Hypercytokinemia, found in SARS-CoV-2 infection, contributes to multiple organ dysfunctions with acute respiratory distress syndrome, shock etc. The aim of this study was to describe cytokine storm signatures in patients with acute COVID-19 and to investigate their influence on severity of the infection. Plasma levels of 47 cytokines were investigated in 73 patients with moderate and severe COVID-19 (41 and 32, respectively) and 11 healthy donors (HD). The most elevated levels comparing patients and the HD were observed for seven pro-inflammatory cytokines (IL-6, IL-8, IL-15, IL-18, IL-27, IFNγ, TNFα), three chemokines (GROα, IP-10, MIG), two anti-inflammatory cytokines (IL-1RA, IL-10), and two growth factors (G-CSF, M-CSF). The patients with severe disease had significantly higher levels of FGF-2/FGF-basic, IL-1β, and IL-7 compared to the HD. The two groups of patients differed from each other only based on the levels of EGF, eotaxin, and IL-12 p40. Pneumonia lung injury, characterized by computer tomography, positively correlated with levels of EGF, IP-10, MCP-3 levels and negatively with IL-12 p40. Pro-inflammatory factors including IL-6, TNFα, and IP-10 negatively correlated with the frequency of the circulating T-helper17-like cells (Th17-like) and follicular Th cells that are crucial to develop SARS-CoV-2-specific plasma cells and memory B cells. Obtained data on the cytokine levels illustrate their influence on progression and severity of COVID-19.

Background: The immunological changes associated with COVID-19 are largely unknown. Methods: Patients with COVID-19 showing moderate (n = 18; SpO2 > 93%, respiratory rate > 22 per minute, CRP > 10 mg/L) and severe (n = 23; SpO₂ < 93%, respiratory rate >30 per minute, PaO₂/FiO₂ ≤ 300 mmHg, permanent oxygen therapy, qSOFA > 2) infection, and 37 healthy donors (HD) were enrolled. Circulating T- and B-cell subsets were analyzed by flow cytometry. Results: CD4+Th cells were skewed toward Th2-like phenotypes within CD45RA+CD62L− (CM) and CD45RA–CD62L− (EM) cells in patients with severe COVID-19, while CM CCR6+ Th17-like cells were decreased if compared with HD. Within CM Th17-like cells “classical” Th17-like cells were increased and Th17.1-like cells were decreased in severe COVID-19 cases. Circulating CM follicular Th-like (Tfh) cells were decreased in all COVID-19 patients, and Tfh17-like cells represented the most predominant subset in severe COVID-19 cases. Both groups of patients showed increased levels of IgD-CD38++ B cells, while the levels of IgD+CD38− and IgD–CD38− were decreased. The frequency of IgD+CD27+ and IgD–CD27+ B cells was significantly reduced in severe COVID-19 cases. Conclusions: We showed an imbalance within almost all circulating memory Th subsets during acute COVID-19 and showed that altered Tfh polarization led to a dysregulated humoral immune response.

New investigation results point to the potential participation of extracellular vesicles (EVs) in the pathogenesis of coronavirus infection, its progression, and mechanisms of the therapy effectiveness. This dictates the necessity to transfer scientific testing technologies to medical practice. Here, we demonstrated the method of phenotyping and quantitative analysis of plasma EVs based on differential centrifugation, immunostaining, and high-sensitivity multicolor flow cytometry. We used EV markers that were potentially associated with SARS-CoV-2 dissemination via vesicles and cell-origination markers, characterizing objects from different cell types that could influence clinical manifestation of COVID-19. Plasma levels of CD235a+ and CD14+ EVs in patients with moderate infection were significantly increased while CD8+ and CD19+ EVs were decreased comparing with HD. Patients with severe infection had lower levels of CD4+, CD19+, and CD146+ EVs than HD. These findings demonstrate that EV concentrations in COVID-19 are severity related. Moreover, the three-point dynamic assessment demonstrated significant loss of CD63+ and CD147+ plasma EVs. The used method can be a convenient tool for vital infection pathogenesis investigation and for COVID-19 diagnostics.

