sTNFR1 Levels Research Articles

Exploration of CCL11 and sTNFR2 as potential biomarkers for the efficacy of lymphocyte immunotherapy in women with unexplained recurrent spontaneous abortion

Objective: To explore biomarkers for the efficacy of lymphocyte immunotherapy (LIT) treating women with unexplained recurrent spontaneous abortion (URSA). Methods: Serum samples from 24 URSA potients who received LIT were collected at Peking University Third Hospital from December 2014 to June 2015. Semiquantitative sandwich-based antibody arrays containing 40 cytokines were used to screen target immune cytokines in the peripheral blood of URSA patients before and after LIT. Multifactor quantitative microsphere flow cytometry detection validated the levels of target cytokines. Based on the final pregnancy outcome after LIT, 24 URSA patients were divided into the full-term delivery group (15 cases) and the abortion group (9 cases). Furthermore, linear regression analysis were applied to evaluate the relationship between target cytokines and pregnancy outcomes. Results: Semiquantitative sandwich-based antibody arrays suggested that, among all 24 URSA patients included in this study, the intensities of the fluorescence signal were significantly lower post-LIT versus pre-LIT for the following cytokines: interleukin-15 (IL-15), monokine induced by γ-interferon (MIG), C-C motif chemokine ligand (CCL) 1 (all P<0.05). In the full-term delivery group, the intensities of the fluorescence signal post-LIT were significantly lower than pre-LIT for the following cytokines: IL-15, CCL1, macrophage inflammatory protein (MIP) 1α (all P<0.05). In the abortion group, the intensities of the fluorescence signal post-LIT were significantly lower than pre-LIT for the following cytokines: MIG, MIP-1δ (all P<0.05). Linear regression analysis showed that the intensity of the fluorescence signal of CCL11 was increased and the intensity of the fluorescence signal of soluble tumor necrosis factor receptor 2 (sTNFR2) was decreased in the full-term delivery group after LIT, the differences were statistically significant (P=0.012, 0.029). Validation results of multifactor quantitative microsphere flow cytometry detection showed that the level of CCL11 was significantly increased (P=0.001) and the level of sTNFR2 was significantly decreased (P=0.001) in the full-term delivery group after LIT. Conclusion: CCL11 and sTNFR2 maybe serve as potential biomarkers that could predict pregnancy outcomes after LIT in women with URSA.

Role of plasma suPAR, sTNFR1, and sTNFR2 levels in risk stratification and outcome prediction of complicated acute kidney injury in elderly patients with coronavirus disease 2019

ObjectiveThe aim of this study is to investigate the early prognostic efficacy of plasma soluble urokinase-type plasminogen activator receptor (suPAR), soluble tumor necrosis factor receptor 1 (sTNFR1), and soluble tumor necrosis factor receptor 2 (sTNFR2) in complicated acute kidney injury (AKI) in patients with coronavirus disease 2019 (COVID-19), and to analyze the relevant factors contributing to complicated AKI in these patients. MethodsPatients with COVID-19 hospitalized at the Affiliated Baiyun Hospital of Guizhou Medical University from March 2022 to March 2024 were selected as study participants. A total of 589 patients met the inclusion and exclusion criteria, 68 patients complicated with AKI were classified as AKI group, and the remaining 521 cases were divided into proportion sampling method and randomly selected 200 samples, which were classified as non-AKI group. Additionally, 50 healthy controls were enrolled as the control group. Logistic regression analysis was conducted to identify the relevant factors associated with complicated AKI in patients with COVID-19. Receiver operating characteristic (ROC) curves were plotted to evaluate the prognostic efficacy of plasma suPAR, sTNFR1, and sTNFR2 indicators for complicated AKI in patients with COVID-19. ResultsAmong the patients with COVID-19 in the AKI group, 43 were males (63.20 %), with a median age of 79.00 (interquartile range: 75.00, 83.00) years, while the non-AKI group comprised 83 males (41.50 %), with a median age of 73.00 (interquartile range: 60.00, 80.75) years. Comparison of the sex and age between the two groups indicated that males and elderly patients had increased risks of complicated AKI (P < 0.05). Plasma levels of suPAR, sTNFR1, and sTNFR2 in the AKI group were significantly higher than those in the non-AKI group (P < 0.05). Logistic regression analysis indicated that suPAR and sTNFR2 were independent factors influencing complicated AKI in patients with COVID-19 (P < 0.05). The ROC curve for a single indicator showed that suPAR had the highest prognostic efficacy for complicated AKI, with an area under the curve (AUC) of 0.813, a sensitivity of 79.4 %, and a specificity of 74.0 %. The combined use of suPAR and sTNFR2 for risk assessment yielded the highest AUC of 0.838, with a sensitivity of 66.2 % and a specificity of 87.5 %. The combined risk assessment using all three indicators (suPAR, sTNFR1, and sTNFR2) had an AUC of 0.837, with a sensitivity of 64.7 % and a specificity of 89.0 %. ConclusionElderly patients had increased risks of complicated AKI. Indicators such as suPAR, sTNFR1, and sTNFR2 can assist in assessing the risk in patients with COVID-19 complicated AKI, with suPAR demonstrating the highest prognostic efficacy as a single indicator. The combined detection of suPAR, sTNFR1, and sTNFR2 offers greater prognostic value than using any single indicator.

