scholarly journals Longitudinal characterization of circulating neutrophils uncovers distinct phenotypes associated with disease severity in hospitalized COVID-19 patients

2021 ◽  
Author(s):  
Thomas J. LaSalle ◽  
Anna L. K. Gonye ◽  
Samuel S. Freeman ◽  
Paulina Kaplonek ◽  
Irena Gushterova ◽  
...  
Keyword(s):  
Disease Severity ◽  
Severe Disease ◽  
Gene Expression Signature ◽  
Suppressor Cell ◽  
Gene Signature ◽  
Neutrophil Extracellular Trap ◽  
Effector Functions ◽  
Neutrophil Degranulation ◽  
Mechanistic Basis

Multiple studies have identified an association between neutrophils and COVID-19 disease severity; however, the mechanistic basis of this association remains incompletely understood. Here we collected 781 longitudinal blood samples from 306 hospitalized COVID-19+ patients, 78 COVID-19- acute respiratory distress syndrome patients, and 8 healthy controls, and performed bulk RNA-sequencing of enriched neutrophils, plasma proteomics, cfDNA measurements and high throughput antibody profiling assays to investigate the relationship between neutrophil states and disease severity or death. We identified dynamic switches between six distinct neutrophil subtypes using non-negative matrix factorization (NMF) clustering. At days 3 and 7 post-hospitalization, patients with severe disease had an enrichment of a granulocytic myeloid derived suppressor cell-like state gene expression signature, while non-severe patients with resolved disease were enriched for a progenitor-like immature neutrophil state signature. Severe disease was associated with gene sets related to neutrophil degranulation, neutrophil extracellular trap (NET) signatures, distinct metabolic signatures, and enhanced neutrophil activation and generation of reactive oxygen species (ROS). We found that the majority of patients had a transient interferon-stimulated gene signature upon presentation to the emergency department (ED) defined here as Day 0, regardless of disease severity, which persisted only in patients who subsequently died. Humoral responses were identified as potential drivers of neutrophil effector functions, as enhanced antibody-dependent neutrophil phagocytosis and reduced NETosis was associated with elevated SARS-CoV-2-specific IgG1-to-IgA1 ratios in plasma of severe patients who survived. In vitro experiments confirmed that while patient-derived IgG antibodies mostly drove neutrophil phagocytosis and ROS production in healthy donor neutrophils, patient-derived IgA antibodies induced a predominant NETosis response. Overall, our study demonstrates neutrophil dysregulation in severe COVID-19 and a potential role for IgA-dominant responses in driving neutrophil effector functions in severe disease and mortality.

scholarly journals ZRC3308 Monoclonal Antibody Cocktail Shows Protective Efficacy in Syrian Hamsters against SARS-CoV-2 Infection

Viruses ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2424
Author(s):  
Pragya D. Yadav ◽  
Sanjeev Kumar Mendiratta ◽  
Sreelekshmy Mohandas ◽  
Arun K. Singh ◽  
Priya Abraham ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Protective Efficacy ◽  
Severe Disease ◽  
Binding Affinities ◽  
Immune Effector ◽  
Syrian Hamsters ◽  
Effector Functions ◽  
Prophylactic Use ◽  
Antibody Cocktail

We have developed a monoclonal antibody (mAb) cocktail (ZRC-3308) comprising of ZRC3308-A7 and ZRC3308-B10 in the ratio 1:1 for COVID-19 treatment. The mAbs were designed to have reduced immune effector functions and increased circulation half-life. mAbs showed good binding affinities to non-competing epitopes on RBD of SARS-CoV-2 spike protein and were found neutralizing SARS-CoV-2 variants B.1, B.1.1.7, B.1.351, B.1.617.2, and B.1.617.2 AY.1 in vitro. The mAb cocktail demonstrated effective prophylactic and therapeutic activity against SARS-CoV-2 infection in Syrian hamsters. The antibody cocktail appears to be a promising candidate for prophylactic use and for therapy in early COVID-19 cases that have not progressed to severe disease.

scholarly journals Potential of Bacillus subtilis for controlling bacterial leaf blight pathogen in rice

