Pharmacological inhibition of C-C chemokine receptor 4 aggravates atherosclerosis through prevention of regulatory T cell recruitment to the lesions

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
H Amin ◽  
N Sasaki ◽  
S Horibe ◽  
S Kawauchi ◽  
K Hirata ◽  
...  
Keyword(s):  
T Cells ◽  
Lymph Nodes ◽  
Chemokine Receptor ◽  
Flow Cytometric Analysis ◽  
Atherosclerotic Lesion ◽  
Plaque Formation ◽  
Atherosclerotic Lesions ◽  
Sites Of Action ◽  
Chemokine Receptor 4 ◽  
Deficient Mice

Abstract Background Regulatory T cells (Tregs) are demonstrated to play a protective role in the development of atherosclerosis. However, their sites of action in atherosclerosis remain unclear. Although C-C chemokine receptor 4 (CCR4) has been shown to contribute to the accumulation of Tregs in inflamed tissues and prevention of experimental autoimmune diseases, the role of CCR4 in Treg migration to atherosclerotic lesions and suppression of plaque formation remains unknown. Methods and results We intraperitoneally injected 8-week-old apolipoprotein E–deficient mice fed a normal diet with vehicle (n=9) or a 4-μg dose of a CCR4 antagonist (n=10) 3 times weekly for 8 weeks and evaluated atherosclerotic lesions at 16 weeks old. Administration of the CCR4 antagonist significantly aggravated atherosclerotic plaque formation (aortic sinus plaque area: 2.91±0.87×104 μm2 versus 5.41±0.98×104 μm2 in control vehicle-treated and CCR4 antagonist-treated mice, respectively; P<0.05), associated with increased accumulation of macrophages and CD4+T cells in the plaques. Flow cytometric analysis revealed a decrease in Foxp3+ Tregs in the para-aortic lymph nodes and thoracoabdominal aortas of CCR4 antagonist-treated mice, along with a tendency toward increase in CD44highCD62Lloweffector T cells in para-aortic lymph nodes, indicating CCR4-dependent migration of Tregs to atherosclerotic lesions and their possible atheroprotective role. We observed no changes in splenic Foxp3+ Tregs and effector T cells following CCR4 antagonist treatment. We also investigated the effect of CCR4 blockade on advanced atherosclerosis using LDL receptor–deficient mice fed a high-cholesterol diet. Although 8-week treatment with the CCR4 antagonist led to a decrease in Foxp3+ Tregs in the atherosclerotic lesions, atherosclerotic lesion formation was not significantly affected, suggesting that CCR4-dependent Treg accumulation in atherosclerotic lesions is not critical for prevention of advanced atherosclerosis. Conclusions Our findings indicate an important role for CCR4 in promotion of Treg recruitment into atherosclerotic lesions and subsequent prevention of early atherosclerosis and suggest CCR4 as a novel therapeutic target for atherosclerosis. Funding Acknowledgement Type of funding source: None

scholarly journals Interleukin-12p35 Deficiency Reverses the Th1/Th2 Imbalance, Aggravates the Th17/Treg Imbalance, and Ameliorates Atherosclerosis in ApoE-/- Mice

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Ying Huang ◽  
Haiying Hu ◽  
Ling Liu ◽  
Jing Ye ◽  
Zhen Wang ◽  
...  
Keyword(s):  
T Cells ◽  
Epstein Barr Virus ◽  
Critical Role ◽  
Plaque Formation ◽  
Barr Virus ◽  
Atherosclerotic Lesions ◽  
Atherosclerotic Plaque Formation ◽  
Epstein Barr ◽  
Deficient Mice ◽  
Recombinant Mice

