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1.
Int J Mol Sci ; 25(15)2024 Jul 27.
Artículo en Inglés | MEDLINE | ID: mdl-39125764

RESUMEN

NIBV is an acute and highly contagious virus that has a major impact on the poultry industry. Wogonin, as a flavonoid drug, has antiviral effects, but there have been no reports indicating its role in renal injury caused by NIBV infection. The aim of this study is to investigate the antiviral effect of wogonin against NIBV. Renal tubular epithelial cells were isolated and cultured, and divided into four groups: Con, Con+Wog, NIBV and NIBV+Wog. We found that wogonin significantly inhibited the copy number of NIBV and significantly alleviated NIBV-induced cell apoptosis and necrosis. Moreover, wogonin inhibited the reduction in mitochondrial membrane potential and the aberrant opening of mPTP caused by NIBV. In conclusion, wogonin can protect renal tubular epithelial cells from damage by inhibiting the replication of NIBV and preventing mitochondrial apoptosis and necroptosis induced by NIBV.


Asunto(s)
Apoptosis , Pollos , Células Epiteliales , Flavanonas , Túbulos Renales , Necroptosis , Animales , Flavanonas/farmacología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/virología , Células Epiteliales/metabolismo , Necroptosis/efectos de los fármacos , Apoptosis/efectos de los fármacos , Túbulos Renales/virología , Túbulos Renales/efectos de los fármacos , Túbulos Renales/citología , Túbulos Renales/patología , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Antivirales/farmacología , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Infecciones por Coronavirus/virología , Infecciones por Coronavirus/tratamiento farmacológico , Enfermedades de las Aves de Corral/virología , Enfermedades de las Aves de Corral/tratamiento farmacológico , Replicación Viral/efectos de los fármacos , Células Cultivadas
2.
Poult Sci ; 103(7): 103774, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38669820

RESUMEN

Goose astrovirus genotype 2 (GAstV-2) mainly causes gout in goslings; therefore, it is a major pathogen threatening to goose flocks. However, the mechanisms underlying host-GAstV-2 interactions remain unclear because host cells suitable for GAstV-2 replication have been unavailable. We previously noted that GAstV-2 is primarily located in goose renal epithelial cells, where it causes kidney damage. Therefore, here, we derived goose primary renal tubular epithelial (RTE) cells (GRTE cells) from the kidneys of goose embryos after collagenase I digestion. After culture in Dulbecco's modified Eagle medium/Nutrient mixture F-12 with 10% fetal bovine serum (FBS), the isolated cells had polygonal with roadstone-like morphology; they were identified to be epithelial cells based on the presence of cytokeratin 18 expression detected through immunofluorescence assay (IFA). GAstV-2 infection in GRTE cells led to no obvious cytopathic effects; the maximum amounts of infectious virions were observed 48 h post infection through IFA and quantitative PCR. Next, RNA-seq was performed to identify and map post-GAstV-2 infection differentially expressed genes. The downregulated pathways were mainly related to metabolism, including tryptophan metabolism, drug metabolism by cytochrome P450, xenobiotic metabolism by cytochrome P450, retinol metabolism, butanoate metabolism, starch and sucrose metabolism, ascorbate and aldarate metabolism, and drug metabolism by other enzymes and peroxisome. In contrast, the upregulated pathways were mostly related to the host cell defense and proliferation, including extracellular matrix-receptor interaction, complement and coagulation cascades, phagosome, PI3K-Akt signaling pathway, human T-lymphotropic virus 1 infection, lysosome, and tumor necrosis factor signaling pathway. In conclusion, we developed a GRTE cell line for GAstV-2 replication and analyzed the potential host-GAstV-2 interactions through RNA-seq; our results may aid in further investigating the pathogenic mechanisms underlying GAstV-2 infection and provide strategies for its prevention and control.


