Search

Objective: Giant cell arteritis (GCA) is characterized by granulomatous inflammation of the medium‐ and large‐sized arteries accompanied by remodeling of the vessel wall. Fibroblast activation protein alpha (FAP) is a serine protease that promotes both inflammation and fibrosis. Here, we investigated the plasma levels and vascular expression of FAP in GCA. Methods: Plasma FAP levels were measured with enzyme‐linked immunosorbent assay in treatment‐naive patients with GCA (n = 60) and polymyalgia rheumatica (PMR) (n = 63) compared with age‐ and sex‐matched healthy controls (HCs) (n = 42) and during follow‐up, including treatment‐free remission (TFR). Inflamed temporal artery biopsies (TABs) of patients with GCA (n = 9), noninflamed TABs (n = 14), and aorta samples from GCA‐related (n = 9) and atherosclerosis‐related aneurysm (n = 11) were stained for FAP using immunohistochemistry. Immunofluorescence staining was performed for fibroblasts (CD90), macrophages (CD68/CD206/folate receptor beta), vascular smooth muscle cells (desmin), myofibroblasts (α‐smooth muscle actin), interleukin‐6 (IL‐6), and matrix metalloproteinase‐9 (MMP‐9). Results: Baseline plasma FAP levels were significantly lower in patients with GCA compared with patients with PMR and HCs and inversely correlated with systemic markers of inflammation and angiogenesis. FAP levels decreased even further at 3 months on remission in patients with GCA and gradually increased to the level of HCs in TFR. FAP expression was increased in inflamed TABs and aorta of patients with GCA compared with control tissues. FAP was abundantly expressed in fibroblasts and macrophages. Some of the FAP+ fibroblasts expressed IL‐6 and MMP‐9. Conclusion: FAP expression in GCA is clearly modulated both in plasma and in vessels. FAP may be involved in the inflammatory and remodeling processes in GCA and have utility as a target for imaging and therapeutic intervention. [ABSTRACT FROM AUTHOR]

Objectives Evidence from temporal artery tissue and blood suggests involvement of CD8+ T cells in the pathogenesis of GCA, but their exact role is poorly understood. Therefore, we performed a comprehensive analysis of circulating and lesional CD8+ T cells in GCA patients. Methods Circulating CD8+ T cells were analysed for differentiation status (CD45RO, CCR7), markers of activation (CD69 and CD25) and proliferation (Ki-67) in 14 newly diagnosed GCA patients and 18 healthy controls by flow cytometry. Proliferative capacity of CD8+ T cells upon anti-CD3 and anti-CD3/28 in vitro stimulation was assessed. Single-cell RNA sequencing of peripheral blood mononuclear cells of patients and controls (n  = 3 each) was performed for mechanistic insight. Immunohistochemistry was used to detect CD3, CD8, Ki-67, TNF-α and IFN-γ in GCA-affected tissues. Results GCA patients had decreased numbers of circulating effector memory CD8+ T cells but the percentage of Ki-67-expressing effector memory CD8+ T cells was increased. Circulating CD8+ T cells from GCA patients demonstrated reduced T cell receptor activation thresholds and displayed a gene expression profile that is concurrent with increased proliferation. CD8+ T cells were detected in GCA temporal arteries and aorta. These vascular CD8+ T cells expressed IFN-γ but not Ki-67. Conclusion In GCA, circulating effector memory CD8+ T cells demonstrate a proliferation-prone phenotype. The presence of CD8+ T cells in inflamed arteries seems to reflect recruitment of circulating cells rather than local expansion. CD8+ T cells in inflamed tissues produce IFN-γ, which is an important mediator of local inflammatory responses in GCA. [ABSTRACT FROM AUTHOR]

