interleukin-17A Concentrations Research Articles

Altered serum interleukin-17A and interleukin-23A levels may be associated with the pathophysiology and development of generalized anxiety disorder

In recent times, the pathogenesis of generalized anxiety disorder (GAD) and the influence of pro- and anti-inflammatory cytokines on it have garnered considerable interest. Cytokine research, especially Th-17 cytokine research on GAD patients, is limited. Here, we aim to assess the role of interleukin-17A (IL-17A) and interleukin-23A (IL-23A) in the pathophysiology and development of GAD. This investigation included 50 GAD patients and 38 age-sex-matched healthy controls (HCs). A psychiatrist diagnosed patients with GAD and assessed symptom severity using the DSM-5 and the GAD-7 scales. The serum concentrations of IL-17A and IL-23A were determined using commercially available ELISA kits. GAD patients exhibited elevated levels of IL-17A (77.14 ± 58.30 pg/ml) and IL-23A (644.90 ± 296.70 pg/ml) compared to HCs (43.50 ± 25.54 pg/ml and 334.40 ± 176.0 pg/ml). We observed a positive correlation between disease severity and cytokine changes (IL-23A: r = 0.359, p = 0.039; IL-17A: r = 0.397, p = 0.032). These findings indicate that IL-17A and IL-23A may be associated with the pathophysiology of GAD. ROC analysis revealed moderately higher AUC values (IL-23A: 0.824 and IL-17A: 0.710), demonstrating their potential to discriminate between patients and HCs. Also, the sensitivity values of both cytokines were relatively higher (IL-23A: 80.49% and IL-17A: 77.27%). According to the present findings, there may be an association between peripheral serum levels of IL-17A and IL-23A and the pathophysiology and development of GAD. These altered serum IL-17A and IL-23A levels may play a role in directing the early risk of developing GAD. We recommend further research to ascertain their exact role in the pathophysiology and their performance as risk assessment markers of GAD.

Amiloride lowers plasma TNF and interleukin-6 but not interleukin-17A in patients with hypertension and type 2 diabetes.

Interleukin (IL)-17A contributes to hypertension in preclinical models. T helper 17 and dendritic cells are activated by NaCl, which could involve the epithelial Na+ channel (ENaC). We hypothesized that the ENaC blocker amiloride reduces plasma IL-17A and related cytokines in patients with hypertension. Concentrations of IL-17A, IFN-γ, TNF, IL-6, IL-1β, and IL-10 were determined by immunoassays in plasma from two patient cohorts before and after amiloride treatment: 1) patients with type 2 diabetes mellitus (T2DM) and treatment-resistant hypertension (n = 69, amiloride 5-10 mg/day for 8 wk) and 2) patients with hypertension and type 1 diabetes mellitus (T1DM) (n = 29) on standardized salt intake (amiloride 20-40 mg/day, 2 days). Plasma and tissue from ANG II-hypertensive mice with T1DM treated with amiloride (2 mg/kg/day, 4 days) were analyzed. The effect of amiloride and benzamil on macrophage cytokines was determined in vitro. Plasma cytokines showed higher concentrations (IL-17A ∼40-fold) in patients with T2DM compared with T1DM. In patients with T2DM, amiloride had no effect on IL-17A but lowered TNF and IL-6. In patients with T1DM, amiloride had no effect on IL-17A but increased TNF. In both cohorts, blood pressure decline and plasma K+ increase did not relate to plasma cytokine changes. In mice, amiloride exerted no effect on IL-17A in the plasma, kidney, aorta, or left cardiac ventricle but increased TNF in cardiac and kidney tissues. In lipopolysaccharide-stimulated human THP-1 macrophages, amiloride and benzamil (from 1 nmol/L) decreased TNF, IL-6, IL-10, and IL-1β. In conclusion, inhibition of ENaC by amiloride reduces proinflammatory cytokines TNF and IL-6 but not IL-17A in patients with T2DM, potentially by a direct action on macrophages.NEW & NOTEWORTHY ENaC activity may contribute to macrophage-derived cytokine release, since amiloride exerts anti-inflammatory effects by suppression of TNF and IL-6 cytokines in patients with resistant hypertension and type 2 diabetes and in THP-1-derived macrophages in vitro.

Establishment of reference intervals for plasma IL-2, IL-4, IL-5, and IL-17A in healthy adults from the Jiangsu region of eastern China using flow cytometry: A single-center study

BackgroundCytokines are of utmost importance in both the physiological and pathological immune responses of the human body. This study utilized flow cytometry to measure the levels of plasma interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-5 (IL-5) and interleukin-17A (IL-17A) and established their reference intervals, aiming to provide data support for the diagnosis and treatment of clinical diseases. MethodsAccording to the inclusion and exclusion criteria, a total of 728 reference individuals were included in this study from January 2023 to June 2023. The Kolmogorov–Smirnov test was used to analyse the distributions of plasma IL-2, IL-4, IL-5 and IL-17A. The reference intervals of plasma IL-2, IL-4, IL-5 and IL-17A were established by the unilateral percentile method (95th percentile) based on the guidelines of C28-A 3 and WS/T 402–2012. ResultsIn this study, the levels of plasma IL-2, IL-4, IL-5 and IL-17A were nonnormally distributed. The concentrations of plasma IL-2, IL-4, IL-5 and IL-17A in healthy adults were not significantly different by sex or age (all P > 0.05). Therefore, all the reference individuals were combined into one group, and the reference intervals of plasma IL-2, IL-4, IL-5 and IL-17 were established by flow cytometry (IL-2 ≤ 10.25 pg/mL, IL-4 ≤ 9.87 pg/mL, IL-5 ≤ 3.36 pg/mL and IL-17A ≤ 9.46 pg/mL). ConclusionsWe first established the reference intervals of plasma IL-2, IL-4, IL-5 and IL-17A in healthy adults based on a single-center population in the Jiangsu region in eastern China, which will provide an important reference value for evaluating human immune status and the diagnosis and treatment of clinical diseases.

