scholarly journals Overlapping but Disparate Inflammatory and Immunosuppressive Responses to SARS-CoV-2 and Bacterial Sepsis: An Immunological Time Course Analysis

2021 ◽  
Vol 12 ◽  
Author(s):  
Tyler J. Loftus ◽  
Ricardo Ungaro ◽  
Marvin Dirain ◽  
Philip A. Efron ◽  
Monty B. Mazer ◽  
...  
Keyword(s):  
Clinical Outcomes ◽  
Immune Suppression ◽  
Bacterial Infections ◽  
Time Course ◽  
Ex Vivo ◽  
Bacterial Sepsis ◽  
Ifn Gamma ◽  
Hla Dr ◽  
Secondary Infections ◽  
Biomarker Analysis

Both severe SARS-CoV-2 infections and bacterial sepsis exhibit an immunological dyscrasia and propensity for secondary infections. The nature of the immunological dyscrasias for these differing etiologies and their time course remain unclear. In this study, thirty hospitalized patients with SARS-CoV-2 infection were compared with ten critically ill patients with bacterial sepsis over 21 days, as well as ten healthy control subjects. Blood was sampled between days 1 and 21 after admission for targeted plasma biomarker analysis, cellular phenotyping, and leukocyte functional analysis via enzyme-linked immunospot assay. We found that circulating inflammatory markers were significantly higher early after bacterial sepsis compared with SARS-CoV-2. Both cohorts exhibited profound immune suppression through 21 days (suppressed HLA-DR expression, reduced mononuclear cell IFN-gamma production), and expanded numbers of myeloid-derived suppressor cells (MDSCs). In addition, MDSC expansion and ex vivo production of IFN-gamma and TNF-alpha were resolving over time in bacterial sepsis, whereas in SARS-CoV-2, immunosuppression and inflammation were accelerating. Despite less severe initial physiologic derangement, SARS-CoV-2 patients had similar incidence of secondary infections (23% vs 30%) as bacterial sepsis patients. Finally, COVID patients who developed secondary bacterial infections exhibited profound immunosuppression evident by elevated sPD-L1 and depressed HLA-DR. Although both bacterial sepsis and SARS-CoV-2 are associated with inflammation and immune suppression, their immune dyscrasia temporal patterns and clinical outcomes are different. SARS-CoV-2 patients had less severe early inflammation and organ dysfunction but had persistent inflammation and immunosuppression and suffered worse clinical outcomes, especially when SARS-CoV-2 infection was followed by secondary bacterial infection.

scholarly journals 1758. Impact of Accelerate Pheno™ Rapid Blood Culture Detection System on Laboratory and Clinical Outcomes in Bacteremic Patients

2018 ◽  
Vol 5 (suppl_1) ◽  
pp. S61-S61 ◽  
Author(s):  
Ryan Dare ◽  
Kelsey McCain ◽  
Katherine Lusardi ◽  
Kay Daniels ◽  
Jacob Painter ◽  
...  
Keyword(s):  
Blood Culture ◽  
Clinical Outcomes ◽  
Bacterial Infections ◽  
Detection System ◽  
Positive Culture ◽  
Retrospective Chart Review ◽  
Improve Patient Care ◽  
Standard Of Care ◽  
Historical Cohort ◽  
Stewardship Program

