Analysis Of Immunological Parameters Research Articles

Loss-of-phosphorylation of IKZF1 results in gain-of-function associated with immune dysregulation

BackgroundImmune dysregulation often presents as autoimmunity, inflammation and/or lymphoproliferation. Several germline genetic defects have been associated with immune dysregulation including heterozygous gain-of-function (GOF) mutations in IKZF1, an essential transcription factor for hematopoiesis containing zinc finger domains (ZFs). However, in a large part of patients the genetic origin of their immunedysregulation remains undetermined. ObjectiveA family with two members presenting immune dysregulation signs was studied to identify the genetic cause of their disease. MethodsWhole exome sequencing, analysis of immunological parameters, functional assays including western blotting, EMSA during cell cycle and T helper differentiation were performed. ResultsThe two patients carried a novel heterozygous mutation in IKZF1 (IKZF1T398M). IKZF1 heterozygous mutations have been previously shown to be responsible of several distinct human immunological diseases by directly impacting the ability of ZFs to bind to DNA or to dimerize. Herein, we showed that the IKZF1T398M which is outside the ZFs caused impaired phosphorylation of IKZF1 resulting in enhanced DNA-binding ability at the S phase of cell cycle, reduction of the G1-S phase transition and decreased proliferation. Confirming these data, similar functional alterations were observed with IKZF1T398A, but not with IKZF1T398D mimicking dephosphorylation and phosphorylation, respectively. In T lymphocytes, expression of IKZF1T398M led to T helper (TH) differentiation skewed towards TH2. Thus, our data indicate that the IKZF1T398M behaves as a GOF variant underlying immune dysregulation. ConclusionDisturbed IKZF1 phosphorylation represents a novel GOF mechanism (gain-of-function by loss-of-phosphorylation; GOF-LOP), associated with immune dysregulation, highlighting the regulatory role of IKZF1 during cell cycle progression through phosphorylation.

Early expansion of activated adaptive but also exhausted NK cells during acute severe SARS-CoV-2 infection.

The analysis of immunological parameters during the course of a SARS-CoV-2 infection is of great importance, both to identify diagnostic markers for the risk of a severe course of COVID-19, and to better understand the role of the immune system during the infection. By using multicolor flow cytometry we compared the phenotype of Natural Killer (NK) cells from hospitalized COVID-19 patients during early SARS-CoV-2 infection with samples from recovered and SARS-CoV-2 naïve subjects. Unsupervised high-dimensional analysis of 28-color flow cytometric data revealed a strong enrichment of NKG2C expressing NK cells in response to the acute viral infection. In addition, we found an overrepresentation of highly activated NK cell subsets with an exhausted phenotype. Moreover, our data show long-lasting phenotypic changes within the NK cell compartment that did not completely reverse up to 2 months after recovery. This demonstrates that NK cells are involved in the early innate immune response against SARS-CoV-2.

Cellular and Humoral Disorders of the Immune System at Osteoarthritis with Comorbidity of Exocrine Pancreatic Insufficiency

The objective: to study the change in immunological parameters in patients with primary osteoarthritis (OA) under conditions of comorbidity with the digestive tract (DT) diseases, accompanied by exocrine pancreatic insufficiency (EPI). Materials and methods. The study included 304 patients with primary OA in comorbidity with DT diseases, accompanied by EPI out of exacerbation. The comparison group included 30 almost healthy persons. The patients with primary OA were comparable by age, gender and course of primary OA and were divided into 5 groups by type of comorbid pathology of DT organs, accompanied by EPI: the I group (n=62) – patients with primary OA without comorbid pathology, the 2d group (n=59) – patients with primary OA in comorbidity with chronic pancreatitis (CP), the 3d group (n = 60) – patients with primary OA in comorbidity with chronic noncalculous cholecystitis and functional diseases of gallbladder and biliary system, the 4th group (n=61) – patients with primary OA and chronic gastroduodenitis, the 5th group (n=61) – patients with primary OA and type 2 diabetes mellitus (T2DM). Results. The analysis of immunological parameters in patients with primary OA indicated on the statistically significant changes in the studied indicators in all groups compared to the control group (p<0.001), which specifies immunological disorders in the patients. A statistically significant increase in the level of T-lymphocytes in all studied groups was revealed compared to the control group (p<0.001), which indicates the activation of the T-cell link in immunity in patients with primary OA. The presence of negative effects of comorbid gastroenterological diseases in primary OA on the formation of immunodeficiency was determined. Conclusions. A statistically higher level of autoimmune disorders in patients with primary osteoarthritis (OA) has been established under conditions of comorbidity with gastroenterological diseases, accompanied by exocrine pancreatic insufficiency. It is proved that the deepening and progression of autoimmune disorders in primary OA is affected by the comorbidity of the digestive tract pathology.