Asunto(s)
Infecciones por Astroviridae , Células Epiteliales , Gansos , Genotipo , Enfermedades de las Aves de Corral , Animales , Gansos/virología , Células Epiteliales/virología , Enfermedades de las Aves de Corral/virología , Infecciones por Astroviridae/veterinaria , Infecciones por Astroviridae/virología , Análisis de Secuencia de ARN/veterinaria , Túbulos Renales/virología , Túbulos Renales/citología , Avastrovirus/fisiología , Avastrovirus/genética , Células Cultivadas
3.
BMC Nephrol ; 22(1): 278, 2021 08 10.
Artículo en Inglés | MEDLINE | ID: mdl-34376184

RESUMEN

BACKGROUND: The recent COVID-19 pandemic has raised concerns about patient diagnosis and follow-up of chronically ill patients. Patients suffering from chronic illnesses, concomitantly infected by SARS-CoV-2, globally tend to have a worse prognosis and poor outcomes. Renal tropism and acute kidney injury following SARS-CoV-2 infection has recently been described in the literature, with elevated mortality rates. Furthermore, patients with pre-existing chronic kidney disease, infected by SARS-CoV-2, should be monitored carefully. Here, we report the case of a 69-year-old patient with splenic marginal zone lymphoma, suffering from longstanding chronic kidney disease following SARS-CoV-2 infection. CASE PRESENTATION: A 69-year-old male patient previously diagnosed with pulmonary embolism and splenic marginal zone lymphoma (Splenomegaly, Matutes 2/5, CD5 negative and CD23 positive), was admitted to the hospital with shortness of breath, fever and asthenia. A nasopharyngeal swab test was performed in addition to a CT-scan, which confirmed SARS-CoV-2 infection. Blood creatinine increased following SARS-CoV-2 infection at 130 µmol/l, with usual values at 95 µmol/l. The patient was discharged at home with rest and symptomatic medical treatment (paracetamol and hydration), then readmitted to the hospital in August 2020. A kidney biopsy was therefore conducted as blood creatinine levels were abnormally elevated. Immunodetection performed in a renal biopsy specimen confirmed co-localization of SARS-CoV2 nucleocapsid and protease 3C proteins with ACE2, Lewis x and sialyl-Lewis x antigens in proximal convoluted tubules and podocytes. Co-localization of structural and non-structural viral proteins clearly demonstrated viral replication in proximal convoluted tubules in this chronically ill patient. Additionally, we observed the co-localization of sialyl-Lewis x and ACE2 receptors in the same proximal convoluted tubules. Reverse Transcriptase-Polymerase Chain Reaction test performed on the kidney biopsy was negative, with very low Ct levels (above 40). The patient was finally readmitted to the haematology department for initiation of chemotherapy, including CHOP protocol and Rituximab. CONCLUSIONS: Our case emphasizes on the importance of monitoring kidney function in immunosuppressed patients and patients suffering from cancer following SARS-CoV-2 infection, through histological screening. Further studies will be required to decipher the mechanisms underlying chronic kidney disease and the putative role of sialyl-Lewis x and HBGA during SARS-CoV-2 infection.


Asunto(s)
COVID-19/complicaciones , Túbulos Renales/virología , Insuficiencia Renal Crónica/virología , SARS-CoV-2/fisiología , Replicación Viral , Anciano , Enzima Convertidora de Angiotensina 2/análisis , Biopsia , COVID-19/sangre , COVID-19/diagnóstico , Proteínas de la Nucleocápside de Coronavirus/análisis , Creatinina/sangre , Humanos , Riñón/química , Riñón/patología , Riñón/virología , Túbulos Renales/química , Túbulos Renales/patología , Antígeno Lewis X/análisis , Linfoma de Células B de la Zona Marginal/complicaciones , Masculino , Insuficiencia Renal Crónica/patología , Antígeno Sialil Lewis X/análisis , Neoplasias del Bazo/complicaciones
4.
Nat Commun ; 12(1): 2506, 2021 05 04.
Artículo en Inglés | MEDLINE | ID: mdl-33947851

RESUMEN

It is unclear whether severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can directly infect human kidney, thus leading to acute kidney injury (AKI). Here, we perform a retrospective analysis of clinical parameters from 85 patients with laboratory-confirmed coronavirus disease 2019 (COVID-19); moreover, kidney histopathology from six additional COVID-19 patients with post-mortem examinations was performed. We find that 27% (23/85) of patients exhibited AKI. The elderly patients and cases with comorbidities (hypertension and heart failure) are more prone to develop AKI. Haematoxylin & eosin staining shows that the kidneys from COVID-19 autopsies have moderate to severe tubular damage. In situ hybridization assays illustrate that viral RNA accumulates in tubules. Immunohistochemistry shows nucleocapsid and spike protein deposits in the tubules, and immunofluorescence double staining shows that both antigens are restricted to the angiotensin converting enzyme-II-positive tubules. SARS-CoV-2 infection triggers the expression of hypoxic damage-associated molecules, including DP2 and prostaglandin D synthase in infected tubules. Moreover, it enhances CD68+ macrophages infiltration into the tubulointerstitium, and complement C5b-9 deposition on tubules is also observed. These results suggest that SARS-CoV-2 directly infects human kidney to mediate tubular pathogenesis and AKI.


