RESUMO
BACKGROUND: BK virus (BKV) is a significant cause of nephropathy in kidney transplantation. The goal of this study was to characterize the course and source of BKV in kidney transplant recipients. METHODS: We prospectively collected pretransplant plasma and urine samples from living and deceased kidney donors and performed BKV polymerase chain reaction (PCR) and immunoglobulin G (IgG) testing on pretransplant and serially collected posttransplant samples in kidney transplant recipients. RESULTS: Among deceased donors, 8.1% (17/208) had detectable BKV DNA in urine prior to organ procurement. BK viruria was observed in 15.4% (6/39) of living donors and 8.5% (4/47) of deceased donors of recipients at our institution (P = .50). BKV VP1 sequencing revealed identical virus between donor-recipient pairs to suggest donor transmission of virus. Recipients of BK viruric donors were more likely to develop BK viruria (66.6% vs 7.8%; P < .001) and viremia (66.6% vs 8.9%; P < .001) with a shorter time to onset (log-rank test, P < .001). Though donor BKV IgG titers were higher in recipients who developed BK viremia, pretransplant donor, recipient, and combined donor/recipient serology status was not associated with BK viremia (P = .31, P = .75, and P = .51, respectively). CONCLUSIONS: Donor BK viruria is associated with early BK viruria and viremia in kidney transplant recipients. BKV PCR testing of donor urine may be useful in identifying recipients at risk for BKV complications.
Assuntos
Vírus BK/isolamento & purificação , Nefropatias/virologia , Transplante de Rim/efeitos adversos , Infecções por Polyomavirus/virologia , Infecções Tumorais por Vírus/virologia , Adulto , Feminino , Humanos , Imunoglobulina G/sangue , Rim/virologia , Nefropatias/sangue , Nefropatias/urina , Doadores Vivos , Masculino , Pessoa de Meia-Idade , Infecções por Polyomavirus/sangue , Infecções por Polyomavirus/urina , Estudos Prospectivos , Transplantados , Infecções Tumorais por Vírus/sangue , Infecções Tumorais por Vírus/urina , Viremia/sangue , Viremia/urina , Viremia/virologiaRESUMO
BACKGROUND: HIV-positive persons bear an excess burden of chronic kidney disease (CKD); however, conventional methods to assess kidney health are insensitive and non-specific for detecting early kidney injury. Urinary biomarkers can detect early kidney injury, and may help mitigate the risk of overt CKD. METHODS: Cross-sectional study of HIV-positive persons in the Multicenter AIDS Cohort Study and the Women's Interagency HIV Study. We measured levels of 14 biomarkers, capturing multiple dimensions of kidney injury. We then evaluated associations of known CKD risk factors with urine biomarkers using separate multivariable adjusted models for each biomarker. RESULTS: Of the 198 participants, one third were on HAART and virally suppressed. The vast majority (95%) had preserved kidney function as assessed by serum creatinine, with a median eGFR of 103 ml/min/1.73 m2 (interquartile range (IQR): 88, 116). In our multivariable analyses, the associations of each CKD risk factor with urinary biomarker levels varied in magnitude. For example, HIV viral load was predominantly associated with elevations in interleukin(IL)-18, and albuminuria, while higher CD4 levels were associated with lower monocyte chemoattractant protein-1 (MCP-1) and ß2-microglobulin. In contrast, older age was significantly associated with elevations in α1-microglobulin, kidney injury marker-1, clusterin, MCP-1, and chitinase-3-like protein-1 levels, as well as lower epidermal growth factor, and uromodulin levels. CONCLUSIONS: Among HIV-positive persons, CKD risk factors are associated with unique and heterogeneous patterns of changes in urine biomarkers levels. Additional work is needed to develop parsimonious algorithms that integrate multiple biomarkers and clinical data to discern the risk of overt CKD and its progression.
