Pediatric Sepsis Markers: Interleukins and Others

Dysregulation of cytokine expression in tubulointerstitial nephritis associated with murine malaria

Balance between inflammatory and anti-inflammatory cytokines may be important in malaria presentation and outcome. To clarify cytokine interactions that produce pathology of malaria and control infection, C57BL/6 mice were infected with 10(4) parasitized RBCs from a non-lethal strain of Plasmodium yoelii. Kinetics was monitored showing the course of parasitemia, and cytokines were determined by RT-PCR from liver and spleen tissues. Inflammatory cytokines such as interferon-γ (IFNγ), interleukin (IL)-12, IL-6, tumor necrosis factor-α (TNFα) and anti-inflammatory cytokines, including IL-4 and IL-10, were investigated as key molecules that interact with immune cells in the activation of the immune responses. The production of IFNγ mRNA was found to be higher on day 7 than on day 21 after infection, and IL-12 and IL-6 showed higher expression in the liver than in the spleen. Though TNFα was highly expressed on day 14 after infection and on day 21 in the liver, such expression was decreased on day 21 in the spleen. Anti-inflammatory cytokines showed high expression in both the liver and spleen. The results suggest that a relative balance between inflammatory and anti-inflammatory cytokines is crucial and that the increase of inflammatory cytokine levels during the acute phase of malaria may reflect an early and effective immune response.The counteraction effect of anti-inflammatory cytokines is thought to play a role in limiting progression from uncomplicated malaria to severe life-threatening complications.

BackgroundBeyond their crucial role in hemostasis, platelets possess the ability to regulate inflammation and combat infections through various mechanisms. Stringent control of macrophage activation is essential during innate immune responses in sepsis. Macrophages are considered crucial phagocytic cells that aid in the elimination of pathogens. Platelet interactions with monocytes-macrophages are known to be significant in the response against bacterial infections, but the primary mediator driving these interactions remains unclear. EGFR plays critical role in the regulation of inflammation and infection through various mechanisms.ResultsThe overexpression of platelets by thrombopoietin (TPO) leads to the sequestration of both pro-inflammatory (IL-6/IL-1) and anti-inflammatory (IL-10) cytokines in the organ tissue of septic mice. Epidermal growth factor receptor (EGFR) is critical for platelet activation in sepsis. EGFR-licensed platelets enhance macrophage immune function, including the production of reactive oxygen species (ROS) and the clearance of bacteria. Platelet EGFR also induces M1 macrophage polarization by increasing the expression of inducible nitric oxide synthase (iNOS) and CD64.ConclusionEGFR can activate platelet immune function. Moreover, activated platelets efficiently regulate bacterial phagocytosis and pro-inflammatory function of macrophages through an EGFR-dependent pathway.

Research objective: to determine the level of serum cytokines in the pregravidar period in women with chronic diseases of the hepatobiliary system and reproductive losses in the first trimester in the anamnesis.Materials and methods. 90 patients of reproductive age with chronic pathology of the hepatobiliary system (liver steatosis and non-alcoholic steatohepatitis) were examined. All patients had spontaneous miscarriages in the early stages of gestation in anamnesis. The research was conducted during the pregravid preparation period. The laboratory research included an additional study of the interleukins (ІL-2, ІL-6, ІL-8, ІL-10) and tumor necrosis factor (TNFα) in the blood serum using enzyme immunoferment assay. 8 female donors were examined to determine the reference norm of cytokines in the blood serum. Results. The cytokine status in patients with chronic pathology of the hepatobiliary system against the background of early miscarriage was characterized by an increase in proinflammatory cytokines (TNFα in 2,29 times more, ІL-2 in 1,46 times, ІL-6 in 1,84 times and ІL-8 in 1,50 times) and anti-inflammatory cytokines (ІL-10 in 1,31 times). The integral indices that are characterizing the oppositional cytokines ratio increased (TNFα/ІL-10, ІL-2/ІL-10 and ІL-6/ІL-10) which indicates an inflammatory process in the body. Minimal changes were registered in patients with signs of steatosis – a moderate increase in proinflammatory cytokines (TNFα is 1,34 times more, ІL-2 in 1,18 times, ІL-6 in 1,33 times and ІL-8 in 1,13 times) and preservation within the reference norm of ІL-10. The maximum imbalance of serum cytokines was observed in the exacerbation of non-alcoholic steatohepatitis – a significant increase in proinflammatory cytokines (TNFα is 3,66 times more, ІL-2 in 1,95 times, ІL-6 in 2,57 times and ІL-8 in 2,18 times) against the background of a moderate increase in ІL-10 (1,30 times). In patients with clinical and laboratory signs of remission of non-alcoholic steatohepatitis a moderate increase in proinflammatory (TNFα is 1,87 times more, ІL-2 in 1,20 times, ІL-6 in 1,61 times and ІL-8 in 1,20 times) and anti-inflammatory (ІL-10 in 1,51 times) cytokines was diagnosed. The analysis of the integral indices of oppositional cytokines showed a tendency to increase their values in patients with liver steatosis and non-alcoholic steatohepatitis in the phase of clinical and laboratory remission. During the period of the non-alcoholic steatohepatitis exacerbation a considerable increase in the studied coefficients was revealed. Conclusions. Changes in the cytokine status are noted in patients with early spontaneous miscarriages in anamnesis and chronic diseases of the hepatobiliary system, which are characterized by the severity of inflammatory reactions and progression of the pathological process in liver. Obtained results can be used to predict miscarriage and be taken into account in the program of drug preparation in the pregravid period.