Interleukin- (IL-) 35, a novel functional cytokine of regulatory T cells (Treg) comprised of the IL-12p35 subunit and the other subunit Epstein-Barr virus-induced gene 3 (EBI3), regulates the activity of CD4+ T cells and macrophages, thereby playing a critical role in inflammatory and autoimmune diseases. Previous studies demonstrated that both recombinant mice and human IL-35 attenuated atherosclerosis in ApoE-/- mice. Additionally, EBI3 deficiency enhanced the activation of macrophages and increased atherosclerotic lesions in LDLR-/- mice. This study generated double-deficient mice for ApoE and IL-12p35 (ApoE-/- IL-12p35-/- mice) and investigated the effect of IL-12p35 deficiency on atherosclerosis. IL-12p35 deficiency alleviated Th1/Th2 imbalance, aggravated Th17/Treg imbalance, and attenuated atherosclerotic plaque formation in ApoE-/- mice. Additionally, exogenous rIL-35 treatment reversed the imbalance of Th17/Treg and attenuated atherosclerosis in ApoE-/- mice. These findings suggest that IL-12p35 deficiency ameliorates atherosclerosis in ApoE-/- mice, partially, via attenuating the Th1/Th2 imbalance, although IL-12p35 deficiency aggravates the Th17/Treg imbalance.

Abstract 584: Smooth Muscle Cell-derived IL-17C Plays an Atherogenic Role via the Recruitment of Pro-inflammatory IL-17A + T Cells to the Aorta

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Matthew J Butcher ◽  
Tayab C Waseem ◽  
Elena V Galkina
Keyword(s):  
T Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cell ◽  
Muscle Cell ◽  
Th17 Cells ◽  
Ex Vivo ◽  
Flow Cytometric Analysis ◽  
Atherosclerotic Lesion ◽  
Atherosclerotic Lesions

Atherosclerosis is characterized by frequent communication between infiltrating leukocytes and vascular cells, through chemokine and cytokine networks. IL-17 cytokine family members, including IL-17C, are detectable within atherosclerotic plaques, however the potential involvement of these cytokines have not been examined. Thus we sought to investigate the role of IL-17C in atherosclerosis. The expression of IL-17 cytokines was profiled within atherosclerotic Apoe -/- aortas and Il17c expression was elevated. Flow cytometry experiments revealed a major population of aortic IL-17C-producing smooth muscle cells. To determine the role of IL-17C in atherosclerosis, we generated Il17c -/- Apoe -/- mice and compared atherosclerotic lesions between western diet-fed Apoe -/- and Il17c -/- Apoe -/- mice. Atherosclerotic lesion and collagen content was diminished within WD-fed Il17c -/- Apoe -/- aortas and aortic roots in comparison to Apoe -/- controls, and IL-17C treated Apoe -/- aortas up-regulated Col1A1 expression ex vivo . Flow cytometric analysis of Il17c -/- Apoe -/- aortas revealed a proportional reduction in aortic leukocytes, macrophages, neutrophils, T cells, Th1, and T regulatory cells, without corresponding changes in the peripheral immune composition. Examination of aortic IL-17A + TCRγδ T cells and Th17 cells demonstrated a stark reduction in the percentage and number of these subsets within Il17c -/- Apoe -/- mice versus Apoe -/- controls. Explanted 12 week WD Apoe -/- aortas treated with IL-17C resulted in the induction of multiple vascular chemokines and cytokines, and short-term homing experiments revealed diminished recruitment of Th17 cells to the aorta of Il17c -/- Apoe -/- recipients. Smooth muscle cell-derived IL-17C plays a pro-atherogenic role by supporting the recruitment of Th17 cells to atherosclerotic lesions.

scholarly journals Tc17 CD8+ T cells accumulate in murine atherosclerotic lesions, but do not contribute to early atherosclerosis development

2020 ◽  
Author(s):  
Janine van Duijn ◽  
Maaike J M de Jong ◽  
Naomi Benne ◽  
Romain J T Leboux ◽  
Marieke E van Ooijen ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Cd8 T Cells ◽  
Cell Subset ◽  
Atherosclerotic Lesion ◽  
Interleukin 17 ◽  
Pronounced Increase ◽  
Atherosclerotic Lesions ◽  
Tc17 Cells ◽  
Deficient Mice