Notch signaling regulates Th17 cells differentiation through PI3K/AKT/mTORC1 pathway and involves in the thyroid injury of autoimmune thyroiditis.

Autoimmune Thyroiditis (AIT) is the most common thyroid disease; however, there were no measures to prevent the progression of the disease. The present study attempts to identify that Notch signaling regulates the differentiation of T helper 17 (Th17) cells by activating downstream Phosphatidylinositol-3 kinase/protein kinase/mechanistic target of rapamycin complex 1 (PI3K/AKT/mTORC1) pathway participating in the thyroid injury of the experimental autoimmune thyroiditis (EAT). In vivo experiments, mice were randomly divided into 4 groups: a control group, an EAT group, and two groups with LY294002 treatment (pTg plus 25mg/kg or 50mg/kg LY294002, respectively). The degrees of thyroiditis were evaluated, and the percentage of Th17 cells, expression of interleukin-17A (IL-17A), and the main components of the Notch-PI3K signaling pathway were detected in different groups. In vitro experiments, two different dosages of LY294002 (25 and 50μM) were used to intervene splenic mononuclear cells (SMCs) from EAT mice to further evaluate the regulatory effect of Notch-PI3K pathway on Th17 cells. Our data demonstrate that the infiltration of Th17 cells and the expressions of IL-17A, Notch, hairy and split 1 (Hes1), p‑AKT (Ser473), p‑AKT (Thr308), p‑mTOR (Ser2448), S6K1, and S6K2 increased remarkably in EAT mice. After PI3K pathway was blocked, the degrees of thyroiditis were significantly alleviated, and the proportion of Th17 cells, the expression of IL-17A, and the above Notch-PI3K pathway-related molecules decreased in a dose-dependent manner. Additionally, the proportion of Th17 cells was positively correlated with the concentration of serum thyroglobulin antibody (TgAb), IL-17A, and Notch-PI3K pathway-related molecules mRNA levels. Notch signal promotes the secretion of IL-17A from Th17 cells by regulating the downstream PI3K/AKT/mTORC1 pathway through Hes-Phosphatase and tensin homolog (PTEN) and participates in thyroid autoimmune damage, and the PI3K pathway inhibitor may play important effects on AIT by affecting Th17 cells differentiation.

Evaluation of Interleukin-17 A Levels in Patients with Breast Carcinoma

Breast cancer is a highly common form of cancer that impacts a considerable proportion of women on a global scale. Interleukin 17A (IL-17A) is a cytokine that has both anti-tumor and pro-tumor effects, which can vary depending on the specific tumor microenvironment. The aim of this study was to determine whether IL-17A can be used as a biomarker for diagnosis of breast cancer. Therefore, we compared concentrations of serum IL-17A in patients suffering from breast carcinoma and normal control women by an enzyme-linked immunosorbent assay (ELISA). This study included 86 women, 44 patients that were diagnosed with breast carcinoma, and 42 normal control women. Serum IL-17A levels in both case and control groups were measured by sandwich ELISA kits. The IL-17A serum level was significantly higher among patients with breast carcinoma than in the control group (p<0.001). The serum IL-17A concentration was significantly higher in estrogen receptor-positive cases than in estrogen receptor-negative cases (p=0.033). The highest levels of IL-17A were detected in patients with stage 2 breast carcinoma rather than stage 3 with no significant correlation. There was no correlation between IL-17A level and tumor size, lymph node invasion, or metastasis in patients with breast cancer. In conclusion, a high level of IL-17A in breast carcinoma patients compared to the control group was detected in our study. It indicates that IL-17A could be a promising biomarker for diagnosis of breast cancer and may play a role in tumor development. High levels of IL-17A were not a predictor of poor prognosis in breast cancer patients as it was not related to tumor size, lymph node invasion, or metastasis.

IL-17A neutralization fails to attenuate airway remodeling and potentiates a proinflammatory lung microenvironment in diacetyl-exposed rats.

