Concentrations Of IFN Research Articles

Inflammatory biomarkers and growth factors in patients with chronic cerebrovascular disease after coronavirus infection

Objective: to study the level of inflammatory biomarkers and growth factors in the peripheral blood of patients with chronic cerebrovascular disease (CCVD) and anamnesis of coronavirus infection COVID-19.Material and methods. The study included patients with CCVD (n=41), 26 of them had a documented anamnesis of coronavirus infection within 4 months before inclusion in the study, 15 people did not have COVID-19. The control group consisted of 20 apparently healthy individuals of the same age. Neuroimaging was performed using a Philips Achieva 1.5 T device. Pro-inflammatory cytokines were determined in blood serum – tumor necrosis factor α (TNF α ), interleukin 6 (IL6), IL18, interferon γ (IFN γ ); chemokines – monocytic chemoattractant protein 1 (MCP-1), IL8; growth factors – vascular endothelial growth factor type A (VEGF-A), transforming growth factor β 1 (TGF β 1).Results. In patients with CCVD, compared with the control group, an increase in the level of proinflammatory cytokines (TNF α , IL6, IL18), chemokines (MCP1 and IL8), a decrease in the concentration of IFN γ , and divergent changes in the content of growth factors (VEGF-A and TGF β 1) were noted. Patients who recovered from COVID-19 showed an increase in the level of IL6 and a decrease in the level of IFN γ compared with those who had not been ill, which indicates a persistently high activity of immunoinflammatory processes and an insufficient humoral immune response.Conclusion. Postponed coronavirus infection COVID-19 aggravates the existing endothelial dysfunction and intravascular inflammation in patients with CCVD, which may probably require changes in their treatment and prevention strategies in the future.

Development and Evaluation of the Biological Adjuvant Bivalent Vaccine for Preventing Newcastle Disease and Infectious Bursal Disease

Newcastle disease (ND) and infectious bursal disease (IBD) are both one of the most economically important infectious diseases, which cause high infection rate and mortality of chickens and restrict the development of the poultry industry. Currently, vaccination is the most effective method to prevent ND and IBD. In this study, the biological adjuvant bivalent vaccine (rClone30-VP2L-chGM-CSF) was developed for preventing ND and IBD. Furthermore, the immune and protective effects of rClone30-VP2L-chGM-CSF were evaluated in specific-pathogen-free chickens. The experimental results indicated that chickens immunized with rClone30-VP2L-chGM-CSF vaccine significantly improved anti-IBDV and anti-NDV antibody titer; the level of CD4+ T, CD8+ T, B+, MHC-II+, and monocyte/macrophagocyte+ cells; the concentrations of IL-1β, IL-4, IL-17, IFN-α, IFN-β, and IFN-γ; and the splenocyte proliferation rate. Anti-NDV antibody titer reached the theoretical protection value of 4log2 at 7 days after immunization in chickens of rClone30-VP2L-chGM-CSF group; however, which at 14 days after immunization in chickens of rClone30-VP2L and rClone30. These results showed that chickens immunized with rClone30-VP2L-chGM-CSF stimulated stronger immune response than those with the rClone30-VP2L and rClone30. Chickens were challenged with virulent IBDV BC6/85, which were protected in the rClone30-VP2L-chGM-CSF group. Furthermore, IBDV RNA was not measured, and there appeared to be little apoptosis in the bursa of Fabricius in the rClone30-VP2L-chGM-CSF group. Therefore, rClone30-VP2L-chGM-CSF is a promising vaccine candidate against infectious bursal disease virus (IBDV) and newcastle disease virus (NDV), and it provides an idea for developing other poultry vaccines.

Safety, Tolerability, Bioavailability, and Biological Activity of Inhaled Interferon-α2b in Healthy Adults: The IN2COVID Phase I Randomized Trial.

Interferons have been identified as a potential treatment alternative for coronavirus disease 2019. This study assessed the safety, tolerability, bioavailability, and biological activity of inhaled interferon-α2b (IFN)-α2b in healthy adults. A double-blind, randomized, phase I clinical trial was conducted with two cohorts of healthy subjects aged 18-50 years. The first cohort received 2.5 MIU of inhaled IFN-α2b twice daily for 10 days (n = 6) or placebo (n = 3); the second cohort received 5.0 MIU of inhaled IFN-α2b in a similar scheme (n = 6) or placebo (n = 3). The first two doses were administered in an emergency department, then participants completed their treatment at home. Safety was measured through vital signs, new symptoms, and laboratory tests. Tolerability was measured as participants' treatment acceptability. Bioavailability and biological activity were measured from serum IFNα concentrations and real-time quantitative polymerase chain reaction of interferon-induced genes in blood before and after treatments. Exposure to inhaled IFN-α2b at 2.5-MIU or 5-MIU doses did not produce statistically significant changes in participant vital signs, or elicit new symptoms, and standard hematological and biochemical blood measurements were comparable to those recorded in individuals who received placebo. A total of 58 adverse events were observed. All were mild or moderate and did not require medical care. All participants reported very high tolerability towards a twice-daily nebulized treatment for 10 days (98.0, 97.0, and 97.0 in the placebo, 2.5-MIU, and 5-MIU groups, respectively, on a 0- to 100-mm visual analog scale). A dose-dependent mild increase in serum IFN-α concentrations and an increase in serum RNA expression of IFN-induced genes were observed 11 days after treatment (p < 0.05 for all between-group comparisons). Inhaled IFN-α2b was preliminarily safe and well tolerated, and induced systemic biological activity in healthy subjects. The trial was registered in ClinicalTrials.gov (NCT04988217), 3 August, 2021.