identify the nature of anti-inflammatory and pro-inflammatory cytokine regulation in different periods of plasma cell myeloma (PCM) natural history with evaluation of its role as a prognostic criterion for the disease course in the Chornobyl NPP (ChNPP) accident survivors. Levels of pro-inflammatory (IL-6, TNF-α) and anti-inflammatory (IL-10) cytokines both with their relationship were studied in the stage I-II and stage III PCM patients (n = 74) in different periods of the disease natural history i.e. remission/stabilization and progression. Study groups included the ChNPP accident survivors (n = 35) and non-irradiated subjects (n = 39). Immunoenzymatic method was applied using the Vector-Best CJSC commercial kits. There was a unidirectional increase in the levels of IL-6, TNF-α, and IL-10 in irradiated persons, and an elevation of IL-6 and TNF-α concentration but with a decreased level of IL-10 in non-irradiated subjects compared to control at the time of PCM diagnosis. Period of the disease remission/stabilization in PCM stage I-II patients featured a decrease in IL-6 concentration regardless of the exposure to ionizing radiation, while TNF-α content remained at the level of the control group. There was a significant increase in IL-6 concentration in both study groups during the disease relapse, while TNF-α level remained unchanged compared to stabilization phase of the disease. According to the obtained data a certain contribution of radiation exposure to the PCM pathogenesis as a possible predictor of the exacerbated disease course cannon be excluded. Determining the serum level of pro-inflammatory and anti-inflammatory cytokines (IL-6, TNF-α and IL-10 respectively) provides advancement in assessment of the PCM course and predict the effectiveness of administration of therapy protocols.

