Flow Cytometric Analysis of Macrophages and Cytokines Profile in the Bronchoalveolar Lavage Fluid in Patients with Lung Cancer.

Macrophages play an important role in the suppression and activation of immune anti-cancer response, but little is known about dominant macrophage phenotype in the lung cancer environment, evaluated by bronchoalveolar lavage fluid (BALF). The aim of this study was to characterize macrophages in BALF from a lung affected by cancer (cBALF) and a healthy lung (hBALF) of the same patient regarding their individual macrophage polarization and selected cytokines profile. A total of 36 patients with confirmed lung cancer were investigated. Macrophages markers: CD206 CD163 CD80 CD86 CD40 CD45, Arginase-1, and CD68 were evaluated by flow cytometry. Cytokines (IL-1 RA, IL-6, IL-10, TNF-α, IL-1ß, IL-12, IL-23, and TGF-ß) profile was analyzed. There was higher median proportion of macrophages in Cbalf than in Hbalf. The population of macrophages presented immunophenotype: Ccd68+bright CD206+bright CD163+bright CD80+ CD86+ CD40+bright CD45+ cArginase+. We observed some trends in the expression of the analyzed antigens in clBALF and hlBLAF. The highest concentrations of IL-1RA and IL-6 were in Cbalf and Hbalf supernatant. There were the correlations between pro- and anti-inflammatory cytokines. The findings showed that macrophages include a diverse and plastic group with the presence of different antigens and cytokines, and determining the target phenotype is a complex and variable process.

Neuron-Specific Enolase and S100B: The Earliest Predictors of Poor Outcome in Cardiac Arrest.

Background: Proper prognostication is critical in clinical decision-making following out-of-hospital cardiac arrest (OHCA). However, only a few prognostic tools with reliable accuracy are available within the first 24 h after admission. Aim: To test the value of neuron-specific enolase (NSE) and S100B protein measurements at admission as early biomarkers of poor prognosis after OHCA. Methods: We enrolled 82 consecutive patients with OHCA who were unconscious when admitted. NSE and S100B levels were measured at admission, and routine blood tests were performed. Death and poor neurological status at discharge were considered as poor clinical outcomes. We evaluated the optimal cut-off levels for NSE and S100B using logistic regression and receiver operating characteristic (ROC) analyses. Results: High concentrations of both biomarkers at admission were significantly associated with an increased risk of poor clinical outcome (NSE: odds ratio [OR] 1.042 per 1 ng/dL, [1.007−1.079; p = 0.004]; S100B: OR 1.046 per 50 pg/mL [1.004−1.090; p < 0.001]). The dual-marker approach with cut-off values of ≥27.6 ng/mL and ≥696 ng/mL for NSE and S100B, respectively, identified patients with poor clinical outcomes with 100% specificity. Conclusions: The NSE and S100B-based dual-marker approach allowed for early discrimination of patients with poor clinical outcomes with 100% specificity. The proposed algorithm may shorten the time required to establish a poor prognosis and limit the volume of futile procedures performed.

Time-dependent effect of desensitization with wasp venom on selected parameters of the immune system.

The emergence of tolerance during Hymenoptera venom immunotherapy (VIT) is a complex process. The main goal of VIT is to induce a change from proinflammatory Th2 response to the Th1 response. However, the immune mechanism of acquiring rapid tolerance during VIT has not yet been fully understood. Therefore, we have analyzed (in 4-time points: 0, 2, 6, and 24 weeks after the initiation phase of VIT) the concentration of complement C3, C4, and C5 components, lymphocyte subpopulations (flow cytometry), as well as histamine and tryptase serum concentrations of 43 patients with wasp venom allergy (III and IV Müller grade) classified to ultra-rush treatment and 18 volunteers as the control group (CG). We observed that VIT affected the immune system by inducing changes in the complement system (decreased C3 and C4 compartment protein concentrations) and "normalized" the percentage of lymphocytes and neutrophils in the peripheral blood. Moreover, a significant increase in the percentage of nTreg in the blood of patients treated with VIT was observed. On the other hand, there were no changes in histamine or tryptase concentrations in the blood. Increased percentage of nTreg cells is a well-known mechanism by which VIT affects the immune system. Finally, VIT also modulated the concentrations of the complement components, which may be a previously unknown VIT mechanism of action.

A 63-Year-Old Man with a Diagnosis of Re-Infection with SARS-CoV-2 Nine Weeks After an Initial Hospital Admission with COVID-19 Pneumonia.

