Cellular miR-150-5p may have a crucial role to play in the biology of SARS-CoV-2 infection by regulating nsp10 gene.

The role for circulating miRNAs as biomarkers of the COVID-19 disease remains uncertain. We analysed the circulating miRNA profile in twelve COVID-19 patients with moderate-severe disease. This analysis was conducted by performing next generation sequencing (NGS) followed by real-time polymerase chain reaction (RT-qPCR). Compared with healthy controls, we detected significant changes in the circulating miRNA profile of COVID-19 patients. The miRNAs that were significantly altered in all the COVID-19 patients were miR-150-5p, miR-375, miR-122-5p, miR-494-3p, miR-3197, miR-4690-5p, miR-1915-3p, and miR-3652. Infection assays performed using miRNA mimics in HEK-293 T cells determined miR-150-5p to have a crucial role in SARS-CoV-2 infection and this was based on the following data: (i) miR-150-5p mimic lowered in vitro SARS-CoV-2 infection; (ii) miR-150-5p inhibitor reversed the effects of miR-150-5p mimic on SARS-CoV-2 infection of cells; and (iii) a novel miRNA recognition element (MRE) was identified in the coding strand of SARS-CoV-2 nsp10, the expression of which could be inhibited by miR-150-5p mimic. Our findings identified crucial miRNA footprints in COVID-19 patients with moderate-severe disease. A combination of co-transfection and Western blotting experiments also determined the ability of miR-150-5p to inhibit SARS-CoV-2 infection via directly interacting with MRE in the coding strand of nsp10. Our investigation showed that a sharp decline in the miR-150-5p plasma levels in COVID-19 patients may support enhanced SARS-CoV-2 infection. Furthermore, this study provides insight into one possible mechanism by which COVID-19-induced changes to miR-150-5p levels may promote SARS-CoV-2 infection via modulating nsp10 expression.

Racial differences in incident de novo donor-specific anti-HLA antibody among primary renal allograft recipients: results from a single center cohort study.

Controversy exists as to whether African American (AA) transplant recipients are at risk for developing de novo donor-specific anti-human leucocyte antigen (HLA) antibody (dnDSA). We studied 341 HLA-mismatched, primary renal allograft recipients who were consecutively transplanted between 3/1999 and 12/2010. Sera were collected sequentially pre- and post-transplant and tested for anti-HLA immunoglobulin G (IgG) via single antigen bead assay. Of the 341 transplant patients (225 AA and 116 non-AA), 107 developed dnDSA at a median of 9.2 months post-transplant. AA patients had a 5-year dnDSA incidence of 35%. This was significantly higher than the 5-year dnDSA incidence for non-AA patients (21%). DQ mismatch (risk) and receiving a living-related donor (LRD) transplant (protective) were transplant factors associated with dnDSA. Within the AA patient cohort, HLA-DQ mismatch, not-receiving a LRD transplant, nonadherence and BK viraemia were the most common factors associated with early dnDSA (occurring <24 months post-transplant). Nonadherence and pretransplant diabetes history were the strong precursors to late dnDSA. Despite the higher rates of dnDSA in the AA cohort, post-dnDSA survival was the same in AA and non-AA patients. This study suggests that DQ matching, increasing LRD transplantation in AA patients and minimizing under-immunosuppression will be key to preventing dnDSA.

Incidence and impact of anti-HLA-DP antibodies in renal transplantation.