Bronchiolitis obliterans (BO) is a devastating lung disease that can develop following inhalation exposure to certain chemicals. Diacetyl (DA) is one chemical commonly associated with BO development when inhaled at occupational levels. Previous studies in rats have shown that repetitive DA vapor exposures increased lung CD4+CD25+ T cells and bronchoalveolar (BAL) interleukin-17A (IL-17A) concentrations concurrent with the development of airway remodeling. We hypothesized that IL-17A neutralization would attenuate the severity of airway remodeling after repetitive DA vapor exposures. Sprague-Dawley rats were exposed to 200 parts-per-million DA vapor or filtered air (RA) for 6 h/day × 5 days and monitored for 2 wk postexposure. Treatment with IL-17A neutralization (αIL-17A) or IgG (control) began immediately following exposures and continued twice weekly until study's end. Lungs were harvested for histology, flow cytometry, and BAL analyses. Survival, oxygen saturations, and percent weight change decreased significantly in DA-exposed versus RA-exposed rats, but did not differ significantly between DA + αIL-17A versus DA + IgG. Similarly, the number nor severity of airway lesions did not differ significantly between DA + αIL-17A versus DA + IgG rats despite the percentage of lung regulatory T cells increasing with decreased BAL IL-17A concentrations. Ashcroft scoring of the distal lung parenchyma suggested worse parenchymal remodeling in DA + αIL-17A versus DA + IgG rats with increased expression of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and nuclear factor-kappa B (NF-κB). Collectively, IL-17A neutralization in DA-exposed rats failed to attenuate airway remodeling with increased expression of pro-inflammatory cytokines TNF-α, IL-1β, and NF-κB.NEW & NOTEWORTHY Interleukin-17A (IL-17A) neutralization has shown benefit previously in preclinical models of transplant-associated bronchiolitis obliterans (BO), yet it remains unknown whether IL-17A neutralization has similar benefit for other forms of BO. Here, IL-17A neutralization fails to prevent severe airway remodeling in rats exposed repetitively to the flavoring chemical diacetyl, and instead, promotes a proinflammatory microenvironment with increased expression of TNF-α, IL-1β, and NF-κB within the lung.

Impact of IL-17-producing γδ T cells on chronic otitis media induced by nontypeable Haemophilus influenzae in a mouse model.

Nontypeable Haemophilus influenzae (NTHi) is considered a major pathogen underlying middle ear infection. This study aimed to investigate the impact of IL-17 on chronic otitis media (COM) induced by NTHi in mice. NTHi was inoculated into the tympanic bulla with eustachian tubal obstruction. Middle ear effusions (MEEs) and tissues were collected on Days 3, 14 and at 1, 2 and 6 months after injection. The expression of interleukin-17A (IL-17A) in MEEs was significantly elevated compared to that in the control group at the translational and transcriptional levels during the experiments. The quantities of IL-17-producing γδ T cells were significantly increased compared to that in the control group during COM, but that of Th17 cells did not. Depletion of γδ T cells by anti-γδ T-cell receptor (TCR) monoclonal antibody (mAb) administration significantly decreased the bacteria counts and the concentrations of IL-1β, IL-6, IL-17A, TNF-α, and IL-10 in MEEs. Our results suggest that IL-17 may play an important role in prolonging the inflammation in the middle ear in COM, and that IL-17-producing γδ T cells may contribute to the exacerbated inflammatory response in the middle ear. In this study, anti-γδ TCR mAb administration was found to improve chronic middle ear inflammatory conditions.

Notch Signaling Pathway Promotes Th17 Cell Differentiation and Participates in Thyroid Autoimmune Injury in Experimental Autoimmune Thyroiditis Mice.

To investigate whether the Notch signaling pathway participates in the occurrence and development of experimental autoimmune thyroiditis (EAT) by affecting the differentiation and function of Th17 cells. Experimental mice were randomly divided into a control group, an EAT-A group (porcine thyroid immunoglobulin- (pTg-) treated mice) and an EAT-B group (treated with the DAPT γ-secretase inhibitor before pTg). HE staining, IHC staining, flow cytometry, RT-qPCR, and ELISA were used to evaluate the degrees of thyroiditis, detect the percentage of Th17 cells and measure the expression of retinoic acid-related orphan receptor gamma t (RORγt), interleukin-17A (IL-17A), and the main components of the Notch signaling pathway. The degrees of thyroiditis, the proportions of Th17 cells, and the expression of RORγt and IL-17A were significantly decreased in the EAT-B group after blocking the Notch signaling pathway by DAPT, and these parameters were significantly increased in the EAT-A group compared to the control group (all P < 0.05). Additionally, the Th17 cell percentages and IL-17A concentrations in spleen mononuclear cells (SMCs) from EAT-A mice decreased in a dose-dependent manner after DAPT treatment in vitro (all P < 0.01). Correlation analyses revealed that the Th17 cell percentages were positively correlated with the serum TgAb titers, Notch pathway-related mRNA expression levels, and IL-17A concentrations in EAT mice (all P < 0.05). The expression of Notch signaling pathway components was upregulated in EAT mice, but blockade of the Notch signaling pathway alleviated the degree of thyroiditis, decreased the Th17 cell proportions, and downregulated the IL-17A effector cytokine both in vivo and in vitro. These findings suggested that the Notch signaling pathway may be involved in the pathogenesis of thyroid autoimmune injury in EAT mice by promoting the differentiation of Th17 cells.

Biomarkers of infectious pneumonia in naturally infected calves.