Background & Objectives: Stress response to extensive surgeries in cancer patients results in an imbalance between the pro-inflammatory and anti-inflammatory cytokines, besides activating an inflammatory cascade. We investigated the comparative effects of intravenous infusion of dexmedetomidine and lidocaine on proinflammatory cytokines and the stress reaction.
 Methodology: A prospective randomized double blind trial was performed in National Cancer Institute. 54 patients scheduled for pelvi–abdominal cancer surgery under general anesthesia were randomly allocated to three equal groups. Lidocaine group: patients received 1.5 mg/kg loading dose of lidocaine followed by 1.5 mg/kg/h infusion; Dexmedetomidine group: received a loading dose of dexmedetomidine 1 µg/kg followed by infusion of 0.5 µg/kg/h; or Saline group: received 50 ml of normal saline followed by infusion at a rate of 10 ml/h till the end of the surgery. The primary outcome was a postoperative target level of IL–6 after 24 h.
 Results: Immediate and 24 h postoperative IL–6 and TNFα were significantly lower in both dexmedetomidine and lidocaine groups compared to the control group (P < 0.001). Dexmedetomidine group had a significantly lower IL–6 and TNFα levels compared to lidocaine group at immediate postoperative and 24 h postoperative period (P < 0.001). Dexmedetomidine and lidocaine groups had a significantly lower lactate and insulin levels compared to the control group immediately postoperatively as well as 24 h postoperatively (P < 0.001). The dexmedetomidine group had a significantly lower lactate levels compared to the lidocaine group immediately postoperatively only. The lidocaine group had lower insulin level than the control group regarding immediate postoperative period only.
 Conclusion: Intraoperative infusion of dexmedetomidine or lidocaine, both attenuate the postoperative levels of proinflammatory cytokines and stress response in patients undergoing pelvi–abdominal cancer surgeries.
 Clinical trial registration: The study was registered at clinicaltrials.gov (NCT 04148599).
 Key words: Lidocaine; Dexmedetomidine; Interleukins; Tumor necrosis factor-α; Stress response; Pelvi–abdominal cancer; Surgery
 Abbreviations: TNF - Tumor necrosis factor; IL – Interleukins;
 Citation: Hassan MM, Saleh RG, Abdalla NO, Radwan NH, Abdelghfar EM. Effect of lidocaine infusion compared to dexmedetomidine infusion on proinflammatory cytokines and stress response in pelvi–abdominal cancer surgeries: a randomized clinical trial. Anaesth. pain intensive care 2021;26 (1):44-52.
 DOI: 10.35975/apic.v26i1.1765
 Received: February 15, 2021, Reviewed: December 02, 2021, Accepted: December 05, 2021

The staphylococcal superantigen toxic shock syndrome toxin 1 (TSST-1) induces massive cytokine production, which is believed to be the key factor in the pathogenesis of TSS. The temporal sequence and kinetics of both proinflammatory and anti-inflammatory cytokines induced by TSST-1 in human peripheral blood mononuclear cells were investigated. A panel of loss-of-function single-amino-acid-substitution mutants of TSST-1, previously demonstrated to be defective in either major histocompatibility complex (MHC) class II binding (G31R) or T-cell receptor (TCR) interaction (H135A, S14N), was studied in parallel to further elucidate the mechanisms of cytokine secretion. Wild-type recombinant (WT r) TSST-1 induced a biphasic pattern of cytokine secretion: an early phase with rapid release of proinflammatory cytokines (especially gamma interferon, interleukin-2 [IL-2], and tumor necrosis factor alpha [TNF-alpha]) within 3 to 4 h poststimulation, and a later phase with more gradual production of both proinflammatory (IL-1beta, IL-12, and TNF-beta) and anti-inflammatory (IL-6, IL-10) cytokines within 16 to 72 h poststimulation. G31R, which is defective in MHC class II binding, induced a cytokine profile similar to that of WT rTSST-1, except that secretion of the early-phase proinflammatory cytokines was delayed and production of IL-1beta and IL-12 was markedly reduced. In contrast, mutant toxins defective in TCR interaction either demonstrated complete absence of any cytokine secretion during the entire observation period (H135A) or resulted in complete abolishment of IL-2 and other early-phase proinflammatory cytokines, while secretion of IL-10 appeared unaffected (S14N). Neither WT rTSST-1 nor the mutant toxins induced IL-4 or transforming growth factor beta. Our data indicate that effective TCR interaction is critical for the induction of the early-phase proinflammatory cytokine response, thus underscoring the importance of T-cell signaling in TSS.

Intestinal infections are associated with Gram-negative bacteria like Escherichia coli. When eliminated by treatments during replication, E. coli release lipopolysaccharides (LPS) that can activate the intestinal immune system and increase the expression of cytokines, such as interleukin (IL)-8, IL-18, tumor necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ), by the intestinal epithelium under pathological conditions. This study aimed to evaluate the addition of Bacillus subtilis to the duodenal gene expression of pro-inflammatory and anti-inflammatory cytokines in broilers exposed to LPS from E. coli. RNA was extracted using the Zymo Research total RNA commercial kit, according to the manufacturer's specifications, from the intestinal tissue of the duodenum previously resuspended in the lysis buffer of the kit. The expression of the cytokines of interest was measured using the QuantiNova SYBR green real-time polymerase chain reaction kit (Qiagen). Transcript quantification was performed by the ΔΔC(t) method using glyceraldehyde 3-phosphate dehydrogenase as a normalizing constitutive gene. For the measurement of pro-inflammatory (IL-8, IL-18, TNF-α, and IFN-γ) and anti-inflammatory (IL-10) cytokines, there was no statistically significant difference (p > 0.05) between the basal diet and the diet with antibiotic (avilamycin). There was a statistical difference (p < 0.05) between diets with LPS. The diet with B. subtilis presented the lowest expression; the results differed on each sampling day (days 14, 28, and 42). A decrease in the expression of pro-inflammatory cytokines (IL-8, IL-18, TNF-α, and IFN-γ) and an increase in IL-10 (anti-inflammatory) was observed; in this way, a balance of the inflammatory response to bacterial infection is achieved, suggesting that the use of B. subtilis as an additive in a broiler diet has a similar effect to that produced with antibiotic growth promoter.

