Serum sPD-1 Levels Research Articles

Soluble Programmed Cell Death 1 Protein Is a Promising Biomarker to Predict Severe Liver Inflammation in Chronic Hepatitis B Patients.

Background and Aims: Liver inflammation is important in guiding the initiation of antiviral treatment and affects the progression of chronic hepatitis B(CHB). The soluble programmed cell death 1 protein (sPD-1) was upregulated in inflammatory and infectious diseases and correlated with disease severity. We aimed to investigate the correlation between serum sPD-1 levels and liver inflammation in CHB patients and their role in indicating liver inflammation. Methods: 241 CHB patients who underwent liver biopsy were enrolled. The correlation between sPD-1 levels and the degree of liver inflammation was analyzed. Univariate and multivariate logistic regression analyses were performed to analyze independent variables of severe liver inflammation. Binary logistic regression analysis was conducted to construct a predictive model for severe liver inflammation, and the receiver operating characteristic curve (ROC) was used to evaluate the diagnostic accuracy of the predictive model. Results: sPD-1 was highest in CHB patients with severe liver inflammation, which was higher than that in CHB patients with mild or moderate liver inflammation (P < 0.001). Besides, sPD-1 was weakly correlated with AST (r = 0.278, P < 0.001). Multivariable analysis showed that sPD-1 was an independent predictor of severe liver inflammation. The predictive model containing sPD-1 had areas under the ROC (AUROCs) of 0.917 and 0.921 in predicting severe liver inflammation in CHB patients and CHB patients with ALT ≤ 1× upper limit of normal (ULN), respectively. Conclusions: Serum sPD-1 level is associated with liver inflammation in CHB patients, and high levels of sPD-1 reflect severe liver inflammation. Serum sPD-1 is an independent predictor of severe liver inflammation and shows improved diagnostic accuracy when combined with other clinical indicators.

Decreased serum soluble programmed cell death ligand-1 level as a potential biomarker for missed miscarriage.

Can maternal serum levels of soluble programmed cell death-1 (sPD-1) and its ligand (sPD-L1) serve as biomarkers for missed miscarriage (MM)? Serum sPD-L1 levels are significantly decreased in MM patients and may serve as a potential predictive biomarker for miscarriage. Programmed cell death-1 (PD-1) and its ligand (PD-L1) comprise important immune inhibitory checkpoint signaling to maintain pregnancy. Their soluble forms are detectable in human circulation and are associated with immunosuppression. Three independent cohorts attending tertiary referral hospitals were studied. The first (discovery) cohort was cross-sectional and included MM patients and healthy pregnant (HP) women matched on BMI. The second validation cohort contained MM patients and women with legally induced abortion (IA). The third prospective observational study recruited subjects requiring IVF treatment. In the discovery cohort, we enrolled 108 MM patients and 115 HP women who had a full-term pregnancy at 6-14 weeks of gestation. In the validation cohort, we recruited 25 MM patients and 25 women with IA. Blood samples were collected at the first prenatal visit for HP women or on the day of dilatation and curettage surgery (D&C) for MM and IA subjects to determine serum sPD-1 and sPD-L1 levels. Placenta samples were harvested during the D&C within the validation cohort to measure gene and protein expression. The prospective cohort collected serial blood samples weekly from 75 volunteers with embryo transfer (ET) after IVF. Circulating sPD-L1 levels were reduced by 50% in patients with MM (55.7 ± 16.04 pg/ml) compared to HP controls (106.7 ± 58.46 pg/ml, P < 0.001) and the difference remained significant after adjusting for maternal age and gestational age, whereas no significant differences in sPD-1 level were observed. Likewise, serum sPD-L1 was lower in MM patients than in IA subjects and accompanied by downregulated PD-L1-related gene expression levels in the placenta. In the IVF cohort, applying the changing rate of sPD-L1 level after ET achieved a predictive performance for miscarriage with receiver operating characteristics = 0.73 (95% CI: 0.57-0.88, P < 0.01). The study was mainly confined to East Asian pregnant women. Further large prospective pregnancy cohorts are required to validate the predictive performance of sPD-L1 on miscarriage. Reduced circulating sPD-L1 level and downregulated placental PD-L1 expression in miscarriage indicate that dysfunction in PD-L1 signals is a potential underlying mechanism for pregnancy loss. Our findings further extend the importance of the PD-L1 axis in pregnancy maintenance in early pregnancy. This study was financially supported by grants from the Subject Innovation Team of Shaanxi University of Chinese Medicine (2019-Y502), General Research Fund (14122021), and Key Laboratory of Model Animal Phenotyping and Basic Research in Metabolic Diseases (2018KSYS003). The authors declare that they have no competing interests to be disclosed. N/A.