To investigate the clinical value of selected biomarkers for evaluation of systemic inflammatory response and pulmonary inflammation and damage pattern in calves with different pneumonia forms. 16 calves with fibrinous pneumonia (FP group; infected with Mannheimia haemolytica or Pasteurella multocida), 12 calves with caseonecrotic pneumonia (CNP group; infected with Mycoplasma bovis), and 10 healthy calves (C group) based on results of bronchoalveolar lavage fluid (BALF) examination. Blood and BALF samples were collected. Annexin-A2 (Anx-A2), endothelin-1 (ET-1), calgranulin B (S100A9), transforming growth factor-β1 (TGF-β1), tumor necrosis factor-α (TNF-α), interleukin-17A (IL-17A), haptoglobin (Hp), lipopolysaccharide-binding protein (LBP), and albumin (Alb) concentrations and lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) activities were measured and compared across groups. Serum concentrations of Anx-A2, S100A9, TGF-β1, TNF-α, IL-17A, Hp, and LBP and activities of LDH and ALP were higher and Alb concentrations were lower for the pneumonia groups versus C group. BALF concentration ratios of S100A9:Alb, LBP:Alb, LDH:Alb, and ALP:Alb were higher for the pneumonia groups versus C group (P < .05). BALF concentration ratios of Anx-A2:Alb, TGF-β1:Alb, and IL-17A:Alb were higher for the FP group versus other groups (P < .05). Results indicated that serum Anx-A2, S100A9, TGF-β1, TNF-α, IL-17A, Hp, LBP, and Alb concentrations were useful in determining the systemic inflammatory response and that BALF concentration ratios of S100A9:Alb, TGF-β1:Alb, LBP:Alb, and ALP:Alb were useful biomarkers in determining pulmonary inflammation and damage. Measurements of BALF concentration ratios of Anx-A2:Alb, TGF-β1:Alb, and IL-17A:Alb could be beneficial to defining fibrinous characterization of pulmonary inflammation.

Interleukin 17A infusion has no acute or long-term hypertensive action in conscious unrestrained male mice.

Interleukin 17A (IL-17A) is a candidate mediator of inflammation-driven hypertension, but its direct effect on blood pressure is obscure. The present study was designed to test the hypothesis that systemic IL-17A concentration-dependently increases blood pressure and amplifies ANGII-induced hypertension in mice. Blood pressure was measured by indwelling chronic femoral catheters before and during IL-17A infusion w/wo angiotensin II (ANGII, 60ng/kg/min) in male FVB/n mice. Baseline blood pressure was recorded, and three experimental series were conducted: (1) IL-17A infusion with increasing concentrations over 6 days (two series with IL-17A from two vendors, n = 11); (2) ANGII infusion with IL-17A or vehicle for 9 days (n = 11); and (3) acute bolus infusions with four different concentrations (n = 5). Plasma IL-17A and IL-6 concentrations were determined by ELISA. Mean arterial and systolic blood pressures (MAP, SBP) decreased significantly after IL-17A infusion while heart rate was unchanged. In these mice, plasma IL-17A and IL-6 concentrations increased up to 3500- and 2.4-fold, respectively, above baseline. ANGII infusion increased MAP (~ 25 mmHg) and co-infusion of IL-17A attenuated ANGII-induced hypertension by 4.0 mmHg. Here, plasma IL-17A increased 350-fold above baseline. Acute IL-17A bolus infusion did not change blood pressure or heart rate. IL-17A receptor and IL-6 mRNAs were detected in aorta, heart, and kidneys of mice after IL-17A infusion. Nonphysiologically high concentrations of IL-17A reduce baseline blood pressure and increase IL-6 formation in male FVB/n mice. It is concluded that IL-17A is less likely to drive hypertension as the sole cytokine mediator during inflammation in vivo.

Heparin-binding hemagglutinin as a composition antigen of tuberculosis vaccine exerts protective immune effects by inducing IL-17

To explore the protective immune effect induced by mucosal delivery heparin-binding hemagglutinin (HBHA)-a candidate vaccine antigen of Mycobacterium tuberculosis. Female C57BL/6 mice were between 6 and 8 weeks of age before experimental use. Thirty mice received different immunization strategies and were randomly divided into the control group, the early secreting antigen target-6 (ESAT-6) intranasal immunization group, the HBHA intranasal immunization group, the BCG priming PBS control group, or BCG priming HBHA boost group, 6 mice in each group. In order to analyzed the immune effect, the concentrations of plasma Interleukin-17A (IL-17A) and other cytokines were measured by ELISA. Quantitative real-time PCR analyses were performed to detect the relative quantity (RQ) mRNA of IL-17A in the lung. The lung tissue sections were stained to detect the formation of the tertiary lymphoid structures. The chemokines contributed to formation of the tertiary lymphoid structures were also measured. Flow cytometry was used to detect the frequency of Th1 and Th17 cells in the system. Sixty mice in the BCG priming PBS control group and the BCG priming HBHA boost group were sacrificed at different time points after infection to count the lung bacterial burden. The concentrations of plasma IL-17A and relative quantity of lung IL-17A mRNA were highest in the BCG priming HBHA boost group [(14.76±4.73) pg/mL,RQ (12.27±6.71)], which was significantly higher than the control group [(5.57±2.95) pg/mL,RQ (1.30±0.97)] (t=4.213, P<0.001; t=5.984, P<0.001), and also significantly higher than the BCG priming PBS control group [(6.81±2.18) pg/mL,RQ (1.44±1.16)] (t=3.646 P=0.001; t=6.185 P<0.001). Compared with the BCG priming PBS control group (0.38±0.38)% the frequency of spleen Th17 cells were also significantly increased (t=-0.280, P=0.048) in the BCG-primary HBHA boost group (1.02±0.34)%. In addition, HBHA boosting could promote better formation of the tertiary lymphoid structures in the lung, and decrease the bacterial load on the early stage after BCG challenge. Collectively, mucosal delivery of HBHA can effectively enhance the protective effect after BCG vaccination, and it is a potential candidate vaccine component.