High mobility group box 1 protein (HMGB1) is a highly conserved, ubiquitous protein in the nuclei and cytoplasm of nearly all cell types. HMGB1 is secreted into the extracellular milieu and acts as a proinfl ammatory cytokine. In this article we reviewed briefl y the cellular immune response mediated by HMGB1 in infl ammation and sepsis. This systemic review is mainly based on our own work and other related reports. HMGB1 can actively affect the immune functions of many types of cells including T lymphocytes, regulatory T cells (Tregs), dendritic cells (DCs), macrophages, and natural killer cells (NK cells). Various cellular responses can be mediated by HMGB1 which binds to cell-surface receptors [e.g., the receptor for advanced glycation end products (RAGE), Toll-like receptor (TLR)2, and TLR4]. Anti-HMGB1 treatment, such as anti-HMGB1 polyclonal or monoclonal antibodies, inhibitors (e.g., ethyl pyruvate) and antagonists (e.g., A box), can protect against sepsis lethality and give a wider window for the treatment opportunity. HMGB1 is an attractive target for the development of new therapeutic strategies in the treatment of patients with septic complications.

Parthenium dermatitis is a common airborne allergic contact dermatitis induced by exposures to the weed Parthenium hysterophorus. The disease manifests as itchy erythematous papules, papulovesicular and plaque lesions on exposed areas of the body. The aim of this study was to show the alterations in pro/anti-inflammatory cytokines in parthenium dermatitis. The study included 50 patients with parthenium dermatitis confirmed by patch testing using aqueous extracts of P. hysterophorus and 50 age-matched healthy controls. The levels of pro-inflammatory [tumour necrosis factor-α (TNF-α), interleukin (IL)-6, IL-8, and IL-17] and anti-inflammatory (IL-4 and IL-10) cytokines were estimated by commercially available high sensitivity enzyme-linked immunosorbent assay (ELISA) kits. All the dermatitis patients showed significantly (P < 0.001) elevated levels of TNF-α, IL-6, IL-8, and IL-17 levels as compared to healthy controls. In contrast, the anti-inflammatory cytokine IL-4 showed an insignificant decrease (P < 0.217) and a decrease in level of IL-10 was statistically significant (0.001) compared with controls. The present study suggests the involvement of pro-inflammatory cytokines in the pathogenesis of parthenium dermatitis. A decrease in levels of anti-inflammatory cytokines was demonstrated, which could not downregulate pro-inflammatory cytokines in parthenium dermatitis.

It is necessary to search for new risk factors that contribute to atherogenesis in presence of an unfavorable trend of increasing primary morbidity of ischemic heart disease (IHD). Objective. To study the features of cytokine status and its relationship with the severity of atherosclerotic coronary bed lesion in middle-aged men with chronic coronary syndrome and acute coronary syndrome without ST-segment elevation. Material and methods. A number of men equal 192 were examined (mean age 55.1±3.4 years), who were divided into 3 groups: the 1st group — patients with acute coronary syndrome without ST-segment elevation (n=93; mean age 55.37±3.14 years); the 2nd group — patients with chronic coronary syndrome (n=63; mean age 55.12±3.50 years) and the 3rd group — control, which consisted of conditionally healthy subjects without cardiac pathology (n=36; mean age 52.8±4.2 years). Levels of proinflammatory cytokines (interleukin (IL) IL-8, tumor necrosis factor-α — TNF-α), anti-inflammatory cytokine (IL-4) in the blood serum were determined in all participants of the study. Instrumental examination of patients with IHD included electrocardiography, echocardiography and selective coronary angiography. Results. The highest levels of IL-8 (93.62±13.04; 7.34±2.26; 5.86±1.13 pg/ml; F=38.12; p&lt;0.001), TNF-α (12.44±2.04; 4.34±1.21; 3.53±0.96 pg/ml; F=77.73; p&lt;0.001) and IL-4 (289.83±30.61; 3.70±0.16; 1.87±0.11 pg/ml; F=75.12; p&lt;0.001) have been determined in patients with acute coronary syndrome without ST-segment elevation compared to patients with chronic coronary syndrome and subjects of control group. Correlations between the levels of all studied cytokines and the number of hemodynamically significant stenoses (p&lt;0.05), the total number of the coronary bed sections affected by atherosclerosis, degree of stenosing of different sections of the coronary bed according to coronary angiography results (p&lt;0.05) have been established. The most significant positive correlation was between TNF-α and the number of coronary bed lesions detected by coronary angiography (R=0.54; p&lt;0.05). The cytokine TNF-α level &gt;4.41 pg/ml was related to the probability of acute coronary syndrome without ST-segment elevation in middle-aged men with IHD. The value of the IL-4/IL-8 coefficient &gt;3.4 was associated with the probability of IHD exacerbation in middle-aged males. Conclusions. The development of acute coronary syndrome without ST-segment elevation is associated with increase of both proinflammatory (IL-8, TNF-α) and anti-inflammatory cytokines (IL-4) levels. The imbalance between levels of proinflammatory and anti-inflammatory cytokines contributes to the progression of coronary atherosclerosis in patients with ischemic heart disease.

The continuous growth of cardiovascular diseases is one of the important challenges facing the modern medicine. They are the primary cause of death both in developed countries and inUkrainein particular. The aim of the study is to perform analysis of modern literary sources related to the role of cytokine gene polymorphism in the development of acute myocardial infarction. The basis for the search for candidate genes was study of individual characteristics and identifying genetic polymorphisms that increase the risk of triggering mechanisms of atherosclerotic lesions of the coronary vessels and the consequent acute myocardial infarction development. Cytokine genes feature a very high level of polymorphisms. The obtained results of the study are quite contradictory, in addition there is no consensus on their application. One of these candidate genes is the gene encoding interleukin-6 production. This is one of the proinflammatory cytokines, elevated levels of which are associated with the development and course of coronary heart disease, as well as with the processes of the atherosclerotic plaque destabilization. Polymorphic marker in interleukin-6 (-174-G/C) gene promoter is associated with IL-6 gene expression level and the level of Interleukin-6 inplasma, and can affect the course of ischemic heart disease. The tumor necrosis factor-α is a multifunctional proinflammatory cytokine which is produced mainly by monocytes and macrophages. Concentration of tumor necrosis factor-α in blood plasma is constantly increased in patients after acute myocardial infarction with an increased risk of recurrent coronary events. These data support the hypothesis that stable patients are at risk of constant inflammatory instability. Increased production of tumor necrosis factor-α is considered an important cause of destabilization of atherosclerotic plaques. The gene cluster of tumor necrosis factor-α is within the region of class III genes of highly polymorphic major histocompatibility complex, located on human chromosome 6 at the position 6p21.1–21.3. Another candidate gene encodes interleukin-10, being an anti-inflammatory cytokine is an inhibitor of inflammation located on chromosome 1 at the position 1q31–q32. The research identified a number of IL-10 gene polymorphisms at positions -592, -627, -1082. Conclusions. Analysis of the above literature allows us to make a conclusion that: first, the concentration of cytokines depends on the genetic characteristics of an individual; secondly, cytokine gene polymorphism may be of importance in regard to the course of coronary heart disease.

