Lower Levels Of IFN-γ Research Articles

NK cell activity and methylated HOXA9 ctDNA as prognostic biomarkers in patients with non-small cell lung cancer treated with PD-1/PD-L1 inhibitors

BackgroundPD-1/PD-L1 inhibitors have improved survival for patients with non-small cell lung cancer (NSCLC). We evaluated natural killer cell activity (NKA) and methylated HOXA9 circulating tumor DNA (ctDNA) as prognostic biomarkers in NSCLC patients treated with PD-1/PD-L1 inhibitors.MethodsPlasma was prospectively collected from 71 NSCLC patients before treatment with PD-1/PD-L1 inhibitors and before cycles 2–4. We used the NK Vue® assay to measure the level of interferon gamma (IFNγ) as a surrogate for NKA. Methylated HOXA9 was measured by droplet digital PCR.ResultsA score combining NKA and ctDNA status measured after one treatment cycle had a strong prognostic impact. Group 1 had IFNγ < 250 pg/ml and detectable ctDNA (n = 27), group 2 consisted of patients with either low levels of IFNγ and undetectable ctDNA or high levels of IFNγ and detectable ctDNA (n = 29), group 3 had IFNγ ≥250 pg/ml and undetectable ctDNA (n = 15). Median OS was 221 days (95% CI 121–539 days), 419 days (95% CI 235–650 days), and 1158 days (95% CI 250 days—not reached), respectively (P = 0.002). Group 1 had a poor prognosis with a hazard ratio of 5.560 (95% CI 2.359–13.101, n = 71, P < 0.001) adjusting for PD-L1 status, histology, and performance status.ConclusionsCombining NKA and ctDNA status after one cycle of treatment was prognostic in patients with NSCLC treated with PD-1/PD-L1 inhibitors.

Secondary cues alter ISG protein repertoires to unlock antiviral subprograms triggered by interferons

Abstract Immune defenses are triggered by cytokines like interferon-α (IFN-α) and IFN-γ in diverse cells. Both IFNs induce a class of factors termed IFN-stimulated genes (ISGs). Many ISG proteins have antiviral functions. There is a growing appreciation that ISG repertoires are cell-type specific and that this bias impacts infection outcomes. However, the cues and mechanisms driving these variable responses are ill-defined. Given key roles for immunometabolism in immune cell activation, we hypothesized that local cues like metabolites shape tissue specific ISG defenses. To model the impact of metabolic cues on host defenses in non-immune cells, we infected human cells with two unrelated viruses: vaccinia virus, the prototypical poxvirus, and herpes simplex virus-1 (HSV-1). Unexpectedly, we find that cells favoring glycolysis, but not OXPHOS, catalyze potent IFN-γ restriction of vaccinia and HSV-1 that attenuate viral-gene and protein expression as well as infectious virus production. This response is specific as modulating IFN-α signaling by supplementation with either glycolysis or OXPHOS cues has no effect on viral replication. This interferon-induced subprogram is independent of differences in canonical IFN-γ signal transduction, occurs at low levels of IFN-γ, and induced by physiological levels of glucose. RNA-seq reveals no marked changes in the “usual suspects” (e.g. ISGs). Instead, we find secondary cues regulate ISG protein levels. Deletion of a single ISG protein, which is differentially stabilized by secondary cues, is sufficient to rescue vaccinia replication. These data indicate that secondary cues can alter infection outcomes in the same cell-type by unlocking subprograms embedded in interferon catalyzed responses.

Maternal imprinting and determinants of neonates' immune function in the SEPAGES mother-child cohort.

Immune function in pregnancy is influenced by host-specific and environmental factors. This may impact fetal immune development, but the link between maternal and neonatal immune function is still poorly characterized. Here, we investigate the relationship between maternal and neonatal immune function, and identify factors affecting the association between maternal and child cytokine secretion. In the French prospective cohort SEPAGES, blood samples were obtained from pregnant women (n=322) at gestational week 20 ± 4 and from their child at birth (n=156). Maternal and cord blood cytokine and chemokine (CK) levels were measured at baseline in all subjects and after T cell or dendritic cell activation with phytohemagglutinin or R848 (in total 29 and 27 measures in maternal and cord blood samples, respectively). Associations between environmental, individual factors and CK level were estimated by linear regression modeling. The maternal-cord blood CK relations were assessed by Pearson correlation and regression models. We observed that pregnant women and neonates displayed specific CK secretion profiles in the innate and adaptive compartments at baseline and upon activation. Activation of T cells in cord blood induced high levels of IL-2, but low levels of IFNγ, IL-13 or IL-10, in comparison to maternal blood samples. Elsewhere, neonatal innate immune responses were characterized by low production of IFNα, while productions of IL-1β, IL-6, IL-8, IL-10 and TNFα were higher than maternal responses. Strong correlations were observed between most CK after activation in maternal and cord blood samples. Strikingly, a statistical association between global mother and child cytokine profiles was evidenced. Correlations were observed between some individual CK of pregnant women and their children, both at baseline (MCP1, RANTES) and after activation with R848 (IL-6, IL-8 and IL-10). We looked for factors which could influence cytokine secretion in maternal or cord blood, and found that leucocyte counts, maternal age, pre-conception BMI, smoking and season were associated with the levels of several CK in mothers or children. Our study reveals in utero immune imprinting influencing immune responses in infants, opening the way to investigate the mechanisms responsible for this imprinting. Whether such influences have long lasting effects on children health warrants further investigation.