BACKGROUND This report describes a 63-year-old Polish man presenting with COVID-19 (Coronavirus Disease 2019) pneumonia in early 2020, before vaccines to prevent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection were available. Nine weeks following recovery from the initial infection, he tested positive again for SARS-CoV-2. CASE REPORT Man, age 63, was admitted to the Military Institute of Medicine on March 12, 2020, with body temperature 40°C, a cough, and breathlessness. On March 12, 2020, SARS-CoV-2 RNA was found in a nasopharynx smear. A chest X-ray (RTG) showed discrete areas of interstitial densities. On June 13, 2020, after 32 days of hospitalization and 2 negative real-time polymerase chain rection (RT-PCR) test results, patient was released home in good general condition. On July 23, 2020 he reported to the emergency room with fever of 39°C and general weakness. A nasopharynx smear confirmed SARS-CoV-2 infection. On admission, the patient was in moderately good condition with auscultatory changes typical for pneumonia on both sides of the chest. On the seventh day of hospitalization, the patient was transported to the Intensive Care Unit (ICU) due to drastic deterioration in respiratory function. Respiratory support with non-invasive high-flow oxygen therapy (Opti-Flow) was used. On August 20, 2020, after negative RT-PCR test results, he was discharged in good general condition. CONCLUSIONS This case of COVID-19 pneumonia presented early in the COVID-19 pandemic of 2020, and the laboratory diagnosis of the initial and subsequent SARS-CoV-2 infection relied on the laboratory methods available at that time. However, several cases of repeat SARS-CoV-2 infection have been described before the development of vaccines in late 2020.

Clinical significance of plasma PAF acetylhydrolase activity measurements as a biomarker of anaphylaxis: Cross-sectional study.

INTRODUCTION: Platelet-activating factor (PAF) has a direct role as a mediator in the pathogenesis of various disorders with an inflammatory component, including those with allergic aetiology. The peripheral blood concentration of PAF is dynamically regulated by plasma PAF acetylhydrolase (PAF-AH). Previous research suggest that low activity of plasma PAF-AH could be a predictive marker for increased severity of some types of allergic hypersensitivity reactions-especially anaphylaxis. The purpose of the study was to evaluate the association between plasma PAF-AH activity and severity in patients with anaphylactic reactions following a wasp or bee sting. MATERIALS AND METHODS: The study group of 89 patients was divided into two subgroups depending on the increasing severity of the most severe anaphylactic reaction in the past, which was assessed according to the Müller's scale. The first subgroup included participants with a history of hypersensitivity reactions up to grade II. The second subgroup consisted of patients who have experienced at least one grade III or IV reactions in the past. A control group of 20 people was established. Plasma PAF-AH activity was measured using a colorimetric method. RESULTS: It has been observed that plasma activity of platelet-activating factor acetylhydrolase was significantly lower in patients with anaphylaxis history compared to the control group with negative atopic history (on average 21.38 nmol/min/ml for the control group, 9.47 nmol/min/ml for the first subgroup and 10.16 nmol/min/ml for the second subgroup, in both cases p < 0.0001). CONCLUSION: The plasma activity of PAF-AH is a promising parameter that can help to distinguish a group of patients not threatened with development of anaphylaxis and not requiring laborious or expensive prophylactic procedures.

Inflammatory Markers During Continuous High Cutoff Hemodialysis in Patients with Septic Shock and Acute Kidney Injury.

High cut-off (HCO) continuous veno-venous hemodialysis (CVVHD) is one of the renal replacement therapies which nonselectively removes inflammatory mediators. This study seeks to examine the association between the inflammatory background and the need for catecholamine treatment in hemodynamically instable patients having septic shock and acute kidney injury during HCO-CVVHD. There were 38 patients (F/M; 16/22, mean age 63 ± 16 years) included in the study. The initial content of the cytokines IL-4, IL-12, IL-17, and TNFα, C-reactive protein, and the score of the Sequential Organ Failure Assessment (SOFA) were assessed. The receiver operating characteristic (ROC) plot showed that a combination consisting of IL-17 × SOFA ≤22.3 was a reliable predictive factor of the need for catecholamine treatment during HCO-CVVHD, with 82% sensitivity and 90% specificity, with the area under curve (AUC) of 0.843; p < 0.001. On the other side, SOFA ≤14.0 predicted catecholamine treatment or its discontinuation when started, with both specificity and sensitivity 83% (AUC = 0.899; p < 0.001). In conclusion, the immune system activation, assessed from the initial level of IL-17, and the clinical SOFA evaluation are of practical help in predicting the need for catecholamine treatment or the probability of a reduction thereof in patients on veno-venous hemodialysis due to septic shock.