Abstract Aims CD8+ T cells can differentiate into subpopulations that are characterized by a specific cytokine profile, such as the Tc17 population that produces interleukin-17. The role of this CD8+ T-cell subset in atherosclerosis remains elusive. In this study, we therefore investigated the contribution of Tc17 cells to the development of atherosclerosis. Methods and results Flow cytometry analysis of atherosclerotic lesions from apolipoprotein E-deficient mice revealed a pronounced increase in RORγt+CD8+ T cells compared to the spleen, indicating a lesion-specific increase in Tc17 cells. To study whether and how the Tc17 subset affects atherosclerosis, we performed an adoptive transfer of Tc17 cells or undifferentiated Tc0 cells into CD8−/− low-density lipoprotein receptor-deficient mice fed a Western-type diet. Using flow cytometry, we showed that Tc17 cells retained a high level of interleukin-17A production in vivo. Moreover, Tc17 cells produced lower levels of interferon-γ than their Tc0 counterparts. Analysis of the aortic root revealed that the transfer of Tc17 cells did not increase atherosclerotic lesion size, in contrast to Tc0-treated mice. Conclusion These findings demonstrate a lesion-localized increase in Tc17 cells in an atherosclerotic mouse model. Tc17 cells appeared to be non-atherogenic, in contrast to their Tc0 counterpart.

Abstract 252: Interleukin-27 Signaling Regulates Myeloid Cells Activation in Atherosclerosis

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Iuliia Peshkova ◽  
Aliia Fatkhullina ◽  
Ekaterina Koltsova
Keyword(s):  
T Cells ◽  
T Cell ◽  
Inflammatory Cytokines ◽  
Antigen Presenting Cells ◽  
Atherosclerotic Lesions ◽  
Mediators Of Inflammation ◽  
Antigen Presenting ◽  
Deficient Mice ◽  
Pro Inflammatory Cytokines

Atherosclerosis is a lipid-driven inflammatory disease characterized by the progressive plaque growth in the vessels. Cytokines are important mediators of inflammation and atherosclerosis. While pro-inflammatory cytokines were extensively investigated, little is known about the role of anti-inflammatory cytokines as to their ability to control vascular inflammation. We tested whether immunoregulatory IL-27R signaling is important to control inflammation in mouse models of atherosclerosis. We found that atherosclerosis-prone mice with hematopoietic deficiency of IL-27R ( Ldlr -/- mice reconstituted with bone marrow from Il27ra -/- ) or global deficiency ( Il27ra -/- x Apoe -/- ) developed significantly larger atherosclerotic lesions compared to controls. Atherosclerotic lesions in IL-27R deficient mice contained more CD45 + leukocytes and CD4 + T cells, which produced pro-atherogenic cytokines IL-17A and TNF-α. These cytokines normally suppressed by IL-27, regulated the expression of CCL2 and other chemokines, which in turn led to accumulation of myeloid CD11b + and CD11c + cells in atherosclerotic aortas. Using two-photon microscopy, we found enhanced interactions between antigen presenting cells and T cells in the aortas of IL-27R deficient mice accompanied by enhanced CD4 T cell proliferation. Moreover, macrophages in Il27ra -/- aortas also demonstrated enhanced ability to produce pro-inflammatory cytokines, including IL-1. The blockade of IL-1R signaling, however, strongly suppressed atherosclerosis progression in IL-27R deficient but not control mice, suggesting an important role of IL-27 in the regulation of IL-1 production in atherosclerosis. Overall, our data demonstrate that IL-27R signaling in atherosclerosis is required to control function of antigen presenting cells modulating subsequent T cell activation in the aortas. Moreover, it controls macrophage activation and pro-inflammatory myeloid cell-derived cytokine production. These mechanisms altogether curb pathogenic T cell lineage differentiation and, thus, atherosclerosis, suggesting potent anti-atherogenic role of IL-27.

