scholarly journals Neutrophil Maturation, Reactivity and Granularity Research Parameters to Characterize and Differentiate Convalescent Patients from Active SARS-CoV-2 Infection

Cells ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 2332
Author(s):  
Iwona Kwiecień ◽  
Elżbieta Rutkowska ◽  
Katarzyna Kulik ◽  
Krzysztof Kłos ◽  
Katarzyna Plewka ◽  
...  
Keyword(s):  
Immune Response ◽  
Activation Parameters ◽  
Innate Immune ◽  
Study Group ◽  
Healthy Controls ◽  
Rt Pcr ◽  
Absolute Count ◽  
Active Infection ◽  
Immature Granulocyte ◽  
Pcr Test

Studying the dynamics changes of neutrophils during innate immune response in coronavirus 2019 (COVID-19) can help understand the pathogenesis of this disease. The aim of the study was to assess the usefulness of new neutrophil activation parameters: Immature Granulocyte (IG), Neutrophil Reactivity Intensity (NEUT-RI), Neutrophil Granularity Intensity (NEUT-GI), and data relating to granularity, activity, and neutrophil volume (NE-WX, NE-WY, NE-WZ) available in hematology analyzers to distinguish convalescent patients from patients with active SARS-CoV-2 infection and healthy controls (HC). The study group consisted of 79 patients with a confirmed positive RT-PCR test for SARS-CoV2 infection, 71 convalescent patients, and 20 HC. We observed leukopenia with neutrophilia in patients with active infection compared to convalescents and HC. The IG median absolute count was higher in convalescent patients than in COVID-19 and HC (respectively, 0.08 vs. 0.03 vs. 0.02, p < 0.0001). The value of the NEUT-RI parameter was the highest in HC and the lowest in convalescents (48.3 vs. 43.7, p < 0.0001). We observed the highest proportion of NE-WX, NE-WY, and NE-WZ parameters in HC, without differences between the COVID-19 and convalescent groups. New neutrophil parameters can be useful tools to assess neutrophils’ activity and functionalities in the immune response during infection and recovery from COVID-19 disease.

scholarly journals DNA Methylation Patterns of Interferon Gamma Gene Promoter and Serum Level in Pulmonary Tuberculosis: Their Role in Prognosis

2019 ◽  
Vol 16 (1) ◽  
pp. 0178
Author(s):  
Zayr Et al.
Keyword(s):  
Dna Methylation ◽  
Immune Response ◽  
Interferon Gamma ◽  
Gene Promoter ◽  
Innate Immune ◽  
Healthy Controls ◽  
Healthy Control ◽  
Ifn Γ ◽  
Methylation Patterns ◽  
Normal Controls

Tuberculosis (TB) still remains an important medical problem due to high levels of morbidity and mortality worldwide. A series of innate immune mechanisms that create a cytokine network control the pathogenesis of tuberculosis and this response has the capacity to modify the host genomic DNA structure through epigenetic mechanisms such as DNA methylation which could constantly alter the local gene expression pattern that can modulate the metabolism of the tissues and the immune-response. Interferon-gamma (IFN-γ) is an important pro-inflammatory cytokine regulator of the innate immune response to TB. This study aims to determine DNA methylation patterns of INF-γ gene promoter and measure serum IFN- γ level in newly diagnosed TB patients, relapse TB patients, and healthy control, in order to study the possibility of using these as a biomarker for the prognosis of TB stages in patients. The current case-control study included 66 patients with TB and 33 healthy control subjects. DNA was extracted from peripheral blood(PB) of included subjects and modified using sodium bisulfate specific kit. DNA methylation patterns of IFN-γ gene promoter was determine by using methylation specific polymerase chain reaction(MS-PCR).Serum IFN-γ level  was determined using enzyme linked immune-sorbent assay(ELISA). Results showed that percentages of DNA methylation patterns in normal controls, newly diagnostic TB patients and relapse TB patients were (63.3%, 18.2% and 21.2% respectively). Also, higher significant differences (P≤0.0001) of  un-methylated  IFN-γ gene promoter patterns in newly diagnostic TB patients  than  relapse TB patients comparison with healthy controls. The percentage of un-methylated DNA patterns in healthy controls, newly diagnostic TB patients and relapse TB patients were (9.9%, 39.4% and 51.5%, respectively). The mean of serum IFN-γ levels (pg/ml) for normal controls, newly diagnostic TB patients and relapse TB patients were (59.3± 13.8,75.8±24.3 and 69.6±18.7,respectively).In conclusion, there is a relative association between methylation of IFN-γ gene promoter and predisposing to TB progression.