P-478 The decreases in serum soluble programmed cell death ligand −1 level are associated with missed miscarriage in humans

Abstract Study question The perturbations in serum levels of soluble forms of programmed cell death-1 (sPD-1) and its ligand (sPD-L1) in missed miscarriage (MM) are unknown. Summary answer Dysfunction in PD-L1 signal has been demonstrated in MM as evidenced by a lower circulating sPD-L1 level and declined expressions of PD-L1 in placenta. What is known already Programmed cell death-1 (PD-1) and its ligand (PD-L1) comprise important immune inhibitory checkpoint signaling to maintain pregnancy. During pregnancy, PD-L1 is highly expressed in the trophoblasts of the placenta. The reduced placental PD-L1 expression is associated with the risk of miscarriage. Therefore, disturbance of PD-1/PD-L1 axis in the fetomaternal interface is susceptible to be an important pathogenesis for pregnancy loss. Interestingly, the soluble form of these immune checkpoint molecules is detectable and associated with immunosuppression, it raises the diagnostic potential to monitor their level to monitor pregnancy health. Study design, size, duration In the discovery cohort, we enrolled 108 patients with MM and 115 body mass index (BMI)-matched, healthy women with a full-term pregnancy at 6 to 13 weeks of gestation. Another cohort of 25 MM patients and 25 subjects with induced abortion was recruited for validation. Participants/materials, setting, methods Blood samples were collected at the first prenatal visit for HP women or on the day of dilatation and curettage surgery (D&amp;C) for subjects with MM or IA to measure serum sPD-1 and sPD-L1 levels. Placenta samples were harvested during the D&amp;C within cohort 2 to examine related gene and protein expression. Main results and the role of chance Our results showed that circulating sPD-L1 levels were reduced by 50% in patients with MM (55.71 ± 1.54 pg/mL) compared to HP controls (107.27 ± 5.48 pg/mL, P&amp;lt;0.001) and remained significant after adjusting for maternal age and gestational age, whereas no significant differences in sPD-1 were observed between the two groups. Applying serum sPD-L1 levels achieved an area under the receiver operating characteristics curve (AUROC) of 0.83 (95% CI: 0.77 to 0.88, P&amp;lt;0.001) with the optimal cut-off value of 81.52 pg/ml to detect MM. Our data in the validation cohort further demonstrated that sPD-L1 was lower in MM patients relative to IA subjects. Likewise, placental PD-L1-related gene expressions were downregulated at both mRNA and protein levels in miscarriage samples relative to samples in IA groups and were positively associated with sPD-L1 levels in maternal circulation Limitations, reasons for caution This study was conducted in East Asian women, further studies are required for the evaluation of the serum PD-1 and sPD-L1 levels in different ethnic groups. Secondly, due to the cross-sectional nature of the study design, whether the sPD-L1 levels could utilize as a predictive biomarker is questioned. Wider implications of the findings The maternal serum sPD-L1 level has the potential to be a diagnostic aid for pregnancy maintenance. Moreover, insufficient PD-L1 expression in the feto-maternal interface may act as one of the potential pathogenic events to trigger early pregnancy loss. Trial registration number not applicable

Soluble PD-L1 in blood correlates positively with neutrophil and negatively with lymphocyte mRNA markers and implies adverse sepsis outcome

Sepsis causes a myriad of immunological reactions that result in life-threatening alterations in the human body. Immunosuppression in sepsis is partly attributed to the programmed death receptor (PD-1) and its associated ligand (PD-L1) via the regulation of lymphocytes and neutrophils. Although the soluble forms of these proteins (i.e., sPD-1 and sPD-L1, respectively) are recognized as possible sepsis biomarkers, their functional implications are yet to be elucidated. Our research assessed the correlation between sPD-1 and sPD-L1 and blood mRNA markers and sepsis outcome. Blood samples of septic patients of urogenital origin versus control patients (both groups: n = 18) were analyzed. Blood serum sPD-1 and sPD-L1 levels were determined using the enzyme-linked immunosorbent assay (ELISA). The whole blood mRNA concentrations of PD-1, PD-L1, neutrophil markers (CEACAM8 and MPO), and T-lymphocyte markers (TCRβ, CD4 and CD8) were determined via reverse transcriptase quantitative PCR (RT-qPCR). sPD-L1 levels were significantly increased in septic patients when compared to the controls, whereas sPD-1 levels were unaltered. Patients with high sPD-L1 levels, as dichotomized to the median, had a significantly shorter survival rate than those with low sPD-L1 levels. The sensitivity/specificity characteristics of sPD-L1 proved significant for sepsis detection. Furthermore, sPD-L1 correlated with the mRNA concentrations of PD-L1, CEACAM, and MPO, as well as major inflammatory markers (C-reactive protein and procalcitonin). However, sPD-L1 negatively correlated with TCRβ, CD4, and CD8 mRNAs. sPD-L1 was found to be significantly increased in septic patients. Notably, sPD-L1 correlated with PD-L1 mRNA and neutrophil markers and was indicative of adverse outcomes.