BACKGROUND: The role of anti-HLA-DP antibodies in renal transplantation is poorly defined. This study describes the impact of donor (donor-specific antibody [DSA]) and non-donor-specific antibodies against HLA-DP antigens in renal transplant patients. METHODS: Of 195 consecutive patients transplanted between September 2009 and December 2011, 166 primary kidney recipients and their donors were typed (high-resolution) for DP antigens. Sera taken pre-transplant and at 1, 3, 6, 9, and 12 months, and annually post-transplant were retrospectively tested for anti-DP antibodies using single-antigen beads. RESULTS: In 81 (49%) patients, anti-DP antibodies were found; 64% (n=52) of patients were positive in the pre-transplant samples and 36% (n=29) were positive exclusively post-transplant. The median time from transplantation to antibody was 20.9 months. Fifty-five percent (n=16) of the de novo anti-DP antibodies were accompanied by another de novo DSA. Anti-DP antibody-positive patients had a higher rate of rejection (compared with anti-DP antibody-negative patients, P=.01). The estimated glomerular filtration rate declined more with anti-DP antibodies (-5.5% vs +26%). CONCLUSIONS: Antibodies against HLA-DP antigens are common. De novo anti-DP antibodies commonly appear after acute rejection and accompany DSA, which makes it difficult to determine whether anti-DP antibodies are the cause or the consequence of graft injury.

Cardiovascular Health of North Carolina Undergraduates.

BACKGROUND: Cardiovascular disease is highly prevalent in Eastern North Carolina (ENC). In this study, we investigated cardiometabolic risk in young adults of ENC by sampling entrant undergraduates at East Carolina University (ECU). METHODS: From June to October of 2010, 525 undergraduates were screened for elevated body mass index (BMI), blood pressure, lipids, blood glucose, inactivity, smoking, history of diabetes or hypertension, and family history of coronary disease. Participants were classified as high-risk if they had 3 or more cardiovascular risk factors or as "MetS" if they satisfied the criteria for metabolic syndrome. RESULTS: Forty-four percent of those screened had 2 or more risk factors, 12.5% had 3 or more risk factors, and 1.3% met criteria for MetS. Low levels of high-density lipoprotein (27.6%), overweight status (27.2%), and inactivity (27.1%) were leading risks. Females had an increased risk of inactivity compared to males (relative risk [RR] = 1.81; 95% CI, 1.3-2.52). Blacks had a 4-fold higher risk of metabolic syndrome (RR = 4.21; 95% Cl, 1.0-18.4), and black females had a high risk for obesity (RR = 5.7; 95% CI, 2.5-13) and systolic blood pressure elevation (RR = 4.8; 95% Cl, 1.5-15). Students recognized cardiovascular disease as a valid risk to their well-being. CONCLUSION: ECU undergraduates have a high prevalence of multiple cardiovascular risk factors. High-risk and MetS students recognize cardiovascular disease as a significant health risk, but they mistakenly maintain the self-perception that they are healthy. Efforts to understand risk perception and personal strategies of risk application are needed for this population of young adults.

Health Screening Among a Rural, Hourly Workforce: North Carolina SERVIRE Project.

INTRODUCTION: This study demonstrates the feasibility of a novel, business-partnered, and worksite-based approach to healthcare access to facilitate chronic disease screening and diagnosis among rural hourly workers. The prevalence of undiagnosed and untreated diabetes and hypertension among screening participants was determined. METHODS: From February 2021 to June 2023, investigators partnered with 29 businesses to screen 1,114 workers. Health screenings included a demographic questionnaire, A1c testing for prediabetes (A1c of 5.7-6.4) and diabetes (A1c≥6.5), hypertension (Stage 1: systolic blood pressure of 130-139 mmHg; Stage 2: systolic blood pressure ≥140 mmHg), kidney disease (estimated glomerular filtration rate <60; urine protein ≥1+), and questionnaire assessment of stroke (CHA2DS2-VASc) and sleep apnea (STOP-bang) risk. RESULTS: Of the 1,114 individuals screened (n=632, 56.7% male; n=497, 44.6% Black)), 388 (36%) screened positive for prediabetes or diabetes. Diabetes was previously undiagnosed in 273 (70.4%) of these participants. More than half of the participants (n=680, 62.4%) had an elevated blood pressure reading during the screening, and the majority of these participants (n=445, 65.4%) had not been previously diagnosed with hypertension. In addition, 241 (21.6%) participants were at an increased risk of stroke (CHA2DS2-VASc≥2), and 182 (23.7%) had a STOP-Bang score ≥4, indicating an increased risk of obstructive sleep apnea. CONCLUSIONS: By partnering with local businesses to deliver worksite-based health screenings, high rates of undiagnosed and uncontrolled diabetes and hypertension were identified among the rural, hourly workforce. This worksite-based approach to healthcare access could facilitate early detection of chronic disease, improve patient engagement in the healthcare system, and ultimately yield better long-term public health outcomes.