Abstract Background Molecular-based automated systems for the rapid diagnosis of bacterial infections have potential to improve patient care. The Accelerate Pheno™ blood culture detection system (ACCEL) is an FDA approved platform that allows for identification (ID) and antimicrobial susceptibility testing (AST) 8 hours following growth in routine culture. Methods This is a single-center retrospective chart review of bacteremic adult inpatients before and after implementation of ACCEL. Laboratory and clinical data were collected February–March 2018 (intervention) and compared with a January–April 2017 historical cohort (standard of care). Standard of care ID and AST were performed using VITEK® MS (MALDI-TOF MS) and VITEK®2, respectively. An active antimicrobial stewardship program was in place during both study periods. Patients with polymicrobial cultures, off-panel isolates, previous positive culture, or who were discharged prior to final AST report were excluded. Primary outcome was length of stay (LOS). Secondary outcomes were inpatient antibiotic duration of therapy (DOT) and time to optimal therapy (TTOT). Nonparametric unadjusted analyses were performed due to non-normal distributions. Statistics were performed using SAS 9.4. Results Of the 143 positive cultures performed on ACCEL during intervention, 118 (83%) were identified as on-panel organisms. Seventy-five (64%) of these 118 cultures and 79 (70%) of 113 reviewed standard of care cultures met inclusion criteria. Patient comorbidities (P = NS), MEWS severity score (P = 0.10), source of bacteremia (P = NS), and pathogen detected (P = 0.30) were similar between cohorts. Time from collection to ID (28.2 ± 12.7 hours vs. 53.8 ± 20.9 hours; P < 0.001) and AST (31.9 ± 11 hours vs. 71.8 ± 20 hours; P < 0.001) were shorter in the intervention arm. Conclusion Compared with standard of care, ACCEL shortens laboratory turn-around-time and improves clinical outcomes. The use of this system has resulted in decreased mean antibiotic DOT, TTOT, and LOS. Further studies are needed to verify these findings. Disclosures All authors: No reported disclosures.

scholarly journals Uptake and Distribution of Administered Bone Marrow Mesenchymal Stem Cell Extracellular Vesicles in Retina

Cells ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 730
Author(s):  
Biji Mathew ◽  
Leianne A. Torres ◽  
Lorea Gamboa Gamboa Acha ◽  
Sophie Tran ◽  
Alice Liu ◽  
...  
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Extracellular Vesicles ◽  
Time Course ◽  
Ganglion Cells ◽  
Ex Vivo ◽  
Dependent Manner ◽  
Paracrine Effects

Cell replacement therapy using mesenchymal (MSC) and other stem cells has been evaluated for diabetic retinopathy and glaucoma. This approach has significant limitations, including few cells integrated, aberrant growth, and surgical complications. Mesenchymal Stem Cell Exosomes/Extracellular Vesicles (MSC EVs), which include exosomes and microvesicles, are an emerging alternative, promoting immunomodulation, repair, and regeneration by mediating MSC’s paracrine effects. For the clinical translation of EV therapy, it is important to determine the cellular destination and time course of EV uptake in the retina following administration. Here, we tested the cellular fate of EVs using in vivo rat retinas, ex vivo retinal explant, and primary retinal cells. Intravitreally administered fluorescent EVs were rapidly cleared from the vitreous. Retinal ganglion cells (RGCs) had maximal EV fluorescence at 14 days post administration, and microglia at 7 days. Both in vivo and in the explant model, most EVs were no deeper than the inner nuclear layer. Retinal astrocytes, microglia, and mixed neurons in vitro endocytosed EVs in a dose-dependent manner. Thus, our results indicate that intravitreal EVs are suited for the treatment of retinal diseases affecting the inner retina. Modification of the EV surface should be considered for maintaining EVs in the vitreous for prolonged delivery.

scholarly journals Cytokines in chronic inflammatory arthritis. IV. Granulocyte/macrophage colony-stimulating factor-mediated induction of class II MHC antigen on human monocytes: a possible role in rheumatoid arthritis.

1989 ◽  
Vol 170 (3) ◽  
pp. 865-875 ◽  
Author(s):  
J M Alvaro-Gracia ◽  
N J Zvaifler ◽  
G S Firestein
Keyword(s):  
Rheumatoid Arthritis ◽  
Class Ii ◽  
Tnf Alpha ◽  
Blood Monocytes ◽  
Ifn Gamma ◽  
Normal Peripheral Blood ◽  
Gm Csf ◽  
Hla Dr ◽  
Class Ii Mhc ◽  
Hla Dq

