Non-viremic Patients Research Articles

Detection of JCV or BKV viruria and viremia after kidney transplantation is not associated with unfavorable outcomes.

Studies analyzing the relationship between BK polyomavirus (BKV) or JC polyomavirus (JCV) infection and kidney transplant (KT) long term clinical outcomes are scarce. Therefore, we evaluated this relationship in a single-center retrospective cohort of 288 KT patients followed for 45.4(27.5; 62.5) months. Detection of BKV viremia in two consecutive analyses led to discontinuation of antimetabolite and initiation of mammalian target of rapamycin inhibitor. Outcome data included de novo BKV and/or JCV viremia and/or viruria after KT, death-censored graft survival and patient survival. BKV viruria and viremia were detected in 42.4% and 22.2% of KT recipients, respectively. BKV viremic patients had higher urinary BKV viral loads at the onset of viruria, when compared to nonviremic patients (7 log10 vs. 4.9 log10 cp/mL, p < 0.001). JCV viruria was identified in 38.5% of KT patients; the 5.9% of KT recipients who developed JCV viremia had higher JCV urinary viral loads at the onset of viruria, when compared to non-viremic patients (5.3 vs. 3.7 log10 cp/mL, p = 0.034). No differences were found in estimated glomerular filtration rate at the end of follow up, when comparing BKV or JCV viruric or viremic patients with nonviremic patients. No association was found between JCV or BKV viruria or viremia and death/graft failure. Therefore, higher BKV urinary viral loads at the onset could serve as an early maker of over immunosuppression. JCV and BKV replication was not associated with inferior clinical outcomes in KT patients with the above-mentioned immunosuppression strategy.

Viremia as a predictor of absence of serious bacterial infection in children with fever without source

Most children with fever without source (FWS) require diagnostic laboratory tests to exclude a serious bacterial infection (SBI), often followed by admission and empirical antibiotics. As febrile children with a viral infection are less likely to have a SBI, identifying patients with systemic viral infection could contribute to exclude SBI. We evaluated whether the presence of virus in the blood could be used as a biomarker to rule out SBI. Children < 3 years old with FWS were prospectively enrolled and had real-time (reverse-transcription) PCR performed on the blood for adenovirus, enterovirus, parechovirus, and HHV6. 20/135 patients had SBI, and in 47/135, at least one virus was detected in the blood. Viremia had a higher sensitivity and negative predictive value (90% and 96%) to rule out SBI compared to CRP (65% and 93%) and PCT (55% and 90%). The odds ratio (OR) for the presence of SBI among non-viremic patients was 5.8 (p = 0.0225), compared to 5.5 for CRP ≥ 40 mg/l (p = 0.0009) and 3.7 for PCT ≥ 0.5 ng/mL (0.0093). This remained significant after adjusting for CRP and PCT (OR 5.6 and 5.9, respectively; p = 0.03 for both). Area under the ROC curve for CRP and PCT were 0.754 and 0.779, respectively, but increased to 0.803 and 0.832, respectively, when combined with viremia.Conclusion: The presence of viremia had a better performance than commonly used biomarkers to rule-out SBI and could potentially be used in conjunction with CRP and/or PCT in the evaluation of children with FWS. Larger studies should evaluate the role of point-of-care testing of viruses by (revere-transcription) PCR in the plasma in management algorithms of children with FWS.What is Known:• Most children with FWS have a viral infection, but up to 15% have a SBI; most require laboratory tests, and many admission and empirical antibiotics.• Children with a viral infection are less likely to have a SBI.What is New:• Children with a systemic viral infection are less likely to have an SBI.• Viremia is a better predictor of absence of SBI than commonly used biomarkers and could potentially be used in conjunction with CRP and/or PCT in the evaluation of children with FWS.

Achieving tolerance modifies cancer susceptibility profiles in liver transplant recipients.