Short- and Long-Term Humoral and Cellular Immune Responses to SARS-CoV-2 Vaccination in Patients with Multiple Sclerosis Treated with Disease-Modifying Therapies.

This study aimed to evaluate short- and long-term humoral and T-cell-specific immune responses to SARS-CoV-2 vaccines in patients with multiple sclerosis (MS) treated with different disease-modifying therapies (DMTs). Single-center observational longitudinal study including 102 patients with MS who consecutively received vaccination against SARS-CoV-2. Serum samples were collected at baseline and after receiving the second dose of the vaccine. Specific Th1 responses following in vitro stimulation with spike and nucleocapsid peptides were analyzed by quantifying levels of IFN-γ. Serum IgG-type antibodies against the spike region of SARS-CoV-2 were studied by chemiluminescent microparticle immunoassay. Patients undergoing fingolimod and anti-CD20 therapies had a markedly lower humoral response than those treated with other DMTs and untreated patients. Robust antigen-specific T-cell responses were detected in all patients except those treated with fingolimod, who had lower IFN-γ levels than those treated with other DMTs (25.8 pg/mL vs. 868.7 pg/mL, p = 0.011). At mid-term follow-up, a decrease in vaccine-induced anti-SARS-CoV-2 IgG antibodies was observed in all subgroups of patients receiving DMTs, although most patients receiving induction DMTs or natalizumab and non-treated patients remained protected. Cellular immunity was maintained above protective levels in all DMT subgroups except the fingolimod subgroup. SARS-CoV-2 vaccines induce robust and long-lasting humoral and cell-mediated specific immune responses in most patients with MS.

Role of serum procalcitonin in the diagnosis and monitoring of treatment response in treatment-naïve subjects with chronic pulmonary aspergillosis

BackgroundInterferon-gamma (IFN-γ) down-regulates plasma procalcitonin (PCT), marker of inflammation. Chronic pulmonary aspergillosis (CPA) is associated with low IFN-γ levels. Thus, plasma PCT may be elevated in CPA and could have a role in diagnosing and monitoring treatment response in CPA. Herein, we investigate the diagnostic performance of plasma PCT in CPA. MethodsWe extracted the demographic, clinical, radiological, treatment outcomes, and plasma PCT levels of CPA subjects and controls (previously treated pulmonary tuberculosis with radiological abnormalities on CT chest [diseased controls] and treatment naïve active pulmonary tuberculosis [PTB]). We treated CPA subjects with six months of oral itraconazole. We took 0.25 ng/mL as a cut-off value for PCT. The study’s primary objective was to ascertain the diagnostic performance of PCT in diagnosing CPA. The key secondary outcome was to study the change in the plasma PCT levels after itraconazole therapy. ResultsWe included 190 CPA cases and 40 controls (diseased controls [n = 20] and active PTB [n = 20]). PCT was elevated (≥0.25 ng/mL) in only 7 (3.7%) subjects with CPA. The sensitivity and specificity of PCT (≥0.25 ng/mL) were 3.7% (1.5–7.4%) and 100 (91.2–100%), respectively. The area under the curve for plasma PCT was 0.48 (95% confidence interval, 0.39–0.58). The plasma PCT values were available in 93 subjects at six months. There was a significant decline in the median plasma levels of PCT after treatment; however, the PCT levels either increased or remained the same in 45% of the subjects. ConclusionPlasma procalcitonin has poor performance in diagnosing and following subjects with CPA.

Characteristics of humoral and cellular responses to coronavirus disease 2019 (COVID-19) inactivated vaccine in central China: A prospective, multicenter, longitudinal study.

