Urinary Cytokine Levels Research Articles

Evaluating Single-Nucleotide Polymorphisms in Inflammasome Proteins and Serum Levels of IL-18 and IL-1β in Kidney Interstitial Damage in Anti-Neutrophilic Cytoplasmic Antibody-Associated Vasculitis.

The inflammasome regulates the innate inflammatory response and is involved in autoimmune diseases. In this study, we explored the levels of IL-18 and IL-1β in serum and urine and the influence of various single-nucleotide polymorphisms (SNPs) on kidney lesions at diagnosis in patients with ANCA-associated vasculitis (AAV) and their clinical outcomes. Ninety-two patients with renal AAV were recruited, and blood and urine were collected at diagnosis. Serum and urine cytokine levels were measured by ELISA. DNA was extracted and genotyped using TaqMan assays for SNPs in several inflammasome genes. Lower serum IL-18 (p = 0.049) and the IL-18 rs187238 G-carrier genotype (p = 0.042) were associated with severe fibrosis. The IL-18 rs1946518 TT genotype was associated with an increased risk of relapse (p = 0.05), whereas GG was related to better renal outcomes (p = 0.031). The rs187238 GG genotype was identified as a risk factor for mortality within the first year after AAV diagnosis, independent of the requirement for dialysis or lung involvement (p = 0.013). We suggest that decreased cytokine levels could be a surrogate marker of scarring and chronicity of the renal lesions, together with the rs187238 GG genotype. If our results are validated, the rs1946518 TT genotype predicts the risk of relapse and renal outcomes during follow-up.

Inflammatory markers for improved recurrent UTI diagnosis in postmenopausal women.

Recurrent urinary tract infection (rUTI) severely impacts postmenopausal women. The lack of rapid and accurate diagnostic tools is a major obstacle in rUTI management as current gold standard methods have >24-h diagnostic windows. Work in animal models and limited human cohorts have identified robust inflammatory responses activated during UTI. Consequently, urinary inflammatory cytokines secreted during UTI may function as diagnostic biomarkers. This study aimed to identify urinary cytokines that could accurately diagnose UTI in a controlled cohort of postmenopausal women. Women passing study exclusion criteria were classified into no UTI and active rUTI groups, and urinary cytokine levels were measured by immunoassay. Pro-inflammatory cytokines IL-8, IL-18, IL-1β, and monocyte chemoattractant protein-1 were significantly elevated in the active rUTI group, and anti-inflammatory cytokines IL-13 and IL-4 were elevated in women without UTI. We evaluated cytokine diagnostic performance and found that an IL-8, prostaglandin E2, and IL-13 multivariable model had the lowest misclassification rate and highest sensitivity. Our data identify urinary IL-8, prostaglandin E2, and IL-13 as candidate biomarkers that may be useful in the development of immunoassay-based UTI diagnostics.

Evaluating symptom severity and urinary cytokine levels in interstitial cystitis/bladder pain syndrome patients, with and without Hunner's lesions.

Interstitial cystitis/bladder pain syndrome (IC/BPS) is a condition characterized in part by urinary urgency, frequency, and pain. There is a strong interest in gathering more data to compare and assess the differences in characteristics based on the presence of Hunner's lesions in patients with IC/BPS. Using a nationwide crowdsource effort, we collected surveys and urine samples from patients with a history of IC/BPS. Participants completed the Interstitial Cystitis Symptom Index (ICSI) and Problem Index (ICPI), Overactive Bladder questionnaire (OABq SF), and pain scores. In addition, participants reported any co-morbidities and lifestyle modifications. Urinary cytokine levels were measured and compared to symptom severity. 491 participants enrolled: 119 with history of ulcerative Hunner's lesions (UIC), 372 reported no lesions (NHIC), and 2 unknowns. 96.3% were female, and prevalence of UIC was equal for both genders. Average age was higher for UIC vs. NHIC group (P = 0.011), as was the duration since diagnosis (P < 0.001). Symptom scores were elevated in UIC patients (P < 0.001). Both groups widely implemented lifestyle modifications, with dietary changes being most prevalent (70.1%), followed by prescription medication usage (63.1%). More UIC compared to NHIC participants experienced co-morbidities (P = 0.010). Urine samples were analyzed for GRO, IL-6, IL-8, and MCP-1. MCP-1 levels were significantly higher in UIC patients (P = 0.044). Weak positive correlation was found between cytokines and symptom scores. Patients with UIC and NHIC from across the United States displayed distinct phenotypic and urine biological characteristics. These findings contribute to increased understanding of IC/BPS and may aid in improving our knowledge of the condition.

