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The use of leukemia cells as antigen-presenting cells (APCs) in immunotherapy is critically dependent on their capacity to initiate and sustain an antitumor-specific immune response. Previous studies suggested that pediatric B-cell precursor acute lymphoblastic leukemia (BCP-ALL) cells could be manipulated in vitro through the CD40-CD40L pathway to increase their immunostimulatory capacity. We extended the APC characterization of CD40L-activated BCP-ALL for their potential use in immunotherapy in a series of 19 patients. Engaging CD40 induced the up-regulation of CCR7 in 7 of 11 patients and then the migration to CCL19 in 2 of 5 patients. As accessory cells, CD40L-activated BCP-ALL induced a strong proliferation response of naive T lymphocytes. Leukemia cells, however, were unable to sustain proliferation over time, and T cells eventually became anergic. After CD40-activation, BCP-ALL cells released substantial amounts of interieukin-1 0 (IL-10) but were unable to produce bioactive IL-12 or to polarize T(H)1 effectors. Interestingly, adding exogenous IL-12 induced the generation of interferon-gamma (IFN-gamma)-secreting T(H)1 effectors and reverted the anergic profile in a secondary response. Therefore, engaging CD40 on BCP-ALL cells is insufficient for the acquisition of full functional properties of immunostimulatory APCs. These results suggest caution against the potential use of CD40L-activated BCP-ALL cells as agents for immunotherapy unless additional stimuli, such as IL-12, are provided.

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Traditionally described as a major anti-coagulant system, the protein C (PC) pathway, consisting of thrombomodulin, the endothelial cell protein C receptor and activated PC (APC), is gaining increasing attention as an important regulator of microvascular inflammation. Although they possess several anti-inflammatory and cytoprotective functions, the expression and function of the components of the PC pathway is downregulated during inflammation. Recent evidence suggests that the PC pathway is defective in patients with inflammatory bowel disease (IBD) and that restoring its function has anti-inflammatory effects on cultured intestinal microvascular endothelial cells and in animal models of colitis. Here, we propose that the PC pathway has an important role in governing intestinal microvascular inflammation and might provide a novel therapeutic target in the management of IBD.

Dendritic cells (DCs) comprise a family of cells specializing in antigen capture and presentation to T cells. We have recently shown that DC play an active role in bacterial uptake across mucosal surfaces. Indeed, DC are able to open tight junctions and to sample antigens directly across epithelia, both in vitro and in vivo. Because DC express tight junction proteins, the integrity of the epithelial barrier is preserved. In this study we have analyzed the possible involvement of epithelial cells in controlling DC function. We developed an in vitro model in our laboratory consisting of a three-player system of dendritic cells, epithelial cell monolayers, and bacteria. The crosstalk between epithelial cells and dendritic cells was analyzed, and epithelial cells were tested for their capacity to release cytokines and chemokines that induce the migration and activation of DC. We show that the capacity of epithelial eels to produce cytokines and activate DC is dependent on the invasiveness of the bacteria tested. In particular, invasive bacteria stimulate epithelial cells to release proinflammatory cytokines and to induce the maturation state of DC. By contrast, noninvasive bacteria are unable to stimulate epithelial cells, but can activate DC directly when DC translocate to the apical side. In conclusion, epithelial cells are not simply a barrier to bacteria entering via the oral route, but actively influence the activating properties of bystander DC. © 2004 New York Academy of Sciences.

Dendritic cells (DCs) are able to open the tight junctions between adjacent epithelial cells (ECs) and to take up both invasive and noninvasive bacteria directly from the intestinal lumen. In this study, we describe a tight cross talk between ECs and human monocyte-derived DCs (MoDCs) in bacterial handling across epithelial monolayers. We show that the release of proinflammatory mediators by ECs in response to bacteria is dependent on bacterial invasiveness and on the presence of flagella. This correlates with the capacity of EC-derived factors to modulate MoDC function. MoDCs incubated with supernatants of bacteria-treated ECs are “noninflammatory” as they release interleukin-10 (IL-10) but not IL-12 and can drive only T helper (Th)-2 type T cells. Moreover, noninflammatory MoDCs release chemokines aimed at recruiting Th2 and T-regulatory cells. In contrast, when MoDCs are incubated with ECs and bacteria in a transwell coculture system, and can contact directly the bacteria across stimulated EC monolayers, they are more inflammatory as they release IL-12 and IL-10 and induce both Th1 and Th2 responses. These results suggest that ECs are not simply a barrier to bacteria entering via the oral route, but they actively influence the activating properties of DCs. (Blood. 2005;106:2818-2826)