Monocyte Distribution Width (MDW) in Patients with COVID-19: An Indicator of Disease Severity.

Identifying patients with Coronavirus disease-2019 (COVID-19) who may have a severe illness is essential for timely intervention and decreasing the fatality rate. In the present study, we evaluated the performance of Monocyte Distribution Width (MDW) as a prognostic marker for identifying disease severity in COVID-19 patients. We included 145 patients with PCR-confirmed COVID-19 infection in the study. The performance of MDW was evaluated by calculating the area under the receiver operating characteristic curve (AUC), specificity, sensitivity, negative predictive value, and positive predictive value. Further analysis was conducted for the disease outcome, comparing COVID-19 patients discharged (n = 135) to deceased COVID-19 patients (n = 10). As a marker of disease severity, MDW demonstrated an AUC of 0.702 (95% CI 0.620-0.775) in ROC analysis. If MDW is considered a marker of patient outcome, AUC was 0.916 (95% CI 0.862-0.953), comparing deceased COVID-19 patients vs. those who survived. At a cut-off of > 25.4 on admission, MDW correlates well with poor disease outcomes in COVID-19 patients. MDW can be considered a helpful parameter in predicting the severity of COVID-19 disease and patient outcomes. Its role and incorporation in the standard diagnostic algorithm and management of COVID-19 patients need further validation. Supplementary Information: The online version contains supplementary material available at 10.1007/s12288-023-01665-y.

Prognostic value of cellular population data in patients with COVID-19.

Background and aims: Beckman Coulter hematology analysers identify leukocytes by their volume (V), conductivity (C) and scatter (S) of a laser beam at different angles. Each leukocyte sub-population [neutrophils (NE), lymphocytes (LY), monocytes (MO)] is characterized by the mean (MN) and the standard deviation (SD) of 7 measurements called "cellular population data" (@CPD), corresponding to morphological analysis of the leukocytes. As severe forms of infections to SARS-CoV-2 are characterized by a functional activation of mononuclear cells, leading to a cytokine storm, we evaluated whether CPD variations are correlated to the inflammation state, oxygen requirement and lung damage and whether CPD analysis could be useful for a triage of patients with COVID-19 in the Emergency Department (ED) and could help to identify patients with a high risk of worsening. Materials and method: The CPD of 825 consecutive patients with proven COVID-19 presenting to the ED were recorded and compared to classical biochemical parameters, the need for hospitalization in the ward or ICU, the need for oxygen, or lung injury on CT-scan. Results: 40 of the 42 CPD were significantly modified in COVID-19 patients in comparison to 245 controls. @MN-V-MO and @SD-V-MO were highly correlated with C-reactive protein, procalcitonin, ferritin and D-dimers. SD-UMALS-LY > 21.45 and > 23.92 identified, respectively, patients with critical lung injuries (>75%) and requiring tracheal intubation. @SD-V-MO > 25.03 and @SD-V-NE > 19.4 identified patients required immediate ICU admission, whereas a @MN-V-MO < 183 suggested that the patient could be immediately discharged. Using logistic regression, the combination of 8 CPD with platelet and basophil counts and the existence of diabetes or obesity could identify patients requiring ICU after a first stay in conventional wards (area under the curve = 0.843). Conclusion: CPD analysis constitutes an easy and inexpensive tool for triage and prognosis of COVID-19 patients in the ED.

Can Haematological Parameters Discriminate COVID-19 from Influenza?