In China, the long-term immunogenicity and adverse effects of inactivated vaccines produced by different or the same manufacturer remain unclear. Therefore, the objective of this study was to evaluate the cellular immune responses and neutralizing antibody kinetics of homologous and heterologous administrations of an inactivated coronavirus disease 2019 (COVID-19) vaccine 240 days after the second vaccination. This prospective, multicenter, observational, longitudinal study involved 595 participants with a negative SARS-CoV-2 polymerase chain reaction result who were serologically tested and followed for 8 months after vaccination. Neutralizing antibodies, interferon-gamma (IFN-γ), interleukin (IL)-6, CD4+ T-lymphocyte, and B-lymphocyte counts were evaluated in serum samples after stimulation with 2 μg/mL SARS-CoV-2 spike protein for 16h at follow-up intervals of 2 months. Most participants [582/595; 146 male participants, 449 female participants; mean age 35 (26-50 years)] rapidly developed neutralizing antibodies after two doses of the vaccine administered 3-weeks apart. The positive rate of neutralizing antibodies peaked at 97.7% at 60-90 days, decreased, and stabilized at 82.9% at 181-240 days post-vaccination. Lower antibody concentrations were correlated with older age, longer duration after vaccination, non-health care workers, mixed-manufacturer vaccinations, and intervals of less than 40 days between two doses of vaccination, whereas lower IFN-γ levels and B-lymphocyte counts were associated with older age, blood type A, and non-health care workers. A higher IL-6 level was associated with older age, mixed-manufacturer vaccinations, intervals of less than 40 days between two doses of vaccination, and medical staff. Adverse reactions were mild or moderate and self-limited, with no serious events reported. Two doses of the Chinese inactivated vaccine induced robust and rapid antibody expression and cellular immune responses. Boosting vaccination is considered important, as antibodies and cellular immune responses were reduced in susceptible populations.

Variants of NOD2 in Leishmania guyanensis-infected patients with cutaneous leishmaniasis and correlations with plasma circulating pro-inflammatory cytokines.

Leishmaniases, a group of vector-borne diseases, are caused by the protozoan intracellular parasite Leishmania (L.) and are transmitted by the phlebotomine sandflies. A wide range of clinical manifestations in L- infection is observed. The clinical outcome ranges from asymptomatic, cutaneous leishmaniasis (CL) to severe mucosal leishmaniasis (ML) or visceral leishmaniasis (VL), depending on the L. species. Interestingly, only a fraction of L.-infected individuals progress to disease development, suggesting a key role of host genetics in the clinical outcome. NOD2 plays a critical role in the control of host defense and inflammation. The NOD2-RIK2 pathway is involved in developing a Th1- type response in patients with VL and C57BL/6 mice infected with L. infantum. We investigated whether variants in the NOD2 gene (R702W rs2066844, G908R rs2066845, and L1007fsinsC rs2066847) are associated with susceptibility to CL caused by L. guyanensis (Lg) in 837 patients with Lg-Cl and 797 healthy controls (HC) with no history of leishmaniasis. Both patients and HC are from the same endemic area of the Amazonas state of Brazil. The variants R702W and G908R were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and L1007fsinsC was by direct nucleotide sequencing. The minor allele frequency (MAF) of L1007fsinsC was 0.5% among the patients with Lg-CL and 0.6% in the healthy controls group. R702W genotypes frequencies were similar in both groups. Only 1% and 1.6% were heterozygous for G908R among the patients with Lg-CL and HC, respectively. None of the variants revealed any association with susceptibility to the development of Lg-CL. Correlations of genotypes with the level of plasma cytokines revealed that individuals with the mutant alleles of R702W tend to have low levels of IFN-γ. G908R heterozygotes also tend to have low IFN-γ, TNF-α, IL-17, and IL-8. Variants of NOD2 are not involved in the pathogenesis of Lg-CL.

Peripheral Blood Mononuclear Cells from Patients with Type 1 Diabetes and Diabetic Retinopathy Produce Higher Levels of IL-17A, IL-10 and IL-6 and Lower Levels of IFN-γ-A Pilot Study.

Inflammation is key to the pathogenesis of diabetic retinopathy (DR). This prospective study investigated alterations in inflammatory cytokines in peripheral blood mononuclear cells (PBMCs) in 41 people with type 1 diabetes (T1D), sub-grouped into mild non-proliferative DR (mNPDR; n = 13) and active and inactive (each n = 14) PDR. Age/gender-matched healthy controls (n = 13) were included. PBMCs were isolated from blood samples. Intracellular cytokine expression by PBMCs after 16-h stimulation (either E. coli lipopolysaccharide (LPS), phorbol 12-myristate 13-acetate plus ionomycin, D-glucose or D-mannitol) were assessed by flow cytometry. Cytokine production in plasma, non-stimulated and LPS-stimulated PBMC supernatant was also assessed. Increased BMC IL-10 secretion and reduced expression of IL-6 and IFN-γ in CD3+ cells were observed in mNPDR. Reduced IL-6 and IL-10 secretion, and higher levels of intracellular IL-6 expression, especially in CD11b+ PBMCs, was detected in aPDR; levels were positively correlated with DR duration. Patients with T1D demonstrated increased intracellular expression of IL-17A in myeloid cells and reduced IFN-γ expression in CD3+ cells. Plasma levels of IL-1R1 were increased in mNPDR compared with controls. Results suggest that elevated PBMC-released IL-10, IL-6, in particular myeloid-produced IL-17A, may be involved in early stages of DR. IL-6-producing myeloid cells may play a role in PDR development.