Assuntos
Biomarcadores/urina , Infecções por HIV/urina , Insuficiência Renal Crônica/urina , Terapia Antirretroviral de Alta Atividade , Comorbidade , Creatinina/sangue , Estudos Transversais , Diabetes Mellitus/epidemiologia , Feminino , Taxa de Filtração Glomerular , Infecções por HIV/complicações , Infecções por HIV/tratamento farmacológico , Hepatite C/epidemiologia , Humanos , Hipertensão/epidemiologia , Masculino , Pessoa de Meia-Idade , Insuficiência Renal Crônica/complicações , Insuficiência Renal Crônica/epidemiologia , Fatores de Risco , Sensibilidade e Especificidade , Carga Viral , Viremia/complicações , Viremia/tratamento farmacológico , Viremia/urinaRESUMO
The indirect effects of cytomegalovirus (CMV) viremia can be related to chronic changes in renal allograft structure, but its real impact in early and late graft function remains speculative. A total of 159 patients undergoing renal transplantation using a preemptive therapeutic strategy to prevent CMV disease were included in the present study. The patients were prospectively followed, with serial measurements of urinary retinol-binding protein (uRBP), a marker of proximal tubule injury. uRBP levels and their dynamic performance were compared according to CMV viremia and the 5-year estimated glomerular filtration rate (eGFR), as measured with the modification of diet in renal disease (MDRD) equation. CMV viremia was detected in 79.9% of the patients, with high uRBP levels being detected in 76.0% of these patients (compared with 40.7% in CMV-, P=0.005). High uRBP was associated with male recipients (P=0.02), the number of mismatches (P=0.02) and CMV infection (P=0.001). Five-year eGFR was worse in patients with high uRBP levels (50.3 ± 25.8 compared with 59.8 ± 26.4 ml/min, P=0.04). In a multivariate model, eGFR <60 ml/min was associated with donor age (P<0.001), the number of mismatches (P=0.04), thymoglobulin dose (P=0.02), the presence of and time with delayed graft function (DGF) (P=0.005 and P=0.04), 1-month tacrolimus levels (P=0.03), and uRBP levels after CMV treatment (P=0.01). Patients with CMV viremia in whom uRBP levels were normalized up to 3 months after treatment showed significantly better 5-year eGFR than those in whom uRBP remained high: 61.0 ± 24.2 compared with 42.3 ± 23.9 ml/min, P<0.001. CMV viremia was associated with high uRBP levels, which represent a profile of proximal tubule injury, and the dynamic performance of uRBP after treatment was associated with long-term kidney graft function.
Assuntos
Infecções por Citomegalovirus/urina , Transplante de Rim/métodos , Proteínas de Ligação ao Retinol/urina , Viremia/urina , Adulto , Antivirais/uso terapêutico , Biomarcadores/urina , Infecções por Citomegalovirus/prevenção & controle , Infecções por Citomegalovirus/virologia , Feminino , Ganciclovir/uso terapêutico , Taxa de Filtração Glomerular/efeitos dos fármacos , Sobrevivência de Enxerto/efeitos dos fármacos , Humanos , Túbulos Renais Proximais/efeitos dos fármacos , Túbulos Renais Proximais/patologia , Masculino , Pessoa de Meia-Idade , Fatores de Tempo , Viremia/prevenção & controle , Viremia/virologiaRESUMO
BK virus (BKPyV)-associated nephropathy (BKPyVAN) may affect up to 10% of renal transplant recipients, causing graft failure in the absence of intervention. The dilemma in monitoring BKPyVAN in renal transplant patients has been that only testing urine BK viral load represents higher sensitivity (earlier detection) but lower specificity, while testing plasma BK viral load represents lower sensitivity (later detection) but higher specificity. However, blindly testing both urine and plasma inevitably contributes to unnecessary medical cost. We analyzed 1030 paired urine and plasma BKPyV viral load results and identified a reliable urine BKPyV viral load cutoff (4.0 log IU/mL) that can predict BKPyV viremia with 99.7% negative predictive value (NPV). We propose a cost-effective screening algorithm to first only monitor the urine BKPyV levels until the viral load reaches 4.0 log IU/mL, and then only monitor plasma with higher frequency. This approach ensures 98.7% sensitivity of catching the earliest BKPyV viremia onset, and 100% sensitivity of detecting the critical BKPyV viremia. In addition, we identified a urine BKPyV viral load cutoff of 6.7 log IU/mL as predictive of critical BKPyV viremia (defined as plasma viral load >4.0 log IU/mL) with 100% sensitivity and 100% NPV.
Assuntos
Vírus BK/isolamento & purificação , Nefropatias/diagnóstico , Transplante de Rim/efeitos adversos , Infecções por Polyomavirus/diagnóstico , Infecções Tumorais por Vírus/diagnóstico , Viremia/diagnóstico , Adulto , Vírus BK/fisiologia , Feminino , Humanos , Nefropatias/sangue , Nefropatias/urina , Nefropatias/virologia , Masculino , Pessoa de Meia-Idade , Infecções por Polyomavirus/sangue , Infecções por Polyomavirus/urina , Infecções por Polyomavirus/virologia , Curva ROC , Infecções Tumorais por Vírus/sangue , Infecções Tumorais por Vírus/urina , Infecções Tumorais por Vírus/virologia , Carga Viral , Viremia/urina , Viremia/virologiaRESUMO
BACKGROUND: It remains unclear whether overall degree of immunosuppression or specific effects of individual immunosuppressive agents are causal for increased occurrence of BK polyomavirus (BKPyV) infection in renal transplant recipients (RTR). METHODS: A prospective, multicenter, open-label randomized controlled trial in 361 de novo RTR was performed. A total of 224 RTR were randomized at 6 months into three treatment groups with dual therapy consisting of prednisolone (Pred) plus either cyclosporine (CsA), mycophenolate sodium (MPS), or everolimus (EVL). Primary outcomes were incidence of BK viruria, BK viremia, and BKPyV-associated nephropathy (BKVAN). RESULTS: From 6 months, incidence of BK viruria in the MPS group (43.6%) was significantly higher than in the other groups (CsA: 16.9%, EVL: 19.8%) (P=.003). BKVAN was diagnosed in 3 patients, all treated with MPS (7.8%, P=.001). Longitudinal data analysis showed a lower BKPyV load and a significantly faster clearance of BK viruria in the CsA group compared to the MPS group (P=.03). CONCLUSIONS: Treatment with MPS was associated with an increased incidence of BK viruria. Dual immunosuppressive therapy with CsA and Pred was associated with the lowest rate of BKPyV replication and the fastest clearance of the virus.
