 2001;De Smedt et al, 2001). According to the crucial role of LC migration from epidermis to lymph nodes in the initiation phase of contact hypersensitivity, we explored the mechanism of CCR7 expression after hapten-stimulation of human DC derived from CD34 þ hematopoietic precursors.…”

Implication of the MAPK pathways in the maturation of human dendritic cells induced by nickel and TNF-?

“…The establishment of in vitro models of DC offered the possibility to demonstrate that haptens were able to directly activate cultured human DC derived from peripheral blood monocytes or CD34 + hematopoietic progenitor cells (HPC) (Aiba et al, 1997;Degwert et al, 1997;Rougier et al, 1998). These observations showed the up-regulation of maturation marker (CD83), costimulatory molecules (CD80, CD86, CD40) and the chemokine receptor CCR7 by haptens (Arrighi et al, 2001;Boisleve et al, 2004;Coutant et al, 1999;De Smedt et al, 2001;Rougier et al, 2000;Tuschl et al, 2000).…”

Binding of live conidia of Aspergillus fumigatus activates in vitro-generated human Langerhans cells via a lectin of galactomannan specificity

“…After overnight culture on a plastic surface, adherent cells represented a monocytes‐rich fraction. T lymphocytes were isolated from non‐adherent cells by rosetting with sheep red blood cells, as described previously [15].…”