Nephrotoxicity in the Age of Immune Checkpoint Inhibitors: Mechanisms, Diagnosis, and Management.

Immune checkpoint inhibitors (ICI) revolutionized cancer therapy by augmenting anti-tumor immunity via cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed death-1/programmed death-ligand 1 (PD-1/PD-L1). However, this breakthrough is accompanied by immune-related adverse effects (irAEs), including renal complications. ICI-related nephritis involves complex mechanisms like auto-reactive T cells, auto-antibodies, reactivation of drug-specific T cells, and cytokine-driven inflammation culminating in AKI. ICI-AKI typically manifests weeks to months into treatment, often with other irAEs. Timely detection relies on monitoring creatinine levels and urine characteristics. Biomarkers, like soluble interleukin-2 receptor (sIL-2R) and urine cytokine levels, provide non-invasive insights, while renal biopsy remains the gold standard for confirmation. Management of ICI-AKI requires a balance between discontinuing ICI therapy and prompt immunosuppressive intervention, typically with corticosteroids. Some cases permit ICI therapy resumption, but varying renal recovery rates highlight the importance of vigilant monitoring and effective therapy. Beyond its clinical implications, the potential of irAEs to predict positive treatment responses in certain cancers raises intriguing questions. Data on nephritis-treatment response links are limited, and ongoing research explores this complex interaction. In summary, ICI therapy's transformative impact on cancer treatment is counterbalanced by irAEs, including nephritis. Early recognition and management are vital, with ongoing research refining diagnostic and treatment strategies.

Urine biomarker could be a useful tool for differential diagnosis of a lower urinary tract dysfunction.

A precision diagnosis of lower urinary tract dysfunctions (LUTD) such as bladder outlet obstruction, detrusor overactivity (DO), interstitial cystitis/bladder pain syndrome (IC/BPS), dysfunctional voiding (DV), or detrusor underactivity (DU) needs invasive videourodynamic study. Exploring non-invasive tools to help screening LUTD is necessary for clinicians in their daily practice. This article reviews recently clinical studies of using urinary inflammatory proteins and oxidative stress biomarkers in the identification of specific LUTD among men and women with lower urinary tract symptoms (LUTS). Some important findings have been reported: (1) Using urine chemokines CXCL-1 and interleukin-8 (IL-8), we may discriminate overactive bladder (OAB) symptoms in women between DO and urinary tract infection. (2) Urinary levels of oxidative stress biomarkers such as 8-hydroxydeoxyguanosine (8-OHdG) and 8-isoprostane have a potential being used as a tool to identify women with mixed DO and stress urinary incontinence. (3) Urine levels of total antioxidant capacity (TAC), and prostaglandin E2 (PGE2) are positively correlated with voiding detrusor pressure in patients with DU. (4) Urine levels of brain-derived neurotrophic factor (BDNF) and PGE2 were significantly higher in the DU patients with detrusor function recovery. (5) Women with DV had higher urinary levels of tumor necrosis factor-alpha (TNF-α) and 8-OHdG, and urinary IL-2 level was significantly lower. (6) Urine level of 8-isoprostane was higher in the patients with idiopathic DO and neurogenic DO. (7) Higher urine cytokine levels of monocyte chemoattractant protein-1 (MCP-1), regulated on activation, normal T-cell expressed and secreted (RANTES), CXCL-10, IL-7, and eotaxin-1 in patients with IC/BPS than controls. (8) The urine levels of IL-8, CXCL-10, BDNF, IL-6, and RANTES were significantly higher in patients with Hunner's IC than non-Hunner's IC. (9) Male patients with IC/BPS had a significantly higher level of eotaxin, MCP-1, TNF-α, 8-OHdG, and TAC. Combining a higher eotaxin and a higher TNF-α can provide a satisfactory diagnostic value in discriminating IC/BPS from other LUTD in men. These studies provide evidence that measurement of cluster of urine biomarkers could be used as a diagnostic tool to differentiate different LUTD in patients with similar LUTS.