Long-term survival of transplant recipients is significantly impacted by malignancy. We aimed to determine whether calcineurin inhibitor (CNI)-treated recipients converted to and weaned off molecular target of rapamycin inhibitor (mTOR-I) therapy have favorable changes in their molecular profiles in regard to malignancy risk. We performed gene expression profiling from liver biopsy and blood (PBMC) specimens followed by network analysis of key dysregulated genes, associated diseases and disorders, molecular and cellular functions using IPA software. Twenty non-immune, non-viremic patients were included, and 8 of them achieved tolerance. Two comparisons were performed: (1) tolerance time point vs tacrolimus monotherapy and (2) tolerance time point vs sirolimus monotherapy. Upon achieving tolerance, IPA predicted significant activation of DNA damage response (p=5.40e-04) and inhibition of DNA replication (p=7.56e-03). Conversion from sirolimus to tolerance showed decrease in HCC (p=1.30e-02), hepatic steatosis (p=5.60e-02) and liver fibrosis (p=2.91e-02) associated genes. In conclusion, this longitudinal study of patients eventually achieving tolerance reveals an evolving molecular profile associated with decreased cancer risk and improved hepatic steatosis and liver fibrosis. This provides a biological rationale for attempting conversion to mTOR-I therapy and tolerance following liver transplantation particularly in patients at higher risk of cancer incidence and progression post-transplant.

Circular RNA Profiles in Viremia and ART Suppression Predict Competing circRNA-miRNA-mRNA Networks Exclusive to HIV-1 Viremic Patients.

Since the onset of the HIV-1/AIDS epidemic in 1981, 75 million people have been infected with the virus, and the disease remains a public health crisis worldwide. Circular RNAs (circRNAs) are derived from excised exons and introns during backsplicing, a form of alternative splicing. The relevance of unconventional, non-capped, and non-poly(A) transcripts to transcriptomics studies remains to be routinely investigated. Knowledge gaps to be filled are the interface between host-encoded circRNAs and viral replication in chronically progressed patients and upon treatment with antiviral drugs. We implemented a bioinformatic pipeline and repurpose publicly archived RNA sequence reads from the blood of 19 HIV-1-positive patients that previously compared transcriptomes during viremia and viremia suppression by antiretroviral therapy (ART). The in silico analysis identified viremic patients’ circRNA that became undetectable after ART. The circRNAs originated from a subset of host genes enriched in the HDAC biological pathway. These circRNAs and parental mRNAs held in common a small collection of miRNA response elements (MREs), some of which were present in HIV-1 mRNAs. The function of the MRE-containing target mRNA enriched the RNA polymerase II GO pathway. To visualize the interplay between individual circRNA–miRNA–target mRNA, important for HIV-1 and potentially other diseases, an Interactive Circos tool was developed to efficiently parse the intricately competing endogenous network of circRNA–miRNA–mRNA interactions originating from seven circRNA singled out in viremic versus non-viremic patients. The combined downregulation of the identified circRNAs warrants investigation as a novel antiviral targeting strategy.

Mass cytometry reveals single-cell kinetics of cytotoxic lymphocyte evolution in CMV-infected renal transplant patients

Cytomegalovirus (CMV) infection is associated with graft rejection in renal transplantation. Memory-likenatural killer (NK) cells expressing NKG2C and lacking FcεRIγ are established during CMV infection. Additionally, CD8+ T cells expressing NKG2C have been observed in some CMV-seropositive patients. However, invivo kinetics detailing the development and differentiation of these lymphocyte subsets during CMV infection remain limited. Here, we interrogated the invivo kinetics of lymphocytes in CMV-infected renal transplant patients using longitudinal samples compared with those of nonviremic (NV) patients. Recipient CMV-seropositive (R+) patients had preexisting memory-like NK cells (NKG2C+CD57+FcεRIγ-) at baseline, which decreased in the periphery immediately after transplantation in both viremic and NV patients. We identified a subset of prememory-like NK cells (NKG2C+CD57+FcεRIγlow-dim) that increased during viremia in R+ viremic patients. These cells showed a higher cytotoxic profile than preexisting memory-like NK cells with transient up-regulation of FcεRIγ and Ki67 expression at the acute phase, with the subsequent accumulation of new memory-like NK cells at later phases of viremia. Furthermore, cytotoxic NKG2C+CD8+ T cells and γδ T cells significantly increased in viremic patients but not in NV patients. These three different cytotoxic cells combinatorially responded to viremia, showing a relatively early response in R+ viremic patients compared with recipient CMV-seronegative viremic patients. All viremic patients, except one, overcame viremia and did not experience graft rejection. These data provide insights into the invivo dynamics and interplay of cytotoxic lymphocytes responding to CMV viremia, which are potentially linked with control of CMV viremia to prevent graft rejection.