Granulocyte/macrophage CSF (GM-CSF) has recently been identified in rheumatoid arthritis (RA) synovial effusions. To study a potential role for GM-CSF and other cytokines on the induction of HLA-DR expression on monocytes and synovial macrophages, we analyzed the relative ability of recombinant human cytokines to induce the surface expression of class II MHC antigens on normal peripheral blood monocytes by FACS analysis. GM-CSF (800 U/ml) (mean fluorescence channel 2.54 +/- 0.33 times the control, p less than 0.001) and IFN-gamma (100 U/ml) (5.14 +/- 0.60, p less than 0.001) were the most potent inducers of HLA-DR. TNF-alpha and IL-4 also increased HLA-DR expression, although to a lesser degree [1.31 +/- 0.06 (p less than 0.02) and 1.20 +/- 0.03 (p less than 0.01), respectively]. IL-1 (40 U/ml), IL-2 (10 ng/ml), IL-3 (50 U/ml), IL-6 (100 U/ml), and CSF-1 (1,000 U/ml) did not affect surface HLA-DR density. GM-CSF also increased HLA-DR mRNA expression and surface HLA-DQ expression, but decreased CD14 (a monocyte/macrophage antigen) expression. The effect of GM-CSF on HLA-DR was not mediated by the generation of IFN-gamma in vitro because it was not blocked by anti-IFN-gamma mAb. GM-CSF was additive with IL-4 and low amounts (less than 3 U/ml) of IFN-gamma and synergistic with TNF-alpha. Because we have recently reported that supernatants of cultured RA synovial cells produce a non-IFN-gamma factor that induces HLA-DR on monocytes, we then attempted to neutralize this factor with specific anti-GM-CSF mAb. Four separate synovial tissue supernatants were studied, and the antibody neutralized the HLA-DR-inducing factor in each (p less than 0.01).

scholarly journals Differential effects of gram-positive and gram-negative bacterial products on morphine induced inhibition of phagocytosis

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Jana Ninkovic ◽  
Vidhu Anand ◽  
Raini Dutta ◽  
Li Zhang ◽  
Anuj Saluja ◽  
...  
Keyword(s):  
Bacterial Infections ◽  
Drug Abusers ◽  
Bacterial Clearance ◽  
Chronic Morphine ◽  
Gram Positive ◽  
Bacterial Killing ◽  
Differential Effects ◽  
Secondary Infections ◽  
First Time ◽  
Dependent Signaling Pathway

Abstract Opioid drug abusers have a greater susceptibility to gram positive (Gram (+)) bacterial infections. However, the mechanism underlying opioid modulation of Gram (+) versus Gram (−) bacterial clearance has not been investigated. In this study, we show that opioid treatment resulted in reduced phagocytosis of Gram (+), when compared to Gram (−) bacteria. We further established that LPS priming of chronic morphine treated macrophages leads to potentiated phagocytosis and killing of both Gram (+) and Gram (−) bacteria in a P-38 MAP kinase dependent signaling pathway. In contrast, LTA priming lead to inhibition of both phagocytosis and bacterial killing. This study demonstrates for the first time the differential effects of TLR4 and TLR2 agonists on morphine induced inhibition of phagocytosis. Our results suggest that the incidence and severity of secondary infections with Gram (+) bacteria would be higher in opioid abusers.

scholarly journals Suitability of a Progenitor Cell-Enriching Device for In Vitro Applications

Coatings ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 146
Author(s):  
Antonio Celentano ◽  
Tami Yap ◽  
Giuseppe Pantaleo ◽  
Rita Paolini ◽  
Michael McCullough ◽  
...  
Keyword(s):  
Cell Viability ◽  
Time Course ◽  
Ex Vivo ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Test ◽  
Initial Reduction ◽  
Metal Wear ◽  
Class 1 ◽  
Electron Energy Loss

Rigenera® is a novel class-1 medical device that produces micro-grafts enriched of progenitors cells without ex vivo manipulation of donor tissues. The manufacturer’s protocol has been supported for a wide variety of clinical uses in the field of regenerative medicine. This study aimed to evaluate its potential use for in vitro cell models. Human primary oral fibroblasts were cultured under standard conditions and processed through Rigenera® over a time course of up to 5 min. Cell viability was assessed using a Trypan Blue exclusion test. It is possible to process fibroblasts through Rigenera® although an initial reduction of cell viability was observed. Additionally, debris was evident in the cell suspension of the processed samples. Scanning electron microscopy (SEM) microanalysis of the debris and electron energy-loss spectroscopy confirmed the presence of metal wear possibly due to the processing conditions used in this study. Interestingly, pore sizes within Rigeneracons® grids were found to range between 250–400 μm. This is the first report assessing the suitability of Rigenera® and Rigeneracons® for in vitro applications. Whilst Rigenera® workflow was found to be amenable to laboratory uses, our results strongly suggest that further research and development is necessary to support the utilization of this technology for enrichment of micro-graft derived cells and cell sorting in vitro.