The association between proinflammatory cytokines and kidney function, body composition, and nutritional markers in nondialysis-dependent chronic kidney disease patients with diabetes mellitus type 2
.

BACKGROUND: Chronic kidney disease (CKD) is associated with a subclinical inflammatory state, which contributes to increased mortality in CKD patients. The purpose of this study was to examine the association between chosen cytokines, such as IFN-γ, IL-10, IL-2p70, IL-6, and kidney function as well as the body composition and nutritional markers in patients with CKD and diabetes mellitus type 2. MATERIALS AND METHODS: 21 patients with diabetes mellitus type 2 and CKD stage 3b - 5, with estimated glomerular filtration rate (eGFR) lower than 45 mL/min/1.72m2, not being treated with dialysis were included in the study. Body composition was assessed by bioimpedance spectroscopy (Body Composition Monitor - Fresenius Medical Care). RESULTS: Significant, negative correlations between lean tissue index (LTI) and IFN-γ concentrations (r = -0.52, p = 0.021) as well as IL-6 concentrations (r = -0.46, p = 0.047) were observed. Only the IL-6 levels significantly correlated with kidney function expressed by eGFR (r = -0.47, p = 0.034). We observed a significant positive correlation between IL-6 level and IFN-γ (r = 0.51, p = 0.019) as well as with high-sensitivity C-reactive protein (hsCRP) levels (r = 0.48, p = 0.029). The IL-10 level significantly correlated with hsCRP (r = 0.53, p = 0.015). CONCLUSION: In CKD patients with diabetes mellitus type 2 during conservative treatment, IL-6 levels were associated with kidney function expressed by eGFR. IL-6 levels and IFN-γ levels negatively correlated with the amount of muscle mass. Cytokines did not show any association with the amount of fat tissue this study.

How fast does wasp venom immunotherapy affect a regulatory T cell subpopulation (CD4+ CD25+ Foxp3+) and the synthesis of interleukins 10, 21 and transforming growth factor ß1?

INTRODUCTION: The literature describes the influence of venom immunotherapy (VIT) on the subpopulation of T regulatory cells (CD4+ CD25+ Foxp3+) and the synthesis of IL-10, TGF-ß1 as well as many other cytokines at various times after immunotherapy. AIM: To assess changes in the percentage of cells of CD4+ and CD25+ in peripheral blood and serum concentrations of IL-10, IL-21 and TGF-ß1 in the early stages of VIT. MATERIAL AND METHODS: The study included 18 patients who were allergic to wasp venom and who in the past underwent systemic anaphylactic reaction after stinging, meeting the criteria to qualify for VIT. The immunoenzymatic method (ELISA) was used to assess concentrations of cytokines IL-10, IL-21 and TGF-ß1 and the surface antigens CD4 and CD25 on the cells. The concentrations were determined by flow cytometry method at baseline (before VIT) and after 2.5 and 24 h from the VIT starting point. RESULTS: The mean values of the activity of T lymphocytes CD4+ CD25+ FoxP3+ and concentrations of the cytokines IL-10, IL-21 and TGF-ß1 are shown in table. CONCLUSIONS: A 24-hour activation assessment of serum concentrations of cytokines IL-10, IL-21 and TGF-ß1 during the first day of the Hymenoptera venom immunotherapy by ultra-rush protocol does not show the significant dynamics of change of the examined parameters.

Simvastatin Therapy and Bronchoalveolar Lavage Fluid Biomarkers in Chronic Obstructive Pulmonary Disease.

