We previously demonstrated that infection of primary human thymic dendritic cells (DCs) with laboratory strains of HIV leads to the release of soluble factor(s) which induced thymocyte killing. In the present paper, we extend the characterization of this process. Our results reveal that primary HIV-1 isolates are similarly able to induce the production of cytotoxic factor(s) from thymic DCs and that the release of such factor(s) is dependent on viral infection. Interestingly, we observed that CD4+ and CD8+ purified thymocyte subsets, and activated PBMCs are susceptible to the cytotoxic activity, whereas freshly isolated resting PBMCs are resistant to this effect. Cycloheximide treatment prevents the killing of thymocytes exposed to HIV-infected DC supernatant, revealing that this form of cell death is an active biological process requiring protein synthesis. Finally, our data suggest that FasL and TNF alpha could both participate in the killing process. These in vitro observations provide a plausible model, whereby HIV-infected DCs can play a role in vivo in the induction of uninfected thymocyte killing.
In rodent thymus, associations between dendritic cells (DC) and thymocytes have been suggested to be implicated in differentiation and/or maturation processes. In this study, we report intimate associations formed between human thymic DC and thymocytes in culture and we analyze their ultrastructural cell phenotype. Observations by phase contrast microscopy showed that DC present long and thin dendrites and bind many thymocytes. Transmission (TEM) and scanning electron microscopy (SEM) revealed that both cellular populations were in close connection and tight membrane contact could be observed. The phenotype of DC and attached thymocytes was characterized with a series of monoclonal antibodies by protein A-gold TEM and SEM immunolabelings. Quantitative evaluation of immunolabeling (number of gold granules/microns of cellular membrane) suggests the presence of two subpopulations of CD1+ thymic DC (strong and weak), whereas this discrepancy is not observed in DR+ and CD4+ DC populations. On the other hand, the majority of thymocytes bound to DC strongly express the CD1, CD4, CD8 and CD2 antigens and weakly the CD3 antigen, indicating that they represent double-positive immature thymocytes. Uniform distribution of DC and thymocytes membrane antigens was confirmed with a backscattered SEM study. This morphological and immunolabeling TEM and SEM analysis demonstrates that human thymic DC may form associations with CD4+CD8+CD3weak thymocytes and raises questions about their physiological relationship.
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