Associations of HMGB1, sTNFR-1 and NLR with Premature Delivery Secondary to Infection in Pregnant Women Undergoing Cervical Cerclage

Background: Although cervical cerclage has improved with the rapid development of medical technology, there remains a distinct probability of adverse pregnancy outcomes. To investigate the associations of changes in the high mobility group box 1 (HMGB1), soluble tumor necrosis factor receptor 1 (sTNFR-1) and peripheral blood neutrophil-to-lymphocyte ratio (NLR) with premature delivery secondary to infection in pregnant patients undergoing cervical cerclage. Methods: Sixty-seven pregnant patients with premature delivery after cervical cerclage, who were treated at the Affiliated Matern &amp; Child Care Hospital of Nantong University from January 2022 to October 2023, were enrolled, including 43 with premature delivery secondary to infection (infectious group) and 24 with idiopathic premature delivery (non-infectious group). The pre-delivery serum levels of HMGB1, sTNFR-1 and the peripheral blood level of NLR were compared between the 2 groups. Further, the clinical value of these 3 indicators in predicting premature delivery secondary to infection among pregnant patients undergoing cervical cerclage was assessed by receiver operating characteristic (ROC) curve analysis. Results: The infectious group exhibited significantly higher serum levels of HMGB1, sTNFR-1 and peripheral blood level of NLR compared to those in the non-infectious group, demonstrating significant differences (p &lt; 0.05). Logistic regression analysis revealed that HMGB1 and NLR were independent influencing factors for premature delivery (p &lt; 0.05). According to the ROC curve analysis results, the changes in HMGB1, sTNFR-1 and NLR levels may reflect the risk of premature delivery secondary to infection among pregnant patients undergoing cervical cerclage. The area under the curve (AUC), sensitivity and specificity of combined detection were all markedly higher than those of independent detection. Conclusions: HMGB1, sTNFR-1 and NLR levels are risk factors for third-trimester premature delivery among pregnant patients undergoing cervical cerclage. Timely combined detection of serum HMGB1, sTNFR-1 and peripheral blood NLR during the third trimester can improve the clinical diagnostic acumen, which enables early prevention to help lower the risk of premature delivery.

Plasma levels of sCASP3 predicts carotid intima media thickness progression

Abstract Background Studies on human atherosclerotic plaques have shown that apoptosis is associated with rupture prone plaques and that apoptosis is a key process for fast plaque progression. In line with this, we showed that circulating levels of soluble Caspase-3 (sCaspase-3) predict atherosclerotic events. However, the relationship between circulating levels of apoptosis markers and plaque progression remains to be explored. Purpose Here we aimed to investigate if circulating levels of apoptosis markers could predict mean intima-media thickness in the common carotid artery (IMT-CCA) progression among individuals with atherosclerotic carotid plaques. Methods The Malmö Diet and Cancer (MDC) cardiovascular cohort was used. MDC is a prospective population-based cohort study. Baseline variables (blood, clinical information and carotid ultrasound measurements) were collected between 1991-1994 and the right carotid ultrasound re-examinations were performed between 1994-2001. IMT was measured at by ultrasound at baseline as well as at the re-examinations. The Olink Proximity Extension Assay (SciLifeLab, Uppsala, Sweden) was used to analyze plasma levels of sCaspase-3, sFADD, sCaspase-8, sTRAIL-R2 and sTNFR1 at baseline. Correlation and multiple linear regression analyses, adjusted for age, sex, and cardiovascular risk factors, were employed to evaluate the relationships between apoptosis biomarkers and IMT-CCA as well as average annual change in IMT-CCA. Results In total, 1929 participants (median age 60 years) with an ultrasound confirmed carotid plaque (focal IMT &amp;gt;1.2mm) at baseline were included. Subjects who underwent ultrasound IMT re-examination over a period of &amp;gt;4.5 years were divided into a discovery (n=406) and a validation (n=355) cohort. sCaspase-3 was identified as the marker with the strongest correlation to IMT-CCA at baseline (p=0.02) and upon re-examination (p=0.04), among the five apoptosis markers examined, in both the discovery and validation cohorts. sCaspase-3 also showed a correlation with the average annual change of IMT-CCA (r=0.12, p=8×10-4) in both cohorts. Furthermore, when diving the cohort into tertiles based on plasma levels of sCapase-3, subjects with high levels of sCaspase-3 (3rd tertile) had greater annual changes in IMT-CCA compared to the 1st tertile (p=0.03). Multiple linear regression analyses confirmed that sCapase-3 was associated with the average annual IMT-CCA change in both the discovery (beta=0.14, 95% confidence interval (CI) 0.04–0.24) cohort and the validation cohort (beta=0.11, 95%CI 0.01–0.21). Conclusion Using a large, population-based study with repeated IMT measurements, we demonstrated that circulating levels of sCapase-3 were associated with increasing IMT in the CCA. While further studies are warranted, these results suggest that sCapase-3 levels could serve as potential screening marker to identify individuals at higher risk for plaque progression.