Chronic obstructive pulmonary disease (COPD) is a progressive disease underlain by airway inflammation. Despite trials with new generations of anti-inflammatory drugs to alleviate the disease burden, the effective curative treatment remains elusive. In this context, the aim of this study was to assess the influence of simvastatin, a leading member of the family of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, known to display anti-inflammatory and immunomodulatory activity, on symptoms and lung function, as well as the proportion of inflammatory cells, cytokines, proteolytic enzymes, and surfactant protein D (SP-D) content in bronchoalveolar lavage fluid (BALF) in COPD patients. There were 50 patients with moderate-to-severe airway obstructions included into the study, subdivided into simvastatin-treated (Zocor - MSD; 40 mg daily) and control simvastatin-untreated groups, other treatment being equal. Pulmonary functions tests and bronchofiberoscopy with BALF procedure were performed before and after 3-month-long treatment in both groups. The major finding was that simvastatin treatment caused a distinct increase in the airway content of SP-D. Further effects, albeit smaller in magnitude, consisted of reductions in the proportion of airway neutrophils and in MMP-9 content, all with a benefit of improved score in the disease activity assessment test. There were no appreciable changes noted in lung function or dyspnea perception, which could be ascribed to simvastatin treatment. We conclude that statin's anti-inflammatory and surfactant homeostasis preserving properties may offer promise as an adjunctive treatment in COPD patients. The SP-D content in BALF has a potential to become a marker of COPD inflammatory activity and treatment monitoring.

Removal of Procalcitonin and Selected Cytokines during Continuous Veno-Venous Hemodialysis Using High Cutoff Hemofilters in Patients with Sepsis and Acute Kidney Injury.

INTRODUCTION: The purpose of this study was to evaluate the impact of continuous veno-venous hemodialysis (CVVHD) using high cutoff (HCO) hemofilters on the removal of procalcitonin (PCT), and other inflammatory markers in the treatment of patients during septic shock with acute kidney injury (AKI). MATERIALS AND METHODS: Thirty-six patients with septic shock and AKI were included in the study. Before and after the 24-h HCO-CVVHD, PCT, native C-reactive protein (CRP) and cytokines (interleukin-1ß, interleukin-6, interleukin-12, interleukin-17, tumor necrosis factor-α) in serum and effluent were assessed. RESULTS: After the HCO-CVVHD serum concentrations of PCT, CRP and selected cytokines were significantly lower. The decrease in PCT was bigger than in CRP (p = 0.007). The change in PCT concentration was significantly influenced by PCT and IL-17 clearances (R2 = 0.525; p < 0.001). CONCLUSION: In contrast to the native CRP, monitoring of PCT during HCO-CVVHD is less useful because it reflects the clearance of this marker and anti-inflammatory effectiveness of the method.

Effectiveness of High Cut-Off Hemofilters in the Removal of Selected Cytokines in Patients During Septic Shock Accompanied by Acute Kidney Injury-Preliminary Study.

BACKGROUND In recent years there have been attempts to treat sepsis using various methods of extracorporeal blood purification in order to eliminate selected mediators of inflammation. MATERIAL AND METHODS This retrospective study assessed 28 patients (17 males, 11 females, age 60.3 ± 14.5 years) in septic shock, treated with continuous venovenous hemodialysis (CVVHD). Oligoanuric patients with acute kidney injury were qualified for 24-hour CVVHD using high cut-off (HCO) hemofilter. Before the start of dialysis and after 24 hours of treatment, the concentration levels of selected cytokines (IFN-α, IFN-γ, TNF-α, IL-1ß, IL-2, IL-6, IL-10, IL-12) in serum were assessed. After 12 hours and 24 hours of treatment, the concentration of the same cytokines in the dialysis fluid was assessed. The aim of our study was to evaluate the effectiveness of HCO-CVVHD in the removal of selected cytokines. RESULTS After 24-hour HCO-CVVHD treatment, IL-10 and IL-12 levels in serum were significantly lower. Concentrations of INF-α, IL-1ß and IL-2 in dialysis fluid significantly increased during HCO-CVVHD, which corresponded with the parallel rise of related clearances. Clearance of IL-6 was approximately four times higher than IL-10. The rise of IL-6 during HCO-CVVHD significantly correlated with mortality due to sepsis. CONCLUSIONS Continuous venovenous hemodialysis using high cut-off hemofilter proved to be effective in the removal of IFN-α, IL-1ß, IL-2 and IL-6, IL-10 and IL-12 from serum in patients during septic shock. The rise of IL-6 during HCO-CVVHD seems to be a marker of bad prognosis in septic shock patients.

Evaluation of two distinct cryoprotectants for cryopreservation of human red blood cell concentrates.