Assuntos
Terapia de Imunossupressão/efeitos adversos , Imunossupressores/uso terapêutico , Falência Renal Crônica/cirurgia , Transplante de Rim/efeitos adversos , Infecções por Polyomavirus/epidemiologia , Infecções Tumorais por Vírus/epidemiologia , Adulto , Vírus BK/isolamento & purificação , Quimioterapia Combinada/efeitos adversos , Quimioterapia Combinada/métodos , Feminino , Humanos , Terapia de Imunossupressão/métodos , Incidência , Masculino , Pessoa de Meia-Idade , Infecções por Polyomavirus/sangue , Infecções por Polyomavirus/urina , Infecções por Polyomavirus/virologia , Estudos Prospectivos , Transplantados , Infecções Tumorais por Vírus/sangue , Infecções Tumorais por Vírus/urina , Infecções Tumorais por Vírus/virologia , Carga Viral/efeitos dos fármacos , Viremia/epidemiologia , Viremia/urinaRESUMO
BACKGROUND: Polyomavirus (PV) is a major cause of kidney graft disease. Monitoring by polymerase chain reaction (PCR) on blood is currently recommended. In order to avoid irreversible lesions, we investigated the clinical impact of preemptive reduction of immunosuppression (IS) in kidney transplant recipients (KTR) upon detection of high urinary PV (Upv) load, including BK virus and JC virus. MATERIAL AND METHODS: From 2000 to 2011, in our single center, 789 consecutive KTR were distributed into 4 groups, according to the maximal Upv levels (by PCR) during the first year and the therapeutic option: (A) Upv <104 copies (cp)/mL (n=573), (B) ≥104 Upv <107 cp/mL (n=100), and (C) Upv ≥107 cp/mL (n=116); in group C, the IS drug doses were reduced in subgroup Ca (n=102) only, as 14 patients (subgroup Cb) were at risk for graft rejection. RESULTS: The preemptive reduction of IS (group Ca) increased patient survival as compared with all other groups (P<.05), did not modify graft function, and increased graft survival vs group A (risk ratio: 5.7, confidence interval: 1.8-18.1, P=.003). Differences for risk factors are as follows (groups Ca vs A): incidence of human leukocyte antigen (HLA) immunization (>5% panel reactive antibodies): 3% vs 8% (P=.05), number of HLA mismatches: 2.7 vs 2.5 (P=.049), and incidence of acute rejection: 9.8% vs 24.2% (P=.005). PV-associated nephropathy occurred only in group Ca (2% of total grafts) without effect on patient or graft outcome. CONCLUSION: The reduction of IS in patients with high Upv loads is beneficial for patient survival and does not affect graft survival or graft function.
Assuntos
Vírus BK/efeitos dos fármacos , Rejeição de Enxerto/epidemiologia , Sobrevivência de Enxerto/efeitos dos fármacos , Terapia de Imunossupressão/efeitos adversos , Imunossupressores/administração & dosagem , Vírus JC/efeitos dos fármacos , Transplante de Rim/efeitos adversos , Carga Viral/efeitos dos fármacos , Viremia/urina , Vírus BK/isolamento & purificação , Feminino , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/virologia , Antígenos HLA/imunologia , Humanos , Terapia de Imunossupressão/métodos , Imunossupressores/uso terapêutico , Vírus JC/isolamento & purificação , Nefropatias/epidemiologia , Nefropatias/urina , Nefropatias/virologia , Masculino , Pessoa de Meia-Idade , Infecções por Polyomavirus/epidemiologia , Infecções por Polyomavirus/urina , Infecções por Polyomavirus/virologia , Estudos Retrospectivos , Fatores de Risco , Transplantados/estatística & dados numéricos , Resultado do Tratamento , Infecções Tumorais por Vírus/epidemiologia , Infecções Tumorais por Vírus/urina , Infecções Tumorais por Vírus/virologia , Viremia/virologiaRESUMO
BACKGROUND: Urine is an important source for the detection of infections caused by CMV in stem cell transplant patients. Currently, there is no agreement about the type of urine specimen. In order to investigate which is the better specimen type for quantitative detection of CMV, we compared the results from urine supernatant and sediment from the same patients. METHODS: Seventy urine specimens were collected from patients with hematological disorders or solid tumors. After performing shell vial culture, residual urine specimens were centrifuged. Then, 10 mL of each urine supernatant and sediment were taken and immediately frozen at -70 degrees C. Afterwards, archived urine specimens were thawed at room temperature and CMV-quantitative PCR was performed on both the supernatant and sediment fraction of urine. The results from each patient were reviewed for CMV antigenemia, blood shell vial culture, CMV-IgM or IgG, and clinical symptoms. RESULTS: CMV-qPCR results for the urine sediment fraction revealed a significant difference (p = 0.012) between the active CMV infection group and the latent CMV infection group. In addition, receiver operating characteristic curves for active CMV infection revealed that CMV-qPCR using urine sediment produced more accurate results than urine supernatant. CONCLUSIONS: These findings suggest that the sediment fraction of urine is a more suitable specimen in CMV-qPCR testing.
