H-ferritin and CD68+/H-ferritin+ monocytes/macrophages are increased in the skin of adult-onset Still's disease patients and correlate with the multi-visceral involvement of the disease

Benedetto

Berardicurti

et al. 2020

Sci Rep

Ferritin is an iron-binding molecule, which comprises 24 subunits, heavy (FeH) and light (FeL) subunits, suggested to have a pathogenic role by the 'hyperferritinemic syndrome'. In this work, we tested (1) FeH and FeL in bone marrow (BM) and sera in patients with macrophage activation syndrome (MAS); (2) pro-inflammatory effects of ferritin, FeL, and FeH on macrophages; (3) ability of FeHstimulated macrophages to stimulate the proliferation of peripheral blood mononuclear cells (PBMCs); (4) production of mature IL-1β and IL-12p70 in extracellular compartments of FeH-stimulated macrophages. Immunofluorescence analysis and liquid chromatography mass spectrometry (LC-MS/ MS) based proteomics were performed to identify FeL and FeH in BM and sera, respectively, in the same patients. Macrophages were stimulated with ferritin, FeH, and FeL to assess pro-inflammatory effects by RT-PCR and western blot. The proliferation of co-cultured PBMCs with FeH-stimulated macrophages was tested. Immunofluorescence showed an increased FeH expression in BMs, whereas LC-MS/MS identified that FeL was mainly represented in sera. FeH induced a significant increase of gene expressions of IL-1β, IL-6, IL-12, and TNF-α, more marked with FeH, which also stimulated NLRP3. FeH-stimulated macrophages enhanced the proliferation of PBMCs. The ELISA assays showed that mature form of IL-1β and IL-12p70 were increased, in extracellular compartments of FeHstimulated macrophages. Our results showed FeH in BM biopsies of MAS patients, whereas, LC-MS/ MS identified FeL in the sera. FeH showed pro-inflammatory effects on macrophages, stimulated NLRP3, and increased PBMCs proliferation. Adult-onset Still's disease (AOSD) is an inflammatory disease characterised by high spiking fevers, arthritis, evanescent skin rash, and a typical increase of serum ferritin levels 1,2. AOSD is considered a multigenic autoinflammatory disease at the "crossroads" of autoinflammatory and autoimmune diseases, considering its complex pathogenesis, which involves both arms of the immune system 3. The aberrant activation of the immune system leads to production and release of pro-inflammatory cytokines including interleukin (IL)-1β, IL-6, IL-18, interferon (IFN)-γ and tumor necrosis factor (TNF-α) 1-3 , which represent common therapeutic targets 4,5. Patients with AOSD may experience several life-threatening complications, mostly macrophage activation syndrome (MAS), a hyperinflammatory syndrome, with high mortality rate 6,7. Considering the frequent association between these two diseases, it has been suggested that MAS and AOSD may be anchored to the same disease spectrum, representing more severe and milder form, respectively 8. Continuous high fever, hepatosplenomegaly, severe peripheral blood cytopenia, high serum ferritin levels, and haemophagocytosis by activated macrophages in bone marrow (BM) are typical features of these patients 9. A multi-layer MAS pathogenic model has been suggested,

Section: Discussionsupporting

confidence: 90%

Pro-inflammatory properties of H-ferritin on human macrophages, ex vivo and in vitro observations

Benedetto

Berardicurti

et al. 2020

Sci Rep

“…To our knowledge, this is the first study devoted to investigating the clinical characteristics at the time of diagnosis that may be considered predictive for MAS occurrence, and to understanding whether this complication may be associated with a decreased survival rate, in one of the largest cohorts of patients with AOSD described in literature. The clinical features of our patients did not significantly differ from other available series already published, concerning the prevalence of fever, arthritis, and rash [29][30][31][32][33][34][35][36][37] . Our patients also showed high levels of inflammatory markers, as well as of serum ferritin, thus confirming the systemic inflammatory features of the disease 29,30,31,32 .…”

Section: Discussioncontrasting

confidence: 48%

Macrophage Activation Syndrome in Patients Affected by Adult-onset Still Disease: Analysis of Survival Rates and Predictive Factors in the Gruppo Italiano di Ricerca in Reumatologia Clinica e Sperimentale Cohort

Iacono²,

Ciccia³

et al. 2018

J Rheumatol

“…To date, H-ferritin and CD68 1 /H-ferritin 1 cells were correlated significantly with haematological involvement, serum ferritin and CRP of our MAS patients. These results suggest potentially new biomarkers evaluating severity of the MAS clinical picture, possibly improving outcome in these patients [37,38]. In fact, diagnosis and appropriate treatments may be delayed, due to non-specific findings in the early phases of disease, thus new biomarkers may improve the management of this life-threating syndrome [39][40][41][42].…”

Section: Discussionmentioning

confidence: 98%

H-ferritin and proinflammatory cytokines are increased in the bone marrow of patients affected by macrophage activation syndrome

Clinical and Experimental Immunology

Cipriani

Benedetto

et al. 2017

Self Cite

Macrophage activation syndrome (MAS) is hyperinflammatory life-threatening syndrome, associated typically with high levels of serum ferritin. This is an iron storage protein including heavy (H) and light (L) subunits, categorized on their molecular weight. The H-/L subunits ratio may be different in tissues, depending on the specific tissue and pathophysiological status. In this study, we analysed the bone marrow (BM) biopsies of adult MAS patients to assess the presence of: (i) H-ferritin and L-ferritin; (ii) CD68 /H-ferritin and CD68 /L-ferritin ; and (iii) interleukin (IL)-1β, tumour necrosis factor (TNF) and interferon (IFN)-γ. We also explored possible correlations of these results with clinical data. H-ferritin, IL-1β, TNF and IFN-γ were increased significantly in MAS. Furthermore, an increased number of CD68 /H-ferritin cells and an infiltrate of cells co-expressing H-ferritin and IL-12, suggesting an infiltrate of M1 macrophages, were observed. H-ferritin levels and CD68 /H-ferritin cells were correlated with haematological involvement of the disease, serum ferritin and C-reactive protein. L-ferritin and CD68 /L-ferritin cells did not correlate with these parameters. In conclusion, during MAS, H-ferritin, CD68 /H-ferritin cells and proinflammatory cytokines were increased significantly in the BM inflammatory infiltrate, pointing out a possible vicious pathogenic loop. To date, H-ferritin and CD68 /H-ferritin were associated significantly with haematological involvement of the disease, suggesting biomarkers assessing severity of clinical picture.