Asunto(s)
Lesión Renal Aguda/etiología , COVID-19/complicaciones , Túbulos Renales/virología , SARS-CoV-2/patogenicidad , Lesión Renal Aguda/epidemiología , Lesión Renal Aguda/patología , Lesión Renal Aguda/virología , Adulto , Anciano , Anciano de 80 o más Años , Enzima Convertidora de Angiotensina 2/metabolismo , Antígenos Virales/genética , Antígenos Virales/metabolismo , COVID-19/epidemiología , COVID-19/virología , China/epidemiología , Femenino , Humanos , Inmunidad Innata , Pruebas de Función Renal , Túbulos Renales/metabolismo , Túbulos Renales/patología , Masculino , Persona de Mediana Edad , Pandemias , Estudios Retrospectivos , SARS-CoV-2/genética , SARS-CoV-2/aislamiento & purificación , Proteínas Virales/genética , Proteínas Virales/metabolismo , Adulto Joven
5.
Appl Immunohistochem Mol Morphol ; 29(1): 5-12, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-33086222

RESUMEN

We studied the suitability of commercially available monoclonal antibodies (mAbs) for the immunohistochemical (IHC) detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) in standard archival specimens. Antibodies were screened on HEK293 cells transfected with viral nucleoprotein, S1 subunit and S2 subunit of spike protein and on untransfected cells, as well as a panel of normal tissue. Lung tissue with presence of SARS-CoV2 confirmed by in situ hybridization (ISH) was also used. A total of 7 mAbs were tested: (1) mAb 001 (Sino Biological, 40143-R001), (2) mAb 007 (Sino Biological, 40150-R007), (3) mAb 019 (Sino Biological, 40143-R019), (4) mAb 1A9 (GeneTex, GTX632604), (5) mAb ABM19C9 (Abeomics, 10-10007), (6) FIPV3-70 (Santa Cruz, SC-65653), and (7) mAb 6F10 (BioVision, A2060). Only 2 mAbs, clone 001 to the nucleoprotein and clone 1A9 to the S2 subunit spike protein displayed specific immunoreactivity. Both clones showed strong staining in the acute phase of COVID-19 pneumonia, mostly in areas of acute diffuse alveolar damage, but were not completely congruent. Viral protein was also found in kidney tubules, endothelia of multiple organs and a nasal swab of a patient with persistent SARS-CoV2 infection. The other tested reagents were either poorly reactive or demonstrated nonspecific staining in tissues and lesions not infected by SARS-CoV2. Our study demonstrates that rigid specificity testing is mandatory for the evaluation of mAbs to SARS-CoV2 and that clones 001 to nucleoprotein and 1A9 to S2 subunit spike protein are useful for the in situ detection of SARS-CoV2.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Anticuerpos Antivirales/inmunología , COVID-19/virología , Inmunohistoquímica , SARS-CoV-2/inmunología , COVID-19/inmunología , Endotelio/virología , Células HEK293 , Humanos , Indicadores y Reactivos , Túbulos Renales/virología , Pulmón/virología , Proteínas de la Nucleocápside/análisis , Proteínas de la Nucleocápside/inmunología , Glicoproteína de la Espiga del Coronavirus/análisis , Glicoproteína de la Espiga del Coronavirus/inmunología
6.
AIDS ; 34(11): 1581-1591, 2020 09 01.
Artículo en Inglés | MEDLINE | ID: mdl-32701578