Iron Overload Diminishes the Effectiveness of the Innate Immune Response in Thalassemia Major: a Possible Mechanism for Increased Infection Risk.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4071-4071
Author(s):  
Patrick B Walter ◽  
Paul R Harmatz ◽  
Annie Higa ◽  
David Killilea ◽  
Nancy Sweeters ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Immune Response ◽  
Iron Overload ◽  
Board Of Directors ◽  
Innate Immune Response ◽  
Research Funding ◽  
Innate Immune ◽  
Healthy Controls ◽  
Advisory Committees ◽  
Fluorescent Intensity

Abstract Abstract 4071 Poster Board III-1006 Introduction Infection is the second most common cause of death in thalassemia. The innate immune system provides a first line of defense against infection and specificity depends on pattern recognition receptors (PRRs) specific to microbial pathogens. One class of PRR called the toll-like receptors (TLRs) are important for transducing the signal for bacterial Lipopolysaccharide (LPS), resulting not only in cytokine production, but also in the control of extracellular iron levels through production of neutrophil gelatinase associated Lipocalin (NGAL). However, the exact role that NGAL plays and the expression level of PRRs are unknown in thalassemia. Thus, the goal in these studies is to investigate the relationship of iron overload to the innate immune cell expression of PRRs and NGAL in thalassemia. Patients and Methods Fifteen transfusion dependent thalassemia patients (11 – 29 yrs old) participating in the combination trial of deferasirox (an oral iron chelator) and deferoxamine were enrolled (Novartis sponsored CICL670AUS24T). Fasting blood samples were obtained i) at baseline after a 72 hr washout of chelator, and ii) at 6 and 12 months on study. Five healthy controls (13 - 18 yrs old) were also enrolled. Fresh monocytes were isolated using antibody-linked magnetic microbeads (Miltenyi Biotec Inc). Highly enriched populations of CD14+ monocytes were verified by flow cytometry. The expression of TLR4, also examined by flow cytometry is reported as the mean fluorescent intensity (MFI). In patients with thalassemia, liver iron concentration (LIC) was analyzed by biomagnetic susceptibility (“SQUID”, Ferritometer®). The plasma levels of NGAL were analyzed by ELISA. Results At baseline the expression of monocyte TLR4 (mean 18.8 ± 3.5 MFI) was reduced 30% compared to the healthy controls (mean 26.9 ± 7.6 MFI, p<0.05). The expression of TLR4 over the follow-up period of 52 weeks in patients receiving intensive combination chelator therapy significantly increased 27% / year (7 MFI / year, p=0.005). Interestingly the expression of monocyte TLR4 was negatively correlated with LIC (r=-0.6, p=0.04). Finally, thalassemia patients at baseline have significantly higher levels of NGAL (80 ± 20 ng/ml) compared to controls (42 ± 15 ng/ml, p=0.01). Conclusions These preliminary studies support the hypothesis that iron burden has a negative impact on the innate immune response in thalassemia as demonstrated by the decreased expression of TLR4. After intensive chelation, the levels of TLR4 increased, indicating that decreased iron overload with chelation may improve innate immune responsiveness. Finally, the iron transport protein NGAL is significantly elevated in thalassemia possibly acting to prevent essential iron uptake by pathogenic bacteria. Disclosures: Harmatz: Novartis: Research Funding; Apotex : Membership on an entity's Board of Directors or advisory committees; Ferrokin: Membership on an entity's Board of Directors or advisory committees. Vichinsky:Novartis: Consultancy, Research Funding.

scholarly journals Active Infection of Human Blood Monocytes by Chikungunya Virus Triggers an Innate Immune Response