Willingness to Vaccinate Against COVID-19: Predictors of Vaccine Uptake Among Adults in the US.

The uptake of the COVID-19 vaccine will determine the trajectory for improved population health and economic recovery from the COVID-19 pandemic. Identifying factors associated with vaccine acceptance is imperative as public health officials strategize to improve uptake. In this study, we identified predictors of vaccine willingness and acceptance using univariate logistic regression to model predictors and calculate odds ratios. Participants (N = 946) who reported greater vaccine willingness were male, older, and had a higher level of education and income. Behaviors indicative of reducing the spread of COVID-19 (e.g., testing) and perceived risk of COVID-19 infection were associated with vaccine willingness, as were participants who believed they were "highly likely" to be infected (by a factor of 8). Education tailored to demographic groups with low vaccine uptake should focus on the high degree of communicability associated with COVID-19. Implementing mobile healthcare screenings could remove barriers to healthcare, thereby improving health equity.

Utilization Patterns and Outcomes of People With Diabetes and COVID-19: Evidence From United States Medicare Beneficiaries in 2020.

Objective: Determine differences in utilization patterns, disease severity, and outcomes between patients with and without diabetes mellitus diagnosed with COVID-19 in 2020. Research Design and Methods: We used an observational cohort comprised of Medicare fee-for-service beneficiaries with a medical claim indicating a COVID-19 diagnosis. We performed inverse probability weighting between beneficiaries with and without diabetes to account for differences in socio-demographic characteristics and comorbidities. Results: In the unweighted comparison of beneficiaries, all characteristics were significantly different (P<0.001). Beneficiaries with diabetes were younger, more likely to be black, had more comorbidities, higher rates of Medicare-Medicaid dual-eligibility, and were less likely to be female. In the weighted sample, hospitalization rates for COVID-19 among beneficiaries with diabetes was higher (20.5% vs 17.1%; p < 0.001). Outcomes of hospitalizations were similarly worse among beneficiaries with diabetes: admissions to ICU during hospitalizations (7.78% vs. 6.11%; p < 0.001); in-hospital mortality (3.85% vs 2.93%; p < 0.001); and ICU mortality (2.41% vs 1.77%). Beneficiaries with diabetes had more ambulatory care visits (8.9 vs. 7.8, p < 0.001) and higher overall mortality (17.3% vs. 14.9%, p < 0.001) following COVID-19 diagnosis. Conclusion: Beneficiaries with diabetes and COVID-19 had higher rates of hospitalization, ICU use and overall mortality. While the mechanism of how diabetes impacts the severity of COVID-19 may not be fully understood, there are important clinical implications for persons with diabetes. A diagnosis of COVID-19 leads to greater financial and clinical burden than for their counterparts, persons without diabetes, including perhaps most significantly, higher death rates.

Optimizing tacrolimus therapy in the maintenance of renal allografts: 12-month results.

BACKGROUND: The determination of optimal tacrolimus (TAC) trough levels is needed to prevent adverse effects of calcineurin inhibitors. METHODS: Stable transplant recipients currently receiving cyclosporine (CsA) were assigned randomly (1:1:1) to continue CsA (target trough level of 50-250 ng/mL); or convert to "reduced" TAC (target trough level 3.0-5.9 ng/mL) or "standard" TAC (target trough level 6.0-8.9 ng/mL). RESULTS: At 12 months, there was a significant improvement in renal function in the reduced TAC versus CsA group with lower serum creatinine (P=0.004) and cystatin C (P<0.001), and higher estimated creatinine clearance (P=0.017). However, there were no statistically significant differences in any renal parameter in the standard TAC versus CsA group. Total and low-density lipoprotein cholesterol were significantly reduced in both TAC groups versus the CsA group (P<0.001). Patient and graft survival and episodes of biopsy-confirmed acute rejection were similar for all treatment groups, and no statistically significant differences were observed between groups in the incidence of new-onset diabetes or cardiac conditions, or in the prevalence of hyperglycemia, hypertension, or hyperlipidemia among patients who did not have these conditions at baseline. Alopecia developed more commonly among TAC-treated patients than CsA-treated patients (P<0.001). CONCLUSIONS: Compared with CsA continuation, conversion to reduced TAC target trough concentrations resulted in significantly improved renal function without increasing the risk of rejection. Conversion to TAC, regardless of target concentration, resulted in improved serum lipid profiles in kidney transplant recipients at 12 months.