Immune-Inflammation in Atherosclerosis: A New Twist in an Old Tale

2020 ◽  
Vol 20 (4) ◽  
pp. 525-545 ◽  
Author(s):  
Atefe Ghamar Talepoor ◽  
Hamed Fouladseresht ◽  
Shahdad Khosropanah ◽  
Mehrnoosh Doroudchi
Keyword(s):  
T Cells ◽  
T Cell ◽  
Drug Targets ◽  
Inflammatory Diseases ◽  
Atherosclerotic Lesion ◽  
Preventative Measures ◽  
Atherosclerotic Lesions ◽  
Adaptive Immune Cells ◽  
One Step ◽  
Immune Inflammation

Background and Objective: Atherosclerosis, a chronic and progressive inflammatory disease, is triggered by the activation of endothelial cells followed by infiltration of innate and adaptive immune cells including monocytes and T cells in arterial walls. Major populations of T cells found in human atherosclerotic lesions are antigen-specific activated CD4+ effectors and/or memory T cells from Th1, Th17, Th2 and Treg subsets. In this review, we will discuss the significance of T cell orchestrated immune inflammation in the development and progression of atherosclerosis. Discussion: Pathogen/oxidative stress/lipid induced primary endothelial wound cannot develop to a full-blown atherosclerotic lesion in the absence of chronically induced inflammation. While the primary inflammatory response might be viewed as a lone innate response, the persistence of such a profound response over time must be (and is) associated with diverse local and systemic T cell responses. The interplay between T cells and innate cells contributes to a phenomenon called immuneinflammation and has an impact on the progression and outcome of the lesion. In recent years immuneinflammation, an old term, has had a comeback in connecting the puzzle pieces of chronic inflammatory diseases. Conclusion: Taking one-step back and looking from afar at the players of immune-inflammation may help us provide a broader perspective of these complicated interactions. This may lead to the identification of new drug targets and the development of new therapies as well as preventative measures.

scholarly journals CC Chemokine Receptor 4 Expression on Peripheral Blood CD4+ T Cells Reflects Disease Activity of Atopic Dermatitis

2001 ◽  
Vol 117 (2) ◽  
pp. 188-196 ◽  
Author(s):  
Motoshi Wakugawa ◽  
Koichiro Nakamura ◽  
Takashi Kakinuma ◽  
Kunihiko Tamaki ◽  
Nobuyuki Onai ◽  
...  
Keyword(s):  
T Cells ◽  
Atopic Dermatitis ◽  
Disease Activity ◽  
Chemokine Receptor ◽  
Peripheral Blood ◽  
Cd4 T Cells ◽  
Cc Chemokine ◽  
Chemokine Receptor 4

Abstract 3693: Early Growth Response Gene-1 Deficiency In Bone-marrow-derived Cells Reduces Macrophage Accumulation And Atherosclerotic Lesion Formation In A Hyperlipidemic Mouse Model

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Michael R Preusch ◽  
Claudia Albrecht ◽  
Gotz Hofmann ◽  
Erwin Blessing ◽  
Hugo A Katus ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Atherosclerotic Lesion ◽  
Lesion Development ◽  
Early Growth Response ◽  
Atherosclerotic Lesions ◽  
Early Growth Response Gene ◽  
Bone Marrow Derived Cells ◽  
Deficient Mice ◽  
Atherosclerotic Lesion Development ◽  
Macrophage Accumulation

Early growth response gene-1 (Egr-1), a prototype of a family of zinc-finger transcription factors, is a master regulator of many genes which play important roles in cardiovascular diseases. Within atherosclerotic lesions Egr-1 is expressed in several cell types, such as smooth muscle cells, endothelial cells and monocytes/macrophages. Since macrophages play a pivotal role in atherosclerotic lesion development, this study investigated the effects of Egr-1-deficiency within bone-marrow derived cells on the development of atherosclerosis in a hyperlipidemic mouse model. Therefore we transplanted bone-marrow from Egr-1 deficient mice and wild type controls into lethally irradiated low-density lipoprotein receptor deficient mice. After 20 weeks on a western diet atherosclerotic lesions within the aortic sinus and gene expression of inflammatory genes in the aortas of the recipients were evaluated. Mice receiving Egr-1 deficient bone-marrow had less atherosclerotic lesion development compared with mice receiving wild type bone-marrow (318 736 ± 98 910μm 2 vs. 404 539 ± 92 408μm 2 , p<0.05). The size of the necrotic core within the lesions was also reduced. Immunohistochemistry revealed that mice receiving Egr-1 deficient bone-marrow had less macrophages in comparison to controls. Gene expression analysis in the aortas of the mice demonstrated reduced expression of Vascular Cell Adhesion Molecule (VCAM-1), an important adhesion molecule during the development of atherosclerosis. These results were validated with in vitro studies where Egr-1-deficient peritoneal macrophages revealed less VCAM-1 mRNA expression after stimulation with lipopolysaccharide in comparison to wildtype macrophages. This study demonstrates that bone-marrow derived Egr-1 promotes macrophage accumulation and atherosclerotic lesion development possible over an increased expression of VCAM-1.