2010 ◽  
Vol 184 (10) ◽  
pp. 5903-5913 ◽  
Author(s):  
Zhisheng Her ◽  
Benoit Malleret ◽  
Monica Chan ◽  
Edward K. S. Ong ◽  
Siew-Cheng Wong ◽  
...  
Keyword(s):  
Immune Response ◽  
Innate Immune Response ◽  
Human Blood ◽  
Chikungunya Virus ◽  
Innate Immune ◽  
Blood Monocytes ◽  
Active Infection

scholarly journals SIGIRR and TNFAIP3 Are Differentially Expressed in Both PBMC and TNF-α Secreting Cells of Patients With Major Depressive Disorder

2021 ◽  
Vol 12 ◽  
Author(s):  
Chin-Chuen Lin ◽  
Tiao-Lai Huang
Keyword(s):  
Immune Response ◽  
Major Depressive Disorder ◽  
Depressive Disorder ◽  
Innate Immune ◽  
Differentially Expressed ◽  
Mrna Levels ◽  
Healthy Controls ◽  
Negative Regulators ◽  
Major Depressive ◽  
Tnf Α

Background: Major depressive disorder (MDD) is associated with the activation of the immune/inflammatory system. TNF-α is associated with MDD and poor treatment response. Toll-like receptors (TLR) are responsible in innate immune response, and is associated with MDD and antidepressant response. Some negative regulators of TLR pathway such as SOCS1, TOLLIP, SIGIRR, TNFAIP3, and MyD88s, are reported to be differentially expressed in the peripheral blood samples of patients of MDD.Methods: We recruited patients with MDD and healthy controls, collect their demographic data, and measured their mRNA levels of negative TLR regulators, using peripheral blood mononuclear cells (PBMC) and isolated TNF-α secreting cells. Clinical symptoms were evaluated using Halmiton Depression Rating Scale (Ham-D). Some patients were evaluated again after 4 weeks of antidepressant treatment.Results: Forty-seven patients with MDD and 52 healthy controls were recruited. Between the PBMC samples of 37 MDD patients and 42 controls, mRNA levels of SOCS1, SIGIRR, TNFAIP3, and MyD88s were significantly different. Between TNF-α secreting cells of 10 MDD patients and 10 controls, mRNA levels of SIGIRR and TNFAIP3 were significantly different. Change of Ham-D score only correlated significantly with TOLLIP mRNA level after treatment.Conclusion: SIGIRR and TNFAIP3, two negative regulators of TLR immune response pathways, were differentially expressed in both PBMC and TNF-α secreting cells of patients with MDD as compared to healthy controls. The negative regulations of innate immune response could contribute to the underlying mechanism of MDD.

scholarly journals The duration and specificity of the humoral immune response to SARS-CoV-2

2021 ◽  
pp. 7-12
Author(s):  
E. I. Dubrovskyi ◽  
B. V. Dons’koi
Keyword(s):  
Immune Response ◽  
Quality Control ◽  
Humoral Response ◽  
Threshold Value ◽  
Spike Protein ◽  
Obstetrics And Gynecology ◽  
Study Group ◽  
Specific Antibodies ◽  
Humoral Immune ◽  
Pcr Test

Background. Our assumption that immunity after COVID-19 will persist has been fully confirmed in the researches already conducted. Our work is a continuation of research that demonstrates the results obtained 12 months of determining the humoral response in patients after COVID-19. Materials and methods. The research involved 42 individuals. All subjects had a positive PCR test for COVID-19. At certain intervals, from 40 to 240 days, individuals in the group were tested for IgG SARS-CoV-2. The last step was to check the level of IgG to the COVID-19 nucleocapsid and spike protein in the research group for 360 days from the onset of the disease. A private certified laboratory in Kyiv, the “DNA Laboratory”, was involved. Patients were tested for antibodies to COVID-19 by ELISA using serology COVID-19 test systems VitroTest (Ukraine). The immunological laboratory of the Institute of Pediatrics, Obstetrics and Gynecology was used in parallel for interlaboratory quality control. The results of the research coincided. Results. The level of class G immunoglobulins to nucleocapsid in the subjects has gradually decreased over 8 months. It is noteworthy that in the period from 40 to 150 days in all 42 patients (100 %) antibodies did not disappear. Decreasing of antibodies occurred between 150 and 240 days. However, the data obtained for 360 days significantly changed the picture. In a certain part of the subjects, who had low or even negative levels of antibodies for 8 months, as of 12 months, the level of immunoglobulin (Ig) class G again rose above the threshold value. Thus, we see that from the group of 42 people 92.8 % have positive antibodies to the nucleocapsid, and 7.2 %. Conclusions. The data obtained illustrate that in the study group within 12 months after SARS-CoV-2, the vast majority of individuals remain with specific antibodies to the nucleocapsid and spike-protein.