Reducing Read Time of Point-of-Care Test Does Not Affect Detection of Hepatitis C Virus and Reduces Need for Reflex RNA

Global elimination of hepatitis C virus (HCV) will require increases in diagnosis. Point of care (POC) tests that detect antibodies against HCV can be useful for testing large and difficult to reach populations. The most accurate POC test requires a 20 min read time to identify antibody-positive samples. We investigated whether viremic patients could be identified using a shorter read time, to increase efficiency and reduce the need for reflex tests (a follow-up test for HCV RNA on the same specimen to confirm viremia). Patients with past or current HCV infections provided samples at 2 clinics in Canada for evaluation by the OraQuick HCV Rapid antibody POC test. A community HCV-screening program in Madrid, Spain (real-world cohort) invited people to be tested for HCV with the same OraQuick test. Patients provided samples of whole blood, via finger prick. Fingerprick samples were tested immediately after collection. In the clinic cohort, photographs of the developing test were taken at 15 second intervals, and blinded readers recorded the time to positivity. In the real-world cohort, readers recorded the OraQuick result at 5 minutes, and each minute after, up to 10 minutes, and then again at 20 minutes; viremia was then evaluated using a POC HCV RNA test (GeneXpert HCV Viral Load Assay). Sera from viremic and non-viremic clinic patients were used to quantify antibody titers to investigate the relationship between the time of band appearance and antibody concentration. Fisher's exact test and exact logistic regression were used to determine factors associated with a positive result at 5 minutes. Blood from all viremic patients produced a positive result in the antibody POC test by 5 min. Median time to a positive result for 171 viremic patients was 2.6 min (range, 1.8-4.6 min), vs 4.1 min (range, 2.3-14.4 min) for 108 patients with resolved infection (P < .001). The 5-min threshold identified all viremic cases among 176 HCV antibody-positive patients in the real-world cohort, confirmed by testing for HCV RNA. In the pooled cohorts, antibody positivity at 5 min identified viremic patients with 100% sensitivity (95% CI, 98.4%-100%); the negative predictive value was 100% (95% CI, 94.9%-100%). The positive predictive value at 5 min was 62.0% (95% CI, 56.7%-67.0%) and therefore insufficient alone to detect viremia; an HCV RNA test would still be necessary to confirm active infection. The wait time for the OraQuick HCV Rapid antibody POC blood test can be reduced from 20 min to 5 min and continue to reliably identify patients with HCV infection. Shortening the test time could increase high-throughput screening, reduce loss to follow up, and reduce the need for reflex HCV RNA testing.

Distinct serum HBsAg levels show minimal effect on gene expression profiles of blood leukocyte subpopulations in non-viremic chronic HBV patients

Distinct serum HBsAg levels show minimal effect on gene expression profiles of blood leukocyte subpopulations in non-viremic chronic HBV patients

Exclusion of Hepatitis C Viremia by Early Read of Point-of-Care Antibody Test

Background: To achieve global hepatitis C virus (HCV) elimination, scale-up of testing is required as diagnosis rates remain low. Point-of-care (POC) antibody testing increases access for difficult-to-reach populations, however the current 20-40 minute read time limits utility. We sought to determine whether there was a relationship between time to antibody positivity and presence of viremia. Methods: HCV viremic and non-viremic antibody-positive patients were tested with the OraQuick POC HCV antibody test using whole blood or oral fluid. Results were recorded at 15 second intervals to determine time to positivity. Results were validated in an independent screening cohort of high-risk individuals. Findings: All viremic patients had a positive blood result by 5 minutes. Median time to positivity for 171 viremic patients was 2.6 (1.8-4.6) versus 4.1 minutes (2.3-14.4) for 108 with resolved infection (p<0.001). The 5-minute threshold was confirmed in 176 HCV-antibody-positive individuals in a real-world cohort. Overall, the sensitivity for viremic patients at 5 minutes was 100% (95% CI 98.4-100%) and the negative predictive value was 100% (95% CI 94.9-100%). The positive predictive value at 5 minutes was inadequate to avoid HCV RNA testing 62.0% (95% CI 56.7-67.0%) entirely. Time to positivity was not predictive of viremia in oral fluid. Interpretation: A negative result on the OraQuick antibody test at 5 minutes is adequate to exclude HCV viremia, enabling high throughput screening and reducing the need for reflex HCV RNA testing. Funding Statement: This study was supported by the Viral Hepatitis Care Network (VIRCAN) and 3000 OraQuick HCV antibody kits were received in-kind from OraSure. JVL is supported by a Spanish Ministry of Science, Innovation and Universities Miguel Servet grant (Instituto de Salud Carlos III/ESF, European Union (CP18/00074)). Declaration of Interests: Mia J. Biondi has served as a speaker, a consultant, and/or advisory board member for Gilead, AbbVie, and Specialty Rx; and received research funding from Gilead and AbbVie. David Fletcher has served as an advisory board member for Gilead, AbbVie, and Merck. Hemat Shah has served as a speaker, a consultant and/or an advisory board member for AbbVie, Gilead, Intercept, Lupin, and Merck; and has received research funding from Boehringer-Ingelheim and Gilead. Betttina E. Hansen as served as a speaker, a consultant and/or an advisory board member for Intercept, Cymabay, Albireo, Mirum, ChemomAb, and Calliditas; and has received research funding from Intercept, Cymabay, Albireo and Mirum. Jeffrey V. Lazarus has served as a speaker, a consultant, and/or advisory board member for Gilead and AbbVie; and received research funding from GlaxoSmithKline, Cepheid, and Janssen. Harry L.A. Janssen has served as a speaker, a consultant and/or an advisory board member for Arbutus, Arena, Enyo, Gilead Sciences, GlaxoSmithKline, Janssen, Medimmune, Merck, Roche, Vir-Bio, Viroclinics; and has received research funding from AbbVie, Arbutus, Bristol-Myers Squibb, Gilead Sciences, Janssen, Medimmune, Merck, and Roche. Jordan J. Feld has served as a speaker, a consultant and/or an advisory board member for AbbVie, Enanta, Contravir, Roche, and Abbott; and has received research funding from AbbVie, Gilead, Janssen and Abbott. All other authors declare no conflict of interest. Ethics Approval Statement: All patients provided written informed consent and the protocol was approved by the Research Ethics Board of the University Health Network. The protocol for the Madrid validation cohort was reviewed by El cribado tiene un consentimiento informado y ha pasado el Comite de Etica del Hospital Gregorio Maranon and all individuals provided written informed consent.

