albicans, and T. cruzi infections demonstrate that the entrance of peripheral B and T cells into the thymus, rather than being a pathogen-specific phenomenon is

the consequence of an acute inflammatory process triggered by an early BMS-354825 in vitro production of the Th1 cytokines IL-12 and IL-18. One concern we needed to address is whether or not activated (CD44hi) cells or also naïve T cells are able to reach the thymus in these inflammatory conditions. To examine this question, we adoptively transferred splenocytes from a normal uninfected mouse to a T. cruzi infected mouse and evaluated phenotype of the cells that entered to the thymus. We observed that they are CD44int/hi and CD62Lhi despite the fact that the cells expressed lower levels of CD44 and CD62L before the injection (not shown). Thus, we concluded that because we inject naïve cells into recipient mice that are actively expressing high levels of inflammatory cytokines, naïve cells get activated themselves during the 18 h they reside into the recipient mice. These data support the fact

that only cells with an activated phenotype and expressing CD62L are able to reach the thymus in the context of these inflammatory conditions. Even though we do not describe here what subset of peripheral leukocytes could migrate to the thymus in situations when IL-12 and IL-18 are systemically expressed, it is interesting to note that other investigators PI3K inhibitor have characterized a subset of splenic CD44hi CD8+ T cells that, in the presence of both IL-12 and IL-18, can rapidly secrete IFN-γ in the absence of specific Ag [36, 37]. In vivo, the activation of these cells is triggered by different pathogens such as Listeria monocytogenes [36] or during certain acute viral infections [22]. Based on these reports and our own data with OVA-transgenic mice

that demonstrate that T cells that enter the thymus are not exclusively clones activated by Ags expressed by the pathogen itself, we speculate that in a normal nonimmunized mouse there exists a subset of B and T cells that are able to rapidly respond to IL-12 and IL-18 (or to cytokines induced thereafter), become activated and acquire the capacity to migrate back to the Rebamipide thymus. We still need to determine if these cells originate from the preexisting CD44hi pool or if they derive from naïve CD44lo cells that somehow get activated and upregulate CD44, CD62L, and CCR2 in the presence of inflammatory cytokines and these studies will be the focus of future research. Even though most of the reports that evaluate migration of cells to the thymus use the i.v. route [6-8, 16, 17], we also performed adoptive transfer experiments with splenocytes stained with CFSE and injected i.p. (not shown in this manuscript) and demonstrate that in this case, utilizing a different route other than the bloodstream, peripheral cells migrate in similar proportion to the thymus as when cells were injected intravenously.