Phytotherapy (BNO 1045) of Acute Lower Uncomplicated Urinary Tract Infection in Women Normalizes Local Host Responses

Introduction: Acute lower uncomplicated urinary tract infection (uUTI) affects a large proportion of women. Increased antimicrobial resistance has created an urgent need for novel therapeutics and the phytotherapeutic drug BNO 1045 (Canephron^® N) has previously been shown to be noninferior to standard antimicrobial stewardship. This sub-analysis from a randomized, double-blind, controlled phase III noninferiority clinical trial using BNO 1045 versus fosfomycin to treat uUTI aimed to determine how urine cytokine levels are altered by the two different treatments. Methods: Urine samples from a predefined subset of women diagnosed with uUTI (18–70 years) and treated with BNO 1045 (n = 58) or fosfomycin (n = 69) were analyzed for urine levels of IL-6 and IL-8, using analyte-to-creatinine ratios. Results: BNO 1045 treatment showed similar effects to fosfomycin treatment in reducing both urine IL-6 and IL-8 levels. Mean IL-6 and IL-8 levels were markedly reduced in all patients regardless of treatment. BNO 1045 treatment decreased urine IL-8 significantly (p = 0.0142) and showed a trend toward reduction of urine IL-6 (p = 0.0551). Fosfomycin treatment reduced both IL-6 and IL-8 levels significantly (p = 0.0038, <0.0001 respectively). Conclusion: BNO 1045 is, in addition to reducing symptoms, comparable to fosfomycin treatment in reducing the local inflammatory response associated with uUTI.

Urinary interleukins (IL)-6 and IL-10 in schoolchildren from an area with low prevalence of Schistosoma haematobium infections in coastal Kenya.

Urinary cytokines are gaining traction as tools for assessing morbidity in infectious and non-infectious inflammatory diseases of the urogenital tract. However, little is known about the potential of these cytokines in assessing morbidity due to S. haematobium infections. Factors that may influence the urinary cytokine levels as morbidity markers also remain unknown. Therefore the objective of the present study was to assess how urinary interleukins (IL-) 6 and 10 are associated with gender, age, S. haematobium infections, haematuria and urinary tract pathology and; 2) to assess the effects of urine storage temperatures on the cytokines. This was a cross-sectional study in 2018 involving 245 children aged 5-12 years from a S. haematobium endemic area of coastal Kenya. The children were examined for S. haematobium infections, urinary tract morbidity, haematuria and urinary cytokines (IL-6 and IL-10). Urine specimens were also stored at -20°C, 4°C or 25°C for 14 days before being assayed for IL6 and IL-10 using ELISA. The overall prevalence of S. haematobium infections, urinary tract pathology, haematuria, urinary IL-6 and urinary IL-10 were 36.3%, 35.8%, 14.8%, 59.4% and 80.5%, respectively. There were significant associations between prevalence of urinary IL-6, but not IL-10, and age, S. haematobium infection and haematuria (p = 0.045, 0.011 and 0.005, respectively) but not sex or ultrasound-detectable pathology. There were significant differences in IL-6 and IL-10 levels between urine specimens stored at -20°C and those stored at 4°C (p<0.001) and, between those stored at 4°C and those stored at 25°C (p<0.001). Urinary IL-6, but not IL-10, was associated with children's age, S. haematobium infections and haematuria. However, both urinary IL-6 and IL-10 were not associated with urinary tract morbidity. Both IL-6 and IL-10 were sensitive to urine storage temperatures.

MP39-01 CLASSIFICATION OF INTERSTITIAL CYSTITIS/BLADDER PAIN SYNDROME (IC/BPS) USING A MACHINE LEARNING BASED SYMPTOM SCORE CONSTRUCTED ON INFLAMMATORY URINE CYTOKINES AND PATIENT-REPORTED OUTCOMES IMPROVES DIAGNOSIS