Serum soluble programmed death-1 levels predict the spontaneous HBeAg seroclearance in chronic hepatitis B.

In chronic hepatitis B virus (HBV) infection, earlier seroclearance of hepatitis B e antigen (HBeAg) is associated with more favorable outcomes. Soluble programmed cell death 1 (sPD-1) has been implicated in higher viral load and hepatocellular carcinoma. We investigated the association between sPD-1 levels and spontaneous HBeAg seroclearance. Baseline serum samples from 488 HBeAg-seropositive patients in the REVEAL-HBV cohort were tested for sPD-1 levels. Among them, 329 with available follow-up serum samples were further assayed. Multivariate Cox regression analysis was used to estimate the adjusted rate ratio (aRR) and 95% confidence interval (CI) with adjustment of host and viral factors. The 66th percentile and an annual reduction of ≥ 10% were used as the cut-off point for baseline sPD-1 levels (high/low) and sPD-1 trajectory (decline/no decline), respectively. Lower baseline sPD-1 levels [aRR (95% CI): 2.19 (1.47-3.27)] and long-term decline in sPD-1 levels [aRR (95% CI): 4.08 (2.79-5.97)] were both independent predictors for HBeAg seroclearance. However, further stratification analysis by HBV genotype showed that lower baseline sPD-1 levels were significantly associated with HBeAg seroclearance only in genotype C infection [aRR (95% CI): 4.47 (2.38-8.37)] but not in genotype B infection. On the other hand, long-term decline in sPD-1 levels was predictive for HBeAg seroclearance regardless of HBV genotype with aRR (95% CI) of 4.62 (2.71-7.88) and 2.95 (1.68-5.17), respectively, for genotypes B and C. Serum sPD-1 levels may serve as a novel immunological predictor for spontaneous HBeAg seroclearance in patients with chronic hepatitis B.

Soluble forms of PD-1/PD-L immune checkpoint receptor and ligand in blood serum of breast cancer patients: association with clinical pathologic factors and molecular type of the tumor.

Results of enzyme-linked immunosorbent assay of the soluble forms of PD-1/PD-L immune checkpoint receptor and ligand (sPD-1 and sPD-L1) in pretreatment blood serum of 88 breast cancer patients at various disease stages aged 30-83 years are presented. The control group included 55 practically healthy women aged 19-82 years. Serum sPD-1 and sPD-L1 levels in breast cancer patients highly significantly (p<0.0001) differ from control and these changes are opposite: soluble receptor level is more than 6-fold decreased, while soluble ligand concentration - 5.5 fold increased. Both markers separately, as well as their ratio demonstrate very high sensitivity (94-100%) and specificity (95-100%) in relation to healthy control. No statistically significant associations of sPD-1 and sPD-L1 levels with clinical stage, individual TNM system criteria, tumor histological structure, grade, receptor status, and molecular type were established. In particular, no significant peculiarities of the markers' levels in triple negative breast cancer successfully treated with anti-PD-1/PD-L1 preparations were revealed. Long-term follow-up and dynamic studies of sPD-1 and sPD-L1serum levels in the course of treatment are required for evaluation of their independent from clinical and morphological factors prognostic significance and the possibility of application as low invasive tests for prediction and monitoring of corresponding targeted therapy efficiency.

High levels of soluble programmed death-1 are associated with virological response in chronic hepatitis B patients after antiviral treatment

BackgroundSoluble programmed cell death protein-1 (sPD-1) plays an important role in chronic hepatitis B virus (HBV) infection by counteracting the inhibitory effect of programmed death ligand-1 (PD-L1) on immune cells. Here, we investigated the ability of sPD-1 to predict the virological response (VR) in chronic hepatitis B (CHB) patients undergoing Nucleos(t)ide analogue (NA) therapy. MethodsCHB patients [hepatitis B surface antigen (HBsAg) positive ≥6 months] who initiated NA therapy in March 2007 at Peking University First Hospital (China) were enrolled in this study. Eighty-nine CHB patients were followed-up every 12 weeks for 96 weeks. ResultsSerum sPD-1 levels at baseline were negatively correlated with hepatitis B surface antigen (HBsAg) and HBV DNA. Immune-active CHB patients exhibited higher serum sPD-1 levels at baseline. Patients with VR during the antiviral treatment exhibited higher sPD-1 levels and lower HBsAg levels at baseline. Receiver operating characteristic (ROC) curves were generated to determine the predictive value of sPD-1 and HBsAg for VR in patients who received first-line therapy (entecavir, ETV). The area under ROC (AUROC) values of sPD-1 and HBsAg at baseline were 0.850 (95%CI:0.729–0.971, P = 0.0005) and 0.785 (95%CI: 0.642–0.929, P = 0.005), respectively, and the optimal cut-off values were 459.46 pg/mL and 14,710 IU/mL, respectively. The combination of sPD-1 and HBsAg exhibited a higher AUROC value (0.870,95% CI: 0.748–0.983, P = 0.001) than did sPD-1 or HBsAg alone. In patients administered second-line therapy (lamivudine, LAM/adefovir divipoxil, ADV), baseline sPD-1 levels above 677.2 pg/mL were significantly associated with higher incidence of VR after 96 weeks of antiviral therapy. It is 7.956 times the level of ≤677.2 pg/mL. ConclusionsBy combining sPD-1 and HBsAg, we obtained a biomarker significantly associated with VR in CHB patients. The sPD-1 levels could be used to screen out patients with poor prognosis of antiviral therapy.