Food Research ◽  
2020 ◽  
Vol 4 (S5) ◽  
pp. 124-130
Author(s):  
K.S. Ku Asmah ◽  
Z. Sapak
Keyword(s):  
Bacillus Subtilis ◽  
Disease Severity ◽  
Severe Disease ◽  
Severity Index ◽  
Leaf Blight ◽  
Bacterial Leaf Blight ◽  
In Vivo Studies ◽  
Disease Severity Index ◽  
Rice Plants

Bacterial leaf blight (BLB) of rice is an economically important disease caused by Xanthomonas oryzae pv. oryzae (Xoo) throughout the world. To control this disease, bacterial isolate of Bacillus subtilis UiTMB1 was screened for the antagonistic activity against the pathogen in vitro and in vivo studies. A bacterial assay and detached leaf technique were used to evaluate the potential of the bacterium against BLB pathogen in the laboratory. Meanwhile, the glasshouse study was conducted to further examine the aptitudes of the isolate on the disease control and growth-promoting of rice plants. The findings revealed that B. subtilis UiTMB1 is able to control the disease and enhance the growth of rice plants. Rice plants treated with B. subtilis UiTMB1 before being inoculated with BLB pathogen showed less severe disease symptoms with low disease severity index of 3.43 compared to rice plants without B. subtilis UiTMB1 with high disease severity index of 8.4. Besides controlling the disease, B. subtilis UiTMB1 was also promoting plant height, chlorophyll content, number of tillers and biomass of rice plants.

scholarly journals p53 mutants cooperate with HIF-1 in transcriptional regulation of extracellular matrix components to promote tumor progression

2018 ◽  
Vol 115 (46) ◽  
pp. E10869-E10878 ◽  
Author(s):  
Ivano Amelio ◽  
Mara Mancini ◽  
Varvara Petrova ◽  
Rob A. Cairns ◽  
Polina Vikhreva ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cancer Progression ◽  
Treatment Options ◽  
Hypoxia Inducible Factor ◽  
Gene Expression Signature ◽  
Gene Signature ◽  
Cancer Pathogenesis ◽  
Extracellular Matrix Components

Mutations in the TP53 gene and microenvironmentally driven activation of hypoxia-inducible factor-1 (HIF-1) typically occur in later stages of tumorigenesis. An ongoing challenge is the identification of molecular determinants of advanced cancer pathogenesis to design alternative last-line therapeutic options. Here, we report that p53 mutants influence the tumor microenvironment by cooperating with HIF-1 to promote cancer progression. We demonstrate that in non-small cell lung cancer (NSCLC), p53 mutants exert a gain-of-function (GOF) effect on HIF-1, thus regulating a selective gene expression signature involved in protumorigenic functions. Hypoxia-mediated activation of HIF-1 leads to the formation of a p53 mutant/HIF-1 complex that physically binds the SWI/SNF chromatin remodeling complex, promoting expression of a selective subset of hypoxia-responsive genes. Depletion of p53 mutants impairs the HIF-mediated up-regulation of extracellular matrix (ECM) components, including type VIIa1 collagen and laminin-γ2, thus affecting tumorigenic potential of NSCLC cells in vitro and in mouse models in vivo. Analysis of surgically resected human NSCLC revealed that expression of this ECM gene signature was highly correlated with hypoxic tumors exclusively in patients carrying p53 mutations and was associated with poor prognosis. Our data reveal a GOF effect of p53 mutants in hypoxic tumors and suggest synergistic activities of p53 and HIF-1. These findings have important implications for cancer progression and might provide innovative last-line treatment options for advanced NSCLC.