Assuntos
Infecções por Citomegalovirus/urina , Citomegalovirus/isolamento & purificação , Transplante de Células-Tronco/efeitos adversos , Urinálise/métodos , Viremia/urina , Adolescente , Adulto , Anticorpos Antivirais/urina , Antígenos Virais/urina , Criança , Pré-Escolar , Citomegalovirus/genética , Feminino , Neoplasias Hematológicas/terapia , Neoplasias Hematológicas/urina , Humanos , Imunoglobulina G/urina , Imunoglobulina M/urina , Lactente , Masculino , Pessoa de Meia-Idade , Neoplasias/terapia , Neoplasias/urina , Reação em Cadeia da Polimerase em Tempo Real , Manejo de Espécimes , Adulto JovemRESUMO
Little is known about the urinary virome and how it interacts with the host, particularly in renal transplant diseases. Using metagenomic sequencing, we characterized the urinary virome of 23 kidney transplant recipients longitudinally (11 BKV+ patients and 12 BKV- patients). We applied linear mixed effects models, PERMANOVA, k-means clustering, and MaAsLin2 algorithms to determine virome signatures associated with post-transplant time, BK viremia status, and patient sex. We found that the richness and alpha diversity of urinary virome were significantly different in renal transplant recipients with BKV+ over time in comparison to BKV- (richness P = 0.012, alpha P < 0.0001). Female BKV- patients had significantly higher virome richness than males (P = 0.0063). Virome beta diversity was significantly different between patients by BKV status (P < 0.001). Additionally, we identified underlying interactions between patient sex and BKV status, in terms of virome beta diversity (P = 0.008). BK polyomavirus infections were primarily of subtypes IA, IB1, and IB2. The non-BK dominant samples clustered into six urinary virome community states. BKV- samples had more anelloviruses than BKV+ samples though this difference was not statistically significant. Lastly, we identified specific viruses, associated with BKV+ and time in our samples. Our results indicate that dynamic alterations in the urinary virome over the post-transplant period in kidney transplant recipients can be shaped by BK viremia and patient sex. These findings advance our fundamental understanding of the urinary virome and support a new line of investigation in renal disease and transplantation. IMPORTANCE: The urinary microbiome is increasingly implicated in renal health and disease. While most research focuses on bacteria communities of the microbiome, factors that influence the urinary virome are not understood. Here, we investigated the urinary virome of 23 adult kidney transplant recipients longitudinally over 14 weeks post-transplant. We show that alterations in the urinary virome are associated with kidney transplant recipients with BK polyomavirus viremia that can lead to BK nephropathy and allograft rejection. By modeling the temporal dynamics post-transplant, we delineated specific profiles of the urinary virome associated with patient sex and urinary community states. These findings reveal fundamental aspects of the urinary virome that can be leveraged to better manage kidney diseases.
Assuntos
Vírus BK , Transplante de Rim , Infecções por Polyomavirus , Transplantados , Viremia , Viroma , Humanos , Transplante de Rim/efeitos adversos , Vírus BK/isolamento & purificação , Vírus BK/genética , Masculino , Feminino , Infecções por Polyomavirus/urina , Infecções por Polyomavirus/virologia , Viremia/urina , Pessoa de Meia-Idade , Adulto , Estudos Longitudinais , Infecções Tumorais por Vírus/urina , Infecções Tumorais por Vírus/virologia , Idoso , Fatores Sexuais , Urina/virologiaRESUMO
HIV-1 persists indefinitely in multiple cellular reservoirs despite antiretroviral therapy. We previously demonstrated HIV-1 compartmentalization in kidney and urine. Here, we further characterized viruses in urine and when available, compared them to those present in semen from HIV-1 positive participants with detectable plasma viremia to further understand the viral dynamics in the upper and lower genitourinary tract.Blood and urine samples were obtained from 19 HIV-1 positive participants. Simultaneous semen samples were obtained from 16 of the 19 participants. HIV-1 envelope (env) gene sequences were obtained by single-genome amplification (SGA) and neighbor-joining trees were constructed using the Kimura 2-parameter model.HIV-1 env gene sequences were amplified from blood in 19/19 (100%) participants, urine in 18/19 (95%) participants, and semen in 12/16 (75%). In individuals from which both urine and semen samples were obtained, differences in viral shedding between the 2 sources were observed, where HIV-1 env sequences could only be amplified from either urine or semen. Longitudinal phylogenetic analysis of urine-derived env sequences from 1 participant demonstrated that urine clusters distinct from blood are maintained over time (20 weeks), consistent with viral compartmentalization and local replication. Comparison of urine and semen derived sequences demonstrated either virus compartmentalization or equilibration.Our results demonstrate that when present, viral compartmentalization in urine persists over time. Comparison of timing of viral shedding in urine and semen samples from our cohort suggest different viral kinetics between the upper and lower genitourinary tract and sequence analysis suggests that HIV-1 populations in urine and semen can either be imported from blood or produced locally.