Characterization of Leukocytes From HIV-ART Patients Using Combined Cytometric Profiles of 72 Cell Markers.

Motivation: Mass cytometry is a technique used to measure the intensity levels of proteins expressed by cells, at a single cell resolution. This technique is essential to characterize the phenotypes and functions of immune cell populations, but is currently limited to the measurement of 40 cell markers that restricts the characterization of complex diseases. However, algorithms and multi-tube cytometry techniques have been designed for combining phenotypic information obtained from different cytometric panels. The characterization of chronic HIV infection represents a good study case for multi-tube mass cytometry as this disease triggers a complex interactions network of more than 70 cell markers.Method: We collected whole blood from non-viremic HIV-infected patients on combined antiretroviral therapies and healthy donors. Leukocytes from each individual were stained using three different mass cytometry panels, which consisted of 35, 32, and 33 cell markers. For each patient and using the CytoBackBone algorithm, we combined phenotypic information from three different antibody panels into a single cytometric profile, reaching a phenotypic resolution of 72 markers. These high-resolution cytometric profiles were analyzed using SPADE and viSNE algorithms to decipher the immune response to HIV.Results: We detected an upregulation of several proteins in HIV-infected patients relative to healthy donors using our profiling of 72 cell markers. Among them, CD11a and CD11b were upregulated in PMNs, monocytes, mDCs, NK cells, and T cells. CD11b was also upregulated on pDCs. Other upregulated proteins included: CD38 on PMNs, monocytes, NK cells, basophils, B cells, and T cells; CD83 on monocytes, mDCs, B cells, and T cells; and TLR2, CD32, and CD64 on PMNs and monocytes. These results were validated using a mass cytometry panel of 25 cells markers.Impacts: We demonstrate here that multi-tube cytometry can be applied to mass cytometry for exploring, at an unprecedented level of details, cell populations impacted by complex diseases. We showed that the monocyte and PMN populations were strongly affected by the HIV infection, as CD11a, CD11b, CD32, CD38, CD64, CD83, CD86, and TLR2 were upregulated in these populations. Overall, these results demonstrate that HIV induced a specific environment that similarly affected multiple immune cells.

The impact of hepatitis C viremia status on lung functions in chronic hepatitis c patients.