Multi-omic approach identifies a transcriptional network coupling innate immune response to proliferation in the blood of COVID-19 cancer patients

Clinical outcomes of COVID-19 patients are worsened by the presence of co-morbidities, especially cancer leading to elevated mortality rates. SARS-CoV-2 infection is known to alter immune system homeostasis. Whether cancer patients developing COVID-19 present alterations of immune functions which might contribute to worse outcomes have so far been poorly investigated. We conducted a multi-omic analysis of immunological parameters in peripheral blood mononuclear cells (PBMCs) of COVID-19 patients with and without cancer. Healthy donors and SARS-CoV-2-negative cancer patients were also included as controls. At the infection peak, cytokine multiplex analysis of blood samples, cytometry by time of flight (CyTOF) cell population analyses, and Nanostring gene expression using Pancancer array on PBMCs were performed. We found that eight pro-inflammatory factors (IL-6, IL-8, IL-13, IL-1ra, MIP-1a, IP-10) out of 27 analyzed serum cytokines were modulated in COVID-19 patients irrespective of cancer status. Diverse subpopulations of T lymphocytes such as CD8+T, CD4+T central memory, Mucosal-associated invariant T (MAIT), natural killer (NK), and γδ T cells were reduced, while B plasmablasts were expanded in COVID-19 cancer patients. Our findings illustrate a repertoire of aberrant alterations of gene expression in circulating immune cells of COVID-19 cancer patients. A 19-gene expression signature of PBMCs is able to discriminate COVID-19 patients with and without solid cancers. Gene set enrichment analysis highlights an increased gene expression linked to Interferon α, γ, α/β response and signaling which paired with aberrant cell cycle regulation in cancer patients. Ten out of the 19 genes, validated in a real-world consecutive cohort, were specific of COVID-19 cancer patients independently from different cancer types and stages of the diseases, and useful to stratify patients in a COVID-19 disease severity-manner. We also unveil a transcriptional network involving gene regulators of both inflammation response and proliferation in PBMCs of COVID-19 cancer patients.

Взаимодействие иммунокомпетентных элементов крови при изолированной закрытой тупой травме почки

INTRODUCTION Blunt renal trauma manifest at macro- and microstructural levels as concussions, bruises or lacerations. The pyelonephritis predominates in structure of posttraumatic compli- cations. AIM To study immune changes in patients with renal trauma and their effect on the development of inflammatory post-traumatic complications. MATERIALS AND METHODS Multivariate statistical analysis of immunological parameters in the immediate post-traumatic period in 92 patients with 2-3 degrees of isolated blunt renal trauma, according to the classification OIS/OI AAST, 2018), was carried out. RESULTS three main components were identified: the first was associated with the lymphocyte count; the second component included high indicators of adaptive cellular immunity CD+19, CD+8, while the third component represented indicators of nonspecific humoral immunity. They accounted for 43.7%, 37.5% and 18.9% of the variance of intergroup differences, respectively. A strong negative correlation between neutrophils and lymphocytes and a weak positive correlation between IgM and IgG were revealed, which altogether indicated the preserved adaptive response of the immune system. CONCLUSION The multivariate analysis of immunological reactivity suggests a sufficient degree of mobilization of the immune system for the adequate adaptive response in patients with isolated blunt kidney trauma, which makes the prediction of the development of post-traumatic pyelonephritis doubtful.

The level of the general reactivity of erythrocytes membranes in rats with combined action of infrasound

This study focuses on the level of the general reactivity of erythrocytes membranes in rats with combined action of infrasound. The effect on biological features was investigated through the experiments on the rats. Comparative analysis of immunological parameters, the permeability of erythrocyte membranes and the electrical conductivity of blood in rats under the effects of infrasound at a frequency of 15 Hz and intensity 11 dB through exposure time of 3-3600 seconds allowed proving that infrasound causes nonspecific changes at the cellular and subcellular levels of the organism. There was used many different technologies and methods. Based on the obtained results, direct indirect and combined effect of infrasound immunological parameters had clearly increased the total count of leukocyte index and all parameters of differential count of leukocytes as well as appearance immature products in the peripheral blood.