Assuntos
Infecções por HIV/virologia , HIV-1/genética , Sêmen/virologia , Soro/virologia , Urina/virologia , Viremia/virologia , Adulto , Feminino , Amplificação de Genes , Infecções por HIV/sangue , Infecções por HIV/urina , Humanos , Masculino , Filogenia , Carga Viral , Viremia/sangue , Viremia/urina , Eliminação de Partículas ViraisRESUMO
BACKGROUND: BK virus-associated nephropathy (BKVAN) can be diagnosed only with renal graft biopsy. Definitive diagnosis of BKVAN requires demonstration of BK virus (BKV) replication in renal allograft tissues. Non-invasive analysis of urine and blood is considered essential in screening renal transplant recipients. PATIENTS AND METHODS: This study evaluated prospectively the replication of BKV in plasma and urine with qualitative and quantitative real-time polymerase chain reaction in 32 de novo (group A) and 34 chronic (group B) renal transplant recipients and the long-term impact on graft function. RESULTS: In group A, 456 samples (228 plasma, 228 urine) were examined and BKV was detected in 31 (31/228, 14%) samples of plasma and 57 (57/228, 25%) samples of urine in 20 (20/32, 62.5%) and 23 (23/32, 72%) recipients, respectively. Incidence of viremia and viruria increased during the first 6 months presenting a peak the third postoperative month (viremia: 28% and viruria: 31%). Immune suppressive treatment with tacrolimus showed significant relation with viremia. Renal graft function in de novo renal transplant recipients remained stable throughout the follow-up period without influence of BKV replication. In group B, incidence of viremia and viruria were 3% (1/34) and 9% (3/34) correspondingly, indicating that after the first post-transplant year the risk of BKV re-activation is diminished. CONCLUSION: The highest incidence of BK viremia and viruria is observed the third post-transplantation month, confirming previously published studies in Europe and the United States, and long-term follow up shows that BKV replication decreases significantly after the third post-transplant month and even transient viremia or viruria does not have an impact on renal function.
Assuntos
Vírus BK/isolamento & purificação , Transplante de Rim/efeitos adversos , Infecções por Polyomavirus/epidemiologia , Infecções Tumorais por Vírus/epidemiologia , Viremia/epidemiologia , Replicação Viral , Adulto , Idoso , Vírus BK/genética , Vírus BK/fisiologia , Feminino , Sobrevivência de Enxerto , Humanos , Terapia de Imunossupressão , Incidência , Masculino , Pessoa de Meia-Idade , Infecções por Polyomavirus/sangue , Infecções por Polyomavirus/urina , Infecções por Polyomavirus/virologia , Infecções Tumorais por Vírus/sangue , Infecções Tumorais por Vírus/urina , Infecções Tumorais por Vírus/virologia , Viremia/sangue , Viremia/urina , Viremia/virologia , Ativação Viral , Adulto JovemRESUMO
BACKGROUND: Real-time RT-PCR (Reverse Transcriptase Polymerase Chain Reaction) is considered the gold standard for Zika virus (ZIKV) infection diagnosis, despite its low sensitivity. Diagnosis using recommended serologic cutoffs in co-circulating Flaviviruses areas maybe inadequate due to in-vitro cross-reactivities of Flaviviruses-specific antibodies. We evaluated Zika diagnosis in symptomatic patients using serial RT-PCR and develop a classification model using serial Dengue virus (DENV) and ZIKV serologies. METHODS: A prospective longitudinal multicentric study in Southern Mexico (NCT02831699) enrolled symptomatic and non-symptomatic participants. In the classification model, true positives were symptomatic (using a modified World Health Organization/Pan American Health Organization definition) with RT-PCR positive for ZIKV or DENV. True negatives were non-symptomatic with negative RT-PCR. Serial serology measurements were used to predict disease status. RESULTS: Analyzing ZIKV and DENV RT-PCR at 3 timepoints between days 3 and 13 of symptom onset detected 25% more cases than a single RT-PCR analysis between day 0 and 6. When considering sensitivity and specificity together, the serial serology model predicted all categories of disease and negatives better than manufactures cutoffs. Their cutoffs optimized sensitivity or specificity but not both. CONCLUSIONS: We demonstrated the importance of serial RT-PCR and antibody measurements to diagnose arbovirus infection in symptomatic patients living in regions with co-circulating flaviviruses.