OP32 Pyroptosis Inhibition Prevents the Cytotoxicity Induced by IL-17 Without Impairing Its Beneficial Effects

Abstract Background Th17 cells and their main secreting cytokine interleukin-17A (IL-17) are considered as the main pathogenic factors in inflammatory bowel diseases (IBDs). However, anti-IL-17 neutralizing antibodies, a theoretically curative medication for IBDs, paradoxically aggravated intestinal inflammation. The mechanisms by which it mediates the protective and pathologic effects of IL-17 remain unclear in the intestinal epithelium. Methods The intestinal epithelial responses induced by IL-17 was evaluated using the human small intestinal organoid (enteroid) model. Results Organoid-forming efficiency, cell viability and proliferation of enteroids were decreased in proportion to the concentration of IL-17, which did not differ between the enteroids derived from controls and patients with Crohn’s disease. Bulk RNA-sequencing revealed the enrichment of secretion signaling in IL17-treated enteroids. Among its components, PIGR was up-regulated significantly as the concentration of IL-17 increased, resulting in IgA transcytosis and protective role against pathogens. The IL-17-induced cytotoxicity was predominantly mediated by pyroptosis with activation of CASP1 and cleavage of GSDMD. Single-cell RNA- sequencing identified pyroptosis occurred actively in intestinal stem cells (ISCs) and enterocytes. Anti-IL-17 antibody, izekizumab, completely restored IL-17-induced cytotoxicity, but suppressed mucin secretion and IgA transcytosis. CASP1 inhibitor, Ac-YVAD-cmk, restores cytotoxicity induced by IL-17, without impairing its beneficial effects. Conclusion IL-17 induces pyroptosis of ISCs and enterocytes, as well as mucin secretion and PIGR-induced IgA transcytosis. Paradoxical gastrointestinal effects of IL-17 neutralizing antibodies may be associated with inhibition of mucin secretion and IgA transcytosis. The inhibition of pyroptosis using the CASP1 inhibitor prevents the cytotoxicity induced by IL-17 without compromising its beneficial effects.

Закономірності зміни концентрацій ІЛ-17А, ІЛ-10 та ІЛ-2 у крові людей із гніздовою алопецією до та після лікування збагаченою тромбоцитами плазмою

Alopecia areata is a heterogeneous disease characterized by nonscarring hair loss. It affects 1-2% of the population of both genders and occurs in all age groups. According to the recent studies, helper T cells 1, helper T cells 2, helper T cells 17, and regulatory T cells may be the main possible initiators of alopecia areata. The purpose of the study was to determine the concentrations of serum interleukin-17A, interleukin-10 and interleukin-2 in patients with alopecia areata, and to study the relationship between their levels before and after treatment with platelet-rich plasma. Materials and methods. The study involved 37 patients with alopecia areata aged (35.7±8.9) years and 30 people of the control group aged (37.4±7.9) years. People with alopecia areata were injected with platelet-rich plasma. For this, immediately before administration, venous blood was taken from the patients from the cubital vein into a special tube for the preparation of platelet-rich plasma. Platelet-rich plasma was injected into the lesions at the rate of 0.2 ml/cm2 using an insulin syringe and a 30 G needle, once every 2 weeks for 16 weeks. The patients were examined before the treatment and in 3, 6 and 17 months. The concentrations of interleukin-17A, interleukin-2 and interleukin-10 were determined in blood serum before treatment and after 3, 6 and 17 months of observation by the Sandwich-ELISA method (enzyme-linked immunosorbent assay). Interleukin-17A, interleukin-2 and interleukin-10 in the blood serum in the control group were also determined. The software program SPSS 22.0 was used for the statistical analysis. The normality distribution was tested by Shapiro-Wilk’s method. Independent Student’s t-test for unpaired samples was used to compare mean values between groups. The p-value &lt; 0.05 was considered significant. Results and discussion. The results obtained in our study showed that the concentrations of the pro-inflammatory cytokines interleukin-17A and interleukin-2 were significantly higher in persons with alopecia areata before treatment. After 17 months of therapy with platelet-rich plasma, serum interleukin-17A and interleukin-2 levels approached the control level. Conclusion. There was no difference in the concentration of anti-inflammatory interleukin-10 between patients with alopecia areata and healthy people. In the course of treatment, the concentration of interleukin-10 increased after 3 and 17 months of therapy

Застосування топічного такролімуса у хворих із екземою рук з урахуванням ступеня тяжкості дерматозу, деяких показників цитокінового профілю, вмісту вітаміну д та генетичного поліморфізму гена глюкокортикоїдного рецептора

