Activation of anti-inflammatory cell pathways in skin ulcers upon photodynamic therapy

This study was aimed at assessing the variations in skin histology in ulcers caused by chronic venous insufficiency of the lower extremities, upon photodynamic therapy (PDT). The study was approved by the ethical committee of Azienda Sanitaria Firenze. Patient assessment included clinical history, physical examination and echo-Doppler sonography. Four to five sessions of photodynamic therapy (20% 5-aminolevulinic acid gel application followed by 3 min irradiation at 630 nm, total 180 J/cm2) were administered to 15 patients refractory to previous conventional treatments. Skin biopsies were embedded in freezing tissue medium and quick frozen. Cryosections were post fixed in cold acetone. Sections from each case were stained with hematoxylin and eosin or labelled with primary antibodies against the following antigens: MHC-II class, DC-SIGN, CD68, CD163, BDCA2, CD4, CD25, TNF alpha. In some instances avidin and Ulex europaeus lectin were used to tag mast cells and vessels respectively. Upon treatment, MHC-II signal intensity per positive cell and TNF alpha signal in mast cells increased, as well as the numbers of CD68 positive/CD163 positive cells (M2 macrophages), BDCA2 positive (plasmacytoid dendritic) cells and CD4 positive/CD25 positive (Treg) lymphocytes number. Diffuse tissue TGF beta positivity also increased. DC-SIGN positive cells decreased in number. Mast cells were found in proximity of dendritic cells and of vessels; plasmacytoid dendritic cells were found in proximity of T reg cells. Clinically, mild decrease in ulcer size and granulation at ulcer borders were observed. Therefore treatment apparently led to the activation of cells and of intercellular communication pathways possibly down-regulating the inflammatory response. The same treatment had been shown to increase mast cell expression of basic fibroblast growth factor and fibroblast number (1), potentially responsible for increased production of extracellular matrix. Both types of effects could by synergistically beneficial for ulcer repair. This work was supported by the Italian Ministry of Education, MIUR FIRB 2010 and MIUR PRIN-2009; University and Research, Ente Cassa di Risparmio di Firenze (Grant n. 3681 year 2012) and Foemina Foundation

Blood Cell-Bound C4d as a Marker of Complement Activation in Patients With the Antiphospholipid Syndrome

Antiphospholipid syndrome (APS) is a chronic and disabling condition characterized by recurrent thrombosis and miscarriages mediated by antibodies against phospholipid-binding proteins (aPL), such as beta2glycoprotein I (β2GPI). Complement is involved in APS animal models and complement deposits have been documented in placenta and thrombotic vessels despite normal serum levels. Analysis of circulating blood cells coated with C4d displays higher sensitivity than the conventional assays that measure soluble native complement components and their unstable activation products in systemic lupus erythematosus (SLE). As C4d-coated blood cell count has been reported to be more sensitive than serum levels of complement components and their activation products in systemic lupus erythematosus (SLE) patients, we decided to evaluate the percentage of C4d positive B lymphocytes (BC4d), erythrocytes (EC4d), and platelets (PC4d) in primary APS patients and asymptomatic aPL positive carriers as marker of complement activation in APS. We assessed by flow cytometry the percentages of BC4d, EC4d, and PC4d in primary APS (PAPS; n. 23), 8 asymptomatic aPL positive carriers, 11 APS-associated SLE (SAPS), 17 aPL positive SLE, 16 aPL negative SLE, 8 aPL negative patients with previous thrombosis, 11 immune thrombocytopenia (ITP) patients, and 26 healthy subjects. In addition, we used an in vitro model to evaluate the ability of a monoclonal anti-β2GPI antibody (MBB2) to bind to normal resting or activated platelets and fix complement. EC4d and PC4d percentages were significantly higher in PAPS and aPL carriers as well as aPL positive SLE and SAPS than in aPL negative controls. The highest values were found in PAPS and in SAPS. The EC4d and PC4d percentages were significantly correlated with serum C3/C4 and anti-β2GPI/anti-cardiolipin IgG. In vitro studies showed that MBB2 bound to activated platelets only and induced C4d deposition. The detection of the activation product C4d on circulating erythrocytes and platelets supports the role of complement activation in APS. Complement may represent a new therapeutic target for better treatment and prevention of disability of APS patients

Comparison of injective related reactions following ofatumumab and ocrelizumab in patients with multiple sclerosis: data from the European spontaneous reporting system

IntroductionIn 2021 ofatumumab, a recombinant human anti-CD20 monoclonal antibody (mAb) already authorized for the treatment of chronic lymphocytic leukemia, received the marketing approval for the treatment of relapsing forms of multiple sclerosis (MS). Differently from ocrelizumab, that is administered intravenously, ofatumumab if the first anti-CD20 mAb to be administered subcutaneously without a premedication.Methods and objectivesIn this study we aimed to describe and compare the main characteristics of Individual Case Safety Reports (ICSRs) describing the occurrence of Injective Related Reactions (IRRs) following the treatment with ocrelizumab and ofatumumab reported in the Eudravigilance (EV) database during years 2021–2023.ResultsA total of 860 ICSRs with either ofatumumab and ocrelizumab as suspected drug were retrieved from Eudravigilance, of which 51% associated with ofatumumab and 49% with ocrelizumab. The majority of patients who experienced IRRs following ocrelizumab belonged to the age group of 18–64 years (73%), while the age-group was mostly not specified (55%) in ICSRs reporting ofatumumab as suspected. The distribution of gender was almost similar in the two groups, with the majority of ICSRs related to female patients. “Pyrexia” was the Preferred Term (PT) most reported for ofatumumab, while “Infusion related reaction” were more frequently reported with ocrelizumab. Premedication drugs were reported in 148 ICSRs. Out of 89 ICSRs for which the Time to Event (TTE) was calculated, 74 reported IRRs that occurred the same day of the drug administration.DiscussionBased on the results of this study, although a risk of ofatumumab-induced IRRs cannot be excluded, it should be considered as manageable considering that the drug seems to be mostly associated with the occurrence of fever. Thus, it is important to continue to closely monitor the use of these in clinical practice to improve the knowledge on their long-term safety