Biomarkers of microbial translocation and generalized inflammation are associated with frailty among people with HIV.

Frailty occurs at higher rates and younger ages among people with HIV (PWH) compared to the general population and is often attributed to chronic inflammation and subsequent immune exhaustion. We assessed how inflammatory biomarkers are associated with frailty among PWH. The Centers for AIDS Research Network of Integrated Clinical Systems (CNICS) cohort is comprised of adult PWH in care at 10 sites, and harmonizes demographic, clinical, and patient-reported outcomes (PRO) data. A panel of 13 inflammatory biomarkers was collected from a subset of virally suppressed PWH once per person between 2010-2018. Frailty was measured with a validated PRO phenotype, scored 0-4, from biomarker collection date through July 2022. With adjusted linear mixed models, we estimated longitudinal associations between standard deviation-scaled log2-transformed biomarkers and frailty score. Among 273 PWH, most were male (91%), average age at baseline was 45, 42% were non-Hispanic White while 35% were non-Hispanic Black, and average follow-up time was 5.5 years. Several biomarkers were associated with higher frailty, including those linked to microbial translocation (sCD14, LBP, KT ratio) and systemic inflammation (CRP, IL-6, suPAR, sTNFR1, sTNFR2). Higher IL-6 was associated with a 0.25-point higher frailty score (95%CI:0.12-0.39). Higher sTNFR1 (0.35 [0.13-0.56]), sCD14 (0.21 [0.11-0.31]), and suPAR (0.24 [0.11-0.36]) levels were also associated with higher frailty scores over follow-up. Higher levels of biomarkers linked to microbial translocation and systemic inflammation are associated with higher average frailty scores over time in a cohort of virally suppressed PWH, highlighting these pathways as potential interventional targets for mitigating frailty in PWH.

Regulation of plasma soluble receptors of TNF and IL-1 in patients with COVID-19 differs from that observed in sepsis

IL-1α/β and TNF are closely linked to the pathology of severe COVID-19 and sepsis. The soluble forms of their receptors, functioning as decoy receptors, exhibit inhibitory effects. However, little is known about their regulation in severe bacterial and viral infections, which we aimed to investigate in this study. The circulating soluble receptors of TNF (sTNFR1 and sTNFR2) and IL-1α/β (sIL-1R1, sIL-1R2) were evaluated in the plasma of patients with COVID-19, severe bacterial infections, and sepsis and compared with healthy controls. Additionally, IL1R1, IL1R2, TNFRSF1A, and TNFRSF1B expression was evaluated at the single cell level in PBMCs derived from COVID-19 or sepsis patients. Plasma concentrations of sIL-1R1, sTNFR1, and sTNFR2 were significantly higher in COVID-19 patients compared to healthy subjects. Notably, sIL-1R1 levels were particularly elevated in ICU COVID-19 patients, and transcriptome analysis indicated heightened IL1R1 expression in PBMCs from severe COVID-19 patients. In severe bacterial infections, only sTNFR1 and sTNFR2 exhibited increased levels compared to healthy controls. Sepsis patients had decreased sIL-1R1 plasma concentrations but elevated sIL-1R2, sTNFR1, and sTNFR2 levels compared to healthy individuals, reflecting the heightened expression due to the increased numbers of monocytes present in sepsis. Finally, elevated concentrations of sIL-1R2, sTNFR1, and sTNFR2 were moderately associated with reduced 28-day survival in sepsis patients. Our study reveals distinct regulation of plasma concentrations of soluble IL-1 receptors in COVID-19 and sepsis. Moreover, soluble TNF receptors 1 and 2 consistently rise in all conditions and show a positive correlation with disease severity in sepsis.

A study of the relationship between circulating cytokines (interleukin-2 receptor and tumor necrosis factor receptor 2) and risk of B-cell non-hodgkin lymphoma.

Mature B-cell non-Hodgkin lymphoma (B-NHL) occurs due to uncontrolled B-lymphocyte clonal expansion. Cytokines can directly stimulate B-cell proliferation and prevent B-cell apoptosis. Dysregulation of cytokines may play an important role in the development of B-NHL by enhancing chromosomal translocation, which is the hallmark of B-NHL. Both interleukin 2 and tumor necrosis factor-α are proinflammatory cytokines and play important roles in the growth, differentiation, and apoptosis of B cells.We conducted a prospective case-control study applied to 50 patients with B-NHL at Kasr Al Aini Hospital, Cairo University, and 50 age- and sex-matched controls. Clinical, laboratory and imaging data were collected. In all patients and controls, sIL-2R and sTNF-R2 levels were measured by enzyme-linked immunosorbent assay (ELISA). The Spearman correlation test was used to evaluate the correlation between the studied cytokines and clinical, laboratory and imaging findings. Sensitivity analysis was conducted to detect the cutoff values of the studied cytokines.Serum levels of sIL-2R and sTNF-R2 were significantly higher in patients than in controls. Additionally, their levels were significantly higher in aggressive types and advanced stages of lymphoma. Also, the studied cytokines were significantly correlated with different clinical and laboratory parameters of lymphoma. The level of sIL-2R and sTNF-R2 were closely related to the type of lymphoma (P value ˂ 0.001 and 0.012, respectively), further it was also associated with the natural history of lymphoma (aggressive vs. indolent) (P value ˂0.001 and 0.04 respectively).We concluded that Pretreatment levels of sIL-2R and sTNF-R2 may play a role in the natural history and prognosis of lymphoma. They may be used as a prognostic factor for B-NHL patients and may also help with treatment decisions.