The purpose of the study was to evaluate and compare the therapeutic capacity of 0.1% topical tacrolimus ointment in patients with hand eczema who were resistant to glucocorticoid treatment, taking into account the severity of dermatosis, interleukin-17A and interleukin-2 concentrations, vitamin D and genetic polymorphism of the glucocorticoid receptor gene. Materials and methods. The venous blood of 143 patients with hand eczema aged (42.2±11.1) years was used in the study. During the patients examination the data on age, sex, body mass index (kg/m2), body mass index ≥25 kg/m2 (%), smoking, IgE (iu/ml), interleukin-17A (pg/ml), interleukin-2 (pg/ml), and 1,25(OH)2D3 (ng/ml) were obtained. The Hand Eczema Severity Index was also assessed in each subject. According to the Hand Eczema Severity Index value all patients were divided into three subgroups: mild, moderate and severe eczema. Patients with mild and moderate eczema were appointed topical glucocorticoid 0.1% mometasone furoate cream twice a day for 2 weeks. Patients with severe hand eczema was appointed additionally systemic corticosteroid – a solution of dexamethasone by intramuscular injection at the dose of 8 mg/day No. 3, then another 4 mg/day for other 2 days. After 2 weeks, individuals who did not respond well to glucocorticoid treatment were prescribed tacrolimus 0.1% ointment twice a day for 2 weeks. BclI SNP (rs41423247) of the glucocorticoid receptor gene (NR3C1) was determined using the polymerase chain reaction-restriction fragment length polymorphism method (Fleury et al. 2003). The statistical analysis was performed using the program SPSS 22.0. The quantitative variables were tested for normal distribution by the Shapiro-Wilk method. The comparisons of the means between the two subgroups were performed using Student's t-test for independent samples. The comparison of the frequencies distribution in the subgroups was performed using the Pearson test. The P value &lt;0.05 was considered as significant. Results and discussion. Findings in a previous study showed that not all people with hand eczema responded well to treatment with topical 0.1% mometasone furoate cream and systemic glucocorticoid. After 2 weeks of topical tacrolimus, of 51 patients who were not responsive to glucocorticoid treatment, 39 responded well to therapy. The concentration of interleukin-17A in the blood serum of such people was significantly lower than in people who did not respond to treatment. Conclusion. We did not find any differences in the content of immunoglobulin E, interleukin-2, 1α,25(OH)2D3 and BclI polymorphism of the glucocorticoid receptor gene in the subjects under study

Effects of smoking on the gingival crevicular fluid levels of interleukin-17A, interleukin-17E, and oxidative stress following periodontal treatment process.

How smoking affects periodontal inflammation and healing still needs to be revealed with all its mechanisms. In this study, the gingival crevicular fluid (GCF) levels of: (a) interleukin-17A (IL-17A) and interleukin-17E(IL-17E) with their ratios and (b) oxidative stress by means of total oxidative stress (TOS), total anti-oxidant capacity (TAOC), and their ratios as the oxidative stress index (OSI) were evaluated and compared for smoking and non-smoking periodontitis patients after a periodontitis management process including both the non-surgical and surgical treatments. Fifteen smoker and 15 non-smoker generalized periodontitis patients as 2 distinct groups participated in the study. Conventional clinical and radiographical examinations were utilized for the periodontitis diagnosis. The clinical data and GCF samples were collected at baseline, 4week after non-surgical periodontal treatment (NSPT), and 4weeks after surgical periodontal treatment (SPT). IL-17A, IL-17E, TOS, and TAOC were determined by ELISA and Rel Assay. Clinical parameters in both smokers and non-smokers improved following periodontal treatment (P<.001) and their clinical data were similar for all the examination times (baseline, NSPT, and SPT) (P>.05). Following the treatment phases, the IL-17A concentration decreased and the IL-17E concentration increased in both the smokers and non-smokers (P<.01). The total amount of IL-17A decreased while the total amount of IL-17E increased in smokers throughout NSPT and SPT (P<.01). Such an alteration was seen only at SPT compared to NSPT and baseline in non-smokers (P<.01). The concentration and total amount of IL-17A were higher at baseline, and the concentration and total amount of IL-17E were lower at all examination time points in non-smokers as compared to smokers (P<.01). The 17A/E ratio decreased in both groups following the treatment phases and was higher in smokers at all the examination times (P<.01). TOS were higher and TAOC were lower in smokers versus non-smokers at all the time points, but the differences were significant only for TOS levels (P<.01). Throughout the treatment phases, the concentration and total amount of TOS decreased in smokers(P<.01) and only the total amount of TOS decreased in non-smokers (P<.01). The concentration and total amounts of TAOC increased throughout the treatments in both smokers and non-smokers without significant changes (P>.05). The baseline OSI was higher in smokers, and it decreased only in smokers following the treatment phases (P<.01). Smoking and periodontal inflammation were found to alter IL-17A, IL-17E, and oxidant/anti-oxidant statuses in periodontitis patients. The intra-group assessments in smokers demonstrated more apparent alterations in the oxidant/anti-oxidant statuses and IL-17A and IL-17E levels after periodontitis management.

RELATION BETWEEN GLUCOCORTICOID RESISTANCE AND GLUCOCORTICOID RECEPTOR GENE BclI-POLYMORPHISM AND SOME CYTOKINE PROFILE PARAMETERS IN PATIENTS WITH HAND ECZEMA