You have accessJournal of UrologyCME1 Apr 2023MP39-01 CLASSIFICATION OF INTERSTITIAL CYSTITIS/BLADDER PAIN SYNDROME (IC/BPS) USING A MACHINE LEARNING BASED SYMPTOM SCORE CONSTRUCTED ON INFLAMMATORY URINE CYTOKINES AND PATIENT-REPORTED OUTCOMES IMPROVES DIAGNOSIS Laura Lamb, Joseph Janicki, Nitya Abraham, Melissa Laudano, Christopher Smith, Sarah Bartolone, Elijah Ward, Kenneth Peters, Bernadette Zwaans, Michael, and Chancellor Laura LambLaura Lamb More articles by this author , Joseph JanickiJoseph Janicki More articles by this author , Nitya AbrahamNitya Abraham More articles by this author , Melissa LaudanoMelissa Laudano More articles by this author , Christopher SmithChristopher Smith More articles by this author , Sarah BartoloneSarah Bartolone More articles by this author , Elijah WardElijah Ward More articles by this author , Kenneth PetersKenneth Peters More articles by this author , Bernadette ZwaansBernadette Zwaans More articles by this author , MichaelMichael More articles by this author , and Chancellor Chancellor More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003277.01AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: IC/BPS is a multifactorialsyndrome of severe pelvic and genitalia pain. Diagnosis of IC/BPS currentlyutilizes patient-reported outcomes (PRO) instruments to assess symptoms, howeveroverlapping symptoms with other diseases can delay proper diagnosis and symptomrelief. The objective of this study isto develop an improved IC/BPS risk classification, using machine learningalgorithm to leverage both urine inflammatory cytokines and PROs. METHODS: A national crowdsourcing resulted in 442 clinical urine samples consisting of 153 IC patients (146 female, 7 male), and 289 asymptomatic age-matched controls (155 female, 134 male) with corresponding PRO pain and symptom scores. This included 53 patients with a bladder-centric dysfunction as defined by presence of Hunner’s lesions. An independent validation cohort was also collected. Urinary cytokine levels were determined using Luminex assay. A predictive classification model, termed the Personalized Inflammation Symptom (PIS) Score, was generated from this data into three classification: 1) normal/low IC risk; 2) moderate to high risk for IC with bladder centric dysfunction; and 3) moderate to high risk of IC without bladder centric dysfunction. RESULTS: In the training set, the PIS Score correctly classified all controls, all IC patients without Hunner’s lesions, and 98% (52/53) IC patients with Hunner’s lesions. In the independent validation set, the PIS Score correctly classified 26 of 27 (96%), 14 of 19 (73.7%) IC without Hunner’s lesions, and all IC patient’s with Hunner’s lesions. CONCLUSIONS: In one of the largest collection of urine samples of IC/BPS patients from across all 50 states of US, two key findings emerged: 1) A standardized LUMINEX laboratory urine assay based IC-Risk Score is comparable to ICSI/ICPI or pain VAS in accuracy of diagnosis; 2) PIS, using laboratory-based objective data in combination with PROs, improves the AUC and accuracy of IC/BPS diagnosis. An objective laboratory test for IC/BPS, similar to tests that have been developed to improve cancer diagnosis, may help expand access, accelerate proper diagnosis, and improve care for IC/BPS, a disease with great unmet needs.Figure 1 Source of Funding: This work was supported by the Office of the Assistant Secretary ofDefense for Health Affairs through the Technology/Therapeutic DevelopmentResearch Program under Award No. W81XWH-19-1-0288. The opinions,interpretations, conclusions, and recommendations are those of the authors andare not necessarily endorsed by the Department of Defense. © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e534 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Laura Lamb More articles by this author Joseph Janicki More articles by this author Nitya Abraham More articles by this author Melissa Laudano More articles by this author Christopher Smith More articles by this author Sarah Bartolone More articles by this author Elijah Ward More articles by this author Kenneth Peters More articles by this author Bernadette Zwaans More articles by this author Michael More articles by this author Chancellor More articles by this author Expand All Advertisement PDF downloadLoading ...

Use of urinary cytokine and chemokine levels for identifying bladder conditions and predicting treatment outcomes in patients with interstitial cystitis/bladder pain syndrome

Use of urinary cytokine and chemokine levels for identifying bladder conditions and predicting treatment outcomes in patients with interstitial cystitis/bladder pain syndrome

Protein and functional isoform levels and genetic variants of the BAFF and APRIL pathway components in systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is the prototype of an autoimmune disease. Belimumab, a monoclonal antibody targets BAFF, is the only biologic approved for SLE and active lupus nephritis. BAFF is a cytokine with a key-regulatory role in the B cell homeostasis, which acts by binding to three receptors: BAFF-R, TACI and BCMA. TACI and BCMA also bind APRIL. Many studies reported elevated soluble BAFF and APRIL levels in the sera of SLE patients, but other questions about the role of this system in the disease remain open. The study aimed to investigate the utility of the cytokine levels in serum and urine as biomarkers, the role of non-functional isoforms, and the association of gene variants with the disease. This case–control study includes a cohort (women, 18–60 years old) of 100 patients (48% with nephritis) and 100 healthy controls. We used ELISA assays to measure the cytokine concentrations in serum (sBAFF and sAPRIL) and urine (uBAFF and uAPRIL); TaqMan Gene Expression Assays to quantify the relative mRNA expression of ΔBAFF, βAPRIL, and εAPRIL, and next-generation sequencing to genotype the cytokine (TNFSF13 and TNFSF13B) and receptor (TNFRSF13B, TNFRSF17 and TNFRSF13C) genes. The statistical tests used were: Kruskal–Wallis (qualitative variables), the Spearman Rho coefficient (correlations), the Chi-square and SKAT (association of common and rare genetic variants, respectively). As expected, sBAFF and sAPRIL levels were higher in patients than in controls (p ≤ 0.001) but found differences between patient subgroups. sBAFF and sAPRIL significantly correlated only in patients with nephritis (rs = 0.67, p ≤ 0.001) and βAPRIL levels were lower in patients with nephritis (p = 0.04), and ΔBAFF levels were lower in patients with dsDNA antibodies (p = 0.04). Rare variants of TNFSF13 and TNFRSF13B and TNFSF13 p.Gly67Arg and TNFRSF13B p.Val220Ala were associated with SLE. Our study supports differences among SLE patient subgroups with diverse clinical features in the BAFF/APRIL pathway. In addition, it suggests the involvement of genetic variants in the susceptibility to the disease.