Assuntos
Vírus da Dengue/isolamento & purificação , Dengue/diagnóstico , Infecção por Zika virus/diagnóstico , Zika virus/isolamento & purificação , Adulto , Anticorpos/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Estudos Longitudinais , Masculino , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Viremia/sangue , Viremia/urina , Adulto JovemRESUMO
BACKGROUND: Polyomavirus BK (BKV) encodes two mature miRNAs that regulate the viral life cycle. OBJECTIVES: This study investigated the autoregulatory and immunomodulatory effects of these miRNAs that have been defined in culture systems, but subject to only limited exploration in clinical samples. METHODS: BKV-miR-B1-5p, BKV-miR-BJ1-3p, BKV DNA and BKV VP-1 mRNA levels were measured in 32 paired obtained plasma & urine samples from kidney transplant patients with (a) early stage infection manifesting as viruria, and (b) later stage infections complicated by viremia. RESULTS: All patients showed abundant urine miRNAs (7.84Eâ¯+â¯02-1.91Eâ¯+â¯06 copies/ml, but plasma miRNA was below the limit of detection. There was no statistically significant difference in urinary miRNA levels between viruric and viremic patients. Median 5p miRNA load was 4-6 logs lower than the BKV genomic load. Higher miRNA levels in the urine were associated not with lower but higher urinary viral loads. BKV preferentially used the 3p miRNA for its interactions with host cell mRNAs. The mean ratio of 5p/3p in patients with viruria was 0.09, and 0.03 in patients with viremia. CONCLUSIONS: The data suggest that immune evasion functions of BKV miRNAs over-ride the negative autoregulatory feedback effects in kidney transplant patients with active viral replication.
Assuntos
Vírus BK/genética , Transplante de Rim/efeitos adversos , MicroRNAs/urina , Infecções por Polyomavirus/virologia , Infecções Tumorais por Vírus/virologia , Adolescente , Adulto , Idoso , Vírus BK/fisiologia , Proteínas do Capsídeo/genética , DNA Viral/urina , Humanos , Nefropatias/imunologia , Nefropatias/urina , Nefropatias/virologia , Pessoa de Meia-Idade , Infecções por Polyomavirus/imunologia , Infecções por Polyomavirus/urina , RNA Viral/urina , Infecções Tumorais por Vírus/imunologia , Infecções Tumorais por Vírus/urina , Carga Viral , Viremia/imunologia , Viremia/urina , Viremia/virologia , Replicação Viral , Adulto JovemRESUMO
BACKGROUND: A qualitative highly predictive urinary test for polyomavirus nephropathy (PVN) is the PV-Haufen test. This article evaluates whether a quantitative PV-Haufen analysis, that is, the number of PV-Haufen shed per milliliter urine, predicts PVN disease grades and the severity of intrarenal PV replication. METHODS: Polyomavirus-Haufen were counted in 40 urine samples from patients with biopsy-proven definitive PVN. The number of PV-Haufen was correlated with both histologic PVN disease grades 1 to 3 and the number of SV40-T-expressing cells as indicators of intrarenal PV replication in corresponding renal allograft biopsies (manual counts and automated morphometry). Findings from quantitative PV-Haufen analyses were compared to conventional laboratory test results, that is, BK viremia (quantitative polymerase chain reaction [PCR]) and BK viruria (quantitative PCR and decoy cell counts). RESULTS: Polyomavirus-Haufen counts showed excellent correlation (α0.77-0.86) with the severity of intrarenal PV replication and disease grades. In particular, low PV-Haufen numbers strongly correlated with early PVN grade 1 and minimal intrarenal expression of SV40-T antigen (P < 0.001). In comparison, BK viremia and viruria levels by PCR showed only modest correlations with histologic SV40-T expression (α0.40-0.49) and no significant correlation with disease grades or minimal intrarenal PV replication. No correlations were seen with urinary decoy cell counts. In contrast to conventional quantitative PCR assays or decoy cell counts, quantitative urinary PV-Haufen testing accurately reflects the severity of PV replication, tissue injury, and PVN disease grades. CONCLUSIONS: Quantitative PV-Haufen testing is a novel noninvasive approach to patient management for the diagnosis and prediction of PVN disease grades and monitoring of disease course during therapy.
Assuntos
DNA Viral/urina , Nefropatias/diagnóstico , Nefropatias/virologia , Infecções por Polyomavirus/diagnóstico , Viremia/urina , Viremia/virologia , Antígenos Transformantes de Poliomavirus/química , Biópsia , Sobrevivência de Enxerto , Humanos , Reação em Cadeia da Polimerase , Polyomavirus/fisiologia , Reprodutibilidade dos Testes , Replicação ViralRESUMO
We report a urinary tract infection (UTI) with erythrovirus B19 in an HIV-1-positive homosexual man persisting for more than 7 months after the decline of viremia after a primary infection. During the course of the UTI, the patient complained of soreness in the kidney region and suffered from transient episodes of edema and hematuria. Proteinuria and elevated serum concentrations of creatinine further substantiated the hypothesis of a renal focus of a persistent erythrovirus B19 infection.