BackgroundImmunoregulatory capacity of mesenchymal stem cells (MSC) is triggered by the inflammatory environment, which changes during tissue repair. Macrophages are essential in mediating the inflammatory response after injury and can adopt a range of functional phenotypes, exhibiting pro-inflammatory and anti-inflammatory activities. An accurate characterization of MSC activation by the inflammatory milieu is needed for improving the efficacy of regenerative therapies. In this work, we investigated the immunomodulatory functions of MSC primed with factors secreted from macrophages polarized toward a pro-inflammatory or an anti-inflammatory phenotype. We focused on the role of TNF-α and IL-10, prototypic pro-inflammatory and anti-inflammatory cytokines, respectively, as priming factors for MSC.MethodsSecretion of immunoregulatory mediators from human MSC primed with media conditioned by human macrophages polarized toward a pro-inflammatory or an anti-inflammatory phenotype was determined. Immunomodulatory potential of primed MSC on polarized macrophages was studied using indirect co-cultures. Involvement of TNF-α and IL-10 in priming MSC and of PGE2 in MSC-mediated immunomodulation was investigated employing neutralizing antibodies. Collagen hydrogels were used to study MSC and macrophages interactions in a more physiological environment.ResultsPriming MSC with media conditioned by pro-inflammatory or anti-inflammatory macrophages enhanced their immunomodulatory potential through increased PGE2 secretion. We identified the pro-inflammatory cytokine TNF-α as a priming factor for MSC. Notably, the anti-inflammatory IL-10, mainly produced by pro-resolving macrophages, potentiated the priming effect of TNF-α. Collagen hydrogels acted as instructive microenvironments for MSC and macrophages functions and their crosstalk. Culturing macrophages on hydrogels stimulated anti-inflammatory versus pro-inflammatory cytokine secretion. Encapsulation of MSC within hydrogels increased PGE2 secretion and potentiated immunomodulation on macrophages, attenuating macrophage pro-inflammatory state and sustaining anti-inflammatory activation. Priming with inflammatory factors conferred to MSC loaded in hydrogels greater immunomodulatory potential, promoting anti-inflammatory activity of macrophages.ConclusionsFactors secreted by pro-inflammatory and anti-inflammatory macrophages activated the immunomodulatory potential of MSC. This was partially attributed to the priming effect of TNF-α and IL-10. Immunoregulatory functions of primed MSC were enhanced after encapsulation in hydrogels. These findings may provide insight into novel strategies to enhance MSC immunoregulatory potency.

Counter-stress effects of relaxation on proinflammatory and anti-inflammatory cytokines

Inflammation of the tonsils could be described as acute tonsillitis, mainly due to infection. Recurrent tonsillitis could be defined as 3-7 episodes during the first 3 years of age. Vitamin D, which is a neuro-hormone with pleiotropic biological activities may modulate the immune response by alleviation, and stimulation of Th1 and Th2 cell proliferation, respectively, that influence the stimulation, synthesis, and secretion of both pro and anti-inflammatory cytokines. In this study we aimed to shed light on the levels of vitamin D in children with different episodes of tonsillitis in association with levels of interleukins (TNFα, IL-2, IL-4, IL-10). Blood samples were collected from 48 participants in 3 groups: control, acute tonsillitis (1-2 episodes/year), chronic tonsillitis (more than 7 episodes/year), serum was separated and the levels of Vitamin D, TNFα, IL-2, IL-4 and IL-10 were estimated using ELISA technique. Vitamin D decreased significantly as the episodes of tonsillitis increased, with level of 16.38± 2.41ng/ml in acute and, 14.13± 2.15 ng/ml in chronic tonsillitis as compared to control (30.91± 2.31 ng/ml), while pro-inflammatory cytokines (TNFα and IL-2) significantly increased (46.88± 14.05 and 44.55± 9.24, 1267.25± 111.85 and 1191.72± 121.52 ng/ml, respectively) as compared to control (9.45 and 138.48 ng/ml respectively). Anti-inflammatory (IL-4, IL-10) cytokines in control group were (243.08± 28.72 and 24.27± 1.83 ng/ml, respectively), which increased non-significantly in acute and chronic tonsillitis (302.76± 38.93, 290.12± 44.69 and 28.16± 2.01, 26.29± 1.99 ng/ml, respectively). Significant direct correlation was observed between the levels of vitamin D and anti-inflammatory cytokines in chronic tonsillitis (P&lt;0.05). In conclusion, deficiency of vitamin D may affect the number of episodes of tonsillitis in children by modulation of the secretion of some cytokines.