Previous trials have investigated the effect of hepatitis C on lung functions; however, the role of viral load levels is unclear. The aim of this study was to investigate the effect of HCV viremia status on lung functions. This study was in 60 patients with chronic hepatitis C (CHC). Patients were classified into three groups (non-viremic, low-viremic and high-viremic) based on serum HCV RNA levels. Spirometric parameters (FEV1, FVC, FEV1/FVC) and the proportion of patients with spirometric abnormalities were compared between three groups. High-viremic and low-viremic patients showed a significantly higher prevalance of spirometric abnormality than observed in non-viremic patients (p=0.02). Moreover, there was a significant moderate correlation between viremia level and the percentage of spirometric abnormalities (Cramer's U value=0.452, p=0.002). High-viremic patients were 14.2 times more likely to exhibiting pulmonary dysfunction than non-viremic patients. Additionally, spirometric parameters FEV1 and FVC were significantly reduced in high-viremic and low-viremic patients compared to those in non-viremic patients (p=0.013 and p<0.001 respectively). These results indicate that persistent HCV infection may be associated with reduced pulmonary functions, especially in patients with high viremia levels. Therefore, these patients should be carefully monitored for lung function.

Detection of Occult Hepatitis C Virus Infection (OCI) in Bone Marrow Mononuclear Cells

Background and aim: Occult HCV infection (OCI) diagnosis by cellular-PCR encourages further research for more understanding of extrahepatic tissues affection during non-viremias. OCI among Egyptians recruits more attention because of the convenient diagnostic tools and the availability of direct acting antivirals (DAAs). Our study aims to recognize HCV-infection of bone marrow mononuclear cells (BMMCs) in antiviral IgG-antibody seropositive nonviremic subjects. Subjects and Methodology: Our case control study included 70 subjects who were divided into three groups: - I: non-viremic IgG-seronegative controls (n=25), II: naive HCV-RNA viremic IgG-seropositive controls (n=15), III: naive non-viremic IgG-seropositive patients (n=30). Sera of all study subjects were screened for anti-HCV IgGantibodies by ELISA and for HCV-RNA by PCR. BMMCs and peripheral blood mononuclear cells (PBMCs) of all study populations were tested for HCV-RNA infections by cellular-PCR. Results and conclusions: HCV-RNA viremias were seen when bone-marrow is functioning (P<0.05), while nonviremias were common in bone-marrow failure (P<0.05). There was 100% concordant positivity between serum and cellular HCV-RNA-PCRs in viremic patients. HCV-BMMCs and PBMCs PCRs respectively added extra 20% (P=0.024) and 13.33% (P=0.119) as newly diagnosed infections compared to none by serum-PCR in IgG-antibody seropositive non-viremic subjects. Respectively, non-viremic population (n=55) showed 14.29% and 9.1% positive intra-mononuclear cells HCV-RNA infection in bone marrow (P=0.002) and peripheral blood (P=0.057) compared to serum PCR. In conclusion, consecutive screening of peripheral blood and bone marrow mononuclear cells for HCV-RNA detection by cellular-PCR recognizes naive OCI in IgG-seropositive non-viremic patients. Functioning bone marrow would be required for persistence of HCV-RNA viremia.

Detection of Occult Hepatitis C Virus Infection (OCI) in Bone Marrow Mononuclear Cells

Background and aim: Occult HCV infection (OCI) diagnosis by cellular-PCR encourages further research for more understanding of extrahepatic tissues affection during non-viremias. OCI among Egyptians recruits more attention because of the convenient diagnostic tools and the availability of direct acting antivirals (DAAs). Our study aims to recognize HCV-infection of bone marrow mononuclear cells (BMMCs) in antiviral IgG-antibody seropositive nonviremic subjects. Subjects and Methodology: Our case control study included 70 subjects who were divided into three groups: - I: non-viremic IgG-seronegative controls (n=25), II: naive HCV-RNA viremic IgG-seropositive controls (n=15), III: naive non-viremic IgG-seropositive patients (n=30). Sera of all study subjects were screened for anti-HCV IgGantibodies by ELISA and for HCV-RNA by PCR. BMMCs and peripheral blood mononuclear cells (PBMCs) of all study populations were tested for HCV-RNA infections by cellular-PCR. Results and conclusions: HCV-RNA viremias were seen when bone-marrow is functioning (P<0.05), while nonviremias were common in bone-marrow failure (P<0.05). There was 100% concordant positivity between serum and cellular HCV-RNA-PCRs in viremic patients. HCV-BMMCs and PBMCs PCRs respectively added extra 20% (P=0.024) and 13.33% (P=0.119) as newly diagnosed infections compared to none by serum-PCR in IgG-antibody seropositive non-viremic subjects. Respectively, non-viremic population (n=55) showed 14.29% and 9.1% positive intra-mononuclear cells HCV-RNA infection in bone marrow (P=0.002) and peripheral blood (P=0.057) compared to serum PCR. In conclusion, consecutive screening of peripheral blood and bone marrow mononuclear cells for HCV-RNA detection by cellular-PCR recognizes naive OCI in IgG-seropositive non-viremic patients. Functioning bone marrow would be required for persistence of HCV-RNA viremia.