MO526REAL-WORLD EVIDENCE ON CLINICAL OUTCOMES IN NON-DIABETIC CKD

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Christoph Wanner ◽  
Johannes Schuchhardt ◽  
Chris Bauer ◽  
Stefanie Lindemann ◽  
Meike Brinker ◽  
...  
Keyword(s):  
Clinical Outcomes ◽  
Kidney Failure ◽  
Time Course ◽  
Kidney Injury ◽  
Incidence Rates ◽  
Real World Evidence ◽  
Ckd Stage ◽  
The Us ◽  
Egfr Decline

Abstract Background and Aims Chronic kidney disease (CKD) represents a global public health problem, with significant morbidity and mortality due to cardiovascular disease during CKD progression and due to kidney failure. Although non-diabetic CKD accounts for up to 70% of the global CKD burden, its clinical consequences are poorly understood, and data are needed to help identify individuals at high risk of adverse outcomes. This analysis uses real-world evidence to provide insights into clinical characteristics, care and outcomes in individuals with non-diabetic CKD in routine clinical practice. Method Individual-level data from the US administrative claims database, Optum Clinformatics Data Mart, from January 1, 2008 to December 31, 2018 were analysed. Adults with non-diabetic CKD stage 3 or 4 and ≥365 days continuous insurance coverage were included and followed until insurance disenrollment, end of data availability or death. Individuals with diabetes mellitus, CKD stage 5 or end-stage kidney disease (ESKD) prior to the index date, or who experienced kidney failure (acute or unspecified), kidney transplant or dialysis in the baseline period, were excluded from the analysis. Study outcomes, captured in the database, were defined using common clinical coding systems. Primary outcomes were hospitalisation for heart failure (HHF), a kidney composite of ESKD/kidney failure/need for dialysis, and worsening of CKD stage from baseline. Individual CKD stage was assigned based on estimated glomerular filtration rate (eGFR) values (priority) or the respective International Classification of Diseases code at index and during follow-up. Further prespecified kidney outcomes included individual components of the kidney composite, acute kidney injury, and absolute and relative change in eGFR from baseline. Event-based outcomes were assessed by time-to-first-event analysis. Summary statistics for time-course analysis of metric outcomes were generated on a quarterly basis. Results In total, 504,924 of 64 million individuals in the Optum Clinformatics Data Mart satisfied the selection criteria. Over a median follow-up of 744 (interquartile range 328–1432) days, the incidence rates of primary outcomes of HHF, the kidney composite and worsening of CKD stage from baseline were 3.95, 10.33 and 4.38 events/100 patient-years (PY), respectively. The incidence rates of the components of the kidney composite outcome, namely ESKD/need for dialysis, kidney failure (acute and unspecified) and need for dialysis were 1.78, 9.53 and 0.49 events/100 PY, respectively. Kidney failure events were driven mainly by acute kidney injury, with an incidence of 8.61 events/100 PY. In individuals with at least one available eGFR value at baseline and one value during follow-up (n=295,174), the incidence rates of relative decreases in eGFR of ≥30%, ≥40% and ≥57% from baseline were 1.98, 0.97 and 0.30 events/100 PY, respectively; in this cohort, more rapid eGFR decline was associated with increased risk of HHF and the kidney composite outcome. In individuals with a baseline eGFR value and at least one follow-up eGFR value and an available urine albumin-to-creatinine ratio (n=25,824), time-course analysis of eGFR showed that eGFR decline mostly occurred in individuals with moderately-to-severely increased albuminuria (≥30 mg/g). Conclusion This analysis generates real-world evidence on clinical outcomes in a cohort of individuals with non-diabetic CKD treated in routine clinical practice in the US. Despite known limitations of claims databases (e.g. low availability of some laboratory data, limited individual follow-up time and tactical coding), individuals with moderate-to-severe non-diabetic CKD are shown to be at high risk of serious clinical outcomes. This highlights the high unmet medical need, and urgency for new treatments and targeted interventions for patients with non-diabetic CKD.

Early circulating tumor (ct) DNA dynamics and efficacy of lorlatinib: Analysis from the CROWN study.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. 9011-9011
Author(s):  
Ross A. Soo ◽  
Jean-Francois Martini ◽  
Anthonie J. van der Wekken ◽  
Shunsuke Teraoka ◽  
Alice T. Shaw ◽  
...  
Keyword(s):  
Clinical Outcomes ◽  
Kinase Inhibitor ◽  
Progression Free Survival ◽  
Molecular Response ◽  
Similar Response ◽  
Dna Dynamics ◽  
Paired Samples ◽  
Biomarker Analysis ◽  
Alk Positive ◽  
Ctdna Analysis

9011 Background: Lorlatinib, a third-generation ALK tyrosine kinase inhibitor, significantly improved progression-free survival (PFS) and overall/intracranial responses vs crizotinib in patients (pts) with previously untreated ALK-positive advanced non-small cell lung cancer (NSCLC) in the ongoing randomized Phase 3 CROWN study (NCT03052608). To identify whether additional molecular biomarker analysis correlated with efficacy, we evaluated early ctDNA dynamics compared with clinical outcomes. Methods: Plasma samples were prospectively collected at screening (SC), week 4 (cycle 2, day 1 [C2D1]), week 24 (C7D1), and end of treatment for ctDNA analysis. ctDNA was analyzed using Guardant360CDx (Guardant Health, Inc., Redwood City, CA, USA). Mean variant allele fraction (VAF) of ALK alterations (fusions and/or mutations) and overall detected alterations at each time point and longitudinal mean change (dVAF) as (VAFC2D1) – (VAFSC) were calculated; dVAF <0 indicated decreased ctDNA at week 4. Objective tumor response and PFS were evaluated according to dVAF. These analyses were repeated vs ctDNA results at week 24. Additional correlation analyses between depth of molecular response and/or ctDNA clearance and clinical outcomes are ongoing. Results: Paired samples were available at SC and week 4 from 232 of 255 pts included in the ctDNA analysis: 118/130 (90.8%) in the lorlatinib arm and 114/125 (91.2%) in the crizotinib arm. ALK alterations were detected in 122/232 (52.6%) pts at SC (62/118 [52.5%] from the lorlatinib arm) but only 19/232 (8.2%) at week 4 (8/118 [6.8%] from the lorlatinib arm). Mean VAF of ALK alterations at week 4 was significantly decreased compared with SC in both treatment arms (lorlatinib -1.54, crizotinib -1.25; both P<0.0001; P=0.4239 between arms). In the lorlatinib arm, mean VAF at week 4 was significantly decreased compared with SC in pts with a complete or partial response (dVAF -1.53; n=47; P<0.0001), or stable disease (dVAF -1.37; n=12; P=0.0304). Similar results were observed in the crizotinib arm. In pts with dVAF <0 for ALK alterations, mean percent change from screening in tumor size was -40.8% with lorlatinib (n=59) and -38.7% with crizotinib (n=58). Only 2 pts had dVAF ≥0, both from the crizotinib arm. Median PFS for pts with dVAF <0 for ALK alterations was not reached in the lorlatinib arm (n=62), and was 7.4 months (95% CI, 7.2–9.3) in the crizotinib arm (n=58). Similar response and PFS data were observed in the analysis of dVAF for ALK alterations at week 24. Conclusions: Early ctDNA dynamics may predict lorlatinib efficacy in pts with previously untreated ALK-positive NSCLC. The magnitude of reduction in ctDNA at 4 weeks may be associated with better responses and potentially longer PFS. These findings further support the utility of dynamic ctDNA monitoring in ALK-positive NSCLC. Reference: Shaw AT, et al. N Engl J Med. 2020;383:2018-2029. Clinical trial information: NCT03052608.

Interferon-gamma downregulates CFTR gene expression in epithelial cells

1994 ◽  
Vol 267 (5) ◽  
pp. C1398-C1404 ◽  
Author(s):  
F. Besancon ◽  
G. Przewlocki ◽  
I. Baro ◽  
A. S. Hongre ◽  
D. Escande ◽  
...  
Keyword(s):  
Gene Expression ◽  
Interferon Gamma ◽  
Posttranscriptional Regulation ◽  
Bacterial Infections ◽  
Cftr Gene ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Necrosis Factor Alpha ◽  
Ifn Gamma ◽  
Cl Transport

Cystic fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR) gene, resulting in defective transepithelial Cl- transport. The regulation of CF gene expression is not fully understood. We report that interferon-gamma (IFN-gamma), but not IFN-alpha or -beta, downregulates CFTR mRNA levels in two colon-derived epithelial cell lines, HT-29 and T84, in a time- and concentration (from 0.1 IU/ml)-dependent manner. IFN-gamma has no effect on the transcription rate of the CFTR gene but reduces CFTR mRNA half-life, indicating that it exerts a posttranscriptional regulation of CFTR expression, at least partly, through destabilization of the transcripts. Cells treated with IFN-gamma contain subnormal amounts of 165-kDa CFTR protein. Assays of adenosine 3',5'-cyclic monophosphate-stimulated 36Cl- efflux and whole cell currents show that CFTR function is diminished in IFN-gamma-treated cells. IFN-gamma and tumor necrosis factor-alpha synergistically reduce CFTR gene expression. Our results suggest that production of these cytokines in response to bacterial infections and inflammatory disorders may alter transmembrane Cl- transport.

scholarly journals Randomised controlled trial of GM-CSF in critically ill patients with impaired neutrophil phagocytosis

Thorax ◽  
2018 ◽  
Vol 73 (10) ◽  
pp. 918-925 ◽  
Author(s):  
Emma M Pinder ◽  
Anthony J Rostron ◽  
Thomas P Hellyer ◽  
Marie-Helene Ruchaud-Sparagano ◽  
Jonathan Scott ◽  
...  
Keyword(s):  
Critically Ill ◽  
Critically Ill Patients ◽  
Ex Vivo ◽  
Controlled Trial ◽  
Personalised Medicine ◽  
Common Adverse Event ◽  
Controlled Clinical Trial ◽  
Gm Csf ◽  
Hla Dr ◽  
Increased Risk

BackgroundCritically ill patients with impaired neutrophil phagocytosis have significantly increased risk of nosocomial infection. Granulocyte-macrophage colony-stimulating factor (GM-CSF) improves phagocytosis by neutrophils ex vivo. This study tested the hypothesis that GM-CSF improves neutrophil phagocytosis in critically ill patients in whom phagocytosis is known to be impaired.MethodsThis was a multicentre, phase IIa randomised, placebo-controlled clinical trial. Using a personalised medicine approach, only critically ill patients with impaired neutrophil phagocytosis were included. Patients were randomised 1:1 to subcutaneous GM-CSF (3 μg/kg/day) or placebo, once daily for 4 days. The primary outcome measure was neutrophil phagocytosis 2 days after initiation of GM-CSF. Secondary outcomes included neutrophil phagocytosis over time, neutrophil functions other than phagocytosis, monocyte HLA-DR expression and safety.ResultsThirty-eight patients were recruited from five intensive care units (17 randomised to GM-CSF). Mean neutrophil phagocytosis at day 2 was 57.2% (SD 13.2%) in the GM-CSF group and 49.8% (13.4%) in the placebo group, p=0.73. The proportion of patients with neutrophil phagocytosis≥50% at day 2, and monocyte HLA-DR, appeared significantly higher in the GM-CSF group. Neutrophil functions other than phagocytosis did not appear significantly different between the groups. The most common adverse event associated with GM-CSF was fever.ConclusionsGM-CSF did not improve mean neutrophil phagocytosis at day 2, but was safe and appeared to increase the proportion of patients with adequate phagocytosis. The study suggests proof of principle for a pharmacological effect on neutrophil function in a subset of critically ill patients.

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