RESUMEN

OBJECTIVES: HIV-1 can infect and persist in different organs and tissues, resulting in the generation of multiple viral compartments and reservoirs. Increasing evidence supports the kidney as such a reservoir. Previous work demonstrated that HIV-1 infected CD4 T-cells transfer virus to renal tubule epithelial (RTE) cells through cell-to-cell contact. In addition to CD4 T cells, macrophages represent the other major target of HIV-1. Renal macrophages induce and regulate inflammatory responses and are critical to homeostatic regulation of the kidney environment. Combined with their ability to harbour virus, macrophages may also play an important role in the spread of HIV-1 infection in the kidney. DESIGN AND METHODS: Multiparametric histochemistry analysis was performed on kidney biopsies from individuals with HIV-1 associated nephropathy (HIVAN). Primary monocyte-derived macrophages were infected with a GFP-expressing replication competent HIV-1. HIV-1 transfer from macrophages to RTE cells was carried out in a coculture system and evaluated by fluorescence-microscopy and flow-cytometry. Live imaging was performed to assess the fate of HIV-1 infected RTE cells over time. RESULTS: We show that macrophages are abundantly present in the renal inflammatory infiltrate of individuals with HIVAN. We observed contact-dependent HIV-1 transfer from infected macrophages to both primary and immortalized renal cells. Live imaging of HIV-1 infected RTE cells revealed four different fates: proliferation, hypertrophy, latency and cell death. CONCLUSION: Our study suggests that macrophages may play a role in the dissemination of HIV-1 in the kidney and that proliferation of infected renal cells may contribute to HIV-1 persistence in this compartment.


Asunto(s)
Nefropatía Asociada a SIDA/virología , Células Epiteliales/fisiología , Células Epiteliales/virología , Infecciones por VIH/diagnóstico , Túbulos Renales/virología , Riñón/virología , Macrófagos/virología , Nefropatía Asociada a SIDA/patología , Linfocitos T CD4-Positivos , Proliferación Celular , Humanos , Estudios Retrospectivos , Latencia del Virus , Replicación Viral
7.
J Am Soc Nephrol ; 31(8): 1683-1687, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32371536

RESUMEN

BACKGROUND: A significant fraction of patients with coronavirus disease 2019 (COVID-19) display abnormalities in renal function. Retrospective studies of patients hospitalized with COVID-19 in Wuhan, China, report an incidence of 3%-7% progressing to ARF, a marker of poor prognosis. The cause of the renal failure in COVID-19 is unknown, but one hypothesized mechanism is direct renal infection by the causative virus, SARS-CoV-2. METHODS: We performed an autopsy on a single patient who died of COVID-19 after open repair of an aortic dissection, complicated by hypoxic respiratory failure and oliguric renal failure. We used light and electron microscopy to examine renal tissue for evidence of SARS-CoV-2 within renal cells. RESULTS: Light microscopy of proximal tubules showed geographic isometric vacuolization, corresponding to a focus of tubules with abundant intracellular viral arrays. Individual viruses averaged 76 µm in diameter and had an envelope studded with crown-like, electron-dense spikes. Vacuoles contained double-membrane vesicles suggestive of partially assembled virus. CONCLUSIONS: The presence of viral particles in the renal tubular epithelium that were morphologically identical to SARS-CoV-2, and with viral arrays and other features of virus assembly, provide evidence of a productive direct infection of the kidney by SARS-CoV-2. This finding offers confirmatory evidence that direct renal infection occurs in the setting of AKI in COVID-19. However, the frequency and clinical significance of direct infection in COVID-19 is unclear. Tubular isometric vacuolization observed with light microscopy, which correlates with double-membrane vesicles containing vacuoles observed with electronic microscopy, may be a useful histologic marker for active SARS-CoV-2 infection in kidney biopsy or autopsy specimens.


Asunto(s)
Lesión Renal Aguda/complicaciones , Infecciones por Coronavirus/complicaciones , Túbulos Renales/virología , Neumonía Viral/complicaciones , Lesión Renal Aguda/mortalidad , Disección Aórtica/cirugía , Autopsia , Betacoronavirus , COVID-19 , Infecciones por Coronavirus/mortalidad , Células Epiteliales/patología , Humanos , Túbulos Renales/patología , Túbulos Renales/ultraestructura , Masculino , Persona de Mediana Edad , Nefritis/fisiopatología , Pandemias , Neumonía Viral/mortalidad , Pronóstico , Insuficiencia Respiratoria , Estudios Retrospectivos , SARS-CoV-2
9.
Virus Res ; 263: 164-168, 2019 04 02.
Artículo en Inglés | MEDLINE | ID: mdl-30711577