Use of Hepatitis C Virus Antibody-Positive Donor Livers in Hepatitis C Nonviremic Liver Transplant Recipients

Given the shortage of available liver grafts, transplantation (LTx) of hepatitis C virus antibody-positive, nucleic acid test-negative (HCV Ab+/NAT-) livers into nonviremic HCV recipients can expand the donor pool. Having previously described the sentinel experience of HCV Ab+/NAT- allografts in nonviremic recipients, we report the growth and extended follow-up of this program for 55 patients compared with recipients of Public Health Services (PHS) increased-risk donor HCV Ab-/NAT- allografts. A prospective review of all HCV nonviremic LTx patients receiving HCV Ab+/NAT- organs between March 2016 and August 2018 was performed. All HCV Ab+/NAT- organ recipients underwent HCV testing at 3 months and 1-year post-LTx to determine HCV transmission. Fifty-five HCV nonviremic candidates received HCV Ab+/NAT- organs; 64% male, median age 59 years (range 36 to 69 years) and median Model for End-Stage Liver Disease score of 22.5. Two recipients were excluded due to death before HCV testing. The HCV disease transmission occurred in 5 recipients (9%). Of these, 4 (80%) underwent anti-HCV treatment with eradication of virus. No patient found to be negative at 3 months seroconverted at 1-year follow-up. No patients who received PHS increased-risk donor HCV Ab-/NAT- organs had viremia develop (0 of 57) and there was no difference in graft and renal function, complications, or survival between HCV Ab+/NAT- recipients and PHS increased-risk donor HCV Ab-/NAT- recipients. We report the largest experience with LTx from HCV Ab+/NAT- donors into 55 seronegative recipients with a HCV transmission rate of 9% with no late conversions at 1 year and no difference in function or graft loss compared with PHS increased-risk donor HCV Ab-/NAT- recipients. Due to availability of safe and effective HCV therapies, the use of such organs should be strongly considered to increase the donor organ pool.

Liver transplantation for hepatitis C virus (HCV) non-viremic recipients with HCV viremic donors.

In the context of organ shortage, the opioid epidemic, and effective direct-acting antiviral (DAA) therapy for hepatitis C virus (HCV), more HCV-infected donor organs may be used for liver transplantation. Current data regarding outcomes after donor-derived HCV in previously non-viremic liver transplant recipients are limited. Clinical data for adult liver transplant recipients with donor-derived HCV infection from March 2017 to January 2018 at our institution were extracted from the medical record. Ten patients received livers from donors known to be infected with HCV based on positive nucleic acid testing. Seven had a prior diagnosis of HCV and were treated before liver transplantation. All recipients were non-viremic at the time of transplantation. All 10 recipients derived hepatitis C infection from their donor and achieved sustained virologic response at 12weeks posttreatment with DAA-based regimens, with a median time from transplant to treatment initiation of 43days (IQR 20-59). There have been no instances of graft loss or death, with median follow-up of 380days (IQR 263-434) posttransplant. Transplantation of HCV-viremic livers into non-viremic recipients results in acceptable short-term outcomes. Such strategies may be used to expand the donor pool and increase access to liver transplantation.

Role of Anti-Hepatitis C Virus (HCV) Signal to Cut-off Ratio (S/CO) in Following- up Patients Received Anti-HCV Therapy in Egypt

Background: Hepatitis C Virus (HCV) infection is diagnosed by antibody and RNA based methods. Anti HCV-RNA testing based methods are introduced to confirm viremia in seropositive samples. This study aimed to evaluate the relationship between quantitative anti-HCV (S/CO ratio) and HCV-RNA by PCR as a diagnostic test to identify viremic from non-viremic HCV patients received anti-HCV therapy in Egypt. Subjects and Methods: Patients serum samples used in this study were collected from Al Hussein University hospitals after they had completed their anti-HCV therapy. A total of 172 patients were included in this study 82 were positive RNA, 90 of them were negative RNA their serum samples were assessed for the presence of antibodies to HCV using ELISA method. Results: The results were expressed as the ratio between the signal detected on the sample and the cutoff value of the run (S/CO). Patients with HCV-positive RNA were considered viremic. Receiver operator characteristic (ROC) curve was constructed by plotting sensitivity versus 1 − specificity, using HCV RNA and the S/CO ratio results respectively. Of the 172 patients with HCV infection the mean age was 51.9 ± 7.2 years ranging 35-67 years, 111 (64.5%) were males while 61(35.5%) were females. In the present study there was significant difference in S/CO ratio between viremic and non- viremic subjects. The sensitivity, specificity, negative predictive value, and positive predictive value were 98.78%, 71.11%, 98.46%, and 75.70%, respectively in the S/CO ratio of 8. Area under ROC curve was estimated to be 0.982 (95% confidence interval 0.967 – 0.997). Conclusion: by establishing 8 as cutoff value of the S/CO, it is possible to distinguish between viremic and non-viremic patients without need to use Anti HCV-RNA testing as a confirmatory test.