Assuntos
Infecções por HIV/complicações , HIV-1 , Infecções por Parvoviridae/virologia , Parvovirus B19 Humano/isolamento & purificação , Infecções Urinárias/virologia , Adulto , Sequência de Bases , Doença Crônica , DNA Viral/análise , Humanos , Rim/virologia , Masculino , Infecções por Parvoviridae/complicações , Infecções por Parvoviridae/urina , Parvovirus B19 Humano/química , Parvovirus B19 Humano/classificação , Reação em Cadeia da Polimerase , Infecções Urinárias/complicações , Infecções Urinárias/urina , Viremia/urina , Viremia/virologiaRESUMO
Transmissibility of the D variant of encephalomyocarditis virus (EMC-D) was examined. Eight-week-old ICR:CD-1 male mice inoculated with 10(5) plaque forming units (PFU)/animal of EMC-D intranasally, orally or intraperitoneally showed marked viraemia and prominent pancreatic lesions at 2 days after inoculation (2 DAI), and excreted virus in faeces from 2 to 8 DAI (virus titre: 10(3)-10(5) PFU/g). Only a small proportion of control mice housed with EMC-D-inoculated mice for 10 days developed viraemia and pancreatic lesions.
Assuntos
Vírus da Encefalomiocardite , Infecções por Enterovirus/transmissão , Viremia/transmissão , Animais , Vírus da Encefalomiocardite/isolamento & purificação , Infecções por Enterovirus/microbiologia , Infecções por Enterovirus/urina , Fezes/microbiologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , Viremia/microbiologia , Viremia/urinaAssuntos
Vírus BK , Transplante de Coração , Hospedeiro Imunocomprometido , Insuficiência Renal/diagnóstico , Viremia/diagnóstico , Adulto , Antivirais/uso terapêutico , Diagnóstico Diferencial , Feminino , Humanos , Masculino , Complicações Pós-Operatórias , Insuficiência Renal/sangue , Insuficiência Renal/tratamento farmacológico , Insuficiência Renal/urina , Carga Viral , Viremia/sangue , Viremia/tratamento farmacológico , Viremia/urinaRESUMO
Rotaviruses infect mature, differentiated enterocytes of the small intestine and, by an unknown mechanism, escape the gastrointestinal tract and cause viremia. The neonatal rat model of rotavirus infection was used to determine the kinetics of viremia, spread, and pathology of rotavirus in extraintestinal organs. Five-day-old rat pups were inoculated intragastrically with an animal (RRV) or human (HAL1166) rotavirus or phosphate-buffered saline. Blood was collected from a subset of rat pups, and following perfusion to remove residual blood, organs were removed and homogenized to analyze rotavirus-specific antigen by enzyme-linked immunosorbent assay and infectious rotavirus by fluorescent focus assay or fixed in formalin for histology and immunohistochemistry. Viremia was detected following rotavirus infection with RRV and HAL1166. The RRV 50% antigenemia dose was 1.8 x 10(3) PFU, and the 50% diarrhea dose was 7.7 x 10(5) PFU, indicating that infection and viremia occurred in the absence of diarrhea and that detecting rotavirus antigen in the blood was a more sensitive measure of infection than diarrhea. Rotavirus antigens and infectious virus were detected in multiple organs (stomach, intestines, liver, lungs, spleen, kidneys, pancreas, thymus, and bladder). Histopathological changes due to rotavirus infection included acute inflammation of the portal tract and bile duct, microsteatosis, necrosis, and inflammatory cell infiltrates in the parenchymas of the liver and lungs. Colocalization of structural and nonstructural proteins with histopathology in the liver and lungs indicated that the histological changes observed were due to rotavirus infection and replication. Replicating rotavirus was also detected in macrophages in the lungs and blood vessels, indicating a possible mechanism of rotavirus dissemination. Extraintestinal infectious rotavirus, but not diarrhea, was observed in the presence of passively or actively acquired rotavirus-specific antibody. These findings alter the previously accepted concept of rotavirus pathogenesis to include not only gastroenteritis but also viremia, and they indicate that rotavirus could cause a broad array of systemic diseases in a number of different organs.
Assuntos
Infecções por Rotavirus/imunologia , Rotavirus/imunologia , Viremia/imunologia , Animais , Animais Recém-Nascidos , Antígenos Virais/sangue , Linhagem Celular , Modelos Animais de Doenças , Feminino , Macrófagos/imunologia , Macrófagos/virologia , Camundongos , Especificidade de Órgãos/imunologia , Ratos , Ratos Endogâmicos Lew , Ratos Sprague-Dawley , Infecções por Rotavirus/patologia , Infecções por Rotavirus/urina , Viremia/patologia , Viremia/urinaRESUMO
Immunoglobulin G (IgG), IgA, and IgM antibodies were measured in serum samples from 71 organ donors, 81 kidney transplant recipients at transplantation, and 67 patients during the posttransplant period by using a virus-like particle-based enzyme-linked immunosorbent assay (ELISA). BK virus (BKV) and JC virus DNA were detected in urine and plasma by real-time PCR. IgG antibodies to BKV were demonstrated in the majority (80.3 to 100%) of patients irrespective of clinical category, but titers were highest in patients with active viral replication. IgA antibodies were present with greater frequency (72.7 to 81.3% versus 0 to 23.6%; P < 0.001) and higher titer (mean optical density, 0.11 to 0.15 versus 0.05 to 0.08; P < 0.001) in patients who were BKV DNA positive than those who were BKV DNA negative. IgM antibodies showed a similar pattern of reactivity but lower frequency in the setting of active viral replication (9.1 to 43.7% versus 0 to 1.4%; P < 0.001). A rise in IgG level of >0.577 optical density (OD) units or a rise in IgA or IgM level of >0.041 OD units was strongly associated with active viral replication. Urine viral load showed a positive correlation with IgM titer (r = 0.22) but a negative correlation with IgG titer (r = -0.28) and IgA titer (r = -0.1). Chronic dialysis patients typically did not have serologic or virologic evidence of active BKV infection. Anti-BKV titers did not rise in patients with JC viruria. In conclusion, measurement of anti-BKV antibody titer and class response can be used to detect the onset of viral replication. ELISAs can be quite specific despite considerable sequence homology between BK virus and JC virus.
