Ed and their role in modulating the subsequent repair process is unknown. As Tregs are known to counteract the effect of Th1 it is possible that they may increase neointima formation by inhibiting IFNc producing Th1 cells. Accordingly, it is possible that arterial injury leads to the activation of CD4+ T cells with partially opposing effect on neointima formation and that the lack of effect observed in MHC class II deficient mice is explained by a loss of all of these cells. To determine if a possible effect of Th1 cells on neointima formation was counteracted by a parallel activation of Tregs we treated wild type mice with CD25 blocking antibodies, a well-established approach for deletion of Tregs [17]. Although this treatment was associated with a 80 reduction of Tregs in both draining and contralateral lymph nodes, as well as with signs of Homotaurine price increased activation of remaining CD4+ T cells, it did not influence neointima formation suggesting that neither Tregs nor Th1 T cells influence vascular repair responses in this model. However, our findings need to be interpreted with some caution since it cannot be completely excluded that CD25 antibody treatment also deleted some CD25-expressing CD4+ T effector cells. Moreover, it is also possible that Tregs themselves may have dual effects on neointima formation in the same way as IFNc producing Th1 cells [13]. Treatment with CD25 antibodies increased the expression of ICOS on remaining CD4+ T cells. Transfer of ICOS2/2 bone marrow has previously been shown to aggravate atherosclerosis in LDL receptor deficient mice suggesting a protective role of this costimulatory Bexagliflozin biological activity molecule [18]. The possible role of ICOS in repair responses to arterial injury is unknown but the present findings demonstrate that an up-regulation of ICOS on CD4+ T cells does not appear to influence neointima formation. An interesting and unexpected observation was that carotid injury was associated with a dramatic emigration of Tregs from the spleen. Three days after injury only 25 of the original Treg population remained in the spleen. The splenic emigration of Tregs appeared to be temporary because at day 7 day after injury Treg numbers were almost back to the same level as in uninjured mice. Swirski and coworkers [19] recently reported that splenic pro-inflammatory monocytes in response to ischemic myocardial injury exit the spleen en masse, accumulate in injured tissue, and participate in wound healing. It is likely that the emigration of Tregs in response to carotid injury observed here represents a parallel regulatory response aimed to control inflammation in the injured tissue. We have previously shown that carotid injury isRegulatory T Cells and Carotid Injuryassociated with a local accumulation of monocytes and that these frequently localize to the periadventitial tissue where they promote neointima formation through release of IL-1b [4]. Carotid collar injury is associated with a rapid formation of adventitial granulation tissue (figure 3). It is an interesting possibility that the primary function of the Tregs recruited in response to collar injury is to control inflammation in this tissue rather than the subsequent neointima formation. The finding of increased neointima formation in both T celldepleted and Rag-12/2 mice suggest the existence of a T cell population with inhibitory properties [12]. Dimayuga and coworkers recently reported that transfer of CD8+ T cells inhibits neointima formation in Rag-12/2 mice an.Ed and their role in modulating the subsequent repair process is unknown. As Tregs are known to counteract the effect of Th1 it is possible that they may increase neointima formation by inhibiting IFNc producing Th1 cells. Accordingly, it is possible that arterial injury leads to the activation of CD4+ T cells with partially opposing effect on neointima formation and that the lack of effect observed in MHC class II deficient mice is explained by a loss of all of these cells. To determine if a possible effect of Th1 cells on neointima formation was counteracted by a parallel activation of Tregs we treated wild type mice with CD25 blocking antibodies, a well-established approach for deletion of Tregs [17]. Although this treatment was associated with a 80 reduction of Tregs in both draining and contralateral lymph nodes, as well as with signs of increased activation of remaining CD4+ T cells, it did not influence neointima formation suggesting that neither Tregs nor Th1 T cells influence vascular repair responses in this model. However, our findings need to be interpreted with some caution since it cannot be completely excluded that CD25 antibody treatment also deleted some CD25-expressing CD4+ T effector cells. Moreover, it is also possible that Tregs themselves may have dual effects on neointima formation in the same way as IFNc producing Th1 cells [13]. Treatment with CD25 antibodies increased the expression of ICOS on remaining CD4+ T cells. Transfer of ICOS2/2 bone marrow has previously been shown to aggravate atherosclerosis in LDL receptor deficient mice suggesting a protective role of this costimulatory molecule [18]. The possible role of ICOS in repair responses to arterial injury is unknown but the present findings demonstrate that an up-regulation of ICOS on CD4+ T cells does not appear to influence neointima formation. An interesting and unexpected observation was that carotid injury was associated with a dramatic emigration of Tregs from the spleen. Three days after injury only 25 of the original Treg population remained in the spleen. The splenic emigration of Tregs appeared to be temporary because at day 7 day after injury Treg numbers were almost back to the same level as in uninjured mice. Swirski and coworkers [19] recently reported that splenic pro-inflammatory monocytes in response to ischemic myocardial injury exit the spleen en masse, accumulate in injured tissue, and participate in wound healing. It is likely that the emigration of Tregs in response to carotid injury observed here represents a parallel regulatory response aimed to control inflammation in the injured tissue. We have previously shown that carotid injury isRegulatory T Cells and Carotid Injuryassociated with a local accumulation of monocytes and that these frequently localize to the periadventitial tissue where they promote neointima formation through release of IL-1b [4]. Carotid collar injury is associated with a rapid formation of adventitial granulation tissue (figure 3). It is an interesting possibility that the primary function of the Tregs recruited in response to collar injury is to control inflammation in this tissue rather than the subsequent neointima formation. The finding of increased neointima formation in both T celldepleted and Rag-12/2 mice suggest the existence of a T cell population with inhibitory properties [12]. Dimayuga and coworkers recently reported that transfer of CD8+ T cells inhibits neointima formation in Rag-12/2 mice an.