Use of Urinary Cytokine and Chemokine Levels for Identifying Bladder Conditions and Predicting Treatment Outcomes in Patients with Interstitial Cystitis/Bladder Pain Syndrome.

Background: Interstitial cystitis/bladder pain syndrome (IC/BPS) is a condition causing bladder inflammation. Urinary biomarkers have been assessed as suitable for the diagnosis and treatment. This study aimed at investigating the role of urinary biomarkers in identifying bladder conditions and predicting the treatment outcome of IC/BPS. Methods: A total of 309 patients with IC/BPS and 30 controls were enrolled in this study. All patients underwent a comprehensive urological workup of symptoms, pain severity, and cystoscopic hydrodistention findings including maximal bladder capacity (MBC) and glomerulation grade. Urine samples were collected to investigate the levels of urinary cytokines and chemokines. According to MBC and glomerulation grade, patients with IC/BPS were further classified into the Hunner’s IC (HIC) and non-HIC groups. The urinary biomarkers between IC/BPS and control groups and HIC and non-HIC groups were compared. Moreover, the treatment response was graded according to global response assessment (GRA) scores, and urinary biomarker levels were analyzed based on different GRAs. Results: Patients with IC/BPS had significantly high urinary monocyte chemoattractant protein-1, eotaxin, tumor necrosis factor -alpha (TNF-α), and prostaglandin E2 levels. Significantly higher levels of urinary interleukin-8, C-X-C motif chemokine ligand 10 (CXCL 10), brain-derived neurotrophic factor, eotaxin, and regulated-on-activation, normal T-cell expressed and secreted (RANTES) were noted in HIC than those with non-HIC and controls. Among all biomarkers, TNF-α had the best sensitivity, specificity, positive predictive value, and negative predictive value. There was a significant correlation between biomarker levels and GRA. Conclusions: Significantly higher urine cytokines and chemokine levels were found in patients with IC/BPS. Most urinary biomarkers were significantly associated with MBC, glomerulation grade, and treatment outcome.

Utility of urinary cytokine levels as predictors of the immunogenicity and reactogenicity of AS01-adjuvanted hepatitis B vaccine in healthy adults

Plasma cytokines are useful indicators of the inflammatory response to vaccination, and can serve as potential biomarkers of the systemic reactogenicity and immunogenicity of vaccines. Measurement of cytokines in urine may represent a non-invasive alternative to the blood-based markers. To evaluate whether urinary cytokine levels can help predict vaccine responses to an AS01B-adjuvanted vaccine, we measured concentrations of 24 cytokines in the urine from 30 hepatitis B virus (HBV)-naïve adults following administration of AS01B-adjuvanted HBV surface antigen vaccine (NCT01777295). Levels post-dose 2 were compared with the levels measured following a single placebo (saline) injection, which was administered 1 month before the first vaccination in the same participants. Urine was collected at eight timepoints before or up to 1 week following each treatment. Urinary concentrations were normalized to creatinine levels, and paired with previously reported, participant-matched plasma levels, local and systemic reactogenicity scores, and antibody response magnitudes. Of the urine cytokine panel, only few analytes were detectable: IL-8, IL-18 and IL-6 receptor, each showing no clear changes after vaccination as compared to placebo administration, and MCP-1 (CCL2) and IP-10 (CXCL10), which displayed in most participants transient surges post-vaccination. Urine levels did not correlate with the matched plasma levels. Interestingly, urinary IP-10 levels at 1 day post-second vaccination were significantly correlated (P = 0.023) with the concurrent intensity scores of systemic reactogenicity, though not with the local reactogenicity scores or peak antibody responses. No significant correlations were detected for MCP-1. Altogether, most urinary cytokines have limited utility as a proxy for plasma cytokines to help predict the inflammatory response, the immunogenicity or the reactogenicity of AS01B-adjuvanted vaccine, with the possible exception of IP-10. The utility of urinary IP-10 as a potential complementary biomarker of systemic vaccine reactogenicity needs substantiation in larger studies.