Subsets of natural killer cells in chronic myeloid leukemia and their relation with some inflammatory cytokines

BACKGROUND: As in other malignancies, different subsets of natural killer (NK) cells play a crucial role in the recognition and lysing of malignant cells in chronic myeloid leukemia (CML). OBJECTIVES: This study aims to identify two subsets of NK, cytotoxic (cluster of differentiation [CD] 16+bright) and cytokine-producing NK (CD56+bright) in newly diagnosed CML patients. MATERIALS AND METHODS: This study is conducted on 20 newly diagnosed Iraqi patients (12 males and 8 females) with CML, in chronic phase, at the age range of 17–55 years. Along with patients, 20 healthy subjects (with matched age and gender) were enrolled to act as a control group. To identify NK cells and their subsets in peripheral blood samples, the expression of CD45, CD3, CD56, and CD16 markers was evaluated by flow cytometry technique. Furthermore, the serum level of interferon gamma (IFN-γ) and interleukin (IL)-18 was determined by the enzyme-linked immunosorbent assay technique. RESULTS: The age of patients at the diagnosis of disease is (35.6 ± 12.2 years) and the male: female ratio was 1.5:1. The serum level of IL-18 in newly diagnosed CML patients (30.3 ± 6.5 pg/mL) was significantly (P &lt; 0.0001) higher than those in control group (18.3 ± 7.8 pg/mL), while the serum levels of IFN-γ in newly diagnosed patients are significantly (P = 0.006) dropped down to (89.1 ± 7.2 pg/mL from that in control group (109.4 ± 30.3 pg/mL). The percentage of NK cells in newly diagnosed CML patients is significantly lower than in control group. There is a significant elevation in the cytotoxic NK cells (CD16+bright) subset, and a significant decrease in the cytokine-producing NK subset (CD56+bright) in newly diagnosed patients when compared to those in control group. CONCLUSION: Although there is an elevation in the percentage of cytotoxic NK cells (CD16+bright) subset of CML patients at the first diagnosis, these cells are not able to recognize and attack malignant cells, which may be due to low expression of their activating receptors and needs more investigation. Furthermore, present results found a low percentage of cytokine-producing NK cells (CD56+bright) and a low level of IFN-γ in CML patients, although there is an elevation in IL-18, which indicates that IL-18 may be not the main stimulator to these cells, so activation pathway of this subset of NK cells needs further investigation.

Association between serum levels of 12 different cytokines and short-term efficacy of anti-PD-1 monoclonal antibody combined with chemotherapy in advanced gastric cancer.

Gastric cancer (GC) is characterized by aggressive tumor growth and poor prognosis. The benefits of targeted anti-programmed death receptor 1 (PD-1) monoclonal antibody combined with chemotherapy have not yet been characterized. The tumor microenvironment and circulating factors have garnered interest as possible predictors of response and prognosis. The aim of this study was to evaluate whether cytokine levels in the serum of patients were related to tumor response to anti-PD-1 monoclonal antibody combined with chemotherapy and survival in advanced GC. Preoperative serum samples were collected from patients with GC (n=52) and healthy individuals (n=31). The levels of 12 different cytokines were measured using a multiple microsphere flow immunofluorescence assay. The association between cytokine levels and clinical response was analyzed using nonparametric Wilcoxon matched-pair ranked tests. Progression-free survival (PFS) time for all patients was recorded via evaluation of imaging results and follow-up via telephone. Kaplan-Meier and log-rank tests were used to plot survival curves. The levels of interleukin (IL)-6, IL-1β, interferon (IFN)-γ, IL-17, and IL-12p70 in the control group were significantly lower than those in the GC group (p=0.0002, p=0.0065, p=0.0003, p=0.0303, and p=0.0295, respectively). The level of IL-4 was significantly higher in healthy individuals than that in patients with GC (p=0.0201). The cytokine levels in the good responder group were higher than those in the poor responder group before therapy. Patients treated with immunochemotherapy showed an overall reduction in all cytokine levels after treatment initiation. A high baseline level of IFN-γ was associated with a better prognosis. However, high IL-6 levels in patients after two cycles of immunochemotherapy indicated resistance. High IL-4 levels in patients treated with four cycles of immunochemotherapy were associated with better PFS. Our study suggests that low levels of IFN-γ before immune checkpoint inhibitor treatment may be useful for the detection of a poor immunological status. Hence, a reduction in IL-6 levels is predictive of a longer PFS, and increased IL-4 levels are predictive of a good response. IL-4 and IL-6 may, therefore, serve as promising circulating predictive biomarkers for patients who can benefit from anti-PD-1 monoclonal antibodies administered in combination with chemotherapy.

Predictors for False-Negative Interferon-Gamma Release Assay Results in Hemodialysis Patients with Latent Tuberculosis Infection.