Immunoregulatory Cells and Cytokines Discriminate Disease Activity Score 28-Remission Statuses and Ultrasound Grades in Rheumatoid Arthritis Patients with Non-High Disease Activity.

It is not clear whether immunoregulatory cytokines and cells are associated with Disease Activity Score 28 (DAS28) scores and ultrasound grades/scores. Here, we investigated the relationships between immunoregulatory cytokines or cells and different DAS28 scores or ultrasound grades/scores in patients with rheumatoid arthritis (RA). This study enrolled 50 RA patients (with 147 visits) who had remission/low/moderate DAS28-ESR scores (92% in remission and low disease activity) at baseline. Blood was collected and an ultrasound was performed three times in a year. Percentages of regulatory B cells and T regulatory type 1 cells and M2 macrophage numbers in the blood were examined. Plasma levels of 10 immunoregulatory cytokines IL-4, IL-5, IL-9, IL-10, IL-13, IL-27, IL-35, TGF-β1, sTNF-R1, and sTNF-R2 and monocyte chemotactic protein-1 (MCP-1) were assessed using ELISA assay. The correlations of cytokines and cells with different DAS28 scores and ultrasound grades were investigated, and cytokines and cells were compared between different categories of DAS28 scores and ultrasound grades. Plasma TGF-β1 levels were higher in the DAS28-ESR < 2.6 (remission) subgroup than in the DAS28-ESR ≥ 2.6 (nonremission) subgroup (p = 0.037). However, plasma TGF-β1 levels were higher in the high ultrasound grade subgroup than those in the low ultrasound grade subgroup (p = 0.007). The number of M2 macrophages was lower in the DAS28-MCP-1 < 2.2 subgroup than in the DAS28-MCP-1 ≥ 2.2 subgroup (p = 0.036). The levels of TGF-β1, sTNF-R2, IL-10, and IL-27 were higher in patients with high ultrasound grades than in those with low ultrasound grades. IL-27 was also higher in the nonremission DAS28-ESR subgroup than the remission one (p = 0.025). Moreover, sTNF-R1 levels in the 2011 American College of Rheumatology/European League Against Rheumatism (ACR/EULAR) remission subgroup were significantly lower than in the 2011 ACR/EULAR nonremission subgroup (p = 0.007). This trend was reflected in that lower sTNF-R1 levels correlated with low DAS28-MCP-1 scores (rho = 0.222, p = 0.007). We conclude that high plasma TGF-β1 levels indicate the DAS28-ESR remission (<2.6) subgroup and the high ultrasound grade subgroup. IL-27 probably connects the nonremission DAS28-ESR to high ultrasound grades. Low sTNF-R1 levels probably link low DAS28-MCP-1 scores with the 2011 ACR/EULAR remission subgroup. It suggests that incongruent immuno-inflammatory abnormalities exist between DAS28 scores and ultrasound grades, and are also dissimilar among various DAS28-formula categories. Therefore, this study may provide a basis for further research into individual cytokines and immunoregulatory cells behind each DAS28 formula and ultrasound grades/scores.

Serum DNase1, sTNFR1 and sTNFR2 as Risk Factors for Lupus Nephritis in Systemic Lupus Erythematosus Patients

BACKGROUND: Early detection and management of lupus nephritis (LN) in systemic lupus erythematosus (SLE) are essential to prevent irreversible kidney damage and improve patient outcomes; therefore, identifying reliable biomarkers to predict LN is paramount. However, there are still relatively few studies examining the potential biomarkers for LN in SLE patients. This study was conducted to investigate serum deoxyribonuclease I (DNase), soluble tumor necrosing factor 1 (sTNFR1) and soluble tumor necrosing factor 2 (sTNFR2) as a risk factor for LN in SLE patients.METHODS: A case-control study involving SLE patients aged 20-60 years was conducted. Blood was withdrawn from each subject for the measurement of serum level of DNase1, sTNFR1, and sTNFR2 that was performed using enzyme-linked immunoassay (ELISA) methods. Data was then analyzed using Chi-Square test and logistic regression tests.RESULTS: A total of 22 patients with LN and 22 without LN were included. The cut-off value for DNase1, sTNFR1, and sTNFR2 were 5.05 ng/mL, 6.52 ng/mL, and 7.02 ng/mL, respectively. The risk factors of LN in SLE patients were the low level of serum DNase1 (aOR=6.64; 95%CI: 1.25-35.29; p=0.026), low level of serum sTNFR1 (aOR=8.12; 95% CI: 1.56-42.10; p=0.013), and low level of serum sTNFR2 (aOR=5.57; 95%CI: 1.03-30.11; p=0.046).CONCLUSION: Serum DNase1 lower than 5.05 ng/mL, sTNFR1 lower than 6.52 ng/mL, and sTNFR2 lower than 7.02 ng/mL were risk factors for lupus nephritis in SLE patients. Hence, serum DNase1, sTNFR1 and sTNFR2 could be used as risk factors predictors for LN in SLE patients.KEYWORDS: DNase1, sTNFR1, sTNFR2, SLE, lupus nephritis

