The immune system is strongly involved in atherosclerosis and immune regulation generally leads to attenuated atherosclerosis. B and T lymphocyte attenuator (BTLA) is a novel co-receptor that... Show moreThe immune system is strongly involved in atherosclerosis and immune regulation generally leads to attenuated atherosclerosis. B and T lymphocyte attenuator (BTLA) is a novel co-receptor that negatively regulates the activation of B and T cells, however, there have been no reports of BTLA and its function in atherosclerosis or cardiovascular disease (CVD). We aimed to assess the dominant BTLA expressing leukocyte in CVD patients and to investigate whether BTLA has a functional role in experimental atherosclerosis. Stimulation of the BTLA pathway in Ldlr-/-mice reduces initial lesion development and increases collagen content of established lesions, presumably by shifting the balance between atherogenic follicular B cells and atheroprotective B cells and directing CD4+ T cells towards regulatory T cells. We provide the first evidence that BTLA is a very promising target for the treatment of atherosclerosis.We show that BTLA is primarily expressed on B cells in CVD patients and follicular B2 cells in low-density lipoprotein receptor-deficient (Ldlr-/-) mice. We treated Ldlr-/- mice that were fed a Western-type diet (WTD) with PBS, an isotype antibody or an agonistic BTLA antibody (3C10) for 6 weeks. We report here that the agonistic BTLA antibody significantly attenuated atherosclerosis. This was associated with a strong reduction in follicular B2 cells, while regulatory B and T cells were increased. The BTLA antibody showed similar immunomodulating effects in a progression study in which Ldlr-/- mice were fed a WTD for 10 weeks before receiving antibody treatment. Most importantly, BTLA stimulation enhanced collagen content, a feature of stable lesions, in preexisting lesions. Show less
Duijn, J. van; Elsas, M. van; Benne, N.; Depuydt, M.; Wezel, A.; Smeets, H.; ... ; Slutter, B. 2019
CD8+ T-cells can be atheroprotective in clinically relevant advanced stages of atherosclerosis, as their depletion results in less stable lesions with a more inflammatory phenotype. However, the... Show moreCD8+ T-cells can be atheroprotective in clinically relevant advanced stages of atherosclerosis, as their depletion results in less stable lesions with a more inflammatory phenotype. However, the phenotype and function of these cells in the lesional microenvironment remains to be determined. Here, we address how the atherosclerotic environment affects the functionality of CD8+ T-cells.We compared the cytokine production of CD8+ T-cells derived from spleens and aortas of apoE-/- mice with advanced atherosclerosis by flow cytometry.CD8+ T-cells isolated from atherosclerotic lesions produced lower amounts of IFN-γ and TNF-α than their splenic counterparts. The observed dysfunctional phenotype of the lesion-derived CD8+ T-cells was associated with an increased expression of the ectonucleotidase CD39, which converts inflammatory extracellular ATP into immunomodulatory adenosine. Indeed, pharmacological inhibition of CD39 in apoE-/- mice partly restored cytokine production by CD8+ T-cells. Using a bone-marrow transplantation approach, we showed that induction of CD39 was a consequence of antigen-specific CD8+ T-cell activation via T-cell receptor (TCR) signaling within the lesions. Importantly, analysis of human endarterectomy samples showed a clear microenvironment specific upregulation of CD39 on CD8+ T-cells in the plaques of human patients compared to matched CD8+ T-cells from the blood .Our results indicate that the continuous TCR signaling in the atherosclerotic plaque induces an immune regulatory CD8+ T-cell phenotype that is associated with decreased cytokine production through increased CD39 expression in both a murine atherosclerotic model and in atherosclerosis patients. This provides a new understanding of atheroprotective immune regulation by CD8+ T-cells. Show less
The immune system is strongly involved in atherosclerosis and immune regulation generally leads to attenuated atherosclerosis. B and T lymphocyte attenuator (BTLA) is a novel co-receptor that... Show moreThe immune system is strongly involved in atherosclerosis and immune regulation generally leads to attenuated atherosclerosis. B and T lymphocyte attenuator (BTLA) is a novel co-receptor that negatively regulates the activation of B and T cells, however, there have been no reports of BTLA and its function in atherosclerosis or cardiovascular disease (CVD). We aimed to assess the dominant BTLA expressing leukocyte in CVD patients and to investigate whether BTLA has a functional role in experimental atherosclerosis.\nStimulation of the BTLA pathway in Ldlr-/-mice reduces initial lesion development and increases collagen content of established lesions, presumably by shifting the balance between atherogenic follicular B cells and atheroprotective B cells and directing CD4+ T cells towards regulatory T cells. We provide the first evidence that BTLA is a very promising target for the treatment of atherosclerosis.\nWe show that BTLA is primarily expressed on B cells in CVD patients and follicular B2 cells in low-density lipoprotein receptor-deficient (Ldlr-/-) mice. We treated Ldlr-/- mice that were fed a Western-type diet (WTD) with PBS, an isotype antibody or an agonistic BTLA antibody (3C10) for 6 weeks. We report here that the agonistic BTLA antibody significantly attenuated atherosclerosis. This was associated with a strong reduction in follicular B2 cells, while regulatory B and T cells were increased. The BTLA antibody showed similar immunomodulating effects in a progression study in which Ldlr-/- mice were fed a WTD for 10 weeks before receiving antibody treatment. Most importantly, BTLA stimulation enhanced collagen content, a feature of stable lesions, in preexisting lesions. Show less
Duijn, J. van; Elsas, M. van; Benne, N.; Depuydt, M.A.C.; Wezel, A.; Smeets, H.; ... ; Slütter, B.A. 2019