NSC828779 Alleviates Renal Tubulointerstitial Lesions Involving Interleukin-36 Signaling in Mice.

Renal tubulointerstitial lesions (TILs), a common pathologic hallmark of chronic kidney disease that evolves to end-stage renal disease, is characterized by progressive inflammation and pronounced fibrosis of the kidney. However, current therapeutic approaches to treat these lesions remain largely ineffectual. Previously, we demonstrated that elevated IL-36α levels in human renal tissue and urine are implicated in impaired renal function, and IL-36 signaling enhances activation of NLRP3 inflammasome in a mouse model of TILs. Recently, we synthesized NSC828779, a salicylanilide derivative (protected by U.S. patents with US 8975255 B2 and US 9162993 B2), which inhibits activation of NF-κB signaling with high immunomodulatory potency and low IC50, and we hypothesized that it would be a potential drug candidate for renal TILs. The current study validated the therapeutic effects of NSC828779 on TILs using a mouse model of unilateral ureteral obstruction (UUO) and relevant cell models, including renal tubular epithelial cells under mechanically induced constant pressure. Treatment with NSC828779 improved renal lesions, as demonstrated by dramatically reduced severity of renal inflammation and fibrosis and decreased urinary cytokine levels in UUO mice. This small molecule specifically inhibits the IL-36α/NLRP3 inflammasome pathway. Based on these results, the beneficial outcome represents synergistic suppression of both the IL-36α-activated MAPK/NLRP3 inflammasome and STAT3- and Smad2/3-dependent fibrogenic signaling. NSC828779 appears justified as a new drug candidate to treat renal progressive inflammation and fibrosis.

Strong Associations Between Early Tubular Damage and Urinary Cytokine, Chemokine, and Growth Factor Levels in Elderly Males and Females.

Acute tubular necrosis is associated with high mortality rates and it is important to develop new biomarkers for tubular damage. The aim of this study was to investigate the effect of early tubular damage on a large number of urinary cytokines, chemokines, and growth factors. We selected 90 urine samples from the Prospective Investigation of the Vasculature in Uppsala Seniors Study (41 males and 49 females). The tubular damage markers cystatin C, neutrophil gelatinase-associated lipocalin (NGAL), and kidney injury molecule-1 (KIM-1) were analyzed in the urine samples and urinary cytokine levels were analyzed with 2 multiplex assays (proximity extension assay). After adjustment for sex, body mass index, estimated glomerular filtration rate, smoking, and multiplicity testing using the false discovery rate approach, there remained 26 cytokines that correlated significantly with urine cystatin C, 27 cytokines that correlated with NGAL, and 66 cytokines that correlated with KIM-1. Tubular damage shows a strong association with urinary cytokines, chemokines, and growth factors. Our findings indicate that multiplex proteomics could be a promising new approach to explore the complex effects of tubular damage.

Urinary cytokines correlate with acute kidney injury in critically ill COVID-19 patients

BackgroundAcute kidney injury is common in COVID-19 patients admitted to the ICU. Urinary biomarkers are a non-invasive way of assaying renal damage, and so far, urinary cytokines are not fully investigated. The current study aimed to assess urinary cytokine levels in COVID-19 patients. MethodsUrine was collected from COVID-19 patients (n = 29) in intensive care and compared to a preoperative group of patients (n = 9) with no critical illness. 92 urinary cytokines were analyzed in multiplex using the Olink Target 96 inflammation panel and compared to clinical characteristics, and urinary markers of kidney injury. ResultsThere were strong correlations between proinflammatory cytokines and between urinary cytokines and urinary kidney injury markers in 29 COVID-19 patients. Several cytokines were correlated to kidney injury, 31 cytokines to AKI stage and 19 cytokines correlated to maximal creatinine. ConclusionsUrinary inflammatory cytokines from a wide range of immune cell lineages were significantly upregulated during COVID-19 and the upregulation correlated with acute kidney injury as well as urinary markers of kidney tissue damage.

Adult mouse kidney stem cell‐derived exosomes attenuate blood pressure and improve kidney functions in aged mice

The physiological function of stem cell-derived extracellular vesicles remains elusive. Most recently, we isolated kidney stem cells (KSC) from adult mouse kidneys and studied the role of KSC-derived exosomes in blood pressure regulation in hypertensive aged mice. We treated aged mice with KSC-exosome (~1.26×1011 ± 1.94×109 particles/mouse/day) or PBS by intraperitoneal injection for 5 days. We found that the blood pressure was reduced (from 150.8±12.2 to 128.6±3 mmHg) at day 6 in KSC-exosome-treated aged mice, while the blood pressure remained unchanged in PBS-treated aged mice. Blood pressure continued to decrease to 116±8.2 mmHg at day 14 after KSC-exosome treatment. Hypertension reemerged gradually and relapsed at day 28 after treatment. To assess the kidney functions, we collected 6-hour urine at day 5 of the treatment. KSC-exosome treatment significantly reduced the increases in urine albumin and creatinine in aged mice, indicating that KSC-exosomes improve aging-associated kidney dysfunction. Notably, the glomerular filtration rate (GFR) was significantly increased in KSC-exosome-treated aged mice, suggesting improved glomerular function. Urinary cytokine levels were increased in aged mice which were largely reduced by KSC-exosomes. KSC-exosomes effectively reduced the kidney injury markers TIM1 and adiponectin and inflammation marker NGAL which were confirmed by ELISA. These data suggest that KSC-exosome treatment attenuates aging-associated renal inflammation and kidney injury. To explore the mechanisms by which KSC-exosome attenuates blood pressure and improves kidney functions in aged mice, we performed proteome analysis of KSC-exosomes by mass spectrometry. We found several pathways that involve in the kidney development were upregulated in KSC-exosome compared to renal tubule cells (RTC) derived-exosomes. Interestingly, KSC-exosome induced cell proliferation and cell migration after injury possibly through activation of AKT signaling demonstrated in vitro. In conclusion, our finding provided the first evidence that the adult mouse kidney stem cell-derived exosomes attenuate aging-associated hypertension and kidney dysfunctions. KSC-exosome treatment may be a novel therapeutic strategy for hypertension-associated kidney injuries in aging.

Microbiome Dysbiosis in Obese Adolescents: A Potential Contributor to Inflammation

Introduction Adolescent obesity remains a public health concern, as the CDC estimates that 1 in 5 adolescents is obese. The development of obesity during childhood not only increases the chance of obesity-related death, but also increases the risk of obesity-related chronic diseases such as type 2 diabetes mellitus, hypertension, and dyslipidemia. Increased inflammation is associated with adipose tissue in obesity and is a main contributor to the development of obesity-related disease. Dysbiosis of the gut microbiome may also contribute to obesity-related systemic inflammation. In this study, patients were educated on dietary habits and exercise in an effort to understand the effect of lifestyle changes on inflammatory markers in the urine. Furthermore, we investigated the gut microbiome profiles of obese adolescents to determine if differences existed from that of healthy adolescents. Methods The study was performed at the Department of Pediatrics at TTUHSC, Amarillo. IRB approval was obtained, and informed consent/assent were signed by the parents and adolescents. Twenty-two obese adolescents ages 8-18 (BMI >25) and 16 healthy adolescents (BMI <25) were recruited into the study. At the start of the study, patients were educated on dietary changes and exercise. Urine samples were collected at enrollment, and at 3- and 6-months post-enrollment visits. Urine samples were analyzed for the presence of pro-inflammatory cytokines TNF-a and IL-6, as well as the marker for endothelial dysfunction, ET-1. Stool samples were procured at the time of enrollment and 16s metagenome sequencing was performed to determine microbiome composition. Results Adolescents with obesity exhibited higher levels of pro-inflammatory TNF-α and IL-6 (3.4 and 1.6-fold increase, respectively) in urine compared to healthy controls. Furthermore, we noted a 1.4-fold increase in urinary ET-1 of obese adolescents. Interestingly, TNF-a, IL-6, and ET-1 levels in the urine decreased in obese adolescents over a period of 6 months, suggesting that changes in diet and exercise may have contributed to these changes. Analysis of the 16s metagenome indicated a difference in the presence of 80 bacterial species between the gut microbiomes of healthy and obese adolescents, as healthy adolescents had 63 bacterial species that exhibited at least a 2-fold higher amount than obese adolescents. Conclusions Obesity is a preventable condition that can result in systemic inflammation and chronic disease. In this study, we found that obese adolescents exhibited higher levels of urinary inflammatory cytokines and altered gut microbiome composition compared to healthy controls. Furthermore, we noted that lifestyle changes over 6 months resulted in decreased urinary inflammatory markers in obese adolescents. The results of this study suggest that the altered gut microbiomes of obese adolescents may be responsible for increased urinary inflammatory cytokine levels and that lifestyle changes may attenuate urinary inflammatory cytokines.