The present study aimed to clinically evaluate the effect of T-cell dysfunction in hemodialysis (HD) patients with latent tuberculosis (TB) infection (LTBI) who were false-negatives in the QuantiFERON-TB Gold In-Tube (QFT-GIT) test. Whole blood samples from a total of 20 active TB patients, 83 HD patients, and 52 healthy individuals were collected, and the QFT-GIT test was used for measuring Mycobacterium tuberculosis (MTB)-specific interferon gamma (IFN-γ) level. The positive rate of the IFN-γ release assays (IGRAs) in HD patients was lower than the negative rate. The mean value of MTB-specific IFN-γ level, which determines the positive rate of the IGRA test, was highest in active TB, followed by HD patients and healthy individuals. Among HD patients, phytohemagglutinin A (PHA)-stimulated IFN-γ levels of approximately 40% were 10.00 IU/mL or less. However, there was no low level of PHA-stimulated IFN-γ in the healthy individuals. This reveals that T-cell function in HD patients was reduced compared to healthy individuals, which leads to the possibility that QFT-GIT results in HD patients are false-negative. The clinical manifestations of TB in patients on HD are quite non-specific, making timely diagnosis difficult and delaying the initiation of curative treatment, delay being a major determinant of outcome.

Ex vivo cytokine production in psoriatic disease: Towards specific signatures in cutaneous psoriasis and peripheral psoriatic arthritis.

Psoriatic arthritis (PsA) and cutaneous psoriasis (PsO) are different phenotypes of psoriatic disease (PsD), whose underlying specific mechanisms remain incompletely understood. As cytokines are key elements to induce and tune up immune responses to drive inflammatory diseases, our objective was to assess whether clinical features, disease phenotype and PsA and PsO activity were associated with a particular ex vivo cytokine production profile. Forty-eight patients (37 PsA and 11 PsO) and 11 healthy subjects (HS) were studied. Cytokine production by peripheral blood mononuclear cells (PBMC) that were either unstimulated, or stimulated with LPS or anti-CD3/CD28 antibodies, were analysed by multiplex assay in the culture supernatants. Cytokine signature of PsD includes a high level of TNFα in supernatants of LPS-stimulated PBMC, higher levels of IL-6 and lower levels of IFN-γ and IL-17A after CD3-CD28 stimulation, as well as higher spontaneous IL-1RA and TNFα production compared to HS. High body mass index (BMI) was associated with lower levels of IL-1β, and metabolic syndrome with lower levels of IFN-γ after LPS stimulation. In PsD, dermatological activity was related with higher IL-17A level, while rheumatic activity was linked with lower levels of IFN-γ and TNFα. Comparing each PsD subtype to HS, IL-1β and IL-6 productions are higher when using LPS stimulation in PsO patients with higher levels of IL-1β and IL-1α in peripheral PsA patients after CD3/CD28 stimulation. LPS stimulation induced high levels of IL-17A in peripheral PsA compared to axial PsA. PsA patients with axial PsA share some features with PsO but shows a distinct cytokine pattern compared to peripheral PsA. PsO and the different PsA subtypes exhibit distinct ex vivo cytokine production profiles and common features of the so-called PsD. Analysis of IL-1 cytokine family and IL-6 seems to be of particular interest to distinguish PsO and peripheral PsA since it depends on monocytes in PsO and T-lymphocytes in peripheral PsA. Peripheral cytokine profiles are influenced by rheumatic and dermatological activity of the disease, and also by metabolic syndrome features. Our results highlight the crucial role of immune cell interactions with different patterns of interaction depending on clinical phenotype.

Feeding tannins to dairy cows in different seasons improves the oxidative status of blood plasma and the antioxidant capacity of cheese.

The aim of the present study was to assess the dietary supplementation of tannins to grazing dairy cows in 2 seasons characterized by a good quality pasture (spring) or a poor-quality pasture (summer). The effects of dietary tannins were assessed on plasma antioxidant status and cytokines profile and on the antioxidant properties of cheese and cheese in vitro digestates. Fourteen lactating dairy cows were divided into 2 homogeneous groups (n = 7): a control group (CON), and an experimental group (TAN) receiving 150 g/head per day of tannins supplementation. The experiment was performed twice, in spring and in summer. The animals were free to graze on spontaneous pasture (spring) or on dry stubble (summer). Blood was sampled at the beginning (d 0), at the midpoint (d 11), and at the end (d 22) of the trial. Individual cheese was produced before the beginning (d -1) and at the end (d 22) of the trial from the milk collected from each cow. On blood plasma, the reactive oxygen metabolites (ROM), biological antioxidant potential (BAP), nonesterified fatty acids quantification, and cytokines profile in terms of IL-10, IL-8, IL-1β, and IFN-γ were determined. Data on ROM demonstrated that tannins supplementation lowered oxidative stress both in spring and in summer. Accordingly, TAN diet increased BAP levels compared with the CON during summer trial. Thus, feeding tannins resulted in lower ratio between ROM and BAP (oxidative stress index) in both spring and summer. Cytokines' profile showed lower IL-1β values in TAN group at d 22 during spring season, with a concomitant higher IL-10 level, during summer season. Moreover, TAN group had a lower level of IFN-γ in plasma than CON group, both in spring and in summer. On cheese samples, the in vitro digestion was performed and on cheese and cheese digestates (gastric and intestinal digestate) the free radical scavenging antioxidant activity was evaluated. The intestinal digestate fraction registered the highest antioxidant activity compared with cheese and gastric digestate, in both spring and summer seasons. Furthermore, an improvement of the antioxidant property of cheese and cheese digestates was found. Present data demonstrated that tannins supplementation contributed to reduce the oxidative stress of lactating dairy cows and showed an increase of anti-inflammatory cytokines ratio.