Symptoms of COVID-19 are similar to the influenza virus, but because treatments and prognoses are different, it is important to accurately and rapidly differentiate these diseases. The aim of this study was to evaluate whether the analysis of complete blood count (CBC), including cellular population (CPD) data of leukocytes and automated flow cytometry analysis, could discriminate these pathologies. In total, 350 patients with COVID-19 and 102 patients with influenza were included between September 2021 and April 2022 in the tertiary hospital of Suresnes (France). Platelets were lower in patients with influenza than in patients with COVID-19, whereas the CD16pos monocyte count and the ratio of the CD16pos monocytes/total monocyte count were higher. Significant differences were observed for 9/56 CPD of COVID-19 and flu patients. A logistic regression model with 17 parameters, including among them 11 CPD, the haemoglobin level, the haematocrit, the red cell distribution width, and B-lymphocyte and CD16pos monocyte levels, discriminates COVID-19 patients from flu patients. The sensitivity and efficiency of the model were 96.2 and 86.6%, respectively, with an area under the curve of 0.862. Classical parameters of CBC are very similar among the three infections, but CPD, CD16pos monocytes, and B-lymphocyte levels can discriminate patients with COVID-19.

Monocyte distribution width (MDW): a useful biomarker to improve sepsis management in Emergency Department.

OBJECTIVES: Sepsis is a time-dependent and life-threating condition. Despite several biomarkers are available, none of them is completely reliable for the diagnosis. This study aimed to evaluate the diagnostic utility of monocyte distribution width (MDW) to early detect sepsis in adult patients admitted in the Emergency Department (ED) with a five part differential analysis as part of the standard clinical practice. METHODS: A prospective cohort study was conducted on 985 patients aged from 18 to 96 and included in the study between November 2019 and December 2019. Enrolled subjects were classified into four groups based on sepsis-2 diagnostic criteria: control, Systemic Inflammatory Response Syndrome (SIRS), infection and sepsis. The hematology analyzer DxH 900 (Beckman Coulter Inc.) provides the new reportable parameter MDW, included in the leukocyte 5 part differential analysis, cleared by Food and Drug administration (FDA) and European Community In-Vitro-Diagnostic Medical Device (CE IVD) marked as early sepsis indicator (ESId). RESULTS: MDW was able to differentiate the sepsis group from all other groups with Area Under the Curve (AUC) of 0.849, sensitivity of 87.3% and specificity of 71.7% at cut-off of 20.1. MDW in combination with white blood cell (WBC) improves the performance for sepsis detection with a sensitivity increased up to 96.8% when at least one of the two biomarkers are abnormal, and a specificity increased up to 94.6% when both biomarkers are abnormal. CONCLUSIONS: MDW can predict sepsis increasing the clinical value of Leukocyte 5 Part Differential analysis and supporting the clinical decision making in sepsis management at the admission to the ED.

Interest of the cellular population data analysis as an aid in the early diagnosis of SARS-CoV-2 infection.

INTRODUCTION: Coronavirus disease 2019 (COVID-19) is characterized by a high contagiousness requiring isolation measures. At this time, diagnosis is based on the positivity of specific RT-PCR and/or chest computed tomography scan, which are time-consuming and may delay diagnosis. Complete blood count (CBC) can potentially contribute to the diagnosis of COVID-19. We studied whether the analysis of cellular population data (CPD), provided as part of CBC-Diff analysis by the DxH 800 analyzers (Beckman Coulter), can help to identify SARS-CoV-2 infection. METHODS: Cellular population data of the different leukocyte subpopulations were analyzed in 137 controls, 322 patients with proven COVID-19 (COVID+), and 285 patients for whom investigations were negative for SARS-CoV-2 infection (COVID-). When CPD of COVID+ were different from controls and COVID- patients, we used receiver operating characteristic analysis to test the discriminating capacity of the individual parameters. Using a random forest classifier, we developed the algorithm based on the combination of 4 monocyte CPD to discriminate COVID+ from COVID- patients. This algorithm was tested prospectively in a series of 222 patients referred to the emergency unit. RESULTS: Among the 222 patients, 86 were diagnosed as COVID-19 and 60.5% were correctly identified using the discriminating protocol. Among the 136 COVID- patients, 10.3% were misclassified (specificity 89.7%, sensitivity 60.5%). False negatives were observed mainly in patients with a low inflammatory state whereas false positives were mainly seen in patients with sepsis. CONCLUSION: Consideration of CPD could constitute a first step and potentially aid in the early diagnosis of COVID-19.

Nuclear oncoprotein prothymosin alpha is a partner of Keap1: implications for expression of oxidative stress-protecting genes.