Effects of Methyl Jasmonate on Cylindrocarpon Root Rot Development on Ginseng

2012 ◽  
Vol 610-613 ◽  
pp. 3511-3517 ◽  
Author(s):  
Jia Man Sun ◽  
Jun Fan Fu ◽  
Ru Jun Zhou ◽  
Wei Na Su ◽  
Xue Rui Yan
Keyword(s):  
Methyl Jasmonate ◽  
Disease Severity ◽  
Root Rot ◽  
Severe Disease ◽  
Defense Responses ◽  
Defense System ◽  
Colony Diameter ◽  
Low Concentrations ◽  
Mda Content

Cylindrocarpon root rot, caused by Cylindrocarpon destructans, is a severe disease on ginseng (Panax ginseng C.A. Meyer) in northeast China. Methyl jasmonate (MeJA) has been studied for its ability to induce plant defenses against pathogens. The effects of MeJA on the colony diameter of C. destructans and seedling growth were examined in vitro. Two-year-old ginseng roots were drenched with MeJA (200 mg/L) and then inoculated with conidia suspension of C. destructans (1×106 conidia/ml). Disease severity was assessed on inoculated roots, and the activation of defense responses was also measured. Results showed that MeJA had no effect on the growth of C. destructans and seedlings at low concentrations. The incidence rate and disease severity on MeJA-treated ginseng roots was significantly reduced compared with untreated roots. The proline content was increased but the malondialdehyde (MDA) content decreased in MeJA- treated roots. The activities of phenylalanine amino-lyase (PAL), catalase (CAT), peroxidase (POD) and polyphenoloxidase (PPO) were significantly enhanced in treated roots. It was concluded that low concentrations of MeJA had no effect on C. destructans, but it stimulated the activities of defense enzymes in treated ginseng roots and alleviated the damage of roots from C. destructans. Therefore, the control of Cylindrocarpon root rot by MeJA may involve the activation of the disease-related defense system.

scholarly journals IMMU-10. GENOMIC DIFFERENCES UNDERLIE MYELOID-DERIVED SUPPRESSOR CELL SEXUAL DIMORPHISM IN GLIOBLASTOMA

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii106-ii107
Author(s):  
Defne Bayik ◽  
Yadi Zhou ◽  
Alice Lo ◽  
Chihyun Park ◽  
Changjin Hong ◽  
...  
Keyword(s):  
Sexual Dimorphism ◽  
Primary Source ◽  
Suppressor Cell ◽  
Gene Signature ◽  
Activation Markers ◽  
Myeloid Derived Suppressor Cell ◽  
Immunosuppressive Tumor Microenvironment ◽  
Female Specific ◽  
Proliferation In Vitro

Abstract A potently immunosuppressive tumor microenvironment facilitates progression of glioblastoma (GBM). We previously demonstrated that myeloid-derived suppressor cell (MDSC) subsets promote tumorigenesis in a sex-specific manner, contributing to sexual dimorphism in GBM incidence and prognosis. Our findings indicated that proliferating monocytic MDSCs (mMDSCs) accumulate in tumors of male mice and patients, while female tumor-bearing mice had an increase in circulating granulocytic MDSC (gMDSC) frequency, and a high gMDSC gene signature correlated with worse outcome of female patients. However, the mechanisms underlying sexual dimorphism of MDSC heterogeneity remain understudied and can provide insights for improved immunotherapy response. Using syngeneic mouse glioma models and sequencing approaches, we show that expression of Y-chromosome-linked genes correlates with upregulation of multiple RNA transcription-related pathways specifically in male mMDSCs. Consistently, adoptive transfer of male mMDSCs but not gMDSCs worsened GBM outcome in male recipients, while the transfer of sex-matched mMDSCs did not impact survival of female mice. In contrast to this cell-intrinsic regulatory pathway, sex steroids had no impact on MDSC profile, as castration or ovariectomy failed to alter MDSC subset accumulation patterns in GBM-bearing mice. Correspondingly, IL-1β, which we had identified as a female-specific drug target, was highly expressed in female but not male gMDSCs. Single-cell sequencing revealed that circulating but not tumor-infiltrating gMDSCs were the primary source of IL-1β and that its neutralization provided a female-specific survival advantage by reducing circulating gMDSCs. This was accompanied by declines in tumor infiltration of microglia, microglia activation status and tumor cell proliferation. In vitro, IL-1β inhibition reduced viability and expression of activation markers by primary microglia. These findings highlight a peripheral gMDSC-microglia communication axis mediated by IL-1β signaling in females with GBM and indicate that expression differences in MDSC subsets represent opportunities for improved immunotherapy efficacy while accounting for sex as a biological variable.