Potential role of EBV and Toll-like receptor 9 ligand in patients with systemic lupus erythematosus.

SLE is a multisystem autoimmune disease characterized by multiple immunological abnormalities including production of autoantibodies. While the etiology of SLE is largely unknown, it is generally accepted that both genetic and environmental factors contribute to disease risk and immune dysregulation. Production of IFN-α is important for protecting the host against infections; however, over stimulation of innate immune pathways can induce autoimmune disease. Environmental factors, particularly Epstein-Barr virus (EBV), have been proposed to play an important role in SLE disease. Improper engagement of Toll-like receptor (TLR) pathways by endogenous or exogenous ligands may lead to the initiation of autoimmune responses and tissue injury. EBV is shown to be a potent stimulant of IFN-α by TLR signaling cascades. Given the highlighted role of IFN-α in SLE pathogenesis and potential role of EBV infection in this disease, the present study is aimed at exploring the in vitro effects of EBV infection and CPG (either alone or in combination) on IFN-α. We also examined the expression level of CD20 and BDCA-4 and CD123 in PBMCs in 32 SLE patients and 32 healthy controls. Our results showed PBMCs treated with CPG-induced higher levels of IFN-α and TLR-9 gene expression fold change compared to cells treated with either EBV or EBV-CPG. Moreover, PBMCs treated with CPG produced significantly higher IFN-α concentration in supernatant compared to cells treated with EBV but not EBV-CPG. Our results further highlight the potential role of EBV infection and TLRs in SLE patients although more studies are warranted to ascertain the global imprint that EBV infection can have on immune signature in patients with SLE.

Luteolin: A Phytochemical to Mitigate S. Typhimurium Flagellin-Induced Inflammation in a Chicken In Vitro Hepatic Model.

The use of natural feed supplements is an alternative tool to diminish the damage caused by certain bacteria, improving animal health and productivity. The present research aimed to investigate the proinflammatory effect of flagellin released from the bacterial flagellum of Salmonella enterica serovar Typhimurium and to attenuate the induced inflammation with luteolin as a plant-derived flavonoid on a chicken primary hepatocyte-non-parenchymal cell co-culture. Cells were cultured in a medium supplemented with 250 ng/mL flagellin and 4 or 16 µg/mL luteolin for 24 h. Cellular metabolic activity, lactate dehydrogenase (LDH) activity, interleukin-6, 8, 10 (IL-6, IL-8, IL-10), interferon-α, γ (IFN-α, IFN-γ), hydrogen peroxide (H2O2) and malondialdehyde (MDA) concentrations were determined. Flagellin significantly increased the concentration of the proinflammatory cytokine IL-8 and the ratio of IFN-γ/IL-10, while it decreased the level of IL-10, indicating that the model served adequate to study inflammation in vitro. Luteolin treatment at 4 µg/mL did not prove to be cytotoxic, as reflected by metabolic activity and extracellular LDH activity, and significantly reduced the flagellin-triggered IL-8 release of the cultured cells. Further, it had a diminishing effect on the concentration of IFN-α, H2O2 and MDA and restored the level of IL-10 and the ratio of IFN-γ/IL-10 when applied in combination with flagellin. These results suggest that luteolin at lower concentrations may protect hepatic cells from an excessive inflammatory response and act as an antioxidant to attenuate oxidative damage.

Practical Application and Influence of “Avigen duck” Immunomodulator to White Pekin Ducks Humoral Immune Factors

Factors of non-specific immunity are responsible for protecting birds from a number of viruses and bacteria. Bactericidal proteins are played here lysozyme and complement, interferon, as well as beta-lysine. Adding appropriate active compounds to the bird's diet to activate the innate immune response are subject to studies by many authors. The practical application and influence of the "Avigen Duck". immunomodulator in the cultivation of White Pekin ducks aims to clarify the dynamics of the indicators. It was found that the ducks treated with the immunomodulator had higher non-specific protection expressed in the values of the observed indicators. Complement activity for the experienced group (529.45 ± 17,85 CH50) is significantly higher than that of untrained birds (308.56 ± 10.19 CH50), on the 30th day of their lives (P &amp;lt;0.001). Lysozyme values for the experienced group (6.34 ± 0.86 mg L-1) is more than twice as high as its concentration in control birds - 2.52 ± 0.59 mg L-1. At 30 days of age, the mean concentration of IFN γ in the control group of ducks was 108.86±6.12 pg ml-1, and in the experimental group treated with immunomodulator - 518.06±12.80 pg ml-1, accompanied by an increase in the values of IL-2 and IL-6 and beta-lysine activity. These data show that, despite the short life of the ducks, the concentrations of the investigated factors of non-specific immunity increase significantly and statistically reliably, even after a single treatment with immunomodulator "Avigen Duck". The practical application of the immunomodulator in the duck diet has a strong effect on their non-specific immunity.

Type I interferon score is associated with the severity and poor prognosis in anti-MDA5 antibody-positive dermatomyositis patients.

To investigate the clinical significance of the interferon (IFN) score, especially the IFN-I score, in patients with anti-melanoma differentiation-associated gene 5 (MDA5) antibody-positive dermatomyositis (anti-MDA5+ DM). We enrolled 262 patients with different autoimmune diseases, including idiopathic inflammatory myopathy, systemic lupus erythematosus, rheumatoid arthritis, adult-onset Still's disease, and Sjögren's syndrome, as well as 58 healthy controls. Multiplex quantitative real-time polymerase chain reaction (RT-qPCR) using four TaqMan probes was used to evaluate type I IFN-stimulated genes (IFI44 and MX1), one type II IFN-stimulated gene (IRF1), and one internal control gene (HRPT1), which were used to determine the IFN-I score. The clinical features and disease activity index were compared between the high and low IFN-I score groups in 61 patients with anti-MDA5+ DM. The associations between laboratory findings and the predictive value of the baseline IFN-I score for mortality were analyzed. The IFN score was significantly higher in patients with anti-MDA5+ DM than in healthy controls. The IFN-I score was positively correlated with the serum IFN-α concentration, ferritin concentration, and Myositis Disease Activity Assessment Visual Analogue Scale (MYOACT) score. Compared with patients with a low IFN-I score, patients with a high IFN-I score showed a higher MYOACT score, C-reactive protein concentration, aspartate transaminase concentration, ferritin concentration, plasma cell percentage, and CD3+ T-cell percentage, as well as lower lymphocyte, natural killer cell, and monocyte counts. The 3-month survival rate was significantly lower in patients with an IFN-I score of >4.9 than in those with an IFN-I score of ≤4.9 (72.9% vs. 100%, respectively; P = 0.044). The IFN score, especially the IFN-I score, measured by multiplex RT-qPCR is a valuable tool to monitor disease activity and predict mortality in patients with anti-MDA5+ DM.

Evaluation of the Immuno- Responses to Brucella Melitensis

Brucella melitensis infection remains a serious health threat to humans and livestock in poorer countries in the Middle East. Vaccination with the isolated crude protein protects against B. melitensis infection. In this study, the efficacy of protective and immunogenicity of a crude protein extract of B. melitensis is administered and evaluated in rats. The immunogenicity of crude protein in the animal blood was assessed by SDS-PAGE and Western blot. The results show that certain proteins were found to be more immunogenic; their molecular weights were (21, 23, 24, 25, 28,31, 34.45, 54.66,83 and 157 kDa). ELISA kits were used to measure the concentrations of IL-4 and IFN- in serum. In contrast to IFN-γ, this study found that the group receiving extracted protein had considerably higher serum levels of IL-4 compared to the control healthy and REV1 group.

Assesment of specific T-cell immunity to SARS-CoV-2 virus antigens in COVID-19 reconvalescents

The development of the COVID-19 pandemic has stimulated the scientific research aimed at studying of the mechanisms of formation the immunity against SARS-CoV-2. Currently, there is a need to develop a domestic simple and cost-effective specific method suitable for monitoring of T-cell response against SARS-CoV-2 in reconvalescents and vaccinated individuals. Development of a screening method for evaluation specific T-cell immunity against SARS-CoV-2. Total 40 individuals who had mild to moderate COVID-19 and 20 healthy volunteers who did not have a history of this disease were examined. The presence and levels of IgG and IgM antibodies to SARS-CoV-2 were identified in participants sera by ELISA using the diagnostic kits from JSC Vector-Best (Novosibirsk, Russian Federation). Antigenic stimulation of mononuclear cells was carried out on commercial plates with adsorbed whole-virion inactivated SARS-CoV-2 antigen (State Research Center of Virology and Biotechnology VECTOR Novosibirsk, Russian Federation). The concentration of IFN- was measured in ELISA using the test systems from JSC Vector-Best (Novosibirsk, Russian Federation). The immunophenotyping of lymphocytes was performed on a flow cytometer Cytomics FC500 (Beckman Coulter, USA). Statistical data processing was carried out using the Microsoft Excel and STATISTICA 10 software package. Stimulation of mononuclear cells isolated from the peripheral blood with whole-virion inactivated SARS-CoV-2 antigen fixed at the bottom of the wells of a polystyrene plate showed a significantly higher median response in terms of IFN- production in 40 people who had history of COVID-19 compared to 20 healthy blood donors (172.1 [34.3575.1] pg/ml versus 15.4 [6.925.8] pg/ml, p 0.0001). There was no difference in median IFN- levels in supernatants collected from unstimulated mononuclear cells from COVID-19 reconvalescents and healthy donors (2.7 [0.411.4] pg/ml versus 0.8 [0.023.3] pg/ml, p 0.05). The overall sensitivity and specificity of this method were 73% (95% CI 5888%) and 100% (95% CI 100100%), respectively, at a cut-off of 50 pg/ml. The developed method for assessment of the cellular immune response to SARS-CoV-2 can be used as a screening method for monitoring the T-cell response in a population against a new coronavirus infection in recovered people.

Ovulation Induced by Intrauterine Seminal Plasma Increases Total Protein, PGE2, IL-8, and IL-1β in Uterine Fluid of Llamas (Lama glama)

The establishment of a state of immunotolerance in the female reproductive tract is important for embryo development, implantation and placentation. Llamas are induced ovulators and more than 98% of pregnancies occur in the left uterine horn. The objective of this study was to determine the uterine immune response of llamas in different stages of the reproductive cycle. Adult llamas (n = 20) were examined daily by transrectal ultrasonography to determine follicular growth and then randomly assigned to four groups: Follicular phase (n = 5); Luteal phase induced by an intramuscular administration of 50 ug of GnRH analogue (n = 5); Luteal phase induced by intrauterine infusion of seminal plasma (n = 5); and Luteal phase induced by mating (n = 5). Uterine fluid was collected separately from both uterine horns by non-surgical flushing to determine the presence of cells, total proteins and concentration of IL-1β, IL-6, IL-8, IFN γ, TNF-α and PGE2. Inflammatory cells were not observed in the uterine fluid and total protein pattern and inflammatory mediators did not differ between the left and the right horn amongst groups. Llamas treated with an intrauterine infusion of seminal plasma showed the highest concentration of total proteins, inflammatory cytokines PGE2, IL-8 and IL-1β in the uterine fluid. In conclusion, seminal plasma is made up of significant numbers of signaling molecules that are able to modify the uterine immune response in llamas.

Identification of biomarkers to stratify response to B-cell-targeted therapies in systemic lupus erythematosus: an exploratory analysis of a randomised controlled trial.

Systemic lupus erythematosus (SLE) is a complex autoimmune disease associated with widespread immune dysregulation and diverse clinical features. Immune abnormalities might be differentially associated with specific organ involvement or response to targeted therapies. We aimed to identify biomarkers of response to belimumab after rituximab to facilitate a personalised approach to therapy. In this exploratory analysis of a randomised controlled trial (BEAT-LUPUS), we investigated immune profiles of patients with SLE recruited to the 52-week clinical trial, which tested the combination of rituximab plus belimumab versus rituximab plus placebo. We used machine learning and conventional statistics to investigate relevant laboratory and clinical biomarkers associated with major clinical response. BEAT LUPUS is registered at ISRCTN, 47873003, and is now complete. Between Feb 2, 2017, and March 28, 2019, 52 patients were recruited to BEAT-LUPUS, of whom 44 provided clinical data at week 52 and were included in this analysis. 21 (48%) of 44 participants were in the belimumab group (mean age 39·5 years [SD 12·1]; 17 [81%] were female, four [19%] were male, 13 [62%] were White) and 23 (52%) were in the placebo group (mean age 42·1 years [SD 10·5]; 21 [91%] were female, two [9%] were male, 16 [70%] were White). Ten (48%) of 21 participants who received belimumab after rituximab and eight (35%) of 23 who received placebo after rituximab had a major clinical response at 52 weeks (between-group difference of 13% [95% CI -15 to 38]). We found a predictive association between baseline serum IgA2 anti-double stranded DNA (dsDNA) antibody concentrations and clinical response to belimumab after rituximab, with a between-group difference in major clinical response of 48% (95% CI 10 to 70) in patients with elevated baseline serum IgA2 anti-dsDNA antibody concentrations. Moreover, among those who had a major clinical response, serum IgA2 anti-dsDNA antibody concentrations significantly decreased from baseline only in the belimumab group. Increased circulating IgA2 (but not total) plasmablast numbers, and T follicular helper cell numbers predicted clinical response and were both reduced only in patients who responded to belimumab after rituximab. Serum IgA2 anti-dsDNA antibody concentrations were also associated with active renal disease, whereas serum IgA1 anti-dsDNA antibody and IFN-α concentrations were associated with mucocutaneous disease activity but did not predict response to B-cell targeted therapy. Patients with a high baseline serum interleukin-6 concentration were less likely to have a major clinical response, irrespective of therapy. This exploratory study revealed the presence of distinct molecular networks associated with renal and mucocutaneous involvement, and response to B-cell-targeted therapies, which, if confirmed, could guide precision targeting of advanced therapies for this heterogenous disease. Versus Arthritis, UCLH Biomedical Research Centre, LUPUS UK, and GSK.

Immunological markers of drug resistant epilepsy and its response to immunomodulatory therapy with ACTH in children.

Drug-resistant epilepsy in infancy and childhood is a devastating condition, frequently associated with neuropsychiatric comorbidities. West syndrome is one of the most severe epilepsy syndromes. Adrenocorticotropic hormone (ACTH) treatment is recommended in such cases, but its mechanism of action is still unknown. We prospectively observed levels of selected cytokines in order to identify biomarkers of response to ACTH and the potential mechanism of its antiseizure effect. Fifty-three infants and young children with pharmacoresistant epilepsy receiving ACTH 1-24 were included. There were 2 control groups - children with epilepsy responding to the first medication and children with no history of epilepsy. Blood concentrations of IL-1b, IL-1Ra, IL-6, IL-8, IL-10, TNF-a, IFN-g, MCP-1 and MIP-1a were analyzed at three time points: T0 (before ACTH), T1 (after intensive ACTH treatment) and at T2 (at the end of ACTH withdrawal). The results were correlated with the response to treatment, dose of ACTH and concomitant medications. We found statistically significantly higher concentrations of IL-1, IL-8 and MIP-1a at baseline (T0) in the study group compared to the control groups. ACTH significantly lowered levels of IL-6, IFN-g and MCP from time T0 to T1. This effect was short lasting and no significant changes in cytokine levels were found between T2 and T0. We did not find any differences in immunological markers between the responders and non-responders to ACTH. Our research did not allow us to identify any reliable immunological marker of response to ACTH treatment. We did not observe a positive effect of higher ACTH doses on the response rate in the patients. Our study showed significantly lower concentrations of IL-10 and IFN in the group of patients receiving levetiracetam. The concentration of TNF was higher and the concentration of MIP was lower in the group receiving valproic acid. Our study indicates that increased levels of IL-1, IL-8 and MIP-1a are associated with drug-resistant epilepsy in infants and young children and might be considered immunological markers. IL-6, IFN-g and MCP-1 take part in the effect of ACTH. Immunological mechanisms seem also to be involved in the mechanism of action of classical antiseizure drugs.

Features of mucosal immunity in the development of periodontal diseases in patients with type II diabetes mellitus

Chronic generalized periodontitis takes one of the leading places in the structure of dental diseases. Inflammatory periodontal disease is one of the most common complications of diabetes. An important role in the progression of periodontitis in type II diabetes mellitus (T2DM) belongs to disorders of the immune system that affect the supportive/retaining complex of the tooth. Inflammation of periodontal tissues which occurs in the patients with carbohydrate metabolism disorders is characterized by a more severe course, which worsens the condition and quality of life of patients with T2DM thus leading to tooth loss. In this regard, the immunological aspects of developing periodontal pathology under conditions of carbohydrate metabolism disorders are of great interest. Purpose of our study included comparative assessment of local TNF, TNF, IFN, IL-1, IL- 12, IL-17A levels in patients with periodontitis with and without carbohydrate metabolism disorders (type II diabetes mellitus), as well as in patients with type II diabetes mellitus without signs of periodontitis.&#x0D; 127 patients were examined, aged 30 to 59 years. The patients were divided into 3 groups, i.e., group I included patients suffering from periodontitis of varying severity without known comorbidities (47 persons); Group II consisted of patients with T2DM and periodontitis of varying severity (49 persons); Group III included patients with T2DM without symptoms of periodontitis (30 persons). The control group consisted of 30 practically healthy volunteers, matched to the patients for age and sex. Saliva specimens were used for laboratory studies. The levels of TNF, TNF, IFN, IL-1, IL-12, IL-17А were determined by ELISA sandwich technique, with specific reagents purchased from RD Diagnostics Inc (USA).&#x0D; A significant increase in the IL-1, TNF, IFN and IL-17A levels was found in patients of all groups compared with controls. At the same time, the increased concentrations of IFN, IL-1, IL-17A correlated with increase in periodontitis severity in the patients of both groups. Decreased TNF levels in saliva samples were revealed in the patients from almost all groups, regardless of the periodontitis severity. Significantly increased levels of IL-12 (p40 subunit) were recorded in saliva of the persons from group II and III when compared with controls and the group without diabetes mellitus.

TMEM173 rs7447927 genetic polymorphism and susceptibility to severe enterovirus 71 infection in Chinese children.

This study was designed to explore the association between the TMEM173 polymorphism (rs7447927) and the severity of enterovirus 71 (EV71) infection among Chinese children. The TMEM173 polymorphism was identified in EV71-infected patients (n = 497) and healthy controls (n = 535) using the improved multiplex ligation detection reaction (iMLDR). The interferon-α (IFN-α) serum levels were detected using enzyme linked immunosorbent assay (ELISA). The frequencies of the GG genotype and G allele of TMEM173 rs7447927 in the mild EV71 infection and severe EV71 infection groups were markedly higher than those in the control group. The GG genotype and G allele frequencies in severely infected EV71 patients were significantly higher than those in mildly infected EV71 patients. Severely infected EV71 patients with the GG genotype had higher white blood cell counts (WBC), and C-reactive proteins (CRP), and blood glucose (BG) levels, longer fever duration, higher vomiting frequency, spirit changes, and electroencephalography (EEG) abnormalities. IFN-α serum concentration in severely infected patients was significantly higher than in the mildly infected group. The IFN-α concentration in the GG genotype was significantly higher compared with those in the GC and CC genotypes in severe cases. The TMEM173 rs7447927 polymorphism was associated with EV71 infection susceptibility and severity. The G allele and GG genotype are susceptibility factors in the development of severe EV71 infection in Chinese children.

Association of the cytokine profile and metabolic syndrome components in young patients

The components of metabolic syndrome (MS) are independent factors of cardiometabolic risk and are associated with impaired humoral immunity. However, the literature data on the cytokine profile features in MS are ambiguous. Therefore, associations between cytokine levels and MS components were determined in a group of MS patients of both sexes, as well as indices of visceral adipose tissue function were studied. The work included 149 patients aged 18-45 years. The patients were divided into 2 groups: group 1 (n = 71) included patients without abdominal obesity and MS components (comparison group); group 2 (n = 78), patients with MS. The concentrations of glucose, glycosylated hemoglobin, insulin, total cholesterol, HDL-C, LDL-C, triglycerides, leptin, adiponectin were determined. The indexes of insulin resistance HOMA-IR, Tg/HDL and TyG, as well as marker of visceral adipose tissue dysfunction VAI were calculated. ELISA technique was used to determine the concentration of IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, IFN, IFN, МСР-1 и TNF. Results: In the patients with MS, we have found increased levels of IL-6, IL-10, MCP-1, and decrease of IL-2, IL-4, IFN levels. Correlation analysis established a relationship between glucose levels and MCP-1; glycosylated hemoglobin and IL-6, TNF. Among the indices of lipid metabolism, we have revealed some associations between LDL-C and IFN; HDL-C and IL-2, IL-4, IFN. The levels of triglycerides correlated with MCP-1. A negative relationship between the presence of arterial hypertension and the IL-4 contents was established. A negative correlation of leptin levels with IL-4 and IFN concentrations was also determined. Markers of insulin resistance (Tg/HDL and TyG) were associated with MCP-1 chemokine, thus supporting chronic inflammatory process. The VAI index, which reflects the dysfunction of visceral adipose tissue, showed a correlation with MCP-1. Thus, the results of investigation suggest an involvement of the cytokine system in the disorders of visceral adipose tissue and development of the metabolic syndrome.

PSV-B-16 Correlation between Plasma, Uterine, and Vaginal Cytokine Concentrations in Postpartum Beef Cows Prior to Artificial Insemination

Abstract Cytokines have a vital role in reproductive immune environment during the postpartum period. Previous data evaluated uterine and vaginal cytokine concentrations prior to insemination; however, was unable to correlate these data with plasma cytokine concentrations. The objective of this study was to determine relationships between uterine, vaginal, and plasma pro- and anti-inflammatory cytokine concentrations in postpartum beef cows prior to timed artificial insemination (TAI). Bos indicus-influenced beef cows (n=20) free of any physical, health or reproductive-related issues were subjected to the Bee Synch II protocol 8 days prior (d-8) to TAI (d0). Uterine and vaginal flushes and blood were collected on d-3 and d-1. Pregnancy was determined by transrectal ultrasonography on d28 (Pregnant, n=7; Open, n=13). Using the RayBiotech Quantibody Bovine Cytokine Array 1 the following cytokines concentrations were determined: interferon (IFN)-α, IFN-γ, interleukin (IL)-13, IL-1α, IL-1-F5, IL-21, tumor necrosis factor (TNF)-α, chemokine ligand (CXCL)-9, CXCL-10, and chemokine ligand 4 (CCL4). Concentration data were analyzed using PROC GLM and correlations using Pearson correlation in SAS. For plasma samples, 8 pro-inflammatory cytokines (IFN-α, IFN-γ, IL-13, IL-1α, IL-1-F5, IL-21, CXCL-9, and TNF-α) had greater (P&amp;lt; 0.05) concentrations in open cows compared with pregnant cows. For uterine flushes, 3 pro-inflammatory cytokines (IFN-γ, IL-1α, and IL-21) had greater (P&amp;lt; 0.05) concentrations in open cows compared with pregnant cows. For vaginal flushes, IFN-α concentrations were greater (P&amp;lt; 0.05) in open cows, while CXCL-10 concentrations were greater in pregnant cows. There were no significant correlations between plasma and uterine or vaginal samples (P&amp;gt;0.05). Interestingly, for open cows there were correlations for IFN-α (r=0.46; P=0.02), IFN-γ (r=0.58; P=0.002), and IL-21 (r=0.54; P=0.004) between uterine and vaginal samples; however, no correlations were observed for pregnant cows. These results suggest a greater abundance of pro-inflammatory cytokines within the uterus, vagina, and in peripheral circulation for resulting open cows prior to TAI.

OS05.6.A Modification of the tumor microenvironment in patients with glioblastoma using autologous, genetically modified, hematopoietic stem cell-based therapy: the TEM-GBM STUDY (NCT03866109)

Abstract Background Bone marrow-derived macrophages account for substantial GBM tumor volume and contribute to the local inflammatory tumor microenvironment, disease progression and treatment response. Material and Methods We have developed a genetically modified, autologous hematopoietic stem cell-based platform designed to deliver Interferon-alpha (IFNa), thanks to a transcriptional and post-transcriptional control mechanism mediated by miRNA target sequences, specifically into the tumor microenvironment via Tie-2 expressing monocytes (Temferon). Results As of Feb 2022, 3 escalating doses of Temferon (from 0.5 to 2.0x106/kg) were tested across 15 patients with newly diagnosed, unmethylated MGMT glioblastoma (GBM) assigned to 5 cohorts. The duration of follow-up from surgery is 6 - 28 mo (2 - 25 mo after Temferon). To date, no dose limiting toxicities have been identified. As expected, one month after the administration of the highest tested dose, the hematopoietic system of Temferon-treated patients was composed of up to 30% of CD14+ genetically modified cells, as determined by the presence of vector genomes in the DNA in peripheral blood and bone marrow cells. Temferon-derived progeny persisted, albeit at lower levels, up to 18 months (longest time of analysis). Despite the substantial proportion of engineered cells, very low median concentrations of IFNα were detected in the plasma (D+30, 5.9; D+90, 8.8pg/mL) and in the CSF (D+30, 1.5; D+90, 2.4pg/mL), indicating tight regulation of transgene expression. SAEs were mostly attributed to conditioning chemotherapy (e.g. infections) or disease progression (e.g. seizures). 1 SUSAR (persistent GGT elevation) has occurred. Median OS is 15 mo from surgery (range 6.1-28.4 mo; 10.8 mo post Temferon). Of the 15 pts treated so far, 4 pts belonging to low dose cohorts underwent 2nd surgery. Homing of transduced cells from BM to the tumor site was demonstrated by the presence of gene-marked cells in the specimens collected from 3 of the 4 analyzed pts. Single-cell RNA seq performed on CD45+ cells purified from the TME of Temferon-treated pts compared to recurrent tumors belonging to GBM pts treated as per the current standard of care, highlighted a Temferon signature defined by the induction of markers of IFNa responses and macrophage repolarization. Potential long-term benefit with Temferon was identified in a patient from cohort 3, who had disease progression at D+120 with two distant enhancing lesions, and increased tumor necrosis. One year following Temferon, with no 2nd line therapy added, there was approximately 40% reduction in enhancing tumor volume compared to D+180 with a stable clinical and imaging picture thereafter. Conclusion The results provide initial evidence of Temferon’s potential to modulate the TME of GBM patients, and anecdotal evidence for long lasting effects of Temferon in prevention of disease progression.

284MO Targeting the tumor microenvironment of glioblastoma multiforme using a macrophage-based treatment for the local delivery of immune-therapeutic payload: The TEM-GBM study (NCT03866109)

Bone marrow-derived macrophages account for a substantial Glioblastoma (GBM) tumor volume and contribute to the local inflammatory tumor microenvironment (TME), disease progression & treatment response. We have developed a genetically modified, autologous hematopoietic stem cell-based platform designed to deliver IFNa, specifically into the TME via Tie-2 expressing monocytes (Temferon). TEM-GBM is an open-label, phase I/IIa dose-escalation study evaluating safety and efficacy of Temferon in up to 21 newly diagnosed, unmeth-MGMT GBM patients. The patients will be assigned to 7 cohorts and will receive a single increasing dose of Temferon. As of April 2022, 4 doses of Temferon (0.5-3.0x106/kg) were tested across 16 patients assigned to 6 cohorts. Follow-up from surgery is 6–28mo (2–25mo after Temferon). To date, no DLTs have been identified. Temferon-derived progeny as defined by gene-marked cells were found within the hematopoietic system within 2-weeks of Temferon administration and persisted, albeit at lower levels, up to 18mo (longest time of analysis). Very low concentrations of IFNα were detected in plasma and CSF, indicating tight regulation of transgene expression. SAEs were mostly attributed to conditioning chemotherapy (infections) or disease progression (PD) (seizures). 1SUSAR (persistent GGT elevation) occurred. Median OS is 15mo from surgery. Transduced cells were detected in the 2nd surgery specimens of 3 out 4 pts belonging to low dose cohorts. Single-cell RNA seq of the TME highlighted a Temferon signature associated with the induction IFNa responsive genes and macrophage repolarization. Potential long-term benefit with Temferon was identified in a patient from C3, who had PD at D+120 with 2-distant enhancing lesions, and increased tumor necrosis. 1y following Temferon, with no 2nd-line therapy added, there was approximately 40% reduction in enhancing tumor volume compared to D+180 with a stable clinical and imaging picture thereafter. The results provide initial evidence of Temferon’s potential to modulate GBM TME, and anecdotal evidence for long lasting effects of Temferon in prevention of PD.

Lactoferrin and its digestive peptides induce interferon-α production and activate plasmacytoid dendritic cells ex vivo

Plasmacytoid dendritic cells (pDCs) recognise viral single-stranded RNA (ssRNA) or CpG DNA via Toll-like receptor (TLR)-7 and TLR9, and produce interferon (IFN)-α. Activated pDCs upregulate human leukocyte antigen (HLA)-DR and CD86 expression levels. Ingestion of bovine lactoferrin (LF) activates pDCs, but little is known about its effects. In this study, the effects of LF and its pepsin hydrolysate (LFH) on the production of IFN-α from peripheral blood mononuclear cells (PBMCs) and pDCs were examined. PBMCs were prepared from peripheral blood of healthy adults and incubated with LF, LFH, or lactoferricin (LFcin) in the absence or presence of ssRNA derived from human immunodeficiency virus. The concentration of IFN-α in the supernatant and the expression levels of IFN-α, HLA-DR, and CD86 in pDCs were quantified by enzyme-linked immunosorbent assay and flow cytometry. In the absence of ssRNA, the concentration of IFN-α was negligible and LF had no effect on it. In the presence of ssRNA, IFN-α was detected at a certain level, and LF and LFH significantly increased its concentration. The increase caused by LFH and LFcin were comparable. In addition, LF significantly upregulated the expression levels of IFN-α, HLA-DR, and CD86 in pDCs. LF and its digestive peptides induced IFN-α production and activated pDCs in the presence of ssRNA, suggesting that LF modulates the immune system by promoting pDC activation upon viral recognition.

CLRM-08 TARGETING IMMUNE-PAYLOAD TO THE GLIOBLASTOMA TUMOR MICROENVIRONMENT USING A MACROPHAGE-BASED TREATMENT RELYING ON AUTOLOGOUS, GENETICALLY MODIFIED, HEMATOPOIETIC STEM CELL-BASED THERAPY: THE TEM-GBM STUDY (NCT03866109)

We developed an autologous hematopoietic stem cell-based platform designed to deliver IFNa, by a transcriptional and post-transcriptional control mechanism mediated by miRNA target sequences, specifically into the tumor microenvironment (TME) via Tie-2 expressing monocytes (Temferon). As of Feb 2022, 3 escalating doses of Temferon (0.5-2.0x106/kg) were tested across 15 newly diagnosed, unmethylated MGMT GBM patients assigned to 5 cohorts. Follow-up from surgery is 6–28mo (2–25mo after Temferon). To date, no DLTs have been identified. As expected, 1mo after the administration of the highest tested dose, the hematopoietic system of Temferon-treated patients was composed of up to 30% of CD14+ modified cells. Temferon-derived progeny persisted, albeit at lower levels, up to 18mo (longest time of analysis). Despite the substantial proportion of engineered cells, very low concentrations of IFNα were detected in the plasma and in the CSF, indicating tight regulation of transgene expression. SAEs were mostly attributed to conditioning chemotherapy (infections) or disease progression (seizures). 1SUSAR (persistent GGT elevation) occurred. Median OS is 15mo from surgery. Homing of transduced cells to the tumor was demonstrated by the presence of gene-marked cells in the 2nd surgery specimens of 3 out 4 pts belonging to low dose cohorts. Single-cell RNA seq of the TME highlighted a Temferon signature associated with the induction IFNa responsive genes and macrophage repolarization. Potential long-term benefit with Temferon was identified in a patient from cohort 3, who had PD at D+120 with two distant enhancing lesions, and increased tumor necrosis. 1y following Temferon, with no 2nd-line therapy added, there was approximately 40% reduction in enhancing tumor volume compared to D+180 with a stable clinical and imaging picture thereafter. The results provide initial evidence of Temferon’s potential to modulate the TME of GBM patients, and anecdotal evidence for long lasting effects of Temferon in prevention of disease progression.