Animal cells counteract oxidative stress and electrophilic attack through coordinated expression of a set of detoxifying and antioxidant enzyme genes mediated by transcription factor Nrf2. In unstressed cells, Nrf2 appears to be sequestered in the cytoplasm via association with an inhibitor protein, Keap1. Here, by using the yeast two-hybrid screen, human Keap1 has been identified as a partner of the nuclear protein prothymosin alpha. The in vivo and in vitro data indicated that the prothymosin alpha-Keap1 interaction is direct, highly specific, and functionally relevant. Furthermore, we showed that Keap1 is a nuclear-cytoplasmic shuttling protein equipped with a nuclear export signal that is important for its inhibitory action. Prothymosin alpha was able to liberate Nrf2 from the Nrf2-Keap1 inhibitory complex in vitro through competition with Nrf2 for binding to the same domain of Keap1. In vivo, the level of Nrf2-dependent transcription was correlated with the intracellular level of prothymosin alpha by using prothymosin alpha overproduction and mRNA interference approaches. Our data attribute to prothymosin alpha the role of intranuclear dissociator of the Nrf2-Keap1 complex, thus revealing a novel function for prothymosin alpha and adding a new dimension to the molecular mechanisms underlying expression of oxidative stress-protecting genes.

Apoptosis-related fragmentation, translocation, and properties of human prothymosin alpha.

Human prothymosin alpha is a proliferation-related nuclear protein undergoing caspase-mediated fragmentation in apoptotic cells. We show here that caspase-3 is the principal executor of prothymosin alpha fragmentation in vivo. In apoptotic HeLa cells as well as in vitro, caspase-3 cleaves prothymosin alpha at one major carboxy terminal (DDVD(99)) and several suboptimal sites. Prothymosin alpha cleavage at two amino-terminal sites (AAVD(6) and NGRD(31)) contributes significantly to the final pattern of prothymosin alpha fragmentation in vitro and could be detected to occur in apoptotic cells. The major caspase cleavage at D(99) disrupts the nuclear localization signal of prothymosin alpha, which leads to a profound alteration in subcellular localization of the truncated protein. By using a set of anti-prothymosin alpha monoclonal antibodies, we were able to observe nuclear escape and cell surface exposure of endogenous prothymosin alpha in apoptotic, but not in normal, cells. We demonstrate also that ectopic production of human prothymosin alpha and its mutants with nuclear or nuclear-cytoplasmic localization confers increased resistance of HeLa cells toward the tumor necrosis factor-induced apoptosis.

Sensing prothymosin alpha origin, mutations and conformation with monoclonal antibodies.

To overcome poor immunogenicity of prothymosin alpha, a small and highly acidic nuclear protein involved in cell proliferation, production of anti-prothymosin alpha antibodies in mice immunized with free human prothymosin alpha, with prothymosin alpha coupled to different carriers and with prothymosin alpha fused to green fluorescent protein was assessed. Fusing prothymosin alpha to green fluorescent protein turned out to be the superior approach resulting in production of high titer anti-prothymosin alpha antibodies. From these studies, two highly specific anti-prothymosin alpha monoclonal antibodies recognizing epitopes within the amino terminal (2F11) and middle (4F4) portions of the human prothymosin alpha molecule were obtained and characterized. As expected, the 2F11 antibody displayed broad species specificity, whereas the 4F4 antibody appeared to be species-specific permitting discrimination of human versus rat protein. Furthermore, a combination of point mutations in prothymosin alpha that alter the properties of the protein precluded recognition by the 4F4 antibody. Intramolecular masking of the 4F4 epitope in prothymosin alpha fused to the Tat transduction peptide of human immunodeficiency virus type 1 was observed. The anti-prothymosin alpha antibodies obtained were suitable for precipitation of human prothymosin alpha from HeLa cell lysates and for immunolocalization of the endogenous prothymosin alpha within the cells. Fusion with green fluorescent protein may thus be helpful in raising antibodies against 'problematic' proteins.