BACKGROUND: Cryopreservaton of packed human red blood cells requires the use of cryoprotectants. OBJECTIVE: The study assessed physiological parameters of 40 RBC units frozen with either 40% glycerol or 6.7% HES. METHODS: After thawing, they were suspended in NaCl or in 6% HES. Tests of Hct, Hb, Na+ and K+ ions, ATP, 2,3-DPG, pH and erythrocyte stability were measured 30 minutes and 24 hours after thawing. RESULTS: Hct was lower after thawing but did not differ significantly between two groups. Hb was lower after thawing, but was statistically significant higher in the HES group (43.8 g/unit vs 35.4 g/unit). K+ concentration increased after thawing and was significantly higher after 24 hours in the glycerol group (29.0 mEq/l vs 8.7 mEq/l). ATP concentration in the HES group was significantly lower (2.15 micromol/g) in comparison with the glycerol group (6.30 micromol/g) 24 hours after thawing. 2,3-DPG levels did not differ significantly between the methods. Stability of RBCs frozen in glycerol were better (94.58%) than RBCs frozen in HES (80.75%) measured 24 hours after thawing. ATP is better protected in erythrocytes frozen in glycerol than in HES. CONCLUSION: Erythrocytes frozen with HES preserved more hemoglobin than with glycerol. Membrane permeability for Na+ and K+ ions was preserved better with HES. HES compared to glycerol offered better protection for erythrocytes.

[The treatment of septic shock with continuous venovenous hemodialysis using high cut-off dialyzer in patient after immediate circulatory arrest]. / Ciagla zylno-zylna hemodializa z uzyciem dializatora high cut-off w leczeniu wstrzasu septycznego u chorego po naglym zatrzymaniu krazenia.

The decreased immunity which occurs frequently in severely intoxicated patients may led to sepsis. The sepsis may be caused by bacterial toxins in unconscious patients with toxic coma which generate decreased immunity. Apart from the wide spectrum antibiotic therapy, crystalloids, colloids, vasopressin and corticosteroids, the renal replacement therapy may be useful in treatment of sepsis due to its complexes pathophysiology. Taking into account the role of cytokines in sepsis pathomechanism, the trials of treatment using high cut-off (HCO) membranes were performed in the recent years. These membranes remove molecules with mass up to 60 kDa, including cytokines typical for severe sepsis. The usefulness of continuous veno-venous hemodialysis--CVVHD with HCO dialyzer in the treatment of patient in septic shock and multiorgan damage--including damage caused by cardiac arrest was presented in the study. The concentration of IL-1P, IL-2, IL-4, IL-6, IL-10, IL-12, INF-alpha, INF-gamma, TGF-alpha in blood were determined before and after the 24-hours procedure. After the procedure the most evident decrease was observed for IL-4, 6, 10, 12 (17.3%, 31.8%, 83.4% i 22.3% respectively). During the following days the general status of patient improved gradually. The patient was discharged from the hospital after 20 days of hospitalization. His general condition was good, the values of inflammatory parameters were normal and the renal function was correct. There are very few studies describing HCO membranes effectiveness and they were performed on limited populations of patients. The presented case study may contribute to the discussion on the usefulness of dialysis with HCO membranes in the treatment of severely intoxicated patients complicated by serious sepsis resistant to standard antibiotic therapy.

[TNF alfa polymorphism and course of ulcerative colitis]. / Wplyw polimorfizmu TNF alfa na przebieg wrzodziejacego zapalenia jelita grubego.

UNLABELLED: Inflammatory bowel diseases including ulcerative colitis are associated with prolonged inflammatory process, that is dependent on cytokine production. Among them crucial role plays tumor necrosis factor TNF-alpha. There is proven association between single nucleotyde polimorphism and ability to produce cytokines. AIM: We analyzed association between TNF-alpha (-308) promoter polymorphism and extension of lesions in ulcerative colitis. TNF-alpha (-308) promoter polymorphism. MATERIALS AND METHODS: Analysis was performed using polymerase chain reaction with sequence specific primers method (PCR-SSP) in 48 patients suffering from ulcerative colitis and association between TNF-alpha (-308) promoter polymorphism and ulcerative colitis macroscopic lesions classified according Montreal classification was investigated. RESULTS: No statistically significant association among groups of patients and TNF-alpha (-308) promoter polymorphism was observed. More cases of TNF-alpha (-308) promoter polymorphism associated with low TNF-alpha production were observed in patients with E2 and E3 lesions according to Montreal classification. CONCLUSIONS. There is no direct association between TNF-alpha (-308) promoter polymorphism and ulcerative colitis macroscopic inflammatory lesions evaluated on basis of Montreal classification. There is statistically irrelevant tendency of more cases of pancolitis in group of patient with TNF-alpha (-308) promoter polymorphism associated with low TNF-alpha production.