Soluble tumor necrosis factor receptor type 1 is an alternative marker of urinary albumin-creatinine ratio and estimated glomerular filtration rate for predicting the decline of renal function in subjects with type 2 diabetes mellitus

BackgroundUrinary albumin-creatinine ratio (UACR) and estimated glomerular filtration rates (eGFR) help predict worsening diabetic kidney disease (DKD) but have their limitations. Soluble tumor necrosis factor receptor type 1 (sTNFR1) is a biomarker of DKD. The predictive abilities of sTNFR1 and UACR plus eGFR have not been compared. MethodsThis prospective cohort study included patients with type 2 diabetes (T2D) to identify the risk factors of worsening DKD. Renal events were defined as > 30 % loss in eGFR based on consecutive tests after 6 months. The associations of sTNFR1, UACR, and eGFR levels and the risks of renal events were tested using a Cox regression model and the area under the curve (AUC) was compared between sTNFR1 levels and UACR plus eGFR using receiver-operating characteristic (ROC) analysis. The accuracy of stratification was evaluated using Kaplan–Meier analysis. ResultsLevels of sTNFR1 and UACR were associated with risks of > 30 % decline in eGFR after adjusting for relevant factors. The association between sTNFR1 levels and renal outcomes was independent of UACR and eGFR at baseline. The AUC of sTNFR1 level was comparable with that of combined UACR and eGFR (0.73 vs. 0.71, respectively, p = 0.72) and the results persisted for quartile groups of sTNFR1 and risk categories of Kidney Disease: Improving Global Outcomes (KDIGO) (0.70 vs. 0.71, respectively, p = 0.84). Both stratifications by sTNFR1 levels and KDIGO were accurate. ConclusionsTNFR1 could be an alternative marker for identifying patients with diabetes at risk of declining renal function.

Abstract 3435: History of breastfeeding in relation to circulating inflammatory biomarkers

Abstract Background: History of breastfeeding has been associated with reduced risk of various chronic diseases in which chronic inflammation is involved in disease etiology, including cardiovascular disease, type II diabetes, breast cancer, and ovarian cancer. However, few studies have investigated the long-term impact of breastfeeding on systemic inflammation. Thus, we aimed to examine the associations between breastfeeding history and circulating inflammatory biomarkers at midlife. Methods: We conducted a cross-sectional analysis including 3,448 parous women with plasma inflammatory biomarkers in the Nurses’ Health Study and Nurses’ Health Study II. We examined self-reported history of ever breastfeeding (ever vs. never) and lifetime total duration of breastfeeding (never, &amp;lt; 6 months, 7-11 months, 12+ months) prior to the time of blood draw. We calculated the geometric means of eight inflammatory biomarkers [i.e., high sensitivity C-reactive protein (hsCRP), interleukin-6 (IL6], sIL6Rα, IL8, IL10, sIL2-receptor-α(Rα), soluble tumor necrosis factor α receptor 2 (sTNFR2), B-cell activating factor (BAFF), C-X-C motif chemokine ligand 13 (CXCL13)] by self-reported history of breastfeeding status adjusted for age, body mass index (BMI), smoking status, duration of oral contraceptive use, parity, proinflammatory diet, analgesic use, menopausal status, and cohort. Tests for trend were calculated using the median value of each lifetime breastfeeding duration category and modeling as a continuous variable. We also conducted stratified analyses by menopausal status and BMI (&amp;lt; 25kg/m2 vs. ≥ 25kg/m2) at blood draw. Results: Overall, we did not observe statistically significant differences in circulating inflammatory biomarker levels by history of breastfeeding among parous women. When stratifying by menopausal status at blood draw, total breastfeeding duration was inversely associated with sTNFR2 levels among premenopausal women (P-trend = 0.03) but not among postmenopausal women. Plasma IL8 levels were suggestively inversely associated with breastfeeding duration among postmenopausal women (P-trend = 0.07) but not among premenopausal women. Among women with BMI ≥ 25 kg/m2, plasma BAFF levels were lower in parous women who ever breastfed compared to those who never breastfed [1382.5 (95%CI = 1242.0 - 1539.0) pg/ml vs. 1611.9 (95%CI = 1424.9 - 1823.4) pg/ml; p = 0.03], and suggestively inversely associated with total breastfeeding duration (p-trend = 0.07) but no statistically significant association was observed in women with BMI &amp;lt; 25kg/m2. IL10 was inversely associated with total breastfeeding duration among women with BMI ≥25 kg/m2 (P-trend = 0.007), but not in women with BMI &amp;lt; 25kg/m2. Conclusion: Overall, history of breastfeeding was not associated with inflammatory biomarkers at mid-life, however the association may vary by menopausal status and BMI. Citation Format: Nan Lin, Jennifer M. Mongiovi, Leslie V. Farland, Tianyi Huang, Heather Eliassen, Mary K. Townsend, Shelley S. Tworoger, Kathryn L. Terry, Naoko Sasamoto. History of breastfeeding in relation to circulating inflammatory biomarkers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3435.