Albumin Urinary Excretion Is Associated with Increased Levels of Urinary Chemokines, Cytokines, and Growth Factors Levels in Humans.

The aim of the present study was to study the associations between urine albumin excretion, and a large number of urinary chemokines, cytokines, and growth factors in a normal population. We selected 90 urine samples from individuals without CVD, diabetes, stroke or kidney disease belonging to the Prospective Investigation of the Vasculature in Uppsala Seniors Study (41 males and 49 females, all aged 75 years). Urinary cytokine levels were analyzed with two multiplex assays (proximity extension assays) and the cytokine levels were correlated with urine albumin. After adjustment for sex, body mass index (BMI), estimated glomerular filtration rate (eGFR), smoking and multiplicity testing, 11 biomarkers remained significantly associated with urine albumin: thrombospondin 2, interleukin 6, interleukin 8, hepatocyte growth factor, matrix metalloproteinase-12 (MMP-12), C-X-C motif chemokine 9, tumor necrosis factor receptor superfamily member 11B, osteoprotegerin, growth-regulated alpha protein, C-X-C motif chemokine 6, oncostatin-M (OSM) and fatty acid-binding protein, intestinal, despite large differences in molecular weights. In this study, we found associations between urinary albumin and both small and large urine proteins. Additional studies are warranted to identify cytokine patterns and potential progression markers in various renal diseases.

Changes of Urinary Cytokines in Non-Diabetic Obese Patients After Laparoscopic Sleeve Gastrectomy.

IntroductionAdipose tissues synthesize and secrete various proinflammatory and anti-inflammatory mediators, termed cytokines. This work aims to assess different serum and urinary cytokine levels before and 12 months after laparoscopic sleeve gastrectomy (LSG).PatientsThis prospective study was performed on 61 obese non-diabetic patients who underwent LSG. All patients were followed up postoperatively at 12 months with the assessment of arterial blood pressure, microalbuminuria, urinary and serum levels of inflammatory cytokines (macrophage migration inhibitory factor “MIF,” monocyte chemotactic protein “MCP”-1, chemokine (C-C motif) ligand 15 (CCL-15), and CCL-18), in addition to serum creatinine, total cholesterol, and C-reactive protein (CRP).ResultsMean BMI showed decreased substantially from 44.78 ± 3.59 Kg/m2 to 34.56 ± 2.45. Systolic blood pressure decreased from 147.03 ± 16.89 mmHg to 128.82 ± 12.52 and diastolic blood pressure decreased from 90.51 ± 12.71 mmHg to 79.69 ± 10.39. At one-year of follow-up, there was statistically significant decrease of mean serum creatinine, total cholesterol, CRP, CCL-15, CCL-18, MIF/creatinine ratio, MCP-1/creatinine ratio, CCL-15/creatinine ratio, and CCL-18/creatinine ratio (P value <0.001).ConclusionImprovement of systemic and renal inflammatory states after LSG may positively affect obesity-related renal disease by steering the adipokine levels towards anti-inflammatory profiles.

Elevated urinary IL-36γ in patients with active lupus nephritis and response to treatment

IL-36 is a new member of the IL-1 family with pro-inflammatory properties. Serum levels of IL-36 are elevated in patients with Systemic Lupus Erythematosus (SLE). However, no data is available on urinary levels of IL-36 in Lupus Nephritis (LN). In psoriasis expression of IL-36 is site specific and expressed in skin. Hence, we studied urinary levels of IL-36 cytokines in SLE patients. A total of 196 patients with SLE [97 active LN patients (ALN), 42 inactive LN (ILN) and 57 active lupus patients with no renal involvement (ANR)] and 25 healthy subjects were recruited for the study after obtaining informed consent. Urinary and plasma IL-36α, IL-36γ and IL-36Ra levels were measured by ELISA. Out of 196 patients 178 were females. Urinary IL-36γ levels in SLE patients [0(14.3) pg/ml] were significantly higher than healthy controls [0(0) pg/ml, (P < 0.01)]. Patients with ALN [0(40.6) pg/ml] had significantly higher IL-36γ when compared to ANR [0(0) pg/ml] as well as ILN [0(0) pg/ml]. Urinary IL-36γ levels in ALN patients had a fair correlation with renal SLEDAI (r = 0.26, P = 0.004).The levels reduced significantly post 3 months in patients with ALN. No inverse relationship was noted between IL-36Ra and IL-36α/IL36γ levels. Urinary IL-36γ is produced locally in kidney, correlates with renal disease activity and reduces upon treatment, suggesting that it may have a role in pathogenesis of LN.