scholarly journals Karakter Jamur Ceratocystis sp. Penyebab Penyakit Busuk Batang pada Acacia decurrens dan Status Penyakitnya di Taman Nasional Gunung Merapi, Yogyakarta

2016 ◽  
Vol 9 (2) ◽  
pp. 94
Author(s):  
Sri Rahayu ◽  
Handojo Hadi Nurjanto ◽  
Rahman Gilang Pratama
Keyword(s):  
Disease Severity ◽  
Morphological Feature ◽  
National Park ◽  
Disease Incidence ◽  
Severe Disease ◽  
Stem Rot ◽  
In Vitro Growth ◽  
The Status ◽  
Rot Disease

Acacia decurrens merupakan salah satu jenis tanaman yang tumbuh mendominasi kawasan Taman Nasional Gunung Merapi (TNGM), pasca erupsi Gunung Merapi tahun 2010. Sekitar 80% tegakan A. decurrens di kawasan tersebut menunjukkan gejala busuk batang akibat infeksi jamur Ceratocystis sp. yang umumnya dipicu oleh luka gerekan kumbang dari kelompok ambrosia. Penelitian bertujuan untuk : (1) mendeskripsikan karakter morfologi jamur Ceratocystis sp., serta kemampuannya beradaptasi pada beberapa jenis tanaman hutan, (2) mengevaluasi status penyakit busuk batang oleh jamur Ceratocystis sp. Karakter morfologi dan kemampuan adaptasinya pada inang akasia, melina, jabon, sengon, dan jati dilakukan di Laboratorium Perlindungan dan Kesehatan Hutan, Fakultas Kehutanan UGM. Survei untuk evaluasi status penyakit busuk batang dilakukan pada bulan Februari sampai Agustus 2014 di demplot restorasi pasca erupsi Merapi (luas 8,4 ha), dengan intensitas sampling 8%. Berdasarkan karakter morfologi, terdapat 2 isolat jamur Ceratocystis sp. yaitu asal lembah (L) dan dari bukit (B) dengan warna koloni krem, luas koloni 20-22 cm2 pada umur 14 hari, membentuk konidia menyerupai tong, dan silindris. Sifat lainnya yaitu memiliki kemampuan yang sama untuk tumbuh, mengkolonisasi, dan menginfeksi inang akasia, sengon, jabon, dan melina, tetapi tidak mampu tumbuh pada inang jati. Berdasarkan luas serangan, status penyakit busuk batang berkisar antara sangat umum sampai menyebar luas (luas serangan = 54-100%), dengan tingkat keparahan bekisar antara ringan sampai parah (intensitas penyakit = 15-67%).Kata kunci: Ceratocystis sp., Acacia decurrens, luas serangan, intensitas penyakit, Taman Nasional Gunung Merapi. Characteristic of stem rot diseases caused by Ceratocystis sp. on Acacia decurrens and its status in Gunung Merapi National Park, YogyakartaAbstractMount Merapi National Park (TNGM) has been dominated by Acacia decurrens after the eruption in 2010. Almost 80% of A. decurrens trees showed stem rot diseases caused by Ceratocystis sp. which may associate with stem wound induced by ambrosia beetle and other physical injuries. The research objective were (1) to characterize the morphological feature, in vitro growth, and ability to adapt, colonize as well as to infect akasia, jabon, sengon, melina and jati sedlings, (2) to evaluate the status of stem rot disease in TNGM demonstration plot. Laboratory work was conducted in order to study the morphological feature of the fungus, in vitro growth on PDA media, and to evaluate its compatibility to growth, colonize, and infect on 5th month seedling of akasia, sengon, jati, jabon and melina. Field monitorings were conducted from February to August 2014 at the restoration plot (8.4 ha) at 8% sampling intensity. Disease status was evaluated in order to ascertain the disease incidence and severity of stem rot disease at the demonstration plot area. Two Ceratocistys isolates found from the hill (B) and valley (L) had similar characteristics on morphologic features i.e. cream color, 20 to 22 cm2 colony size at 14 days growth in PDA media, having both cylindrical and barrel shaped conidia. The other characteristics of the Ceratocistys were an ability to growth, to colonize, and to infect akasia, sengon, melina and jabon, except on jati. The status of stem rot disease was ranged from highly common to widespread (disease incidence = 54%-100%) as the disease severity status was ranged from low to severe (disease severity = 15%-67%).

scholarly journals Circulatory neutrophils exhibit enhanced neutrophil extracellular trap formation in early puerperium: NETs at the nexus of thrombosis and immunity?