scholarly journals DNA Methylation Patterns of Interferon Gamma Gene Promoter and Serum Level in Pulmonary Tuberculosis: Their Role in Prognosis

2019 ◽  
Vol 16 (1(Suppl.)) ◽  
pp. 0178
Author(s):  
Zayr Et al.
Keyword(s):  
Dna Methylation ◽  
Immune Response ◽  
Interferon Gamma ◽  
Gene Promoter ◽  
Innate Immune ◽  
Healthy Controls ◽  
Healthy Control ◽  
Ifn Γ ◽  
Methylation Patterns ◽  
Normal Controls

Tuberculosis (TB) still remains an important medical problem due to high levels of morbidity and mortality worldwide. A series of innate immune mechanisms that create a cytokine network control the pathogenesis of tuberculosis and this response has the capacity to modify the host genomic DNA structure through epigenetic mechanisms such as DNA methylation which could constantly alter the local gene expression pattern that can modulate the metabolism of the tissues and the immune-response. Interferon-gamma (IFN-γ) is an important pro-inflammatory cytokine regulator of the innate immune response to TB. This study aims to determine DNA methylation patterns of INF-γ gene promoter and measure serum IFN- γ level in newly diagnosed TB patients, relapse TB patients, and healthy control, in order to study the possibility of using these as a biomarker for the prognosis of TB stages in patients. The current case-control study included 66 patients with TB and 33 healthy control subjects. DNA was extracted from peripheral blood(PB) of included subjects and modified using sodium bisulfate specific kit. DNA methylation patterns of IFN-γ gene promoter was determine by using methylation specific polymerase chain reaction(MS-PCR).Serum IFN-γ level  was determined using enzyme linked immune-sorbent assay(ELISA). Results showed that percentages of DNA methylation patterns in normal controls, newly diagnostic TB patients and relapse TB patients were (63.3%, 18.2% and 21.2% respectively). Also, higher significant differences (P≤0.0001) of  un-methylated  IFN-γ gene promoter patterns in newly diagnostic TB patients  than  relapse TB patients comparison with healthy controls. The percentage of un-methylated DNA patterns in healthy controls, newly diagnostic TB patients and relapse TB patients were (9.9%, 39.4% and 51.5%, respectively). The mean of serum IFN-γ levels (pg/ml) for normal controls, newly diagnostic TB patients and relapse TB patients were (59.3± 13.8,75.8±24.3 and 69.6±18.7,respectively).In conclusion, there is a relative association between methylation of IFN-γ gene promoter and predisposing to TB progression.

Salivary LL-37 Secretion in Individuals with Down Syndrome is Normal

2006 ◽  
Vol 85 (10) ◽  
pp. 933-936 ◽  
Author(s):  
G. Bachrach ◽  
G. Chaushu ◽  
M. Zigmond ◽  
E. Yefenof ◽  
A. Stabholz ◽  
...  
Keyword(s):  
Immune Response ◽  
Down Syndrome ◽  
Innate Immune System ◽  
Salivary Flow ◽  
Innate Immune ◽  
Antimicrobial Protein ◽  
Healthy Controls ◽  
Parotid Saliva ◽  
Immunoglobulin Deficiency ◽  
Secretion Rates

Antimicrobial peptides play an important role in the innate immune response. Deficiency in salivary LL-37 antimicrobial peptide has been implicated in periodontitis in patients with morbus Kostman syndrome. Down syndrome is associated with periodontitis, diminished salivary flow, and salivary immunoglobulin deficiency. In the present study, levels of LL-37 and its hCAP18 precursor were measured in saliva samples from young individuals with Down syndrome and compared with levels in those from age-matched healthy controls. LL-37 and human cathelicidin antimicrobial protein (hCAP18) were detected in whole but not in parotid saliva. hCAP18 was more abundant than LL-37. The concentrations of salivary hCAP18 and LL-37 were found to be higher in individuals with Down syndrome than in healthy controls, but their secretion rates were similar. We concluded that, while the adaptive immunity of individuals with Down syndrome is impaired at the oral mucosa, the secretion rate of the LL-37 component of the innate immune system is normal.

