Wider, David Leppert, Jens Kuhle, Laura N. for Neuro-axonal injury in COVID-19: the role of systemic inflammation and SARS-CoV-2 specific immune response by Cdric Hirzel, Denis Grandgirard, Bernard Surial, Manon F. Wider, David Leppert, Jens Kuhle, Laura N. Walti, Joerg C. Schefold, Thibaud Spinetti, Franziska Suter-Riniker, Ronald Dijkman and Stephen L. Leib in Therapeutic Improvements in Neurological Disorders Abstract Background: AT7519 trifluoroacetate In coronavirus disease-2019 (COVID-19) patients, there is increasing evidence of neuronal injury by the means of elevated serum neurofilament light chain (sNfL) levels. However, the role of systemic inflammation and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)Cspecific immune response with regard to neuronal injury has not yet been investigated. Methods: In a prospective cohort study, we recruited patients with mildCmoderate (assessments were used to assess the associations between dichotomous COVID-19 severity (mildCmoderate and severe), cytokine levels, antibody concentrations, CMI (SFU/250,000 cells), and sNfL concentrations. To account for the potential for confounding by age, corticosteroid use, and time from symptom onset to sample measurement, we repeated the analyses using multiple linear regression adjusted for those covariates. Variables of interest were log-transformed where appropriate to approximate a normal distribution. We compared outcomes between individuals with severe and mildCmoderate COVID-19 using adjusted imply values, and calculated imply differences for untransformed and ratio of means for log-transformed variables. 31 Cytokine levels, antibody concentrations, CMI (SFU/250,000 cells), and sNfL concentrations in paired samples over time were compared using Wilcoxon signed-rank test. Because sNfL levels increase with age, 18 which is also a major risk factor for severe COVID-19, we further processed our analyses by associating sNfL changes (sNfL: sNfL at follow-up minus sNfL at baseline; sNfL fold-change: sNfL at follow-up divided by sNfL at baseline) with the immune response. We used univariable and multivariable linear regression adjusted for age, corticosteroid use, and time from symptom onset to sample measurement to estimate the association between changes in sNfL levels and cytokine concentrations, antibody levels, and SFUs. sNfL fold-changes were modeled using log-transformation, and complete sNfL differences were modeled using a cube-root VPS33B transformation to account for the long tails in addition to positive and negative values. 32 Statistical significance was defined as a value .05. In this exploratory study, values and widths of 95% confidence intervals were not adjusted for multiplicity.33,34 Statistical analyses were performed using Stata software version 16.0 (Stata Corp., College Station, TX, USA) and R version 4.1.1 (R Core Team 2021, R: A language and environment for statistical computing, R Foundation for Statistical Computing, Vienna, Austria, URL https://www.R-project.org/). Figures were created using R and GraphPad, version 8.0 (LaJolla, CA, USA). Results Demographics We enrolled 55 patients with symptomatic COVID-19 and excluded two patients from analysis due to the presence of an active neuroinflammatory disorder (one AT7519 trifluoroacetate patient with GuillainCBarr syndrome; one patient with a facial palsy due to a suppurative otitis). Patients with moderate to moderate disease (valuevalues ( 0.05) are in strong. Cytokine response All patients experienced baseline serum samples, and follow-up samples were available for 49/53 patients (2 patients died, 1 patient refused follow-up blood work). In unadjusted analysis, median serum cytokine concentrations at baseline were higher in patients with severe COVID-19 compared with those with mildCmoderate disease (IL-6: 167.5 3.3 pg/ml, 14.7 pg/ml, 0.257 pg/ml, 15.1 pg/ml, 1.5 pg/ml, 10.2 pg/ml, 0.176 pg/ml, 10.3 pg/ml, mildCmoderate COVID-19valuevalues ( 0.05) are in strong. Antibody response At baseline, most patients were seronegative for anti-spike IgG antibodies (severe COVID-19: 57.1% (6/14), mildCmoderate COVID-19: 84.6% (33/39), severe) was not associated with the quantity of IFN- positive SFUs (unadjusted analyses: Figure 2(a) and (?(b);b); adjusted analyses: Table 2). Open in a separate window Physique 2. Cell-mediated immunity in relation to COVID-19 severity. (a) SFUs per 250,000 PBMCs upon activation with the SNMO peptide pool at baseline and day 28(7) in patients with severe (reddish) and mildCmoderate (blue) AT7519 trifluoroacetate COVID-19. (b) SFUs per 250,000 PBMCs upon activation with the S2N peptide pool at baseline and day 28(7) in patients with severe (reddish) and mildCmoderate (blue) COVID-19. Black horizontal bars show median values. Whiskers show interquartile ranges. COVID-19, coronavirus disease-2019; IFN-, interferon-gamma; IL-2, interleukin-2; PBMCs, peripheral blood mononuclear cells; SFUs, spot forming models; SNMO, spike protein, nucleocapsid protein, membrane protein, open reading frame proteins; S2N, spike-2 protein, nucleocapsid protein. IL-2 SFUs significantly increased from baseline to follow-up upon activation with the S2N and SNMO peptide pool (Physique 2(a) and (b)). AT7519 trifluoroacetate Upon cell activation with the SNMO peptide pool, patients with severe COVID-19 had significantly more IL-2-positive SFUs compared with individuals with mildCmoderate disease at follow-up (unadjusted analyses: Physique 2(a); adjusted analyses: Table 2). IFN-/IL-2 double-positive spots significantly increased from baseline to follow-up after cell activation with the AT7519 trifluoroacetate SNMO peptide pool and in unadjusted analyses; patients with severe disease had more IFN-/IL-2 double-positive SFU at follow-up compared.
Wider, David Leppert, Jens Kuhle, Laura N