Objectives GCA is a large vessel vasculitis in which metabolically active immune cells play an important role. GCA diagnosis is based on CRP/ESR and temporal artery biopsies (TABs), in combination with 18F-fluorodeoxyglucose ([18F]FDG)-PET/CT relying on enhanced glucose uptake by glycolytic macrophages. Here, we studied circulating Pyruvate Kinase M2 (PKM2), a glycolytic enzyme, as a possible systemic marker of vessel wall inflammation in GCA. Methods Immunohistochemical detection of PKM2 was performed on inflamed (n  = 12) and non-inflamed (n  = 4) TABs from GCA patients and non-GCA (n  = 9) patients. Dimeric PKM2 levels were assessed in plasma of GCA patients (n  = 44), age-matched healthy controls (n  = 41), metastatic melanoma patients (n  = 7) and infection controls (n  = 11). CRP, ESR and macrophage markers calprotectin and YKL-40 were correlated with plasma PKM2 levels. To detect the cellular source of plasma PKM2 in tissue, double IF staining was performed on inflamed GCA TABs. [18F]FDG-PET scans of 23 GCA patients were analysed and maximum standard uptake values and target to background ratios were calculated. Results PKM2 is abundantly expressed in TABs of GCA patients. Dimeric PKM2 plasma levels were elevated in GCA and correlated with CRP, ESR, calprotectin and YKL-40 levels. Elevated plasma PKM2 levels were downmodulated by glucocorticoid treatment. PKM2 was detected in both macrophages and T cells at the site of vascular inflammation. Circulating PKM2 levels correlated with average target to background ratios PET scores. Conclusion Elevated plasma PKM2 levels reflect active vessel inflammation in GCA and may assist in disease diagnosis and in disease monitoring. [ABSTRACT FROM AUTHOR]

Objective: Macrophages mediate inflammation, angiogenesis, and tissue destruction in giant cell arteritis (GCA). Serum levels of the macrophage‐associated protein YKL‐40 (chitinase 3–like protein 1), previously linked to angiogenesis and tissue remodeling, remain elevated in GCA despite glucocorticoid treatment. This study was undertaken to investigate the contribution of YKL‐40 to vasculopathy in GCA. Methods: Immunohistochemistry was performed on GCA temporal artery biopsy specimens (n = 12) and aortas (n = 10) for detection of YKL‐40, its receptor interleukin‐13 receptor α2 (IL‐13Rα2), macrophage markers PU.1 and CD206, and the tissue‐destructive protein matrix metalloproteinase 9 (MMP‐9). Ten noninflamed temporal artery biopsy specimens served as controls. In vitro experiments with granulocyte–macrophage colony‐stimulating factor (GM‐CSF)– or macrophage colony‐stimulating factor (M‐CSF)–skewed monocyte‐derived macrophages were conducted to study the dynamics of YKL‐40 production. Next, small interfering RNA–mediated knockdown of YKL‐40 in GM‐CSF–skewed macrophages was performed to study its effect on MMP‐9 production. Finally, the angiogenic potential of YKL‐40 was investigated by tube formation experiments using human microvascular endothelial cells (HMVECs). Results: YKL‐40 was abundantly expressed by a CD206+MMP‐9+ macrophage subset in inflamed temporal arteries and aortas. GM‐CSF–skewed macrophages from GCA patients, but not healthy controls, released significantly higher levels of YKL‐40 compared to M‐CSF–skewed macrophages (P = 0.039). In inflamed temporal arteries, IL‐13Rα2 was expressed by macrophages and endothelial cells. Functionally, knockdown of YKL‐40 led to a 10–50% reduction in MMP‐9 production by macrophages, whereas exposure of HMVECS to YKL‐40 led to significantly increased tube formation. Conclusion: In GCA, a GM‐CSF–skewed, CD206+MMP‐9+ macrophage subset expresses high levels of YKL‐40 which may stimulate tissue destruction and angiogenesis through IL‐13Rα2 signaling. Targeting YKL‐40 or GM‐CSF may inhibit macrophages that are currently insufficiently suppressed by glucocorticoids. [ABSTRACT FROM AUTHOR]