The Krüppel-Like Factor 4 (KLF4) Modulates Survival and Tissue Distribution of Natural Killer Cells.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 282-282
Author(s):  
Chun Shik Park ◽  
Ping-Hsien Lee ◽  
Takeshi Yamada ◽  
Maksim Mamonkin ◽  
H. Daniel Lacorazza
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Lymph Nodes ◽  
Nk Cells ◽  
Tissue Distribution ◽  
Natural Killer ◽  
Peripheral Blood ◽  
Nk Cell ◽  
Wild Type ◽  
Deficient Mice

Abstract Abstract 282 Natural Killer (NK) cells are important mediators of the innate immune system that could be targeted therapeutically to treat hematologic malignancies and to prevent graft-versus-host disease. Hence, a better understanding of NK cell survival and tissue trafficking at steady state is vital to develop cell-based therapies. Genes that control proliferation are often involved in tissue distribution of lymphocytes, such as KLF2 in T cells. KLF4, another member of the Krüppel-like factor family, can activate and repress genes involved in diverse cellular processes. We recently reported that KLF4 is part of a novel inhibitory pathway that prevents proliferation of naïve T cells during homeostasis and retain memory T cells in lymph nodes (Yamada et al., Nature Immunology, 2009). In this work, we studied the role of KLF4 in the development and maintenance of NK cells by deleting Klf4-floxed gene (fl/fl) using the Mx1-Cre system. The percentage of NK1.1+TCR- cells is significantly reduced in peripheral blood of Klf4-deficient (▪/▪) mice (fl/fl: 3.4±1.1 versus ▪/▪: 1.2±0.1, n=9) and also absolute numbers in spleen (▪/▪: 1.1±1.3 ×106, n=6) due to increased percentage of Annexin V positive cells (fl/fl: 9.2±3.2 versus ▪/▪: 22.9±15.5, n=15). The number of CD49d+TCR- cells was also significantly reduced in peripheral blood and spleen of Klf4-deficient mice. In contrast, the number of NK cells in bone marrow and lymph nodes of Klf4-deficient mice was similar to controls. Deletion of Klf4 gene led to reduced numbers of NK1.1+TCR-CD27+CD11b+ and NK1.1+TCR-CD27-CD11b+ cells, which correlated with increasing apoptosis of these subsets. Yet, the percentages of these NK cell subsets were normal in bone marrow ruling out a developmental defect in this tissue. Transplant of wild type and Klf4-deficient bone marrow cells into wild type mice suggested environmental rather than cell intrinsic defects. NK cells (NK1.1+TCR-) isolated from spleen of Klf4-deficient mice showed to be functional in a cytotoxicity assay using a mixture of differentially CFSE-labeled RMA-S (target) and EL4 (control). In summary, KLF4 plays a key role in the maintenance of mature NK cells in peripheral blood and spleen. Disclosures: No relevant conflicts of interest to declare.

A Gain-of-Function CCR4 Mutations in Adult T-Cell Leukemia/Lymphoma (ATLL) Enhance the Chemotactic Abilities and PI3K/AKT Activation

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 3566-3566
Author(s):  
Masao Nakagawa ◽  
Roland Schmitz ◽  
Wenming Xiao ◽  
Carolyn K Goldman ◽  
Weihong Xu ◽  
...  
Keyword(s):  
T Cells ◽  
Lymph Nodes ◽  
Chemokine Receptor ◽  
Carboxy Terminus ◽  
Coding Region ◽  
Gain Of Function ◽  
Akt Activation ◽  
Whim Syndrome ◽  
Selective Growth Advantage ◽  
Chemotactic Ability