Correlation of Increased Soluble Tumor Necrosis Factor Receptor 1 with Mortality and Dependence on Treatment in Non-Small-Cell Lung Cancer Patients: A Longitudinal Cohort Study

Background: Tumor necrosis factor (TNF) is a multipotent cytokine involved in inflammation and anti-tumor activity. TNF-α exerts its function upon binding to TNF-receptor 1 (TNF-R1) and TNF-receptor 2 (TNF-R2). This study investigates the relationship of soluble (s) TNF-R1 levels in non-small-cell lung cancer (NSCLC) patients with treatment and overall survival. Methods: In total, 134 NSCLC patients treated at the Medical Faculty of Martin Luther University Halle-Wittenberg between 2017 and 2019 were included in this study. Serum levels of sTNF-R1 were measured via ELISA at baseline and during and after treatment. A linear mixed-effects model was used to assess sTNF-R1 changes over time. Linear regression was applied to investigate the association between clinical characteristics and changes in sTNF-R1. Cox regression models were used to estimate associations with overall mortality. Results: The estimated average sTNFR-1 at baseline was 2091.71 pg/mL, with a change of 6.19 pg/mL per day. Cox models revealed that the individual change in sTNF-R1 was more strongly associated with mortality than its baseline value, especially after adjusting for covariates. Conclusions: This study provides evidence that the individual change in sTNF-R1 levels during and after treatment were associated with the risk of mortality, suggesting the use of the sTNF-R1 trajectory as a prognostic marker.

Genetically engineered macrophages derived from iPSCs for self-regulating delivery of anti-inflammatory biologic drugs.

In rheumatoid arthritis, dysregulated cytokine signaling has been implicated as a primary factor in chronic inflammation. Many antirheumatic and biological therapies are used to suppress joint inflammation, but despite these advances, effectiveness is not universal, and delivery is often at high doses, which can predispose patients to significant off-target effects. During chronic inflammation, the inappropriate regulation of signaling factors by macrophages accelerates progression of disease by driving an imbalance of inflammatory cytokines, making macrophages an ideal cellular target. To develop a macrophage-based therapy to treat chronic inflammation, we engineered a novel induced pluripotent stem cell (iPSC)-derived macrophage capable of delivering soluble TNF receptor 1 (TNFR1), an anti-inflammatory biologic inhibitor of tumor necrosis factor alpha (TNF-α), in an auto-regulated manner in response to TNF-α. Murine iPSCs were differentiated into macrophages (iMACs) over a 17-day optimized protocol with continued successful differentiation confirmed at key timepoints. Varying inflammatory and immunomodulatory stimuli demonstrated traditional macrophage function and phenotypes. In response to TNF-α, therapeutic iMACs produced high levels of sTNFR1 in an autoregulated manner, which inhibited inflammatory signaling. This self-regulating iMAC system demonstrated the potential for macrophage-based drug delivery as a novel therapeutic approach for a variety of chronic inflammatory diseases.

Effect of whole-body vibration stimulation on plasma soluble TNF receptors in elderly with sarcopenia: a randomized controlled trial.

Sarcopenia is a pathology resulting from a progressive and severe loss of muscle mass, strength, and function in the course of aging, which has deleterious consequences on quality of life. Among the most widespread studies on the issue are those focused on the effect of different types of physical exercise on patients with sarcopenia. This randomized controlled study aimed to compare the effects of a whole-body vibration exercise (WBV) session on the inflammatory parameters of non-sarcopenic (NSG, n=22) and sarcopenic elderly (SG, n=22). NSG and SG participants were randomly divided into two protocols: intervention (squat with WBV) and control (squat without WBV). After a one-week washout period, participants switched protocols, so that everyone performed both protocols. Body composition was assessed by dual-energy radiological absorptiometry (DXA) and function through the six-minute walk test (6MWD) and Short Physical Performance Battery (SPPB). Plasma soluble tumor necrosis factor receptors (sTNFR) were determined by enzyme-linked immunosorbent assay (ELISA) and measured before and immediately after each protocol. After exercise with WBV, there was an increase in sTNFR2 levels in the NSG (P<0.01; d=-0.69 (-1.30; -0.08) and SG (P<0.01, d=-0.95 (-1.57; -0.32) groups. In conclusion, an acute session of WBV influenced sTNFr2 levels, with sarcopenic individuals showing a greater effect. This suggested that WBV had a more pronounced impact on sTNFr2 in those with loss of muscle strength and/or physical performance. Additionally, WBV is gaining recognition as an efficient strategy for those with persistent health issues.

Endostatin, soluble tumour necrosis factor receptor 1 and soluble tumour necrosis factor receptor 2 cannot predict new onset of microalbuminuria in patients with type 2 diabetes.