Though glucocorticoids are widely used in dermatological practice, some patients with hand eczema may have resistance to glucocorticoids, even when they are taken in heavy doses. Glucocorticoids mediate their actions through glucocorticoid receptors. Polymorphism of the glucocorticoid receptor gene (NR3C1) can inhibit the cellular response to glucocorticoids and lead to reduced response to the therapy. Some cytokines can affect the production of various glucocorticoid receptor subunits, modulating the cell response to glucocorticoids. However, there is still need in detailed study of pathogenetic mechanisms and the detection of highly specific predictors of glucocorticoid resistance. The aim of this work was to investigate the possible relation between glucocorticoid resistance in patients with hand eczema, rs41423247 SNP, and blood concentration of interleukin-17A and interleukin-2. The venous blood of 143 patients with hand eczema (42% of women and 58% of men) mean age 42.2 ± 11.1 years was taken for the study. During the patients examination the data on age, sex, body mass index (kg/m2), body mass index ≥ 25 (kg/m2(%)), the habit of smoking, concemtration of immunoglobulin E (iu/ml), interleukin-17A (pg/ml) and interleukin-2 (pg/ml) were obtained. The еczema аrea and severity index was assessed in each subject before the therapy and in two weeks since the therapy started. According to index value, all the patients were divided into three subgroups: mild eczema, moderate eczema, and severe eczema. Patients with mild and moderate eczema were prescribed to apply topical glucocorticoid 0.1% mometasone furoate cream twice a day for 2 weeks. The patients with severe hand eczema were prescribed to receive additional systemic corticosteroid, a solution of dexamethasone by intramuscular injection in a dose of 8 mg / day, then 4 mg / day for another 2 days. BclI SNP (rs41423247) of the glucocorticoid receptor gene (NR3C1) was determined using PCR-RFLP method. The quantitative variables were tested for normal distribution by the Shapiro-Wilk test. The comparisons of the means between the two subgroups were performed by Student's t-test for independent samples. The comparison of the frequencies distribution in the subgroups was calculated by using the Pearson test. The P value &lt; 0.05 was considered as significant. Thus, the obtained results revealed that insensitivity to glucocorticoids in patients with hand eczema is related to NR3C1 gene Bcl-1 polymorphism, eczema severity and plasma level of interleukin-17, interleukin-2. The plasma content of interleukin-17 and interleukin-2 in patients with glucocorticoid resistance was significantly higher compared to hormone-sensitive subjects. There were significantly more people with the C/G- and G/G genotypes in the group that did not have a clinical response to therapy.

Study on mechanisms of interleukin-17A regulating the expressions of interleukin-1β and interleukin-23 in mouse keratinocytes

Objective: To investigate the mechanisms of interleukin-17A (IL-17A) regulating the expressions of IL-1β and IL-23 in mouse keratinocytes (KCs). Methods: Primary KCs were isolated from the skin of 400 newborn male and female wild type C57BL/6 mice and cultured in 24-well plates with Roswell Park Memorial Institute 1640 medium containing fetal bovine serum in the volume fraction of 10% for the following experiments. (1) The cells were divided into phosphate buffer solution (PBS) control group and IL-17A stimulation group according to the random number table (the same grouping method below), which were cultured with 10 μL PBS or 10 μL IL-17A in the mass concentration of 100 ng/mL for 6 hours, respectively. The expression levels of IL-1β and IL-23 mRNA in cells were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR), with 3 samples in each group. (2) The cells were divided into dimethyl sulfoxide (DMSO) control group, IL-17A+ DMSO group, IL-17A+ nuclear factor κB (NF-κB) inhibitor group, IL-17A+ signal transduction and activator of transcription 3 (STAT3) inhibitor group, IL-17A+ extracellular signal-regulated kinase 1 (ERK1) inhibitor group, IL-17A+ ERK2 inhibitor group, and IL-17A+ c-Jun N-terminal kinase (JNK) inhibitor group. The reagents were added to cells in corresponding groups respectively and cultured for 6 hours. The volume of each reagent was 10 μL, the mass concentration of IL-17A was 100 ng/mL, and the molarity concentrations of NF-κB, STAT3, ERK1, ERK2, JNK signal pathway inhibitors PDTC, S3I-201, SCH772984, SCH772984, SP600125 were 5 μmol/L, 100 μmol/L, 4 nmol/L, 1 nmol/L, and 10 μmol/L, respectively. The expression levels of IL-1β mRNA and IL-23 mRNA in cells were detected by real-time fluorescence quantitative RT-PCR, with 3 samples in each group. (3) The cells were grouped and treated the same as those in experiment (1). The levels of NF-κB phosphorylation, STAT3 phosphorylation, ERK phosphorylation, and JNK phosphorylation were detected by Western blotting, with 3 samples in each group. Data were statistically analyzed with two-tailed Student t test, one-way analysis of variance, t test, and Bonferroni correction. Results: (1) After culture of 6 hours, compared with those in PBS control group, the expression levels of IL-1β and IL-23 mRNA in cells in IL-17A stimulation group were significantly increased (t=13.46, 6.72, P<0.01). (2) After culture of 6 hours, the expression levels of IL-1β and IL-23 mRNA in cells in DMSO control group, IL-17A+ DMSO group, IL-17A+ NF-κB inhibitor group, IL-17A+ STAT3 inhibitor group, IL-17A+ ERK1 inhibitor group, IL-17A+ ERK2 inhibitor group, and IL-17A+ JNK inhibitor group were 1.00±0.11, 4.01±0.32, 0.32±0.06, 1.76±0.43, 3.62±0.24, 3.80±0.43, 4.26±0.74 and 1.03±0.29, 4.08±0.34, 4.76±0.38, 4.70±0.21, 1.06±0.42, 0.92±0.21, 0.39±0.05, respectively. Compared with those in DMSO control group, the expression levels of IL-1β and IL-23 mRNA in cells in IL-17A+ DMSO group were significantly increased (t=9.24, 12.60, P<0.01). Compared with that in IL-17A+ DMSO group, the expression level of IL-1β mRNA was significantly decreased in cells in IL-17A+ NF-κB inhibitor group and IL-17A+ STAT3 inhibitor group (t=11.34, 6.91, P<0.01). Compared with that in IL-17A+ DMSO group, the expression level of IL-23 mRNA was significantly decreased in cells in IL-17A+ ERK1 inhibitor group, IL-17A+ ERK2 inhibitor group, and IL-17A+ JNK inhibitor group (t=12.44, 13.03, 15.21, P<0.01). (3) After culture of 6 hours, compared with those in PBS control group, the levels of NF-κB phosphorylation, STAT3 phosphorylation, ERK phosphorylation, and JNK phosphorylation in cells in IL-17A stimulation group were significantly increased. Conclusions: IL-17A promotes the transcription of IL-1β in mouse KCs through the phosphorylation of NF-κB and STAT3 pathways and IL-23 through the phosphorylation of ERK and JNK pathways.