Platelet adhesion testing may predict early hemodialysis arteriovenous graft and fistula failure in end-stage renal disease patients.

Vascular access thrombosis (VAT) is the most morbid and costly complication in end-stage renal disease (ESRD) patients. Although hypercoagulability is a major risk factor for VAT, in most patients, the cause of hypercoagulability cannot be identified despite clinical suspicion. In this study, platelet hyperreactivity was investigated for a possible role in the hypercoagulability of ESRD and VAT in 42 patients with arteriovenous (AV) grafts or fistulas. Platelet adhesion, platelet aggregation, and the history of VAT were assessed. The statistics included a nonparametric 2-factor ANOVA, a Mann-Whitney analysis, and a Kaplan-Meier analysis of hemodialysis angioaccess survival to examine platelet hyperadhesiveness as a predictor of access survival. The study showed a significant correlation between increased platelet adhesiveness and shortened survival of the primary hemodialysis angioaccess. Collagen-induced platelet aggregation reflected a significantly higher response in those with shortened access survival. These findings may have significant clinical implications for risk assessment and prevention of VAT.

Improvement in 3-month patient-reported gastrointestinal symptoms after conversion from mycophenolate mofetil to enteric-coated mycophenolate sodium in renal transplant patients.

BACKGROUND: The benefit of conversion from mycophenolate mofetil (MMF) to enteric-coated mycophenolate sodium (EC-MPS) in terms of gastrointestinal symptom burden has been evaluated previously using patient-reported outcomes. However, data are lacking concerning the sustained effect of conversion over time, and the potential impact of concomitant calcineurin inhibitor. METHODS: In this 3-month, prospective, multicenter, longitudinal, open-label trial, MMF-treated renal transplant patients with gastrointestinal symptoms receiving cyclosporine or tacrolimus were converted to equimolar doses of EC-MPS. Change in gastrointestinal symptom burden was evaluated using a validated Gastrointestinal Symptom Rating Scale (GSRS). RESULTS: A significant improvement in GSRS score was observed from baseline (2.61, 95% CI 2.54-2.68) to month 1 (1.87, 95% CI 1.81-1.93) after conversion to EC-MPS and was sustained to month 3 (1.81, 95% CI 1.74-188; both P<0.0001 versus baseline). The mean change in overall GSRS score from baseline to month 1 was -0.74 overall (cyclosporine: -0.73 and tacrolimus: -0.74; all P<0.0001 versus baseline), with a slight further improvement (-0.79) at month 3 (cyclosporine: -0.82 and tacrolimus: -0.78; all P<0.0001 versus baseline). A significant improvement in GSRS subscale scores was also observed in the total population regardless of calcineurin inhibitor at month 1, sustained to month 3 (all P<0.0001 versus baseline). The improvement in GSRS score postconversion was similar in African-American and non-African-American patients, and in diabetic and nondiabetic patients. CONCLUSIONS: This exploratory study in 728 patients demonstrates that following conversion from MMF to EC-MPS, regardless of concomitant calcineurin inhibitor, GSRS is improved and sustained over 3 months.

Risk Stratification of Human Leukocyte Antigen Class II Donor Specific Antibody Positive Patients by Immunoglobulin G Subclasses.