Abstract High expression of CC chemokine receptor 4 (CCR4) has been identified as a hallmark gene in ATLL, an aggressive peripheral T-cell neoplasm. CCR4 is a chemokine receptor, which has a critical role in immune cell trafficking including Th2, T-reg and skin-homing memory T-cells. CCR4 ligands, CCL17 and CCL22, were produced in lymph nodes and skin from dendritic cells, macrophages and Langerhans cells. Most ATLL cases express surface CCR4 (90%) and infiltrate to lymph nodes and skin. These observations suggest that CCR4 could have a role in ATLL biology, but it is still unclear whether dysregulation of CCR4 function contributes to ATLL pathogenesis. We performed RNA-Seq for two primary ATLL cases and discovered recurrent non-sense mutations in CCR4. Though an extended analysis using Sanger sequencing, CCR4 mutations were detected in 14/53 ATLL samples (26%) and consisted exclusively of nonsense or frameshift mutations that truncated the coding region at C329, Q330 or Y331 in the carboxy-terminus. We noticed that the location of the CCR4 mutations in ATLL were reminiscent of mutations affecting the chemokine receptor CXCR4 in the WHIM syndrome, a human immunodeficiency disease. Most CXCR4 mutations in the WHIM syndrome are nonsense or frameshift, resulting in carboxy-terminal truncation of the protein and conferring a gain-of-function phenotype with respect to chemotaxis towards the CXCR4 ligand SDF1. We therefore hypothesized that mutant CCR4 isoforms might enhance chemotaxis of the affected cells to CCR4 ligands. Chemotaxic assay using 32Db, a mouse myeloid cell line, and ED40515(+), an ATLL cell line, clarified that the ectopic expression of CCR4-Q330* enhanced the chemotactic ability of the transduced cells toward CCL17 and CCL22 rather than CCR4-WT transduced cells with statistical significance. To understand the mechanism of this enhanced chemotactic ability, we studied the change in surface CCR4 levels after CCL22 exposure in CCR4-WT-and CCR4-Q330*-reconstituted ED40515(+) cells. Compared with CCR4-WT, CCR4 internalization in CCR4-Q330*-reconstituted cells was significantly impaired. Thus, the ATLL CCR4 mutants impair desensitization by ligand, which likely contributes to the enhanced chemotaxis of cells bearing these mutants. We explored the influence of the ATLL CCR4 mutants on PI(3) kinase (PI3K)-dependent activation of AKT since it has been reported that binding of CCL22 to CCR4 activates AKT in CEM leukemic T-cells and in human Th2 cells. CCR4-Q330*-reconstituted ED40515(+) showed strong activation of AKT with CCL22 ligation compared with CCR4-WT-reconstituted cell. The AKT activation was cancelled with pan-PI3K inhibitor, BKM120, indicating that CCR4-mediated AKT activation was PI3K dependent. Lastly, we tested whether the acquisition of CCR4 mutations by ATLL cells imparts a selective growth advantage relative to cells with wild type CCR4. CCR4-Q330*-reconstituted cells had a selective growth advantage in the presence of CCL22, supporting at least in part the hypothesis that CCR4 mutation are able to provide the affected cells a positive selection pressure through CCL22 ligation and contributes to ATLL pathogenesis. We discovered for the first time recurrent somatic mutations in CCR4 in ATLL. CCR4 mutations were detected in 14/53 ATLL samples (26%) and consisted exclusively of nonsense or frameshift mutations that truncated the coding region at C329, Q330 or Y331 in the carboxy-terminus. Functionally, the CCR4-Q330* was a gain-of-function since it increased cell migration towards the CCR4 ligands CCL17 and CCL22, in part by impairing receptor internalization. This mutant enhanced PI(3) kinase/AKT activation following receptor engagement by CCL22 in ATLL cells, and conferred a growth advantage in in vitro cultures. Our findings provide a rationale to test whether inhibition of CCR4 signaling might have a therapeutic potential for patients with ATLL. Disclosures No relevant conflicts of interest to declare.

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