2021 ◽  
Author(s):  
Stavros Giaglis ◽  
Chanchal Sur Chowdhury ◽  
Shane Vontelin Van Breda ◽  
Maria Stoikou ◽  
Guenther Shaefer ◽  
...  
Keyword(s):  
Serum Markers ◽  
Late Gestation ◽  
Neutrophil Extracellular Trap ◽  
Coagulation Cascade ◽  
Physiological Alterations ◽  
Fatal Hemorrhage ◽  
Circulating Microparticles ◽  
Neutrophil Degranulation ◽  
Extracellular Trap

Pregnancy is associated with elevated maternal levels of cell-free DNA of neutrophil extracellular trap (NET) origin, as circulatory neutrophils exhibit increased spontaneous NET formation in vitro, mainly driven by G-CSF and finely modulated by sex hormones. The postpartum period, on the other hand, involves physiological alterations consistent with the need for protection against infections and fatal hemorrhage. Our findings indicate that all relevant serum markers of neutrophil degranulation and NET release are substantially augmented postpartum. Neutrophil pro-NETotic activity in vitro is also upregulated particularly in post-delivery neutrophils. Moreover, maternal puerperal neutrophils exhibit a strong pro-NETotic phenotype, associated with increased levels of all key players in the generation of NETs - citH3, MPO, NE, and ROS, compared to non-pregnant and pregnant controls. Intriguingly, post-delivery NET formation is independent of G-CSF in contrast to late gestation and complemented by the presence of TF on the NETs, alterations in the platelet activation status, and activation of the coagulation cascade, triggered by circulating microparticles. Taken together, our results reveal the highly pro-NETotic and potentially procoagulant nature of postpartum neutrophils, bridging an overt immune activation with possible harmful thrombotic incidence.

scholarly journals CMT disease severity correlates with mutation-induced open conformation of histidyl-tRNA synthetase, not aminoacylation loss, in patient cells

2019 ◽  
Vol 116 (39) ◽  
pp. 19440-19448 ◽  
Author(s):  
David Blocquel ◽  
Litao Sun ◽  
Zaneta Matuszek ◽  
Sheng Li ◽  
Thomas Weber ◽  
...  
Keyword(s):  
Disease Severity ◽  
Severe Disease ◽  
Trna Synthetase ◽  
Disease Phenotype ◽  
Mild Disease ◽  
Open Conformation ◽  
Activity Loss ◽  
Aminoacyl Transfer

Aminoacyl-transfer RNA (tRNA) synthetases (aaRSs) are the largest protein family causatively linked to neurodegenerative Charcot–Marie–Tooth (CMT) disease. Dominant mutations cause the disease, and studies of CMT disease-causing mutant glycyl-tRNA synthetase (GlyRS) and tyrosyl-tRNA synthetase (TyrRS) showed their mutations create neomorphic structures consistent with a gain-of-function mechanism. In contrast, based on a haploid yeast model, loss of aminoacylation function was reported for CMT disease mutants in histidyl-tRNA synthetase (HisRS). However, neither that nor prior work of any CMT disease-causing aaRS investigated the aminoacylation status of tRNAs in the cellular milieu of actual patients. Using an assay that interrogated aminoacylation levels in patient cells, we investigated a HisRS-linked CMT disease family with the most severe disease phenotype. Strikingly, no difference in charged tRNA levels between normal and diseased family members was found. In confirmation, recombinant versions of 4 other HisRS CMT disease-causing mutants showed no correlation between activity loss in vitro and severity of phenotype in vivo. Indeed, a mutation having the most detrimental impact on activity was associated with a mild disease phenotype. In further work, using 3 independent biophysical analyses, structural opening (relaxation) of mutant HisRSs at the dimer interface best correlated with disease severity. In fact, the HisRS mutation in the severely afflicted patient family caused the largest degree of structural relaxation. These data suggest that HisRS-linked CMT disease arises from open conformation-induced mechanisms distinct from loss of aminoacylation.