CD8+T-cells have been attributed both atherogenic and atheroprotective properties, butanalysis of CD8+T-cells has mostly been restricted to the circulation and secondary lymphoid organs.... Show moreCD8+T-cells have been attributed both atherogenic and atheroprotective properties, butanalysis of CD8+T-cells has mostly been restricted to the circulation and secondary lymphoid organs. Theatherosclerotic lesion, however, is a complex microenvironment containing a plethora of inflammatory signals,which may affect CD8+T-cell activation. Here, we address how this environment affects the functionality ofCD8+T-cells. We compared the cytokine production of CD8+T-cells derived from spleens and en-zymatically digested aortas ofapoE−/−mice with advanced atherosclerosis byflow cytometry. Aortic CD8+T-cells produced decreased amounts of IFN-γand TNF-αcompared to their systemic counterparts. The observeddysfunctional phenotype of the lesion-derived CD8+T-cells was not associated with classical exhaustion mar-kers, but with increased expression of the ectonucleotidase CD39. Indeed, pharmacological inhibition of CD39 inapoE−/−mice partly restored cytokine production by CD8+T-cells. Using a bone-marrow transplantation ap-proach, we show that TCR signaling is required to induce CD39 expression on CD8+T-cells in atheroscleroticlesions. Importantly, analysis of human endarterectomy samples showed a strong microenvironment specificupregulation of CD39 on CD8+T-cells in the plaques of human patients compared to matched blood samples. Our results suggest that the continuous TCR signaling in the atherosclerotic environment in thevessel wall induces an immune regulatory CD8+T-cell phenotype that is associated with decreased cytokineproduction through increased CD39 expression in both a murine atherosclerotic model and in atherosclerosispatients. This provides a new understanding of immune regulation by CD8+T-cells in atherosclerosis. Show less
Duijn, J. van; Elsas, M. van; Benne, N.; Depuydt, M.; Wezel, A.; Smeets, H.; ... ; Slütter, B. 2019
CD8+T-cells have been attributed both atherogenic and atheroprotective properties, butanalysis of CD8+T-cells has mostly been restricted to the circulation and secondary lymphoid organs.... Show moreCD8+T-cells have been attributed both atherogenic and atheroprotective properties, butanalysis of CD8+T-cells has mostly been restricted to the circulation and secondary lymphoid organs. Theatherosclerotic lesion, however, is a complex microenvironment containing a plethora of inflammatory signals,which may affect CD8+T-cell activation. Here, we address how this environment affects the functionality ofCD8+T-cells. We compared the cytokine production of CD8+T-cells derived from spleens and en-zymatically digested aortas ofapoE−/−mice with advanced atherosclerosis byflow cytometry. Aortic CD8+T-cells produced decreased amounts of IFN-γand TNF-αcompared to their systemic counterparts. The observeddysfunctional phenotype of the lesion-derived CD8+T-cells was not associated with classical exhaustion mar-kers, but with increased expression of the ectonucleotidase CD39. Indeed, pharmacological inhibition of CD39 inapoE−/−mice partly restored cytokine production by CD8+T-cells. Using a bone-marrow transplantation ap-proach, we show that TCR signaling is required to induce CD39 expression on CD8+T-cells in atheroscleroticlesions. Importantly, analysis of human endarterectomy samples showed a strong microenvironment specificupregulation of CD39 on CD8+T-cells in the plaques of human patients compared to matched blood samples. Our results suggest that the continuous TCR signaling in the atherosclerotic environment in thevessel wall induces an immune regulatory CD8+T-cell phenotype that is associated with decreased cytokineproduction through increased CD39 expression in both a murine atherosclerotic model and in atherosclerosispatients. This provides a new understanding of immune regulation by CD8+T-cells in atherosclerosis. Show less
Duijn, J. van; Kritikou, E.; Benne, N.; Heijden, T. van der; Puijvelde, G.H. van; Kröner, M.J.; ... ; Slütter, B. 2018
Human endarterectomy samples analyzed by flow cytometry showed a negative correlation between the percentage of CD8+ T-cells and macrophages, suggesting a possible protective role for these cells... Show moreHuman endarterectomy samples analyzed by flow cytometry showed a negative correlation between the percentage of CD8+ T-cells and macrophages, suggesting a possible protective role for these cells in lesion development. To further test this hypothesis, LDLr-/- mice were fed a Western-type diet (WTD) for 10 weeks to induce atherosclerosis, after which they received CD8α-depleting or isotype control antibody for six weeks. Depletion of CD8+ T-cells in advanced atherosclerosis resulted in less stable lesions, with significantly reduced collagen content in the trivalve area, increased macrophage content and increased necrotic core area compared to controls. Mechanistically, we observed that CD8 depletion specifically increased the fraction of Th1 CD4+ T-cells in the lesions. Treatment of WTD-fed LDLr-/- mice with a FasL-neutralizing antibody resulted in similar changes in macrophages and CD4+ T-cell skewing as CD8+ T-cell depletion. T lymphocytes play an important role in atherosclerosis development, but the role of the CD8+ T-cell remains debated, especially in the clinically relevant advanced stages of atherosclerosis development. Here, we set out to determine the role of CD8+ T-cells in advanced atherosclerosis. These findings demonstrate for the first time a local, protective role for CD8+ T-cells in advanced atherosclerosis, through limiting accumulation of Th1 cells and macrophages, identifying a novel regulatory mechanism for these cells in atherosclerosis. Methods and Results Aims Conclusion Show less
Kritikou, E.; Heijden, T. van der; Swart, M.; Duijn, J. van; Slutter, B.; Smeets, H.; ... ; Bot, I. 2017
