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Development of antibody protection during SARS-CoV-2 infection is a pressing question for public health and for vaccine development. We developed highly sensitive SARS-CoV-2-specific antibody and neutralization assays. SARS-CoV-2 Spike protein or Nucleocapsid protein specific IgG antibodies at titers more than 1:100,000 were detectable in all PCR+ subjects (n = 115) and were absent in the negative controls. Other isotype antibodies (IgA, IgG1-4) were also detected. SARS-CoV-2 neutralization was determined in COVID-19 and convalescent plasma at up to 10,000-fold dilution, using Spike protein pseudotyped lentiviruses, which were also blocked by neutralizing antibodies (NAbs). Hospitalized patients had up to 3000-fold higher antibody and neutralization titers compared to outpatients or convalescent plasma donors. Interestingly, some COVID-19 patients also possessed NAbs against SARS-CoV Spike protein pseudovirus. Together these results demonstrate the high specificity and sensitivity of our assays, which may impact understanding the quality or duration of the antibody response during COVID-19 and in determining the effectiveness of potential vaccines.

Background: COVID-19 can present with lymphopenia and extraordinary complex multiorgan pathologies that can trigger long-term sequela.Aims: Given that inflammasome products, like caspase-1, play a role in the pathophysiology of a number of co-morbid conditions, we investigated caspases across the spectrum of COVID-19 disease. Materials & Methods:We assessed transcriptional states of multiple caspases and using flow cytometry, the expression of active caspase-1 in blood cells from COVID-19 patients in acute and convalescent stages of disease. Non-COVID-19 subject presenting with various comorbid conditions served as controls.Results: Single-cell RNA-seq data of immune cells from COVID-19 patients showed a distinct caspase expression pattern in T cells, neutrophils, dendritic cells, and eosinophils compared with controls. Caspase-1 was upregulated in CD4+ T-cells from hospitalized COVID-19 patients compared with unexposed controls. Post-COVID-19 patients with lingering symptoms (long-haulers) also showed upregulated caspase-1activity in CD4+ T-cells that ex vivo was attenuated with a select pan-caspase inhibitor. We observed elevated caspase-3/7levels in red blood cells from COVID-19 patients compared with controls that was reduced following caspase inhibition. Discussion: Our preliminary results suggest an exuberant caspase response in COVID-19 that may facilitate immune-related pathological processes leading to severe outcomes. Further clinical correlations of caspase expression in different stages of COVID-19 will be needed. Conclusion:Pan-caspase inhibition could emerge as a therapeutic strategy to ameliorate or prevent severe COVID-19.

supplementary data for Table S1, reviewed data of all previous cases. CMB made a critical revision of the manuscript, helped in the acquisition of data, provided supplementary data for Table S1, reviewed data of all previous cases. All authors revised and gave the approval of the final manuscript.

Phylogenetic analyses of partial sequences spanning approximately 450 nucleotides near the 5’end of the 18s rDNA strongly support the monophyly of Apogastropoda and its constituent clades, Caenogastropoda and Heterobranchia. Representatives of the architaenioglossan groups Cyclophoroidea, Ampullariidae and Viviparidae invariably emerge within Caenogastropoda in all analyses. While the Cyclophoroidea and Ampullariidae are monophyletic, the varying position of Viviparidae in all outcomes contradicts its hypothesized sister group relationship with Ampullariidae, and thus the monophyly of Ampullarioidea. Because of the position of Viviparidae, Architaenioglossa does not emerge as a clade in any of our analyses. Campanile consistently emerges between Cyclophoroidea and Cerithioidea, or in a clade with Cyclophoroidea and Ampullariidae, a position not predicted by previous morphological studies. Maximum parsimony analyses of sequence data show Caenogastropoda to comprise a series of sequentially diverging higher taxa. However, maximum likelihood analyses as well as maximum parsimony analyses using only trans‐versions divide Caenogastropoda into two clades, one containing the architaenioglossan taxa, Campaniloidea and Cerithioidea, the other containing the higher caenogastropod taxa included in Eucaenogastropoda (Haszprunar, 1988) [= Hypsogastropoda (Ponder & Lindberg 1997)l. Denser taxon sampling revealed insertions to be present in the 18s rDNA gene of several caenogastropod taxa. Earlier reports (Harasewych et al. 1997b) of reduced sequence divergence levels in Caenogastropoda are shown to be restricted to Hypsogastropoda. Based on a broader taxonomic sampling, divergence levels within Caenogastropoda are comparable to those found within Heterobranchia.

Gaucher disease (GD) patients often present with abnormalities in immune response that may be the result of alterations in cellular and/or humoral immunity. However, how the treatment and clinical features of patients impact the perturbation of their immunological status remains unclear. To address this, we assessed the immune profile of 26 GD patients who were part of an enzyme replacement therapy (ERT) study. Patients were evaluated clinically for onset of GD symptoms, duration of therapy and validated outcome measures for ERT. According to DS3 disease severity scoring system criteria, they were assigned to have mild, moderate or severe GD. Flow cytometry based immunophenotyping was performed to analyze subsets of T, B, NK, NKT and dendritic cells. GD patients showed multiple types of immune abnormalities associated to T and B lymphocytes with respect to their subpopulations as well as memory and activation markers. Skewing of CD4 and CD8 T cell numbers resulting in lower CD4/CD8 ratio and an increase in overall T cell activation were observed. A decrease in the overall B cells and an increase in NK and NKT cells were noted in the GD patients compared to controls. These immune alterations do not correlate with GD clinical type or level of biomarkers. However, subjects with persistent immune alterations, especially in B cells and DCs correlate with longer delay in initiation of ERT (ΔTX). Thus, while ERT may reverse some of these immune abnormalities, the immune cell alterations become persistent if therapy is further delayed. These findings have important implications in understanding the immune disruptions before and after treatment of GD patients.

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Background:The short stability window of several hours from blood collection to measuring basophil activation has limited the use of flow cytometry-based basophil activation assays in clinical settings. We examine if it is possible to extend this