Inflammation and angiogenesis play an important role in the development of early diabetic kidney disease. We investigated the association of soluble Tumour Necrosis Factor Receptor 1 (sTNF-R1), sTNF-R2 and endostatin with new onset microalbuminuria in normoalbuminuric patients with diabetes mellitus type 2. We conducted a case control study to assess serum levels of sTNF-R1, sTNF-R2 and endostatin in 169 patients with new onset microalbuminuria and in 188 matched normoalbuminuric, diabetic controls. Baseline serum samples from participants of the ROADMAP (Randomized Olmesartan and Diabetes Microalbuminuria Prevention) and observational follow-up (ROADMAP-OFU) studies were used. Endostatin and sTNF-R1 but not sTNF-R2 were increased at baseline in patients with future microalbuminuria. In the multivariate analysis, each log2 increment in endostatin levels was associated with an increase of only 6% in the risk of development of microalbuminuria (adjusted HR (95% CI) 1.006 (1.001-1011). sTNF-R1 and sTNF-R2 levels were conversely associated with microalbuminuria, but the results did not reach statistical significance. The respective adjusted HRs (95% CI) were 1.305 (0.928-1.774) and 0.874 (0.711-1.074). sTNF-R1 and sTNF-R2 failed to predict the occurrence of microalbuminuria in normoalbuminuric patients with type 2 diabetes. Likewise, the utility of endostatin in predicting new onset proteinuria is limited.

Ex vivo lung perfusion moderates gene expression differences between cardiac death and brain death donor lungs

Donation after cardiac death (DCD) donor lungs have been shown to express less pro-inflammatory genes than donation after brain death (DBD) lungs, likely due to the absence of brain-death related inflammatory physiology. However, it is unclear whether this difference is clinically significant following reperfusion. To avoid confounding by the recipient immune system and activation state, we utilized ex vivo lung perfusion (EVLP) as a reperfusion-like event and examined the effect of EVLP on the transcriptome of DCD (n=39) and DBD (n=49) lungs. To validate our RNA results, banked EVLP perfusates from a separate cohort of DCD (n=24) and DBD (n=24) cases were assayed for IL-6, IL-8, IL-10, IL-1β, sTNFR1, and sTREM1 protein levels at 15 min intervals for three hours. While DCD lungs demonstrated lower levels of pro-inflammatory transcripts and perfusate cytokine protein levels than DBD lungs prior to EVLP, after EVLP there were no significant gene expression differences or cytokine protein levels between groups. Therefore, while DCD and DBD lungs differ by the amounts of pro-inflammatory cytokines following procurement, the propagation of inflammation becomes limited during EVLP, and DBD and DCD lungs reach a similar plateau of transcript expression, including pro-inflammatory cytokines at the end of perfusion. EVLP may therefore play a pre-conditioning role by dampening the pro-inflammatory state prior to transplant reperfusion.

A comparison of serum inflammatory parameters in progressive forms of multiple sclerosis

IntroductionMultiple sclerosis (MS) is a chronic, inflammatory demyelinating disease of the central nervous system. Primary progressive MS (PPMS) is diagnosed in approximately 10–15 % of MS patients. Disease-modifying therapies (DMT) are less effective in modifying the course of progressive types of MS. It seems that inflammatory processes differ in the MS subtypes. ObjectivesThe objective of this study was to assess differences in the inflammatory parameters between PPMS and other courses of MS. Materials and methodsA total of 84 subjects were included in the study. The study group was divided according to the course of MS into the following categories: PPMS (n = 24); SPMS—secondary progressive multiple sclerosis (n = 14); RRMS—relapsing–remitting multiple sclerosis (n = 46). PPMS patients were further divided into treated with ocrelizumab and treatment-naive groups. The concentrations of serum inflammatory parameters were evaluated. ResultsPPMS and SPMS significantly differed in the serum levels of sCD30, gp130, sIL-6R alpha, osteopontin, pentraxin-3 and sTNF-R1. The serum concentrations of IFN-alpha2, IL-10, IL-20, IL-29 and osteopontin significantly differed between PPMS and RRMS. The serum levels of BAFF, IL-19, IL-20, pentraxin-3, s-TNF-R1 and s-TNF-R2 significantly differed between PPMS treated with ocrelizumab and treatment-naive. ConclusionAlthough inflammatory processes take part in the pathogenesis of all types of MS, they differ between MS courses. Serum inflammatory parameters seem to be promising biomarkers in helping to differentiate courses of MS, and in assessing reactions to DMT treatment. Further investigations on their usage are required.

Alpha-Fetoprotein as a Factor of Differentiation and Functional Activity of Myeloid-Derived Suppressor Cells.

We studied the role of alpha-fetoprotein (AFP) in regulation of differentiation and functional activity of human myeloid-derived suppressor cells (MDSC) in vitro. To obtain MDSC, CD11b+ cells were isolated from the peripheral blood of healthy donors followed by cytokine induction (IL-1β+GM-CSF) into the MDSC phenotype. The cell functions were assessed by the expression of indoleamine 2,3-dioxygenase (IDO) and arginase-1 (Arg1) and cytokine profile of the cell cultures. Native AFP did not affect the total number of MDSC and the percentage of polymorphonuclear MDSC (PMN-MDSC), but increased the number of monocytic MDSC (M-MDSC). AFP did not change the expression of Arg1, but in low concentrations (10 and 50 U/ml) increased the number of IDO-containing cells. AFP modulated the cytokine profile of CD11b+ cells: it reliably decreased the level of IL-19 (50 and100 U/ml) and showed a tendency to decrease the levels of IL-34, MMP-2, sCD163, CHI3L1, OPN and to increase the levels of IL-29, IL-32, APRIL, PTX3, and sTNF-R1. Thus, we have demonstrated a regulatory effect of native AFP at the level of MDSC generated from CD11b+ cells under conditions of cytokine induction in vitro.