Soluble programmed death molecule 1 (sPD-1) as a predictor of interstitial lung disease in rheumatoid arthritis

BackgroundPrevious studies have indicated that the programmed death molecule 1 (PD-1) signaling pathway may play a key role in rheumatoid arthritis (RA). However, the pathogenesis of rheumatoid arthritis-related interstitial lung disease (RA-ILD) is not clear. We examined the serum levels of soluble PD-1 in patients with RA and its relationship with RA-ILD.MethodsBlood samples were obtained from 87 patients with RA (58 with ILD and 29 without ILD) and 45 healthy controls. Serum sPD-1 was measured by Enzyme-Linked Immunosorbent Assay. The pulmonary interstitial disease score was completed by a pulmonary physician and a radiologist through chest high-resolution computed tomography. Patients with RA-ILD were tested for lung function [e.g., forced vital capacity (FVC%), diffusing capacity of lungs for carbon monoxide (DLCO%)]. Associations between ILD and various markers, including sPD-1 and confounding factors, were investigated by logistic regression analysis. Diagnostic values of sPD-1 for the presence of ILD were investigated using receiver operating characteristic curve analysis.ResultsSerum sPD-1 levels were higher in RA patients with ILD than in RA patients without ILD and healthy controls (185.1 ± 109.0 pg/ml vs. 119.1 ± 77.5 pg/ml vs. 52.1 ± 21.7 pg/ml, P < 0.05). Serum sPD-1 levels were positively correlated with RF titer (P = 0.02, r = 0.249), anti-cyclic citrullinated peptide antibody status (P = 0.02, r = 0.243), and serum IgG levels (P < 0.001, r = 0.368), negatively associated with FVC% (P = 0.02, r = − 0.344), forced expiratory volume (FEV1%) (P = 0.01, r = − 0.354), total lung capacity (TLC%) (P = 0.046, r = − 0.302), and was independently associated with the presence of ILD in RA patients by multivariate logistic regression analysis. The sensitivity and specificity of sPD-1 levels for the detection of ILD in RA patients were 58.6% and 75.9%, respectively. The area under the curve was 0.689.ConclusionSerum sPD-1 levels were increased in RA patients with ILD. Increased sPD-1 may be a valuable biomarker to predict the presence of ILD in patients with RA.

Serum soluble PD-1 plays a role in predicting infection complications in patients with acute pancreatitis.

BackgroundMost of acute pancreatitis (AP) are mild and self‐limiting, however, 15%–20% of patients develop severe acute pancreatitis (SAP) or moderately acute pancreatitis (MSAP) with local or systemic complications. Infection complications (ICs) result in 40%–70% morbidity and high mortality rates among SAP and MSAP patients. It is more important to early recognize of ICs of MSAP or SAP. Several studies have indicated that serum soluble programmed cell death protein (sPD‐1) or programmed cell death 1 ligand (sPD‐L1) levels were higher in patients with severe sepsis than in healthy volunteers and have a predictive capacity for mortality. However, the role of serum sPD‐1/sPD‐L1 in AP remains unclear. This study aimed to investigate whether the ICs of AP patients is associated with their sPD‐1 and sPD‐L1 levels, which were determined via enzyme‐linked immunosorbent assay of peripheral blood samples from 63 MSAP and SAP patients and 30 healthy volunteers.ResultsThe serum sPD‐1 levels in AP patients on Days 1, 3, and 10 after onset were significantly increased in a time‐dependent manner compared with that in healthy volunteers. Moreover, the AP patients with ICs had significantly higher serum sPD‐1 levels than the AP without ICs. While serum sPD‐L1 levels in AP were similar to that in healthy volunteers. Besides, serum levels of sPD‐1/sPD‐L1 were negatively correlated with circulating lymphocytes. Univariate and multivariate regression analyses showed that the upregulated serum sPD‐1 level was an independent risk factor for ICs in AP. The area under the receiver operating characteristics curve indicated that combination with Acute Physiology and Chronic Health Evaluation II score and serum sPD‐1 level had a high accuracy in predicting ICs in AP.ConclusionSerum sPD‐1/sPD‐L1 may be involved in the immunosuppressive process in AP. Moreover, the serum sPD‐1 level may be an independent risk factor for predicting ICs in AP patients.