Mutations in the GBA1 gene, encoding the lysosomal enzyme glucocerebrosidase (GCase), cause Gaucher disease (GD) and are the most common genetic risk factor for Parkinson’s disease (PD). Pharmacological chaperones (PCs) are being developed as an alternative treatment approach for GD and PD. To date, NCGC00241607 (NCGC607) is one of the most promising PCs. Using molecular docking and molecular dynamics simulation we identified and characterized six allosteric binding sites on the GCase surface suitable for PCs. Two sites were energetically more preferable for NCGC607 and located nearby to the active site of the enzyme. We evaluated the effects of NCGC607 treatment on GCase activity and protein levels, glycolipids concentration in cultured macrophages from GD (n = 9) and GBA-PD (n = 5) patients as well as in induced human pluripotent stem cells (iPSC)—derived dopaminergic (DA) neurons from GBA-PD patient. The results showed that NCGC607 treatment increased GCase activity (by 1.3-fold) and protein levels (by 1.5-fold), decreased glycolipids concentration (by 4.0-fold) in cultured macrophages derived from GD patients and also enhanced GCase activity (by 1.5-fold) in cultured macrophages derived from GBA-PD patients with N370S mutation (p < 0.05). In iPSC-derived DA neurons from GBA-PD patients with N370S mutation NCGC607 treatment increased GCase activity and protein levels by 1.1-fold and 1.7-fold (p < 0.05). Thus, our results showed that NCGC607 could bind to allosteric sites on the GCase surface and confirmed its efficacy on cultured macrophages from GD and GBA-PD patients as well as on iPSC-derived DA neurons from GBA-PD patients.

Aim. To collect and analyze real-world data on long-term enzyme replacement therapy with Glurazyme® in patients with type I Gaucher disease (GD). Materials and methods. The study included 31 patients with type I GD (21 patients with intact spleen and 10 patients with a history of splenectomy) over 18 years. Efficacy was assessed by the change in hemoglobin level (primary endpoint), platelet count, spleen and liver volumes, prevalence of bone marrow infiltration, and the number of patients with quiet hip disease according to magnetic resonance imaging (secondary endpoints). Safety was assessed by the incidence of treatment-related adverse events and serious adverse events, as well as by the incidence of production of imiglucerase binding and neutralizing antibodies (IgG, IgE). The mean follow-up duration was 54 weeks. Results. The study did not show a statistically significant change in hemoglobin levels, platelet count, or spleen and liver volumes. There was an upward tendency in platelet count in the overall group (p=0.1) and patients after splenectomy (p=0.08). The prevalence of specific bone marrow infiltration and the number of patients with quiet hip disease remained unchanged. During the safety analysis, one adverse reaction, a mild increase in alanine aminotransferase, was reported, which resolved spontaneously by the end of the study. The immunogenicity analysis showed the initial presence of anti-drug antibodies (ADAs) in 5 (16.7%) of the 30 examined patients. At the end of the study, ADAs were detected in only 3 (10%) patients. The detected ADAs had a low titer and no neutralizing activity, and they did not affect the treatment effectiveness. Conclusion. Long-term therapy with the imiglucerase biosimilar was associated with a stable course of GD without progression. It was characterized by a good safety profile and low immunogenicity.

Mutations in the GBA1 gene, encoding lysosomal β-glucocerebrosidase (GCase), cause Gaucher disease (GD) and represent the most common genetic risk factor for Parkinson’s disease (PD). To target this pathway, therapeutic approaches for both GD and PD are actively being developed, focusing on two main drug classes: pharmacological chaperones (e.g., ambroxol) and allosteric GCase activators. Here, we characterize a novel allosteric GCase activator, N2, a derivative of the non-inhibitory chaperone NCGC607, and evaluate its therapeutic potential using computational, in vitro, and in vivo approaches. Molecular docking and molecular dynamics simulations indicated that N2 binds within the allosteric site of the mutant p.N370S GCase structure. In patient-derived cells (peripheral blood mononuclear cell-derived macrophages and induced pluripotent stem cell-derived dopaminergic neurons from GD and GBA1-PD patients), N2 enhanced GCase lysosomal translocation, modulated its enzymatic activity and protein level, and reduced substrate accumulation. Furthermore, we demonstrate for the first time that N2 exerts robust neuroprotective effects in an in vivo 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced mouse model of PD. Specifically, N2 treatment attenuated MPTP-induced hexosylsphingosine accumulation in the prefrontal cortex, and ameliorated both tyrosine hydroxylase-positive neuronal loss in the substantia nigra and dopamine depletion in the striatum. Our findings first demonstrated the neuroprotective properties of a GCase allosteric activator on an MPTP-induced mouse model of PD and supported the emerging perspectives of such compounds for the treatment of GCase-related disorders.