Elevated Levels of Serum IL-17A Secondary to Repeated Intravitreal Injections of Aflibercept in Treatment-Naive Patients with Neovascular Age-Related Macular Degeneration

We evaluated the effect of three monthly intravitreal injections of aflibercept on the serum concentration of interleukin 17A (IL-17A), monocyte chemoattractant protein 1 (MCP-1/CCL2), vascular endothelial growth factor (VEGF) and placental growth factor (PlGF) in treatment-naive patients with neovascular age-related macular degeneration (nAMD). Twenty-two eyes of twenty-two patients with nAMD scheduled for the initial loading phase of intravitreal aflibercept (2 mg/0.05 mL) were included. Serum VEGF, PlGF, MCP-1/CCL2 and IL-17A levels were determined four times in each individual—just before the first injection, 2–3 days after the first injection, just before the third injection, and then 2–3 days after the third aflibercept injection. A statistically significant difference was found between the serum PlGF and IL-17A levels measured before the first injection and after the initial loading phase, with a mean value (MV) of 440.884 vs. 302.151 (p = 0.023) for PlGF and MV = 139.088 pg/mL vs. 151.233 pg/mL (p = 0.016) for IL-17A, respectively. There were no statistically significant differences for VEGF and MCP-1/CCL2 between any of the compared measurements. We reveal that repeated injections of aflibercept promote an increase in serum IL-17A concentration, which may lead to a systemic inflammatory response mediated by IL-17A, but not by MCP-1.

The Role of Highly Active Antiretroviral Therapy (HAART) on Interleukin 17A (IL-17A) in Normotensive and Preeclamptic Black South African Women.

Introduction Interleukin 17A has been implicated in the pathophysiology of both human immune deficiency virus and preeclampsia. This study evaluated serum levels of IL-17A based on pregnancy type, gestational age, HIV status, and duration of HAART. Material and Methods. A sample size of 250 was analysed: normotensives (n = 150; N) and preeclamptics (n = 100; PE). Normotensives were further stratified into HIV negative (n = 90), HAART-acute (n = 30), and HAART-chronic (n = 30). The PE group was divided into early onset (n = 50; EOPE) and late onset (n = 50; LOPE). The EOPE and LOPE groups were subdivided into HIV negative (n = 30), HAART-acute (n = 10), and HAART-chronic (n = 10). Analysis of IL-17A was performed using a multiple Bio-Plex immunoassay method. Results Pregnancy type: the levels of IL-17A were increased in PE compared to N (P = 0.0014). Gestational age: the levels of IL-17A were increased in EOPE compared to N group (P = 0.0113). A significant increase in the levels of IL-17A in LOPE compared to N was observed (P = 0.0063). HIV status: the levels of IL-17A were increased in PE compared to N (P = 0.0114) and in EOPE compared to N groups (P = 0.0071). HAART duration: the concentration of IL-17A was increased in HAART-chronic PE compared to N groups (P = 0.0062). There was also an increase in the levels of IL-17A in EOPE compared to N (P = 0.0029). Conclusion The study demonstrates that IL-17A is involved in the pathophysiology of PE and that in the presence of HIV infection, chronic HAART administration predisposes women to the development of EOPE.

The influence of overnight orthokeratology on ocular surface and dry eye-related cytokines IL-17A, IL-6, and PGE2 in children

ObjectiveTo investigate the effect of overnight orthokeratology (OOK) on the ocular surface and dry eye-related cytokines in children. MethodsA non-randomized, prospective pilot study was conducted including sixty myopes treated with OOK and sixty age-matched spectacle wearing participants. The following tests were performed before and after 1, 3, 6 and 12 months: ocular surface disease index (OSDI), noninvasive tear breakup time (NITBUT), tear meniscus height (TMH), corneal fluorescein staining (CFS), meiboscore using noncontact meibography. Then the concentrations of interleukin-17A (IL-17A), interleukin-6 (IL-6), and prostaglandin E2 (PGE2) in tear samples were detected with a multiplex immunobead assay at different time points. ResultsAll parameters had no statistical differences between the two groups prior to treatment. No adverse events were observed except trace to moderate corneal staining and allergic conjunctivitis in the treatment group. NITBUT significantly decreased after 6 and 12 months OOK wearing (P = 0.003 and P = 0.001, respectively). After wearing OOK there was a significant increase in CFS at each follow-up time point compared with baseline (P = 0.023, P = 0.016, P = 0.001, and P < 0.001at 1, 3, 6, and 12 months, respectively). The upper meiboscore and the total meiboscore increased gradually and peaked at 12 months of OOK (both P < 0.001). The concentration of the three cytokines in the treatment group significantly increased after OOK wearing. These increases occurred at different time points: IL-17A increased significantly 3 months after OOK, IL-6 at 6 months, and PGE2 at 12 months (all P < 0.001). However, there were no significant changes in the above parameters in the control group. There were no significant differences in the OSDI or TMH at any follow-up time point compared to baseline in both groups (both P > 0.05). ConclusionsShort-term OOK may reduce the stability of the tear film and increase damage to the corneal epithelium. Long-term OOK could induce ocular inflammation through the disruption of meibomian glands.