Effects of potential probiotic Bacillus subtilis KADR1 and its subcellular components on immune responses and disease resistance in Labeo rohita

The present study was conducted to determine the effects of Bacillus subtilisKADR1 and its subcellular components on immunity and disease resistance in Labeo rohita against Aeromonas hydrophila infection. Fish were fed diet containing different concentrations of live bacterial cells (DI-106, DII-108 and DIII-1010 CFU/g) and another group of fish were immunized intraperitoneally with cellular components (WCPs, CWPs and ECPs) of Bacillus subtilisKADR1. After 4 weeks of trial, fish were challenged intraperitoneally with Aeromonas hydrophila cell suspension and survival percentage was recorded. Significantly higher post-challenge survivability was recorded in fish groups fed 108 CFU/g of KADR1 (80.24%; RPS = 75.76%) or immunized with WCPs (77.77%; RPS = 72.73%), compared with the control (18.51%). Analysis of immunological parameters viz. serum lysozyme, phagocytosis, serum total protein, respiratory burst, serum IgM levels, superoxide dismutase and alternative complement pathway activity reflected significant enhancement (p < .05) of immune response in fish fed 106, 108 and 1010 CFU/g of live cells, or immunized with cellular components of Bacillus subtilisKADR1, with the highest values were observed DII fed group, followed by the group immunized with WCPs. Our results suggest that dietary administration of Bacillus subtilisKADR1 at 108 CFU/g can effectively enhance the immune responses and disease resistance in Labeo rohita against Aeromonas hydrophila.

Immunological parameters in assessing the risk of decompensation in children with mental retardation

To analyze the correlation between clinical and immunological parameters in children with mental retardation (MR) in order to explore the possibilities of using immunological data in assessing the severity of patient's condition and predicting a risk of decompensation, exacerbation of mental disorders comorbid to MR. Seventy-three school children, aged 8-17 years, mean age 12,6±2,4 years, with MR of different genesis and 64 physically and mentally healthy children (control group) of the same age and sex were studied clinically and immunologically. The degree of clinical severity of MR was evaluated by CGI-S scale (the Clinical global impression-severity), the level of intellectual disabilities was evaluated by the Wechsler Intelligence Scale. Enzymatic activity of leukocyte elastase; functional activity of α1-proteinase inhibitor; levels of autoantibodies to neurospecific antigens - S-100b and myelin basic protein were analyzed in the serum of patients and healthy children. The activation of the immune system was associated with comorbid to MR current psychopathological disturbances that were more severe, persistent and required long-term maintenance treatment. Analysis of immunological parameters can be used in children with MR treated in outpatient network as an additional test for the detection of mental state decompensation.

Functional Alteration of Natural Killer Cells and Cytotoxic T Lymphocytes upon Asbestos Exposure and in Malignant Mesothelioma Patients.

Malignant mesothelioma is caused by exposure to asbestos, which is known to have carcinogenic effects. However, the development of mesothelioma takes a long period and results from a low or intermediate dose of exposure. These findings have motivated us to investigate the immunological effects of asbestos exposure and analyze immune functions of patients with mesothelioma and pleural plaque, a sign of exposure to asbestos. Here, we review our knowledge concerning natural killer (NK) cells and cytotoxic T lymphocytes (CTL). NK cells showed impaired cytotoxicity with altered expression of activating receptors upon exposure to asbestos, while induction of granzyme+ cells in CD8+ lymphocytes was suppressed by asbestos exposure. It is interesting that a decrease in NKp46, a representative activating receptor, is common between NK cells in PBMC culture with asbestos and those of mesothelioma patients. Moreover, it was observed that CD8+ lymphocytes may be stimulated by some kind of “nonself” cells in plaque-positive individuals and in mesothelioma patients, whereas CTL in mesothelioma is impaired by poststimulation maintenance of cytotoxicity. These findings suggest that analysis of immunological parameters might contribute to the evaluation of health conditions of asbestos-exposed individuals and to a greater understanding of the pathology of malignant mesothelioma.

Chronic recurrent multifocal ostemyelitis (CRMO) and activity of immune system

CRMO (chronic recurrent multifocal osteomyelitis) is a rare inflammatory disease observed in children. The clinical forms included non-bacterial osteitis acute or chronic, CRMO and SAPHO (synovitis, acne, pustulosis, hyperostosis and osteitis). The effective therapy is based on non-steroid anti-inflammatory drugs, steroids and bisphosphonates. The clinical course characterizes remission and relapses. The purpose of this study was analysis of immunological parameters in 9 children with diagnosis of CRMO in remission. In one patient the co-existance of Crohn's disease was noted. Study included immunoglobulins and C3c, C4 complement molecules level, number of T, B lymphocytes and NK cells, response to mitogen stimulation. The results showed IgG, IgA and IgM level within normal range for age, increased level of C3c molecule of complement in 5 children, C4 only in one child. The number of T lymphocytes was increased in 2 patients, decreased in 3 patients. CD4:CD8 ratio was within normal range in all patients. The number of activated T lymphocytes (CD3/HLA-DR) was increased in 5 patients. The response to mitogens stimulation (PHA, PWM and antiCD3 antibody) was decreased in 3 patients. There were no relations between the initial clinical symptoms, course of disease and immunological disturbances.