INDICATORS OF PARAMETERS OF CYTOKINE STATUS IN BACTERIAL VAGINOSIS ASSOCIATED WITH FEMALE AGE

According to scientific reports, the frequency of bacterial vaginosis (BV) is quite variable and ranges from 30% to 80%, and in 24-50% of cases, it is asymptomatic. In the general population, BV is diagnosed in 20% of women, in 87% - with pathological secretions, in the structure of patients who turned to the primary outpatient appointment - 41%&#x0D; The purpose of the study: to establish the nature of immunological violations of the parameters of local anti-infective protection, which underlie the development and progression of bacterial vaginosis in women of reproductive age.&#x0D; Research materials and methods: to assess the state of local immune protection, a study of women of reproductive age with bacterial vaginosis was conducted. 30 patients under the age of 35 belonged to group I; group 2 consisted of 30 women over the age of 35. The control group included 30 patients of reproductive age with normobiocenosis of the vaginal mucosa (including: 10 women under 35 years of age and 20 women over 35 years of age) without acute gynecological pathology, infertility, endometriosis and background processes of the cervix. Methods of measuring the vagina pH, Gram microscopy, Amsel’s diagnostic criteria, dysbiosis according to the Real-Time PCR "Femoflor" were used; PCR test for antigens of TORCH infections and statistical methods were also applied.&#x0D; Research results and their discussion. In patients of the older age category, women presented clinical complaints in the form of increased secretion and discomfort 1.7 times more often, which accounted for 63.3% of observations. Severe dysbiosis was diagnosed 1.6 times more often in patients of the older age category compared to the data of women of group I with significant statistical differences in the control. A decrease in the level of lactoferrin was revealed in the samples of women of group 2 (by 3.8 times). Indicators of local immune protection were characterized by an increase in the concentration of IgM by 3.4 times, depression of sIg A synthesis (by 2.7 times) in a third of observations (36.7%), a decrease in the level of IL-1β (by 1.7 times), an increase in levels of TNF-α (by 3.8 times), low levels of IFN-γ – by 1.9 times (p&lt;0.05), which demonstrates reduced opportunities to eliminate pathogens and an imbalance of cytokine synthesis. Low indicators of the level of IFN-γ, as well as a decrease in the level of the anti-inflammatory cytokine IL-4 (by 2.5 times compared to control data, p&lt;0.05), as a marker of the immunopathological process, demonstrate the depression of interferonogenesis processes, reduced opportunities for the elimination of pathogens and imbalance cytokine synthesis.&#x0D; Conclusions. In patients with bacterial vaginosis and in those who are older than 35 years, a characteristic feature of local immunity is a pronounced dysbiosis caused by hormonal imbalance and hypoestrogenemia, a decrease in the level of lactoferrin, depression of sIg A synthesis., interferon deficiency and imbalance of pro-inflammatory cytokine synthesis. Clinical assessment of inflammation from the standpoint of the immune response prevents a one-sided stencil approach to the correction of violations of the vaginal micro ecosystem, helps to improve the diagnosis of the phase of immune inflammation, and optimize the use of antibacterial agents.&#x0D; Further studies of the level of parameters of local immune protection, taking into account the comorbidity of gynecological pathology and the violation of the micro ecosystem of the vaginal mucosa, are promising.

Donor peritoneal-derived cells can attenuate graft-versus-host disease after MHC-incompatible bone marrow transplantation in mice