Assuntos
Vírus BK , Imunoglobulina A/imunologia , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Transplante de Rim , Adolescente , Adulto , Idoso , Anticorpos Antivirais/imunologia , Vírus BK/genética , Vírus BK/imunologia , Vírus BK/isolamento & purificação , Vírus BK/fisiologia , DNA Viral/sangue , DNA Viral/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Viremia/urina , Replicação ViralRESUMO
BK viremia and nephritis are increasing problems in renal transplant recipients. The exact cause of the increasing prevalence of this condition remains poorly understood. Increasing prevalence has been correlated with newer immunosuppressive agents and the decline in acute rejection rates in recent years. The clinical manifestation varies from the asymptomatic state of viremia and nephritis to clinical renal dysfunction. The diagnosis of this infection is based on the combination of the presence of urinary decoy cells, virus in the urine/blood, and typical renal histological findings of interstitial nephritis. Routine post-transplant screening for BK viremia and viruria prior to the occurrence of nephritis and the reduction in immunosuppressive therapy for subjects with viremia appear to be attractive approaches. The treatment of BKV nephritis (BKVN) consists of reduction in immunosuppressive therapy and antiviral therapy with cidofovir or leflunomide or a combination of both. Approximately 30-60% of subjects with BKVN experienced irreversible graft failure. However, in recent years, the combinations of early detection, prompt diagnosis, and appropriate reduction in immunosuppressive therapy have been associated with better outcome. The pathogenesis of BK virus infection in renal transplant recipients needs to be explored. The source of BKV infection (donor as opposed to recipient), the role of host humoral, and cellular immunity to BKV, and the role of alloimmune activation in renal graft to the occurrence of nephritis are discussed in this review.
Assuntos
Vírus BK/isolamento & purificação , Transplante de Rim/efeitos adversos , Nefrite , Infecções por Polyomavirus , Infecções Tumorais por Vírus , Anticorpos Antivirais , Antivirais/uso terapêutico , Vírus BK/genética , Vírus BK/imunologia , Vírus BK/fisiologia , Cidofovir , Citosina/análogos & derivados , Citosina/uso terapêutico , DNA Viral/sangue , DNA Viral/urina , Sobrevivência de Enxerto/imunologia , Humanos , Imunidade Celular , Imunossupressores/efeitos adversos , Isoxazóis/uso terapêutico , Rim/patologia , Rim/fisiopatologia , Rim/virologia , Transplante de Rim/imunologia , Leflunomida , Nefrite/sangue , Nefrite/epidemiologia , Nefrite/etiologia , Nefrite/virologia , Organofosfonatos/uso terapêutico , Infecções por Polyomavirus/sangue , Infecções por Polyomavirus/tratamento farmacológico , Infecções por Polyomavirus/epidemiologia , Infecções por Polyomavirus/etiologia , Prevalência , Fatores de Risco , Resultado do Tratamento , Infecções Tumorais por Vírus/sangue , Infecções Tumorais por Vírus/tratamento farmacológico , Infecções Tumorais por Vírus/epidemiologia , Infecções Tumorais por Vírus/etiologia , Viremia/tratamento farmacológico , Viremia/epidemiologia , Viremia/etiologia , Viremia/urinaRESUMO
In a previous study, we performed serial BK virus (BKV), polymerase chain reaction (PCR) and detected active BKV infection in 70 (35.4%) of 198 renal transplant recipients. In the current study, pre-transplant donor and recipient samples were analyzed for BKV antibody titer and HLA alleles. Donor antibody titer was inversely proportional to onset of viruria, p<0.001, directly proportional to duration of viruria, p=0.014 and directly proportional to peak urine viral titer p=0.005. Recipient pairs receiving kidneys from the same donor were concordant for BKV infection, p=0.017, and had matched sequences of segments of the NCCR and VP1 genes that tended to vary among recipients of kidneys from different donors. We did not see an association of HLA A, B, or DR, HLA allele mismatches or total HLA mismatches and BK infection. However, all 11 recipients with sustained BK viremia received kidneys from donors lacking HLA C7, and 10 recipients also lacked C7. These findings derive from the largest and most comprehensive prospective study of BKV infection in renal transplant recipients performed to date. Our data support donor origin for early BKV infection in kidney transplant recipients, and suggest that a specific HLA C locus may be associated with failure to control BKV infection.