High serum sPD-1 and sPD-L1 levels 1 predict outcome in hepatocellular carcinoma patients treated with radiotherapy.

e16672 Background: Accumulating data have shown that soluble (s)PD-1 and sPD-L1 are potential biomarkers for the curative efficacy of chemotherapy in several cancers. However, these markers have not been well established for outcomes in hepatocellular carcinoma (HCC) radiotherapy (RT). Therefore, this study aims to assess sPD-1/sPD-L1 levels before and after RT and their relationship to clinical characteristics and prognoses. Methods: Pre-treatment serum sPD-L1 and sPD-1 levels were measured with an enzyme-linked immunosorbent assay (ELISA) in 281 patients with HCC treated with RT. The association between serum sPD-L1 and sPD-1 levels and prognosis was assessed using survival analysis. Formalin-fixed paraffin-embedded samples were obtained from 40 patients with HCC who underwent surgery. The expression of PD-L1 was evaluated by immunohistochemical analysis. Results: Using multivariate analyses; sPD-L1 and sPD-1 levels were significant independent predictors of disease-free and overall survival. Patients with maximum tumor diameters ≥5 cm and Barcelona clinic liver cancer (BCLC) staging B and C had significantly higher median sPD-1/sPD-L1 serum concentrations compared to patients with smaller tumor sizes and earlier stages. There was a positive correlation (r = 0.377, p &lt; 0.001) between sPD-L1 and sPD-1 levels. High pretreatment sPD-1/sPD-L1 levels were associated with poor prognoses, and increased sPD-1/sPD-L1 levels were associated with disease progression after RT. In post-irradiated tumor tissue, PD-L1 expression was significantly higher than matched, non-irradiated tissue. In addition, higher PD-L1 expression was correlated with decreased cytotoxic T-cell infiltration and poor survival. Therefore, sPD-1/sPD-L1 could be used as noninvasive biomarkers for evaluating HCC malignancy before RT and in predicting the response to RT in HCC patients. Conclusions: High pretreatment serum sPD-L1 and sPD-1 were associated with poor prognoses, and increased serum sPD-L1 levels were associated with the disease progression of HCC patients treated with RT. PD-L1/ PD-1 pathway may be explored as therapeutic targets and potential predictive biomarkers for HCC treated with RT.

Profiles of serum soluble programmed death‐1 and programmed death‐ligand 1 levels in chronic hepatitis B virus‐infected patients with different disease phases and after anti‐viral treatment

Soluble programmed death-1 (sPD-1) and soluble programmed death-ligand 1 (sPD-L1) play a role in immune regulation of chronic hepatitis B virus (HBV) infection. To investigate the profiles of serum sPD-1 and sPD-L1 in chronic HBV-infected patients with different disease phases and after anti-viral treatment. A total of 99 chronic HBV-infected patients were enrolled and divided into HBeAg-positive chronic HBV infection (EPI) group, HBeAg-positive chronic hepatitis B (EPH) group, HBeAg-negative chronic hepatitis B (ENH) group and HBeAg-negative chronic HBV infection (ENI) group. Eleven healthy subjects were included as healthy controls (HCs). Thirty-two EPH patients received anti-viral treatment with nucleos(t)ide analogues and were followed up to 5years. Serum sPD-1 and sPD-L1 levels were detected by Multiplex Immunoassays. Serum sPD-1 and sPD-L1 levels of chronic HBV infected patients were significantly higher than that of HCs (P<0.01). Patients in EPH, ENH and EPI groups had higher serum sPD-1 and sPD-L1 levels than that in HCs (P<0.01). After anti-viral treatment, serum sPD-1 and sPD-L1 levels declined rapidly. EPH patients with HBeAg clearance after 2years of anti-viral treatment showed lower baseline HBeAg and sPD-1 levels compared to those without HBeAg clearance. Serum sPD-1 and sPD-L1 levels varied among chronic HBV infected patients with different disease phases. Lower baseline sPD-1 levels were associated with HBeAg clearance after 2years of anti-viral treatment in EPH patients.