Association between the expression of IL-10 and T cell activation proteins loss in early breast cancer patients

Breast cancer patients may express abnormal cellular immune responses affecting their immunological competence. The analysis of immunological parameters may be useful as indicators of T cell function. To determine the expression of lymphocyte activation proteins and cytokines in tumor and non-metastatic axillary lymph nodes, 30 breast cancer patients were monitored. CD3 polypeptides, PTKs (protein tyrosine kinases) and phosphorylated tyrosines were studied by Western Blot and cytokines mRNA expression was determined by RT-PCR (reverse transcription-polymerase chain reaction). This group of patient had shown high immunohistochemistry expression of IL-10 in tumors. Activation proteins were mainly expressed in involved lymph nodes comparing with their expression in tumors. The differences in expression of CD3 polypeptides and p56(lck) between both locations were significant. There was no statistical association between PTKs and IL-10 in the tumor but more than 50% of cases who express IL-10 lost p56(lck), p59(fyn). A direct association between IL-10 and CD3-polypeptides was observed, however 52.2% of patients who express IL-10 did not express 41 kDa CD3-zeta form. IL-10 mRNA was detected in more than 50% of tumors contrary to the prevalence of type 1 cytokines in regional nodes (40%). The lack of expression of lymphocyte activations proteins and the high expression of IL-10 suggest a downregulation on T cells function in the tumors. These results are useful in order to understand the local immune response that would be key in the control of the tumor progression.

Beneficial effects of protein-bound polysaccharide K plus tegafur/uracil in patients with stage II or III colorectal cancer: Analysis of immunological parameters

Protein-bound polysaccharide K (PSK) increased the 5-year disease-free survival rate and reduced the risk of recurrence in a randomised, controlled study for stage II and III colorectal cancer. In order to elucidate the disease-free survival benefits with PSK and what immunological markers could indicate a PSK responder, serial changes in immunological parameters were monitored in the study. PSK decreased the mean serum immunosuppressive acidic protein (IAP) level, and increased the mean population of natural killer (NK) cells compared with the controls. The 5-year disease-free and overall survival rate for patients with serum IAP values <or=500 microg ml(-1), which represents the normal value, were 75.5% (95% CI: 66.8-84.2%; p=0.016) and 85.1% (95% CI: 77.9-92.3%; p=0.032), respectively, in the PSK group compared with 57.5% (95% CI: 43.3-71.6%) and 70.2% (95% CI: 57.1-83.3%) in the control group. In patients with NK cell population >or=8% at 3 months after surgery, PSK conferred a significantly better (p=0.038) 5-year disease-free survival (86.7%; 95% CI: 74.5-98.8%) compared to the control group (60.0%; 95% CI: 29.6-90.4%). In the proportional hazards model, the presence of regional metastases (relative risk, 3.595; 95% CI: 1.518 to 8.518; p=0.004) and omission of PSK treatment (relative risk, 3.099; 95% CI: 1.202 to 7.990; p=0.019) were significant indicators of recurrence. PSK acts as an immunomodulatory activity and biochemical modulator in stage II or III colorectal cancer. Pre-operative serum IAP values <or=500 microg ml(-1) and an NK cell population >or=8% at 3 months after surgery are possible PSK response predictors.

Protective efficacy of a recombinant protective antigen against Bacillus anthracis challenge and assessment of immunological markers

The efficacy of recombinant Bacillus anthracis Protective Antigen (rPA) produced in Bacillus subtilis and formulated in Alhydrogel or MPL-TDM-CWS (Ribi adjuvant) has been tested and compared to the licensed UK human vaccine in guinea pigs challenged by the aerosol route with the Ames strain of B. anthracis. rPA combined with the Ribi adjuvant was found to be only formulation to provide 100% protection from challenge. Analysis of immunological parameters in the individual animals revealed significant differences between the rPARibi vaccine group and rPA/Alhydrogel and human vaccine groups for antigen specific lymphocyte proliferation, PA neutralisation and antigen specific IgG2 levels, but indicated no significant differences in PA-specific IgG1 levels. rPA formulated in Alhydrogel induced a mainly IgG1 response whilst the rPARibi vaccine produced a predominantly IgG2 response.