BACKGROUND: Human leukocyte antigen (HLA) antibodies are a major cause of graft loss in mismatched transplant recipients. However, the time to graft loss resulting from antibody induced injury is unpredictable. The unpredictable nature of antibodies may be related to the subclass of antibodies. In this study, HLA immunoglobulin G (IgG) subclasses were investigated to determine whether a unique IgG subclass composition could better identify those patients at eminent risk for graft loss. METHODS: The serial serum samples from the 57 patients with post-transplant HLA class II donor specific antibodies (DSA) were tested for the three IgG subclasses (IgG1, IgG3, and IgG4). RESULTS: IgG3 and IgG4 were highly prevalent in failed patients compared to functioning patients (82 % vs. 34%, 45% vs. 20%, respectively). IgG3 development showed a distinct subclass trend between failed and functioning patients with poor graft survival (log rank p=0.0006). IgG1 was almost equally abundant in both groups (100% and 97%, respectively). Of the 5 patterns of IgG subclass combinations observed, IgG1+3+ showed the strongest association with graft failure (hazard ratio 3.14, p=0.007). CONCLUSION: Patients with IgG3 subclass HLA DSA showed lower graft survival. Post-transplant monitoring for IgG subclasses rather than total IgG monitoring may identify patients at risk for graft failure.

Improved Long-Term Survival in Kidney Transplant Recipients with Donor-Specific HLA Antibodies After Mycophenolic Acid Escalation.

The development of donor specific antibodies (DSA) post transplant has been associated with chronic rejection and graft failure. In a longitudinal study, we have shown that increases in DSA precede rejection by months, thus allowing time for intervention. We hypothesized that mycophenolic acid (MPA) dose increases may reduce and/or stabilize DSA strength and also preserve renal function. Thirty stable DSA positive kidney transplant recipients participated in this Institutional Review Board approved, exploratory, open-label, single center study to assess the efficacy of MPA dose escalation in patients with DSA. MPA escalation was well tolerated and most patients were able to take higher doses for at least two years (duration of the study). In addition, MPA escalation is safe and participants had no significant side effects such as cytomegalovirus and BK infections. Long-term allograft survival of the MPA escalation group was superior when compared with the control group (p = 0.018). This pilot study indicates that escalation of MPA is safe and may stabilize DSA. In addition, five-year follow up demonstrates improved long-term survival with MPA escalation compared with DSA positive recipients receiving the standard of care. Additional studies using larger cohorts are warranted.

Impact of IgM and IgG3 anti-HLA alloantibodies in primary renal allograft recipients.

BACKGROUND: With standard IgG donor-specific anti-HLA antibody (DSA) testing, it is unclear which immunoglobulin-G (IgG) DSA positive patients will fail. We looked further into the immune response by studying immunoglobulin-M (IgM) and IgG subclass 3 (IgG3) DSA to determine if these identify the IgG DSA patients at highest risk for allograft loss. METHODS: In 189 consecutively transplanted primary renal allograft recipients, sera were collected sequentially pre- and posttransplant. Of the 189, 179 patients had sera available to retrospectively test for anti-HLA IgG, IgM, and IgG3 antibodies via LABScreen single-antigen bead assay and were included in the study. All patients had a negative crossmatch. Per patient, all DSA (IgM, IgG3, and IgG) refers to the same serologic specificity. RESULTS: Overall, 100 (56%) patients developed an alloimmune response (IgM or IgG DSA positive, or both). Ninety-five patients developed IgM DSA and 47 patients developed IgG DSA. IgM DSA was detected in 42 of 47 patients with IgG DSA. IgM DSA alone did not increase the allograft loss risk, whereas IgG DSA did (P=0.002). Once IgG DSA appeared, IgM DSA persisted in 33 patients and an isotype switch to IgG3 positive DSA occurred in 25 patients. Patients with IgM persistent IgG3 positive DSA (n=19) were more likely to have allograft failure than those without (P=0.02). CONCLUSION: This study shows the evolution of the humoral immune response from IgM to IgG DSA posttransplant. We found that development of IgM persistent IgG3 positive DSA identifies the most dangerous IgG DSA subpopulation.

Changes in successive measures of de novo donor-specific anti-human leukocyte antigen antibodies intensity and the development of allograft dysfunction.

BACKGROUND: Many patients develop de novo donor-specific anti-human leukocyte antigen antibodies (dnDSA) after transplantation. Despite development of dnDSA, not all patients will immediately fail. This study analyzes dnDSA intensity and longitudinal trends as prospective clinical parameters to assess subsequent allograft function. METHODS: Twenty-four patients with dnDSA onset in the first 2 years after transplantation received antibody monitoring by LABScreen single antigen beads. Estimated glomerular filtration rate (eGFR) was recorded at time of dnDSA onset and up to 24 months thereafter. The dnDSA mean fluorescence intensity (MFI) of the stable function patient group (n=8; eGFR decline ≤ 25%) was compared with the impaired function patient group (n=16; eGFR decline>25%) using first year peak MFI (pMFI), eight month MFI change (ΔMFI), and eighteen month MFI trend (MFI slope). RESULTS: Both groups showed similar dnDSA characteristics (time to onset after transplantation, class I/II distribution, and initial MFI). Between groups, MFI trends were analyzed. Impaired patients showed a higher pMFI during the first year (median pMFI, 13,055 vs. 2,397; P=0.007). Longitudinal analysis revealed that ΔMFI was strongly associated with dysfunction. Both a ΔMFI increase greater than 20% as well as a stronger increase (ΔMFI>50%) were followed by graft dysfunction in almost all patients and could significantly differentiate between stable and impaired function patients (P=0.001 and P=0.04, respectively). CONCLUSION: Our study suggests that tracking dnDSA intensity, particularly in the early period after onset, is important to estimate the impact of dnDSA on the allograft and could, therefore, determine help on how best to monitor patients with dnDSA.

Trends and characteristics in early glomerular filtration rate decline after posttransplantation alloantibody appearance.

BACKGROUND: Approximately 7% to 9% of patients with donor-specific anti-human leukocyte antigen (HLA) antibodies (DSA) fail within 1 year post-DSA onset. However, little is known as to how this DSA-associated failure temporally progresses. This longitudinal study investigates DSA's temporal relationship to allograft dysfunction and identifies predictors of allograft function's progressive deterioration post-DSA. METHODS: A cohort of 175 non-HLA identical patients receiving their first transplant between March 1999 and March 2006 were analyzed. Protocol testing for DSA via single antigen beads was done before transplantation and at 1, 3, 6, 9, and 12 months after transplantation then annually. Estimated glomerular filtration rate (eGFR) was analyzed before and after DSA onset. RESULTS: Forty-two patients developed DSA and had adequate eGFR information for analysis. Before DSA onset, the 42 patients had stable eGFR. By 1 year post-DSA, the cohort's eGFR was significantly lower (P<0.001); however, 30 of 42 had stable function. Twelve patients had failure or early allograft dysfunction (eGFR decline >25% from DSA onset). Those who failed early (by 1 year post-DSA) had more antibody-mediated rejection than stable patients (P=0.03). Late failures (after 1 year post-DSA) were predictable with evidence of early allograft dysfunction (eGFR decline >25% by 1 year post-DSA; P<0.001). Early allograft dysfunction preceded late failure by nearly 1 year. CONCLUSIONS: DSA is temporally related to allograft function deterioration. However, in many cases, late allograft failures are preceded by early allograft dysfunction. Therefore, monitoring for early allograft dysfunction provides treating physicians with a window of opportunity for treatment or continued monitoring.

Racial differences in survival among hemodialysis patients after coronary artery bypass grafting.

The aim of this study was to examine racial differences in long-term survival among hemodialysis patients after coronary artery bypass grafting (CABG). To our knowledge this has not been previously addressed in the literature. Black and white hemodialysis patients undergoing first-time, isolated CABG procedures between 1992 and 2011 were compared. Survival probabilities were computed using the Kaplan-Meier product-limit method and stratified by race. Hazard ratios (HR) and 95% confidence intervals (CI) were computed using a Cox regression model. A total of 207 (2%) patients were on hemodialysis at the time of CABG. White (n = 80) hemodialysis patients had significantly decreased 5-year survival compared with black (n = 127) patients (adjusted HR = 1.9, 95% CI = 1.2-2.8). Our finding provides useful outcome information for surgeons, primary care providers, and their patients.

Implications of Hemodialysis in Patients Undergoing Coronary Artery Bypass Grafting.

BACKGROUND: Cardiovascular disease is the leading cause of morbidity and mortality in patients on hemodialysis. To our knowledge, no studies have examined long-term outcomes of hemodialysis patients following coronary artery bypass grafting (CABG) in a predominately rural, low-income, and racially dichotomous population. METHODS: Long-term survival of hemodialysis patients undergoing non-emergent, isolated CABG was compared with non-hemodialysis patients. Survival probabilities were computed using the Kaplan-Meier product limit method and stratified by hemodialysis. Hazard ratios (HR) and 95% confidence intervals (95%CI) were computed using a Cox regression model. RESULTS: Hemodialysis patients (n=220) had shorter long-term survival than non-hemodialysis patients (median survival=3.3 versus 14 years, p<0.0001). The survival difference remained statistically significant after adjusting for clinically relevant variables (HR=5.2, 95%CI=4.4-6.2). CONCLUSION: Hemodialysis patients had significantly shorter long-term survival compared with non-hemodialysis patients after CABG. Further research is needed to address the cost and policy implications of our findings, especially among priority populations.

Novel biomarker combination predicts long-term allograft outcome.

Donor specific human leukocyte antigen (HLA) antibodies (DSA) are a significant cause of allograft failure. However, it has been reported that some DSA negative patients still experience allograft failure. In addition, some DSA positive patients maintain good graft function for >20 years. These findings suggest that while DSA is a cause of failure, it is not the sole risk factor for graft dysfunction and that the presence of DSA alone may not predict the time course of graft failure. Here, we report the predictive value of a proprietary panel of four biomarkers in long-term renal allograft outcome. A total of 310 consecutive patients, who received kidney transplants between 1999 and 2012, were included in this study. Recipient sera was tested for HLA antibodies and biomarkers at 3, 6, 12, 24, and 36 months post-transplant. HLA antibodies were identified using Labscreen single antigen beads. The biomarker combination (BMC) test consisted of a proprietary panel of 4 biomarkers and was performed using Luminex. Sera were defined as positive when any one of the 4 biomarkers became detectable. Sera of normal healthy people were used as negative controls. Graft survival analyses were performed and compared between different patient groups based on the positivity of DSA and BMC. Our results indicate that 57% of DSA negative patients and 54% of DSA positive patients had detectable biomarkers. There was no significant difference in BMC positive patients between the DSA positive and negative groups, which suggests that presence of BMC is not associated with HLA DSA. DSA positive patients had a 10% lower 10-year graft survival rate than patients without DSA, while BMC positive patients had a 25% lower 10-year graft survival rate than patients without detectable BMC. When DSA negative patients were divided into two groups based on the positivity of BMC, BMC positive patients had a 20% lower 10-year graft survival rate compared to BMC negative patients (p<0.05). Similarly, when DSA positive patients were divided into two groups based on the positivity of BMC, BMC positive patients had a 30% lower 10-year graft survival rate compared to BMC negative patients (p<0.01). When both DSA and BMC testing results were considered, DSA and BMC double positive patients had the lowest and double negative patients had the highest graft survival rates. The survival rates for the BMC alone and DSA alone positive groups were in between (p<0.001). Multivariate Cox models confirmed that BMC was an independent risk factor for graft failure, with a higher hazard ratio than DSA (BMC=2.60 versus DSA=1.64). In conclusion, serum BMC is an independent predictor of graft failure. BMC was more significantly associated with graft failure than DSA. In combination with DSA, BMC better predicted graft outcome than DSA or BMC alone.