High Level of Soluble Programmed Death-1 Is Associated with Functional Cure in Chronic Hepatitis B Upon Long-Term Treatment

Background & Aims: Some studies showed that soluble forms of programmed death1(sPD-1) could interact with programmed death ligand-1 (PD-L1) expressed on the membrane and counteract its inhibitory influence on immune cells. This study aimed to investigate the predictive value of sPD-1 in chronic hepatitis B (CHB) for functional cure during Nucleos(t)ide analogues (NAs) therapy. Methods: Eighty-four CHB patients were enrolled and started antiviral treatment in March 2007 at the Department of Infectious Diseases at Peking University First Hospital in Beijing. Findings: After 10 years of follow-up, 44 patients achieved low hepatitis B surface antigen(HBsAg) levels. The results indicated that a baseline sPD-1 level ≥500 pg/mL was associated with a higher incidence of low HBsAg levels (n=30/34, 88.2% at year 10) compared with a level <500 pg/mL (n=14/29, 48.3% at year 10) (HR 2.94; P<0.001). Moreover, sPD-1≥500 pg/mL at baseline (OR 17.33; P=0.001) and age (OR 1.11; P=0.031) could predict low HBsAg levels after 10-year follow-up. Interpretation: Baseline sPD-1≥500 pg/mL could predict low HBsAg levels and was associated with a higher incidence of low HBsAg levels after the 10-year follow-up. High serum sPD-1 levels and age might be used to distinguish patients with a higher incidence of a functional cure. Funding Statement: The 13th Five-Year Plan, No.2018ZX09206005-002，Ministry of Science and Technology of the People’s Republic of China. Declaration of Interests: There are no conflicts of interest to report. Ethical Approval Statement: The study was approved by the Institutional Review Board of the Peking University First Hospital. The study protocol conformed to the ethical guidelines of the Declaration of Helsinki and was approved by the Ethics Committee of Shanghai Jing An Central Hospital (Approval no. 090f51e6809a26e1 v1.0).

Changes and significance of serum soluble programmed death-1 in patients with chronic hepatitis C

Objective To analyze the changes and significance of serum soluble programmed death-1 (sPD-1) in patients with chronic hepatitis C (CHC) and study its role in the progression of CHC. Methods Serum levels of sPD-1 in CHC patients and healthy controls (HC) were measured using ELISA and compared. Correlations of serum sPD-1 with peripheral hepatitis C virus (HCV) RNA, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and liver fibrosis (indicated by APRI) were analyzed. Results The serum sPD-1 level in the CHC group was significantly higher than that in the HC group (P<0.05), and positively correlated with peripheral HCV RNA, ALT and AST (P<0.05). In addition, the serum sPD-1 levels in patients with APRI greater than 1.2 (indicating severe liver fibrosis) were higher than those in patients without or with mild liver fibrosis (P<0.05). Conclusions sPD-1 might be involve in the progression of CHC. Measuring serum sPD-1 in CHC patients would assist the prediction of disease progression and help to make the correct diagnosis and appropriate clinical therapy decision. Key words: Soluble programmed death-1; Chronic hepatitis C; Liver fibrosis

Predictive Value of Soluble Programmed Death-1 for Severe Sepsis and Septic Shock During the First Week in an Intensive Care Unit.

Programmed death-1 (PD-1)/programmed death ligand-1 (PD-L1) exists in both membrane-bound and soluble forms. In this study, we evaluated the predictive value of soluble PD-1 (sPD-1) for severity and 28-day mortality in patients with severe sepsis and septic shock during the first week in an intensive care unit (ICU). In this prospective cohort study, patients were classified into the severe sepsis group or the septic shock group according to the severity of their condition on ICU admission. All patients were also separated into the survivor or nonsurvivor groups according to their 28-day outcomes. Peripheral blood sPD-1 and soluble PD-L1 (sPD-L1) levels, PD-1 expression on CD4 and CD8 T cells, and PD-L1 expression on monocytes were measured and compared between the groups on days 1 and 7 after ICU admission. In all, 45 healthy volunteers and 112 patients were recruited. Serum sPD-1 levels were positively correlated with the severity of sepsis, sPD-L1 levels, PD-1 expression on CD4 or CD8 T cells, and PD-L1 expression on monocytes. The sPD-1 was an independent predictive factor for 28-day mortality both on day 1 and day 7. The area under the curve (AUC) of the sPD-1 on day 7 (0.871) was higher than that on day 1 (0.785) (P < 0.05), and better than the AUC of the percentages of PD-L1 on monocytes (0.770) on day 7 (P < 0.05). Serum sPD-1 shows valuable predictive ability for the severity and 28-day mortality of severe sepsis and septic shock during the first week of ICU treatment.

Changes of serum sPD-1 levels in HBeAg-positive chronic hepatitis B patients with entecavir treatment and correlation with curative effect

Background/aim: This study aimed to assess expression changes of serum sPD-1 levels in HBeAg-positive chronic hepatitis B patients with entecavir treatment and explore the correlation with the curative effect. Materials and methods: A total of 220 HBeAg-positive CHB patients and 207 healthy individuals were included. sPD-1 levels and virological and biochemical responses were assessed at baseline and at 12, 24, and 48 weeks in CHB patients after antiviral therapy. Results: sPD-1 levels in HBeAg-positive CHB patients before treatment were significantly lower than those of healthy controls. After entecavir treatment, sPD-1 levels increased gradually over time. At 48 weeks after entecavir treatment, sPD-1 amounts in the HBeAg seroconversion group were significantly higher than the values of the HBeAg-negative and HBeAg-positive groups. sPD-1 levels in patients with alanine aminotransferase (ALT) returning to normal were significantly higher than in those with ALT not returning to normal. sPD-1 levels in treated patients with no HBV-DNA were significantly higher than in those with HBV-DNA remaining positive. sPD-1 was negatively correlated with ALT and HBV-DNA at 12, 24, and 48 weeks after entecavir treatment. Conclusion: sPD-1 may play a certain role in chronic hepatitis B and has a close relationship with the curative effect of entecavir.

WITHDRAWN: Immune imbalance of PD-1 and 1 PD-Ls in liver tissues from patients with Hepatitis B-related hepatocellular carcinoma

// Yong Liu 1,2 , Guiyang Wang 3 , Rui Huang 3 , Yingying Hao 4 , Haiyan Chang 3 , Chen Tian 3 , Yang Li 3 , Xiang-An Zhao 3 , Juan Xia 3 , Yuxin Chen 3,5 and Chao Wu 3 1 Department of Experimental Medicine, Nanjing Drum Tower Hospital, Nanjing University Medical School, Nanjing, China 2 Jiangsu Key Laboratory for Molecular Medicine, Nanjing University Medical School, Nanjing, China 3 Department of Infectious Diseases, Nanjing Drum Tower Hospital, Nanjing University Medical School, Nanjing, China 4 Department of Intensive Care Units, Nanjing Drum Tower Hospital, Nanjing University Medical School, Nanjing, China 5 Department of Laboratory Medicine, Nanjing Drum Tower Hospital, Nanjing University Medical School, Nanjing, China Correspondence to: Chao Wu, email: dr.wu@nju.edu.cn Yuxin Chen, email: yuxin_chen2015@163.com Keywords : PD-1; PD-L1; PD-L2; microenvironment; hepatitis B-related hepatocellular carcinoma Received: October 27, 2017&nbsp;&nbsp;&nbsp;&nbsp; Accepted: December 05, 2017&nbsp;&nbsp;&nbsp;&nbsp; Epub: January 12, 2018 Abstract 2 Dysregulated immune microenvironment, such as imbalance of programmed death-1 (PD-1) and its ligands (PD-Ls), plays a key role in hepatocarcinogenesis. Here we investigate the key immune checkpoint molecules, PD-1/PD-Ls, in liver tissues from patients with hepatitis B virus-related hepatocellular carcinoma (HBV-related HCC). We show that the characteristic of T cells in peripheral blood does not fully reflect the changes of immune microenvironment in tumor tissues. There is an imbalanced immune microenvironment in liver tissues, characterized by increased CD4/CD8 ratio and high expression of PD-1 on CD8 T cells in tumor tissues. Soluble PD-1 (sPD-1) levels in tumor tissues are increased, whereas sPD-L1 and sPD-L2 levels are decreased. Compared with control subjects, the patients with HBV-related HCC have higher serum sPD-1 and sPD-L1 levels, lower serum sPD-L2 levels. We also show that the altered immune microenvironment is not only found in tumor tissues, but also in non-tumor tissues from HBV-related HCC patients. The correlation between PD-1 and PD-L1 increases successively in hepatocellular carcinomas tissues, paracancerous tissues and adjacent non-tumor tissues. In tumor tissues, sPD-1 levels are negatively correlated with clinicopathological stages of HCC, while serum sPD-L1 levels have positive correlation with the clinicopathological stages. Combined, HBV infection alone may not affect the characteristics of T cell subpopulations, but might be related with the higher PD-1 levels in liver tissues. The immune imbalance of PD-1/PD-Ls in liver tissues may have important role in tumor microenvironment and immune escape, closely related with tumor initiation and progression.

Circulating soluble programmed death-1 levels may differentiate immune-tolerant phase from other phases and hepatocellular carcinoma from other clinical diseases in chronic hepatitis B virus infection.

Programmed death-1 (PD-1) is involved in the immune dysfunction of hepatitis B virus (HBV) infection and hepatocellular carcinoma (HCC). This study analyzed the association of circulating soluble PD-1 (sPD-1) levels with the phases and clinical diseases in chronic HBV infection. Serum sPD-1 levels were determined by enzyme linked immunosorbent assay in patients with different phases and liver diseases of chronic HBV infection. The sPD-1 levels in patients with chronic HBV infection were significantly elevated compared with HBV infection resolvers or healthy controls. According to phases, sPD-1 level in immune-tolerant phase (IT) was significantly lower than in other phases. Multivariate analysis showed that sPD-1 was an independent factor associated with IT. Area under the receiver operating characteristic (ROC) curves (AUC) showed that sPD-1 was significantly discriminative of IT from other phases with a cut-off of 1.535 ng/mL (AUC, 0.984; P<0.001). According to clinical diseases, sPD-1 level in HBV-related HCC was significantly higher than in other clinical diseases. Multivariate analysis showed that sPD-1 was an independent factor associated with HCC. The sPD-1 was significantly discriminative of HCC from other clinical diseases with a cut-off of 6.058 ng/mL (AUC, 0.962; P<0.001). The sPD-1 levels were significantly associated with HCC patients’ overall survival. HCC resection resulted in remarkable reduction in sPD-1 levels. These results demonstrate the involvement of sPD-1 in the disease course of chronic HBV infection and indicate the potential to apply sPD-1 as a biomarker for differentiating IT from other phases and HCC from other disease conditions in chronic HBV infection.

Serum sPD-L1, Upregulated in Sepsis, May Reflect Disease Severity and Clinical Outcomes in Septic Patients.

We aimed to find the correlation between serum sPD-L1 (soluble programmed cell death L-1 ligand) and sepsis. Totally 91 consecutive patients with sepsis were performed in a 15-bed medical intensive care unit (ICU) of the second affiliated hospital, Xi'an Jiaotong University in Xi'an, China, between February 2015 and May 2016. Healthy controls (HC) consisted of 29 healthy volunteer. Baseline demographic data were recorded. Blood samples were collected through an indwelling central venous or by peripheral venipuncture. Serum sPD-L1 and sPD-1 levels were determined with enzyme-linked immunosorbent assay kits (Elabscience Biotechnology Co. Ltd, Wuhan, China). SPSS19.0 software (SPSS Inc., Chicago, Illinois, USA) was used for statistical analysis. Kaplan-Meier survival analysis and Cox regression analysis were also performed. Serum sPD-L1 levels and sPD-1 levels were significantly increased in septic patients compared with HC (P=0.000). Serum sPD-L1 levels were significantly increased in non-survivors compared with survivors (P<0.05), but there was no statistically difference on serum sPD-1 levels between non-survivors and survivors (P>0.05). Serum sPD-L1 levels were correlated with absolute lymphocyte (ALC), platelets and SOFA scores. Serum sPD-L1/sPD-1 levels were negatively correlated with ALC and platelets, and SOFA scores. The prognostic accuracy of the sPD-L1 level to predict 28-day mortality was similar to that of the APACHE-II scores and SOFA scores. Cox regression analysis showed that sPD-L1 was an independent prognostic factor. Serum sPD-L1 is upregulated in sepsis and may reflect disease severity and clinical outcomes in patients. Serum sPD-L1 may be an independent prognostic factor for sepsis.

Serum levels of soluble programmed death-1 and programmed death ligand-1 in systemic sclerosis: Association with extent of skin sclerosis.

The interaction of programmed death-1 (PD-1) with its ligand, programmed death ligand-1 (PD-L1), has been considered to play a key role in the negative regulation of immune responses. Patients with diffuse cutaneous systemic sclerosis (SSc) had higher levels of soluble PD-1 (sPD-1) than those with limited cutaneous SSc and healthy individuals. Serum sPD-1 levels positively correlated with the severity of skin sclerosis. In contrast, serum sPD-L1 levels were significantly increased in patients with SSc compared with healthy individuals. Moreover, serum sPD-L1 levels were not associated with the extent of skin sclerosis and were elevated not only in patients with diffuse cutaneous SSc, but also in those with limited cutaneous SSc. These results suggested that serum sPD-1 levels may increase in patients with SSc and correlate with the severity of skin sclerosis. PD-1/PD-L1 interaction may contribute to the development of skin sclerosis in SSc.

Soluble Programmed Death 1 (PD-1) Is Decreased in Patients With Immune Thrombocytopenia (ITP): Potential Involvement of PD-1 Pathway in ITP Immunopathogenesis.

Immune thrombocytopenia (ITP) is an autoimmune disease characterized by dysregulation of T cells. Programmed death (PD) 1 and programmed death 1 ligand 1 (PD-L1) are cosignaling molecules, and the major role of the PD-1 pathway is the inhibition of self-reactive T cells and to protect against autoimmune diseases. We measured levels of serum soluble PD 1 (sPD-1) and serum soluble PD-L1 (sPD-L1) in 67 patients with ITP (24 newly diagnosed ITP [ndITP], 43 chronic ITP [cITP]) and 21 healthy controls (HCs). We determined decreased serum sPD-1 levels both in patients with ndITP and in patients with cITP when compared to HC. Moreover, there was a positive correlation between sPD-1 levels and platelet counts. The sPD-L1 levels were decreased in patients with ndITP when compared to patients with cITP. This is the first study investigating PD-1 signaling pathway in ITP. Decreased sPD-1 levels may have a role in ITP pathogenesis as without the inhibitory regulation of PD-1, sustained activation of T cells may cause inflammatory responses which is the case in ITP.