Prospective Assessment of Adenovirus Infection in Pediatric Kidney Transplant Recipients.

Adenovirus infection is associated with graft dysfunction and graft loss in pediatric cardiac, lung, and liver transplants in prior retrospective studies, but data in pediatric kidney transplant recipients is limited. We conducted a prospective single-center cohort study of 75 consecutive pediatric kidney transplant recipients who underwent monthly screening for adenovirus viremia and symptom assessment for 2 years posttransplant. Adenovirus viremia was detected in 11 (14.7%) patients at a median onset of 173 days (interquartile range, 109-310 days) posttransplant, 6 (8%) had asymptomatic viremia, and 5 (6.7%) had symptomatic disease (2 with hematuria and 3 with an acute febrile respiratory illness). Viremic patients did not differ from nonviremic patients in age, immunosuppression management, or cytomegalovirus or Epstein-Barr virus serostatus, but were more likely to develop cytomegalovirus viremia during the first 2 years posttransplant. No patient had an increase in creatinine from baseline during the time of adenovirus viremia. In a Cox proportional hazards regression, subclinical adenovirus viremia was not associated with a faster time to a 30% decline in estimated glomerular filtration rate. Adenovirus infection is common among pediatric kidney transplant recipients and frequently causes symptomatic disease; however, symptoms are often mild and are not associated with a decline in graft function. Routine monitoring for adenovirus viremia in pediatric kidney transplant recipients may not be warranted.

Impact of low-level BK polyomavirus viremia on intermediate-term renal allograft function.

BK polyomavirus (BKPyV)-associated nephropathy (PyVAN) is a significant cause of premature renal transplant failure. High-level BKPyV viremia is predictive for PyVAN; however, low-level BKPyV viremia does not necessarily exclude the presence of PyVAN. As data are limited regarding whether or not low-level BKPyV viremia has an effect on intermediate-term graft outcome, this study analyzes the impact of low-level BKPyV viremia on intermediate-term graft function and outcome compared with high-level viremia and non-viremic patients. All renal transplant patients received follow-up examinations at the Department of Nephrology, University Hospital Essen. Patients were screened for BKPyV viremia and stratified into three groups according to their maximum BKPyV load in serum (low-level viremia, high-level viremia, and no viremia). In 142 of 213 (67%) patients, BKPyV was never detected in serum; 42 of 213 (20%) patients were found positive for low-level viremia (≤104 copies/mL); and 29 of 213 (13%) patients showed high-level viremia (>104 copies/mL). No significant differences regarding transplant function and graft failure were observed between patients without BKPyV viremia (delta estimated glomerular filtration rate [eGFR] +0.1mL/min [month 1 vs last visit at month 44]) and patients with low-level BKPyV viremia (delta eGFR -1.7mL/min). In patients with high-level viremia, transplant function was significantly restricted (delta eGFR -6.5mL/min) compared with low-level viremia until the last visit at 44±9.7months after transplantation. Although the graft function and graft loss were worse in the high-level viremia group compared with no viremia (eGFR 37 vs 45mL/min), the difference was not significant. High-level viremia was associated with impaired graft function. In contrast, low-level BKPyV viremia had no significant impact on intermediate-term graft function.