Inflammatory biomarkers at different stages of Sarcopenia in older women

In recent years, studies have found that Sarcopenia alters inflammatory biomarkers. However, the behavior of inflammatory biomarkers at different stages of Sarcopenia is not well understood. This study aimed to compare a broad panel of inflammatory biomarkers in older women at different stages of Sarcopenia. The study included 71 Brazilian community-dwelling older women. Muscle Strength was assessed by using handgrip strength (Jamar dynamometer). The Short Physical Performance Battery (SPPB) was performed to assess the physical performance, and body composition was assessed by DEXA. Sarcopenia was diagnosed and classified according to the EWGSOP2 criteria. Blood was drawn, and inflammatory biomarkers associated with Sarcopenia (IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, TNF, adiponectin, leptin, resistin, BDNF, sTNFr-1 and sTNFr-2) was analysed. After diagnosis and classification of sarcopenia, 45% of women did not present Sarcopenia (NS, N = 32), 23.9% were diagnosed with Sarcopenia Probable (SP, N = 17), 19,7% with Sarcopenia Confirmed (SC, N = 14), and 11.3% with Severe Sarcopenia (SS, N = 8). The analysis of inflammatory biomarkers revealed that the more advanced the stage of Sarcopenia, the higher the levels of BDNF, IL-8, sTNFr-1, and sTNFr-2. The assessment of BDNF, IL-8, sTNFr-1, and sTNFr-2 levels may be an adjuvant tool in diagnosis and severity classification of Sarcopenia in older Brazilian women.

#4713 TUMOR NECROSIS FACTOR RECEPTOR 2 POLYMORPHISM (RS1061622) AND INFLAMMATORY RESPONSE IN END-STAGE RENAL DISEASE PATIENTS UNDER DIALYSIS

Abstract Background and Aims Enhanced levels of soluble tumor necrosis factor receptor 2 (sTNFR2) are known to associate with progressive chronic kidney disease (CKD), and is pointed as a potential biomarker for early detection of CKD; moreover, it has been reported as an independent predictor of all-cause mortality in end-stage renal disease (ESRD) patients under dialysis. Despite the increase in TNFR2 and in other inflammatory markers, recognized as risk factors for mortality in dialysis patients, the hypothesis that genetic polymorphisms of those biomarkers might modulate the inflammatory response and, thereby, the patients’ survival predisposition, has been poorly studied. Concerning TNFR2 genetic variants, a single nucleotide polymorphism in TNFR2 (+ 676 T/G; rs1061622), that results in amino acid change at position 196 (Met/Arg), was associated with higher levels of sTNFR2 in inflammatory conditions. The aim of this study was to determine the allelic frequencies of TNFR2 in ESRD patients and controls, and to evaluate its relationship with the circulating levels of inflammatory biomarkers. Method We studied 277 ESRD patients on dialysis and 32 controls, matched for gender, body mass index, and, as far as possible, for age. Real time PCR TaqMan SNP genotyping assay was used to assess allelic frequencies of TNFR2 (rs1061622). We also evaluated the circulating levels of TNF-alpha, sTNFR2, ferritin, hepcidin, elastase and cell-free DNA (cfDNA). Deaths occurring along 1-year follow-up period were recorded and mortality rates were assessed. Results ESRD patients presented higher levels of all studied biomarkers, as compared to controls; their overall mortality rate was 10.5%. Allelic frequencies in ESRD patients and controls were similar for TNFR2 (rs1061622) considering the homozygous and heterozygous individuals (χ2, p = 0.518). Concerning sTNFR2 values, no significant differences were observed between patients with genotypes TT, GG or TG. The GG genotype patients, compared to TT genotype carriers, presented significantly lower ferritin (p = 0.048), hepcidin (p = 0.038), elastase (p = 0.006) and cfDNA levels (p = 0.016); and compared to TG genotype patients, showed significantly lower ferritin (p = 0.039) and a trend towards lower values of hepcidin (p = 0.097), elastase (p = 0.079) and cfDNA (p = 0.0164). TG genotype patients showed higher TNF-alpha (p = 0.049) and a trend towards lower elastase (p = 0.081) than TT genotype subjects. The GG genotype patients presented a trend towards lower mortality rate (6.3%, 7.5%, and 12.4% for GG, TG and TT, respectively). Conclusion No differences were found in the allelic frequencies between controls and ESRD patients. The GG genotype patients for TNFR2 rs1061622 polymorphism showed decreased levels of inflammation, suggesting a more favorable inflammatory response, which is usually associated to a lower mortality risk in these patients. In accordance with the scientific community, recommending studies on genetic survival predisposition in dialysis patients, the polymorphisms of TNFR2 and of other inflammatory biomarkers deserve further studies. Acknowledgements: This work was financed by CESPU, through the project SNPsCKD-GI2-CESPU-2022; FCT, through the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences—UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB.