Distinct immune signatures discriminate between asymptomatic and presymptomatic SARS-CoV-2pos subjects

AbstractIncreasing numbers of SARS-CoV-2-positive (SARS-CoV-2pos) subjects are detected at silent SARS-CoV-2 infection stage (SSIS). Yet, SSIS represents a poorly examined time-window wherein unknown immunity patterns may contribute to the fate determination towards persistently asymptomatic or overt disease. Here, we retrieved blood samples from 19 asymptomatic and 12 presymptomatic SARS-CoV-2pos subjects, 47 age/gender-matched patients with mild or moderate COVID-19 and 27 normal subjects, and interrogated them with combined assays of 44-plex CyTOF, RNA-seq and Olink. Notably, both asymptomatic and presymptomatic subjects exhibited numerous readily detectable immunological alterations, while certain parameters including more severely decreased frequencies of CD107alow classical monocytes, intermediate monocytes, non-classical monocytes and CD62Lhi CD8+ Tnaïve cells, reduced plasma STC1 level but an increased frequency of CD4+ NKT cells combined to distinguish the latter. Intercorrelation analyses revealed a particular presymptomatic immunotype mainly manifesting as monocytic overactivation and differentiation blockage, a likely lymphocyte exhaustion and immunosuppression, yielding mechanistic insights into SSIS fate determination, which could potentially improve SARS-CoV-2 management.

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Monocyte subsets predict mortality after cardiac arrest

European Heart Journal ◽

10.1093/ehjci/ehaa946.1855 ◽

2020 ◽

Vol 41 (Supplement_2) ◽

Author(s):

K.A Krychtiuk ◽

M Lenz ◽

B Richter ◽

K Huber ◽

J Wojta ◽

...

Keyword(s):

Cardiac Arrest ◽

Spontaneous Circulation ◽

Strong Increase ◽

Funding Source ◽

Monocyte Subset ◽

Male Mortality ◽

National Budget ◽

Classical Monocytes ◽

Intermediate Monocytes ◽

Subset Distribution

Abstract Background After successful cardiopulmonary resuscitation with return of spontaneous circulation (ROSC), many patients show signs of an overactive immune activation. Monocytes are a heterogenous cell population that can be distinguished into three subsets. Purpose The aim of this prospective, observational study was to analyze whether monocyte subset distribution is associated with mortality at 6 months in patients after cardiac arrest. Methods We included 53 patients admitted to our medical ICU after cardiac arrest. Blood was taken on admission and monocyte subset distribution was analyzed by flow cytometry and distinguished into classical monocytes (CM; CD14++CD16-), intermediate monocytes (IM; CD14++CD16+CCR2+) and non-classical monocytes (NCM; CD14+CD16++CCR2-). Results Median age was 64.5 (IQR 49.8–74.3) years and 75.5% of patients were male. Mortality at 6 months was 50.9% and survival with good neurological outcome was 37.7%. Of interest, monocyte subset distribution upon admission to the ICU did not differ according to survival. However, patients that died within 6 months showed a strong increase in the pro-inflammatory subset of intermediate monocytes (8.3% (3.8–14.6)% vs. 4.1% (1.5–8.2)%; p=0.025), and a decrease of classical monocytes (87.5% (79.9–89.0)% vs. 90.8% (85.9–92.7)%; p=0.036) 72 hours after admission. In addition, intermediate monocytes were predictive of outcome independent of initial rhythm and time to ROSC and correlated with the CPC-score at 6 months (R=0.32; p=0.043). Discussion Monocyte subset distribution is associated with outcome in patients surviving a cardiac arrest. This suggests that activation of the innate immune system may play a significant role in patient outcome after cardiac arrest. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): FWF - Fonds zur Förderung der wissenschaftlichen Forschung

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Human T Cell Leukemia Virus Type 1 Infection of the Three Monocyte Subsets Contributes to Viral Burden in Humans

Journal of Virology ◽

10.1128/jvi.02735-15 ◽

2015 ◽

Vol 90 (5) ◽

pp. 2195-2207 ◽

Cited By ~ 28

Author(s):

Maria Fernanda de Castro-Amarante ◽

Cynthia A. Pise-Masison ◽

Katherine McKinnon ◽

Robyn Washington Parks ◽

Veronica Galli ◽

...

Keyword(s):

T Cells ◽

Leukemia Virus ◽

Receptor Expression ◽

Monocyte Subsets ◽

Viral Dna ◽

Cell Leukemia Virus Type ◽

Human T Cell ◽

T Cell Leukemia Virus ◽

Classical Monocytes ◽

Intermediate Monocytes

ABSTRACTBecause the viral DNA burden correlates with disease development, we investigated the contribution of monocyte subsets (classical, intermediate, and nonclassical monocytes) to the total viral burden in 22 human T cell leukemia virus type 1 (HTLV-1)-infected individuals by assessing their infectivity status, frequency, as well as chemotactic and phagocytic functions. All three monocyte subsets sorted from HTLV-1-infected individuals were positive for viral DNA, and the frequency of classical monocytes was lower in the blood of HTLV-1-infected individuals than in that of uninfected individuals, while the expression levels of the chemokine receptors CCR5, CXCR3, and CX3CR1 in classical monocytes were higher in HTLV-1-infected individuals than uninfected individuals; the percentage of intermediate monocytes and their levels of chemokine receptor expression did not differ between HTLV-1-infected and uninfected individuals. However, the capacity of intermediate monocytes to migrate to CCL5, the ligand for CCR5, was higher, and a higher proportion of nonclassical monocytes expressed CCR1, CXCR3, and CX3CR1. The level of viral DNA in the monocyte subsets correlated with the capacity to migrate to CCL2, CCL5, and CX3CL1 for classical monocytes, with lower levels of phagocytosis for intermediate monocytes, and with the level of viral DNA in CD8+and CD4+T cells for nonclassical monocytes. These data suggest a model whereby HTLV-1 infection augments the number of classical monocytes that migrate to tissues and become infected and the number of infected nonclassical monocytes that transmit virus to CD4+and CD8+T cells. These results, together with prior findings in a macaque model of HTLV-1 infection, support the notion that infection of monocytes by HTLV-1 is likely a requisite for viral persistence in humans.IMPORTANCEMonocytes have been implicated in immune regulation and disease progression in patients with HTLV-1-associated inflammatory diseases. We detected HTLV-1 DNA in all three monocyte subsets and found that infection impacts surface receptor expression, migratory function, and subset frequency. The frequency of nonclassical patrolling monocytes is increased in HTLV-1-infected individuals, and they have increased expression of CCR1, CXCR3, and CX3CR1. The viral DNA level in nonclassical monocytes correlated with the viral DNA level in CD4+and CD8+T cells. Altogether, these data suggest an increased recruitment of classical monocytes to inflammation sites that may result in virus acquisition and, in turn, facilitate virus dissemination and viral persistence. Our findings thus provide new insight into the importance of monocyte infection in viral spread and suggest targeting of monocytes for therapeutic intervention.

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A Two-Cohort RNA-seq Study Reveals Changes in Endometrial and Blood miRNome in Fertile and Infertile Women

Genes ◽

10.3390/genes9120574 ◽

2018 ◽

Vol 9 (12) ◽

pp. 574 ◽

Cited By ~ 5

Author(s):

Kadri Rekker ◽

Signe Altmäe ◽

Marina Suhorutshenko ◽

Maire Peters ◽

Juan F. Martinez-Blanch ◽

...

Keyword(s):

Embryo Implantation ◽

Endometrial Receptivity ◽

Small Rna Sequencing ◽

Rna Seq ◽

Recurrent Implantation Failure ◽

Secretory Phase ◽

Blood Samples ◽

Infertile Women ◽

Secretory Endometrium ◽

Fertile Women

The endometrium undergoes extensive changes to prepare for embryo implantation and microRNAs (miRNAs) have been described as playing a significant role in the regulation of endometrial receptivity. However, there is no consensus about the miRNAs involved in mid-secretory endometrial functions. We analysed the complete endometrial miRNome from early secretory (pre-receptive) and mid-secretory (receptive) phases from fertile women and from patients with recurrent implantation failure (RIF) to reveal differentially expressed (DE) miRNAs in the mid-secretory endometrium. Furthermore, we investigated whether the overall changes during early to mid-secretory phase transition and with RIF condition could be reflected in blood miRNA profiles. In total, 116 endometrial and 114 matched blood samples collected from two different population cohorts were subjected to small RNA sequencing. Among fertile women, 91 DE miRNAs were identified in the mid-secretory vs. early secretory endometrium, while no differences were found in the corresponding blood samples. The comparison of mid-secretory phase samples between fertile and infertile women revealed 21 DE miRNAs from the endometrium and one from blood samples. Among discovered novel miRNAs, chr2_4401 was validated and showed up-regulation in the mid-secretory endometrium. Besides novel findings, we confirmed the involvement of miR-30 and miR-200 family members in mid-secretory endometrial functions.

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Flow cytometric analysis of thiazole orange uptake by platelets: a diagnostic aid in the evaluation of thrombocytopenic disorders

Blood ◽

10.1182/blood.v75.1.116.bloodjournal751116 ◽

1990 ◽

Vol 75 (1) ◽

pp. 116-121 ◽

Cited By ~ 5

Author(s):

J Kienast ◽

G Schmitz

Keyword(s):

Fluorescence Enhancement ◽

Flow Cytometric Analysis ◽

Normal Subjects ◽

Thiazole Orange ◽

High Quantum Yield ◽

Specific Test ◽

Blood Samples ◽

Flow Cytometric ◽

Patient Groups ◽

The Absolute

Thiazole orange (TO), a fluorescent dye originally synthesized for reticulocyte analysis, is characterized by a large fluorescence enhancement and high quantum yield on binding to nucleic acids, particularly RNA. In addition, the dye readily permeates live cell membranes. We applied TO staining, followed by fluorescence-activated flow cytometric analysis, to platelets in whole blood samples from hematologically normal subjects and patients with various quantitative platelet disorders. The percentage of TO-positive platelets in 50 control subjects was 8.6 +/- 2.8% (mean +/- SD) ranging from 2.8% to 15.8%. In 21 thrombocytopenic patients whose bone marrow contained normal to increased numbers of megakaryocytes, the percentage of fluorescently labeled platelets was significantly elevated (P less than .0001) to 26.9 +/- 10.9% (range, 13.3% to 57.1%). In contrast, the proportion of positively stained platelets in 23 patients with thrombocytopenia due to impaired platelet production (various conditions with reduced marrow megakaryocytes) did not significantly differ from the controls, whereas the absolute counts of TO-positive platelets were significantly lowered (P less than .0001). Differences in the distributions of the percentage values as well as of the absolute counts for TO-positive platelets between the two patient groups were again highly significant (P less than .0001). Both the sensitivity and the specificity of this method in distinguishing between these categories of thrombocytopenia were greater than or equal to 95%. We conclude that flow cytometric analysis of platelets after staining with TO is a sensitive and specific test that rapidly provides information on the thrombopoietic activity in thrombocytopenic disorders. Our data further suggest that increased amounts of residual RNA characterize platelets released under conditions of “stress thrombopoiesis.”

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Monocytes of Different Subsets in Complexes with Platelets in Patients with Myocardial Infarction

Thrombosis and Haemostasis ◽

10.1055/s-0038-1673342 ◽

2018 ◽

Vol 118 (11) ◽

pp. 1969-1981 ◽

Cited By ~ 7

Author(s):

Marina Loguinova ◽

Natalia Pinegina ◽

Valeria Kogan ◽

Murad Vagida ◽

Anush Arakelyan ◽

...

Keyword(s):

Myocardial Infarction ◽

Annexin V ◽

Published Data ◽

Healthy Controls ◽

New Information ◽

Platelet Antigen ◽

Classical Monocytes ◽

Intermediate Monocytes

AbstractAcute myocardial infarction (AMI) is associated with activation of various cells, including platelets that form monocyte–platelet complexes (MPCs). Here, we analysed MPC in vivo and in vitro and investigated the abilities of different monocyte subclasses to form MPC, the characteristics of the cells involved in MPC formation and MPC changes in AMI. We identified MPC by co-staining for platelet antigen CD41a and monocyte antigens CD14 and CD16. Platelet activation was evaluated from expression of phosphatidylserine as revealed by annexin V. Our results confirm published data and provide new information regarding the patterns of MPC in AMI patients. We found that the patterns of platelet aggregation with monocytes were different in AMI patients and controls: (1) in AMI patients, MPC formed by intermediate monocytes carry more platelets whereas in healthy controls more platelets aggregated with classical monocytes; (2) the numbers of MPC in AMI patients, being already higher than in controls, were further increased if these patients suffered various in-hospital complications; (3) on the basis of the CD41a fluorescence of the antibody-stained MPC, some of the aggregates seem to consist of monocytes and platelet-derived extracellular vesicles (EVs); (4) aggregation of monocytes with platelet EV occurred in in vitro experiments; and (5) these experiments demonstrated that monocytes from AMI patients aggregate with both platelets and platelet EVs more efficiently than do monocytes from controls. MPC in AMI patients may play an important role in this pathology.

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Blood RNA Sequencing Confirms Upregulated BATF2 and FCGR1A Expression in Children with Autism Spectrum Disorder

10.21203/rs.3.rs-1113898/v1 ◽

2021 ◽

Author(s):

Irena Voinsky ◽

Yazeed Zoabi ◽

Noam Shomron ◽

Moria Harel ◽

Hanoch Cassuto ◽

...

Keyword(s):

Autism Spectrum Disorder ◽

Cancer Risk ◽

Rna Sequencing ◽

Whole Blood ◽

Autism Spectrum ◽

Spectrum Disorder ◽

Rna Seq ◽

Blood Samples ◽

Children With Asd ◽

Whole Blood Samples

Abstract Mutations in over 100 genes are implicated in autism spectrum disorder (ASD). DNA mutations and epigenomic modifications also contribute to ASD. Transcriptomics analysis of blood samples may offer clues for pathways dysregulated in ASD. To expand and validate published findings of RNA-sequencing (RNA-seq) studies, we performed RNA-seq of whole blood samples from a discovery cohort of eight children with ASD compared with nine age- and sex-matched neurotypical children. This revealed 10 genes with differential expression. Using real-time PCR, we compared whole blood samples from 35 children with ASD and 21 matched neurotypical children for the 10 dysregulated genes detected by RNA-seq. This revealed higher expression levels of the proinflammatory transcripts BATF2 and FCGR1A, and lower expression levels of the anti-inflammatory transcripts ISG15 and MT2A in the ASD compared to the control group. BATF2 and FCGR1A were recently reported as upregulated in blood samples of Japanese adults with ASD. Coupled with that publication, our findings support involvement of these genes in ASD phenotypes, independent of age and ethnicity. Upregulation of BATF2 and FCGR1A and downregulation of ISG15 and MT2A were reported to reduce cancer risk. Implications of the dysregulated genes for pro-inflammatory phenotypes, immunity, and cancer risk in ASD are discussed.

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Enzyme immunoassay of free thyroxin in dried blood samples on filter paper.

Clinical Chemistry ◽

10.1093/clinchem/31.5.750 ◽

1985 ◽

Vol 31 (5) ◽

pp. 750-753 ◽

Cited By ~ 8

Author(s):

N Hata ◽

K Miyai ◽

M Ito ◽

Y Endo ◽

Y Iijimi ◽

...

Keyword(s):

Enzyme Immunoassay ◽

Filter Paper ◽

Room Temperature ◽

Normal Subjects ◽

Blood Samples ◽

Double Antibody ◽

Coefficients Of Variation ◽

The Mean ◽

Measurable Range

Abstract We describe a double-antibody enzyme immunoassay for determination of free thyroxin (FT4) in dried blood samples on filter paper, with use of a T4-beta-D-galactosidase complex. The measurable range of FT4 concentration in two 3-mm blood discs, each of which contained about 2.7 microL of blood, was 1.9 to 93 ng/L, as determined by comparison with concentrations of FT4 in known serum standards. FT4 in blood samples dried on filter paper was stable for at least four weeks when kept dry at -20 degrees C, room temperature, or 37 degrees C. The mean coefficients of variation were 7.6% (within assay) and 6.4% (between assays). Results for FT4 by this method correlated well with those for serum determined by radioimmunoassay (r = 0.98). The proposed method can be used to differentiate persons with hyper- and hypothyroidism from normal subjects and those with abnormal concentrations of thyroxin-binding globulin. The procedure seems suited for screening studies.

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Aspirin Does Not Affect the Flow Cytometric Detection of Fibrinogen Binding to, or Release of α-Granules or Lysosomes from, Human Platelets

Clinical Science ◽

10.1042/cs0870575 ◽

1994 ◽

Vol 87 (5) ◽

pp. 575-580 ◽

Cited By ~ 70

Author(s):

Nicolas A. F. Chronos ◽

Darren J. Wilson ◽

Sarah L. Janes ◽

Ronald A. Hutton ◽

Nigel P. Buller ◽

...