BackgroundGraft-versus-host disease (GVHD) remains one of the most important barriers to bone marrow transplantation (BMT) and quality of life. Peritoneal-derived cells (PCs) can regulate immune function. MethodsThe GVHD mouse model, which was established by injection with T-cell depleted bone marrow cells and spleen cells, was treated with donor PCs. The survival time, weight, GVHD score, and pathological organ damage were used to assess the efficacy of PCs. Various cytokines (e.g., TNF-α, IFN-γ, IL-2, IL-17, IL-15, IL-4, IL-10, and TGF-β) and immunocytes (e.g., CD4+, CD8+, Treg, and NK cells) were employed to investigate the preliminary mechanism. The cytokine levels were tested via Luminex liquid chips, real-time PCR, and Western blotting, and the proportion of each T-cell subpopulation was detected via flow cytometry. One-way Analysis of Variance (ANOVA) followed by Tukey's post-test was employed to perform multiple comparisons, and Kaplan–Meier method was used to perform survival analysis. ResultsCompared to the saline group, the PC group harbored a significant reduction in pathological damage to the colon and small intestine, a longer survival time, and a lower GVHD score. The mice in the PC group had significantly higher levels of TGF-β, IL-10, and IL-4 and lower levels of IFN-γ, TNF-α, and IL-15 than those in the saline group. In the PC group, CD8+ and Th1 ratios decreased, while CD4+, Th2, NK, and Treg ratios increased. ConclusionsDonor PCs can attenuate GVHD caused by MHC-incompatible BMT by regulating cytokine levels and immunocyte ratios.

Clinical and diagnostic significance of determining the cytokine status in infants with chronic cytomegalovirus infection against the background of hypoxic-ischemic CNS damage

Objective: based on the production of cytokines, to identify the immunological features of the chronic course of cytomegalovirus infection in children of the first year of life against the background of hypoxic-ischemic CNS damage.Research methods:108 newborns with cytomegalovirus infection occurring against the background of perinatal hypoxic-ischemic lesions of the central unequal system were examined. All observed patients immediately after the diagnosis of cytomegalovirus infection underwent an immunological examination, including the determination of the levels of interferon-alpha (IFN-α) and interferon-gamma (IFN-γ), the level of interleukins — 2 and 4 (IL -2 and IL-4) necrosis factor human alpha tumors (TNF-α in blood serum was determined by enzyme immunoassay using a set of reagents ProCon IF2 plus, ProCon Ifgamma, ProCon TNFα (Protein contour LLC, Russia, St. Petersburg). At 1 and 6 months of life .The observation groups consisted of 78 children (72.2%) with an acute course of the disease (Group 1) and 30 children (27.3%) with a chronic course (Group 2). The control group consisted of 15 newborns without herpes virus infection.Results. Of the totality of the studied cytokines, statistically significant for the chronic course of cytomegalovirus infection in children of the first year of life against the background of hypoxic-ischemic CNS damage were found: IL-2, IFN-γ. It was found that in children with a persistent low level of IFN-γ and an increased level of IL-4 in the blood serum at the age of 6 months, there was a chronic course of cytomegalovirus infection against the background of perinatal hypoxic-ischemic CNS damage.A decrease in IFN-γ production indicates a congenital or acquired deficiency of the interferon system and can be considered as an indication for long-term interferon replacement therapy.

Low baseline IFN-γ response could predict hospitalization in COVID-19 patients.

The SARS-CoV-2 infection has spread rapidly around the world causing millions of deaths. Several treatments can reduce mortality and hospitalization. However, their efficacy depends on the choice of the molecule and the precise timing of its administration to ensure viral clearance and avoid a deleterious inflammatory response. Here, we investigated IFN-γ, assessed by a functional immunoassay, as a predictive biomarker for the risk of hospitalization at an early stage of infection or within one month prior to infection. Individuals with IFN-γ levels below 15 IU/mL were 6.57-times more likely to be hospitalized than those with higher values (p<0.001). As confirmed by multivariable analysis, low IFN-γ levels, age >65 years, and no vaccination were independently associated with hospitalization. In addition, we found a significant inverse correlation between low IFN-γ response and high level of IL-6 in plasma (Spearman’s rho=-0.38, p=0.003). Early analysis of the IFN-γ response in a contact or recently infected subject with SARS-CoV-2 could predict hospitalization and thus help the clinician to choose the appropriate treatment avoiding severe forms of infection and hospitalization.

Clinical Correlation of Function and TCR vβ Diversity of MAGE-C2-Specific CD8+ T Cell Response in Esophageal Cancer.

Melanoma-associated Ag (MAGE)-C2, an immunogenic cancer germline (testis) Ag, is highly expressed by various tumor cells, thymic medullary epithelial cells, and germ cells. In this study, we aimed to explore the immunologic properties of MAGE-C2-specific CD8+ T cells and the relationship of its TCR β-chain V region (TCR vβ) subfamily distribution to prognosis of patients with esophageal cancer. PBMCs and tumor-infiltrating lymphocytes expanded by CD3/CD28 Dynabeads and MAGE-C2 peptides in vitro resulted in the induction of lysosome-associated membrane protein-1 (LAMP-1 or CD107a) on the cell surface and the production of IFN-γ by MAGE-C2-specific CD8+ T cells. We found differential TCR vβ subfamily distribution among flow-sorted CD107a+IFN-γ+ and CD107a-IFN-γ- CD8+ T cells. The proportion of CD107a+ and/or IFN-γ+ tetramer+ CD8+ T cells was lower in patients with lymph node metastasis, late tumor stage, and poorly differentiated state (p < 0.05). T-box transcription factor was positively correlated with CD107a and IFN-γ. Kaplan-Meier analysis showed that patients whose MAGE-C2-specific CD8+ T cells expressed high CD107a and/or IFN-γ had a longer survival time when compared with patients whose MAGE-C2-specific CD8+ T cells expressed low levels of CD107a and/or IFN-γ. Moreover, analysis of TCR vβ subfamily distribution revealed that a higher frequency of TCR vβ16 in MAGE-C2-specific CD8+ T cells was positively correlated with a better prognosis. These results suggest that the presence of functional MAGE-C2-specific CD8+ T cells had an independent prognostic impact on the survival of patients with esophageal cancer.

Smoking negatively impacts the clinical, microbiological, and immunological treatment response of young adults with Grade C periodontitis.

This study aimed to investigate the influence of smoking on clinical, microbiological and immunological parameters in young adult with stage III-IV Grade C periodontitis after full-mouth ultrasonic debridement (FMUD) associated with Amoxicillin and Metronidazole (AMX + MTZ), comparing smokers (PerioC-Y-Smk) with non-smokers (PerioC-Y-NSmk). Fifteen PerioC-Y-NSmk and 14 PerioC-Y-Smk patients underwent FMUD associated with AMX + MTZ for 10 days. All parameters were collected at baseline and 3 and 6months after treatment. Plaque index (PI), bleeding on probing (BoP), probing depth (PD), clinical attachment level (CAL)- the primary variable-, and gingival recession (GR) were clinically assessed. The impact of PI on CAL change at 6-month was verified by a regression analysis. Samples of the subgingival biofilm was collected for detection of levels of Aggregatibacter actinomycetemcomitans (A.actinomycetemcomitans), Porphyromonas gingivalis (P.gingivalis), Tannerella forsythia (T.forsythia), and Fusobacterium nucleatum ssp (F.nucleatum), and were analyzed by real-time qPCR; gingival crevicular fluid was collected for detection of levels of interleukin (IL)-1β, IL-4, IL-6, IL-10, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ, which were analyzed using an enzyme immunoassay. PerioC-Y-Smk had significantly higher PI, BOP, and GR at baseline compared to non-smokers (p< .05). PerioC-Y-Smk presented higher PD, CAL, and GR at 3 and 6months (p<.05) compared with PerioC-Y-NSmk in the same periods; PI negatively affected CAL gain in PerioC-Y-NSmk at 6-month follow-up (p=.052) and did not impact on clinical response in PerioC-Y-Smk (p=.882). Lower levels of IFN-γ, IL1-β, and IL-4 were observed at 3months in the PerioC-Y-NSmk (p<.05) compared with PerioC-Y-Smk. Lower proportions of P.gingivalis were observed in PerioC-Y-NSmk at baseline and at 3months (p<.05) and lower proportions of F.nucleatum were observed at 6months, in the PerioC-Y-NSmk (p<.05). PerioC-Y-Smk presents an unfavorable clinical, microbiological, and immunological response after 3 and 6months after FMUD associated with AMX + MTZ. Smoking worsens periodontal condition of young treated adults presenting stage III/IV Grade C periodontitis.

The In Vitro and In Vivo Effect of Lipoxygenase Pathway Inhibitors Nordihydroguaiaretic Acid and Its Derivative Tetra-O-methyl Nordihydroguaiaretic Acid against Brucella abortus 544.

This study investigated the contribution of lipoxygenase (LOX) inhibitors, nordihydroguaiaretic acid (NDGA), tetra-O-methyl nordihydroguaiaretic acid (M4N) and zileuton (ZIL), and thromboxane A2 (TXA2) inhibitor 4,5-diphenylimidazole (DPI) in the proliferation of Brucella abortus infection. None of the compounds affected the uptake of Brucella into the macrophages. We determined the effect of neutralizing leukotriene B4 (LTB4) receptor and showed that the uptake of the bacteria was inhibited at 30 min post-infection. M4N treatment attenuated intracellular survival of Brucella at 2 h post-incubation but it was not observed in the succeeding time points. DPI treatment showed reduced survival of Brucella at 24 h post-incubation while blocking LTB4 receptor was observed to have a lower intracellular growth at 48 h post-incubation suggesting different action of the inhibitors in the course of the survival of Brucella within the cells. Reduced proliferation of the bacteria in the spleens of mice was observed in animals treated with ZIL or DPI. Increased serum cytokine level of TNF-α and MCP-1 was observed in mice treated with M4N or ZIL while a lower IFN-γ level in ZIL-treated mice and a higher IL-12 serum level in DPI-treated mice were observed at 7 d post-infection. At 14 d post-infection, ZIL-treated mice displayed reduced serum level of IL-12 and IL-10. Overall, inhibition of 5-LOX or TXA2 or a combination therapy promises a potential alternative therapy against B. abortus infection. Furthermore, strong ligands for LTB4 receptor could also be a good candidate for the control of Brucella infection.