scholarly journals Circulating Neutrophil Extracellular Traps and Neutrophil Activation Are Increased in Proportion to Disease Severity in Human Malaria

2018 ◽  
Vol 219 (12) ◽  
pp. 1994-2004 ◽  
Author(s):  
Steven Kho ◽  
Gabriela Minigo ◽  
Benediktus Andries ◽  
Leo Leonardo ◽  
Pak Prayoga ◽  
...  
Keyword(s):  
Severe Malaria ◽  
Falciparum Malaria ◽  
Disease Severity ◽  
Severe Disease ◽  
Plasmodium Species ◽  
Neutrophil Extracellular Traps ◽  
Falciparum Infection ◽  
Neutrophil Activation ◽  
Extracellular Traps

AbstractBackgroundNeutrophil activation results in Plasmodium parasite killing in vitro, but neutrophil products including neutrophil extracellular traps (NETs) mediate host organ damage and may contribute to severe malaria. The role of NETs in the pathogenesis of severe malaria has not been examined.MethodsIn Papua, Indonesia, we enrolled adults with symptomatic Plasmodium falciparum (n = 47 uncomplicated, n = 8 severe), Plasmodium vivax (n = 37), or Plasmodium malariae (n = 14) malaria; asymptomatic P falciparum (n = 19) or P vivax (n = 21) parasitemia; and healthy adults (n = 23) without parasitemia. Neutrophil activation and NETs were quantified by immunoassays and microscopy and correlated with parasite biomass and disease severity.ResultsIn patients with symptomatic malaria, neutrophil activation and NET counts were increased in all 3 Plasmodium species. In falciparum malaria, neutrophil activation and NET counts positively correlated with parasite biomass (Spearman rho = 0.41, P = .005 and r2 = 0.26, P = .002, respectively) and were significantly increased in severe disease. In contrast, NETs were inversely associated with parasitemia in adults with asymptomatic P falciparum infection (r2 = 0.24, P = .031) but not asymptomatic P vivax infection.ConclusionsAlthough NETs may inhibit parasite growth in asymptomatic P falciparum infection, neutrophil activation and NET release may contribute to pathogenesis in severe falciparum malaria. Agents with potential to attenuate these processes should be evaluated.

scholarly journals ZRC3308 monoclonal antibody cocktail shows protective efficacy in Syrian hamsters against SARS-CoV-2 infection

2021 ◽  
Author(s):  
Pragya Yadav ◽  
Sanjeev Kumar Mendiratta ◽  
Sreelekshmy Mohandas ◽  
Arun K Singh ◽  
Priya Abraham ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Protective Efficacy ◽  
Severe Disease ◽  
Binding Affinities ◽  
Immune Effector ◽  
Syrian Hamsters ◽  
Effector Functions ◽  
Prophylactic Use ◽  
Antibody Cocktail

We have developed a monoclonal antibody (mAb) cocktail (ZRC-3308) comprising of ZRC3308-A7 and ZRC3308-B10 in the ratio 1:1 for COVID-19 treatment. The mAbs were designed to have reduced immune effector functions and increased circulation half-life. mAbs showed good binding affinities to non-competing epitopes on RBD of SARS-CoV-2 spike protein and were found neutralizing SARS-CoV-2 variants B.1, B.1.1.7, B.1.351, B.1.617.2 and B.1.617.2 AY.1 in vitro. The mAb cocktail demonstrated effective prophylactic and therapeutic activity against SARS-CoV-2 infection in Syrian hamsters. The antibody cocktail appears to be a promising candidate for the prophylactic use and for therapy in early COVID-19 cases which have not progressed to severe disease.

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