scholarly journals Evaluation of Antibody Response in Symptomatic and Asymptomatic COVID-19 Patients and Diagnostic Assessment of New IgM/IgG ELISA Kits

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 161
Author(s):  
Hadeel T. Al-Jighefee ◽  
Hadi M. Yassine ◽  
Maryam A. Al-Nesf ◽  
Ali A. Hssain ◽  
Sara Taleb ◽  
...  
Keyword(s):  
Immune Response ◽  
Severe Acute Respiratory Syndrome ◽  
Antibody Response ◽  
Diagnostic Assessment ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Serum Samples ◽  
Time Intervals ◽  
Low Sensitivity ◽  
Pcr Test

This study aims to study the immune response and evaluate the performances of four new IgM and five IgG enzyme-linked immunosorbent assay (ELISA) kits for detecting anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies against different antigens in symptomatic and asymptomatic coronavirus disease 2019 (COVID-19) patients. A total of 291 samples collected from symptomatic and asymptomatic RT–PCR-confirmed patients were used to evaluate the ELISA kits’ performance (EDI, AnshLabs, DiaPro, NovaLisa, and Lionex). The sensitivity was measured at three different time-intervals post symptoms onset or positive SARS-CoV-2 RT–PCR test (≤14, 14–30, >30 days). The specificity was investigated using 119 pre-pandemic serum samples. The sensitivity of all IgM kits gradually decreased with time, ranging from 48.7% (EDI)–66.4% (Lionex) at ≤14 days, 29.1% (NovaLisa)–61.8% (Lionex) at 14–30 days, and 6.0% (AnshLabs)–47.9% (Lionex) at >30 days. The sensitivity of IgG kits increased with time, peaking in the latest interval (>30 days) at 96.6% (Lionex). Specificity of IgM ranged from 88.2% (Lionex)–99.2% (EDI), while IgG ranged from 75.6% (DiaPro)–98.3% (Lionex). Among all RT–PCR-positive patients, 23 samples (7.9%) were seronegative by all IgG kits, of which only seven samples (30.4%) had detectable IgM antibodies. IgM assays have variable and low sensitivity, thus considered a poor marker for COVID-19 diagnosis. IgG assays can miss at least 8% of RT–PCR-positive cases.

scholarly journals Levels of different cytokines in women and men with asymptomatic genital infection caused by Chlamydia

2019 ◽  
Vol 13 (09) ◽  
pp. 847-850
Author(s):  
Alessandra Bua ◽  
Sara Cannas ◽  
Stefania Zanetti ◽  
Paola Molicotti
Keyword(s):  
Immune Response ◽  
Statistical Analysis ◽  
Chlamydia Trachomatis ◽  
Genital Infection ◽  
Healthy Controls ◽  
Rt Pcr ◽  
Chlamydia Trachomatis Infection ◽  
Molecular Assays ◽  
Significant Difference ◽  
Student Test

Introduction: Immune response to genital Chlamydia trachomatis infection is involved in both immunity and pathology. The cytokine profile during infection has been implicated in the disease outcome, either resolution or severe sequelae. Methodology: In total, 3900 patients were analyzed for presence of genital infections caused by Chlamydia using molecular assays. Interleukins (IL) IL-10, IL-17, IL-6, IL-2 and chemokine IP-10 were estimated by ELISA in urine, cervical swabs and semen samples. Statistical analysis was performed using the T student test. Results: A total of 47 out of 3900 samples (1.2%) were found to be positive for Chlamydia trachomatis based on the Real Time (RT) PCR results. Statistical analysis revealed that the differences between Chlamydia trachomatis positive and negative samples regarding levels of cytokines were not significant. Conclusions: Our results demonstrated that no significant difference in cytokine concentrations exists in Chlamydia trachomatis infected patients when compared to healthy controls. In further study, we aim to test on a greater number of positive samples a greater number of cytokines involved in the immune response to Chlamydia trachomatis infections.

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