Keyword(s):

Flow Cytometry ◽

Arachidonic Acid ◽

Thromboxane A2 ◽

Normal Subjects ◽

Human Platelets ◽

Aspirin Therapy ◽

Flow Cytometric Assay ◽

Blood Samples ◽

Flow Cytometric ◽

Fibrinogen Binding

1. Aspirin inhibits the conversion of arachidonic acid to thromboxane A2 which reinforces the effects of weak agonists such as ADP in platelets. 2. In this study the effect of aspirin (300 mg/day) on platelet agonist response was measured by whole blood flow cytometry of unfixed blood samples from normal subjects (n = 10), an assay that investigates aggregation-independent changes in the platelet. 3. Fibrinogen binding to unstimulated platelets or to platelets stimulated with ADP or thrombin was unaffected by aspirin. 4. Under the conditions of this assay, platelets undergo a partial degranulation of α-granules and lysosomes (evidenced by expression of P-selectin and CD63, respectively) in response to ADP, and full degranulation in response to thrombin. P-selectin expression was paralleled by release of β-thromboglobulin. None of these events was affected by aspirin. 5. Thromboxane formation was totally prevented by the aspirin treatment, as shown by Born aggregometry in which the platelet aggregatory response to arachidonic acid was abolished and secondary aggregation by ADP was inhibited. 6. The flow cytometric assay can therefore be used to investigate platelets in patients, regardless of aspirin therapy. 7. These findings suggest that platelet fibrinogen binding and the release of platelet α-granule and lysosomal contents, in response to stimulation with physiological agonists, can continue in patients despite aspirin therapy. This may help to explain why aspirin is only partially effective in preventing thrombotic events.

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Simplified radioimmunoassay of bradykinin in human plasma.

Clinical Chemistry ◽

10.1093/clinchem/26.3.0429 ◽

1980 ◽

Vol 26 (3) ◽

pp. 429-432

Author(s):

P S Verma ◽

P E Lorenz ◽

G E Sander

Keyword(s):

Molecular Mass ◽

Human Plasma ◽

Normal Values ◽

Normal Subjects ◽

Single Step ◽

Relative Molecular Mass ◽

Blood Samples ◽

Analytical Recovery ◽

Assay Tube ◽

The Mean

Abstract A greatly simplified radioimmunoassay for bradykinin in human plasma is described. Current techniques require multiple chromatographic steps or extraction procedures with analytical recoveries of bradykinin of often less than 60%. We present a method in which bradykinin is separated from components of higher relative molecular mass (including kininogens) in a single step, by use of a column of Sephadex G-25 medium (PD-10). The mean analytical recovery of tritiated bradykinin added to plasma is 85.5% (SD, 3.5%). The sensitivity of this radioimmunoassay is 25 pg per assay tube, equivalent to 125 ng per liter of plasma. Twenty to 30 blood samples may be completely processed and assayed within 6 h. As determined with this technique, concentrations of bradykinin in plasma from apparently normal subjects ranged from 2.5 to 5.2 microgram/L (mean 4.2, SD 1.1 microgram/L); these values are consistent with previously reported normal values.

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Effect of induced fever on serum iron and ferritin concentrations in man

Blood ◽

10.1182/blood.v49.1.147.147 ◽

1977 ◽

Vol 49 (1) ◽

pp. 147-153 ◽

Cited By ~ 99

Author(s):

RJ Elin ◽

SM Wolff ◽

CA Finch

Keyword(s):

Serum Ferritin ◽

Intramuscular Injection ◽

Serum Iron ◽

Normal Subjects ◽

Compound A ◽

Single Episode ◽

Blood Samples ◽

Iron Parameters ◽

Quantitative Changes

Abstract Previous reports have shown that endotoxin decreases serum iron in experimental animals. In this study fever was produced in nine female and nine male normal subjects in order to define the temporal and quantitative changes in serum iron and ferritin concentrations. Six volunteers were randomly given bacterial endotoxin (5 ng/kg) or saline intravenously and received the alternative compound a week later. Serial blood samples were drawn at 4-hr intervals for a 24-hr period, beginning when the compound was administered, for the determination of serum iron and ferritin concentrations. The same study was performed with intramuscular etiocholanolone (0.3 mg/kg) or the vehicle, propylene glycol, as a control, but the first blood sample was obtained 9 hr after the compound was given. In addition, blood samples were obtained at 12-hr intervals in six volunteers for 11 days after an intramuscular injection of etiocholanolone. The results showed a significant increase (p less than 0.005 for etiocholanolone, P less than 0.01 for endotoxin) in serum ferritin and a significant decrease (p less than 0.005 for etiocholanolone, p less than 0.001 for endotoxin) in serum iron for both pyrogenic compounds compared with the control compounds. However, the amount of fever and the changes in the iron parameters were greater with etiocholanolone. One episode of induced fever with etiocholanolone effected changes in serum ferritin and iron concentrations that lasted 10 days. Thus this study demonstrated that a single episode of fever in man produced rapid and prolonged changes in serum iron and ferritin concentrations.

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