25 Pre-emptive therapy for cmv viremia in living donor liver transplant recipients: a single centre experience

Background CMV infection is among the most common complication after liver transplantation which may result in graft loss, survival and increased cost. There are two major strategies for CMV disease prevention after transplantation: preemptive therapy and universal prophylaxis. We present our cohort of recipients of Living Donor Liver Transplant (LDLT) with non-detectable CMV DNA pre-transplant, with moderate risk of CMV viremia (D+/R+, d-/R+), who were preemptively treated. Aim To determine whether preemptive strategy may be feasible approach in patients with moderate risk for CMV viremia in LDLT and its appropriateness for Acute Cellular Rejection(ACR), length of hospital stay and risk of developing other infection and all-cause mortality. Method In this retrospective cohort, 225 adults with moderate risk for CMV viremia who underwent LDLT at Shifa International Hospital from 29/4/2011 to 26/4/2016 were included. All recipients were checked for CMV viremia at day 7, patients with significant viremia (DNA >137 IU/ml) were treated for CMV preemptively. Non-viremic patients on day 7 were only re-checked if had deranged LFT on follow up. Results Out of 225 patients, 83 (36.8%) patients had detectable CMV DNA at day 7. Patients with higher pre-transplant MELD >18 had more chances of developing CMV viremia n=42 (50.6) than those with MELD Conclusions Pre-emptive CMV strategy may be an acceptable approach in patients with moderate risk of CMV viremia in resource constraint setting; however, this needs prospective randomised trials for validation.

Bone Marrow CD34+Progenitor Cells from HIV-Infected Patients Show an Impaired T Cell Differentiation Potential Related to Proinflammatory Cytokines

The impact of HIV infection on the frequency and differentiation capability of CD34+ bone marrow hematopoietic progenitor cells (BM-HPCs) is still debated, having a possible primary role in antiretroviral-induced immunoreconstitution. We investigated the influence of HIV replication or proinflammatory cytokines on lymphopoietic capability of BM-HPCs from seven viremic (VR) and five nonviremic (NVR) HIV-infected patients. We found that BM-HPCs from VR patients were unable to differentiate in vitro toward T cells, and produced proinflammatory cytokines in the absence of viral replication. In contrast, the lymphoid differentiation potential of BM-HPCs was partially restored in successfully antiretroviral therapy-treated patients. We also showed that TLR8 triggering induced BM-HPCs from healthy donors to release proinflammatory cytokines affecting T cell differentiation. These data suggest that in HIV-infected patients, the lymphopoiesis capability of BM-HPCs may be modulated by a virus-driven autocrine mechanism involving proinflammatory cytokines.

Hepatitis C infection and the risk of non-liver-related morbidity and mortality in HIV-positive persons in the Swiss HIV Cohort Study

Hepatitis C virus (HCV) infection has been associated with increased non-liver-related morbidity and mortality. However, studies have yielded inconsistent results.The incidence of clinical events in human immunodeficiency virus (HIV)–infected HCV-seropositive and incidence density–matched HCV-seronegative participants of the Swiss HIV Cohort Study from August 1994 to December 2014 was studied. We compared (1) HCV-seropositive with HCV-seronegative participants and (2) HCV-viremic with successfully treated nonviremic patients. Poisson regression was used to assess differences between these groups.We included 2503 HCV-seropositive participants (540 with spontaneous HCV clearance, 1294 untreated HCV RNA positive, 345 treated with sustained virologic response [SVR], 43 during treatment, and 281 treated without SVR), and 2503 HCV-seronegative controls. After a mean follow-up of 8.2 years, we observed (HCV seropositive and HCV seronegative, respectively) 107 and 18 liver events, 41 and 14 kidney events, 230 and 121 osteoporosis/fractures, 82 and 94 diabetes mellitus, 114 and 129 cardiovascular events, 119 and 147 non-AIDS malignancies, 162 and 126 Centers for Disease Control and Prevention HIV category B/C events, 106 and 10 liver-related deaths, and 227 and 218 non-liver-related deaths. Compared with HCV-negative controls, HCV-seropositive participants had an increased risk of liver events (incidence rate ratio [IRR], 6.29 [95% confidence interval {CI}, 3.52–11.22]), liver-related death (IRR, 8.24 [95% CI, 3.61–18.83]), kidney events (IRR, 2.43 [95% CI, 1.11–5.33]), and osteoporosis/fracture (IRR, 1.43 [95% CI, 1.03–2.01]). Among HCV-seropositive individuals, treated participants without SVR vs those with SVR had a higher risk of liver events (IRR, 6.79 [95% CI, 2.33–19.81]), liver-related death (IRR, 3.29 [95% CI, 1.35–8.05]), and diabetes mellitus (IRR, 4.62 [95% CI, 1.53–13.96]). Similar but not statistically significant differences were found between untreated HCV RNA–positive patients and those with SVR.While HCV exposure was associated with an increased risk of kidney disease and osteoporosis/fracture, this risk did not seem to be dependent of persistent HCV RNA. Successful HCV treatment was associated with a lower incidence of liver disease, liver-related death, and diabetes mellitus, whereas the other conditions studied were less affected.