RESUMEN

A 15-year-old female cockatiel (Nymphicus hollandicus) undergoing long term management for hepatopathy died and underwent necropsy. Microscopic findings were consistent with chronic liver disease characterized by distorted hepatic architecture, fibrosis and biliary proliferation. The additional finding of large intranuclear inclusion bodies within hepatocytes and renal tubular epithelium prompted diagnostic next generation sequencing. The assembled sequences isolated from pooled kidney and liver were related to siadenoviruses. The genus Siadenovirus, within the family Adenoviridae, includes several species of viruses that pathogenically infect avian species including hemorrhagic enteritis virus of turkeys and marble spleen virus of pheasants. Siadenoviruses have previously been reported in seven psittacine species: a plum-headed parakeet (Psittacula cyanocephala), an umbrella cockatoo (Cacatua alba) budgerigars (Melopsittacus undulates), an eastern rosella (Platycercus eximius), a scarlet chested parrot (Neophema splendida), a cockatiel (Nymphicus hollandicus), and a red-crowned parakeet (Cyanoramphus novaezelandiae). This report describes a novel siadenovirus in a cockatiel that is highly identical to budgerigar adenovirus 1 and distinct from PsAdV-2 in cockatiels. We report the clinical pathologic, gross, and histopathologic findings in a cockatiel with chronic hepatitis and a novel siadenovirus, PsAdV-5. The sequencing data is presented with a phylogenetic analysis.


Asunto(s)
Infecciones por Adenoviridae/veterinaria , Enfermedades de las Aves/virología , Cacatúas , Hepatitis Viral Animal/virología , Siadenovirus/clasificación , Siadenovirus/aislamiento & purificación , Infecciones por Adenoviridae/virología , Animales , Enfermedades de las Aves/patología , Femenino , Hepatitis Viral Animal/patología , Secuenciación de Nucleótidos de Alto Rendimiento , Histocitoquímica , Túbulos Renales/patología , Túbulos Renales/virología , Hígado/patología , Hígado/virología , Filogenia , Homología de Secuencia , Siadenovirus/genética
10.
BMC Infect Dis ; 18(1): 645, 2018 Dec 12.
Artículo en Inglés | MEDLINE | ID: mdl-30541481

RESUMEN

BACKGROUND: Hemorrhagic fever with renal syndrome (HFRS) caused by pathogenic hantaviruses in Europe and Asia is often characterized by acute kidney injury (AKI) with massive proteinuria. Renal filtration depends on the integrity of epithelial and endothelial monolayers in the tubular and glomerular apparatus. Tubular and glomerular cells represent target cells of hantavirus infection. However, the detailed mechanisms of renal impairment induced by hantaviruses are not well understood. METHODS: We analyzed the cellular consequences of hantavirus infection by measuring adhesion and migration capacity of human renal cells infected with Puumala (PUUV) or Hantaan (HTNV) virus. The impact of hantaviral nucleocapsid proteins (N proteins) on motility was examined by transfection of podocytes. RESULTS: Infection of kidney cells with hantavirus species PUUV and HTNV causes a significant reduction of migration capacity. The impaired motility depends on viral replication and transfection of podocytes with N protein of PUUV or HTNV reveals that the expression of N protein alone is sufficient to deteriorate podocyte function. The cellular effects are more pronounced for the more pathogenic HTNV than for PUUV that causes a milder form of HFRS. CONCLUSIONS: The direct impairment of migration capacity of renal cells by hantaviral N proteins may contribute substantially to proteinuria observed in the clinical picture of hantavirus infection.


Asunto(s)
Movimiento Celular/fisiología , Células Epiteliales/fisiología , Células Epiteliales/virología , Infecciones por Hantavirus/patología , Riñón/fisiología , Riñón/virología , Orthohantavirus/fisiología , Animales , Células Cultivadas , Chlorocebus aethiops , Células Epiteliales/patología , Orthohantavirus/patogenicidad , Humanos , Riñón/patología , Glomérulos Renales/patología , Glomérulos Renales/fisiología , Glomérulos Renales/virología , Túbulos Renales/patología , Túbulos Renales/fisiología , Túbulos Renales/virología , Podocitos/patología , Podocitos/fisiología , Podocitos/virología , Virus Puumala/fisiología , Células Vero , Replicación Viral/fisiología
11.
Transpl Infect Dis ; 20(6): e12974, 2018 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-30102820

RESUMEN

BACKGROUND: Polyomavirus-associated nephropathy is associated with high risk of kidney allograft loss. Whether the cause of native end-stage renal disease influences the risk of BK infection is unclear. METHODS: A retrospective, single-center study of 2741 adult kidney transplant recipients between 1994 and 2014 was performed. Recipients had end-stage renal disease due to polycystic kidney disease (PKD, n = 549), diabetes mellitus (DM, n = 947), hypertension (HTN, n = 442), or glomerulonephritis (GN, n = 803). RESULTS: A total of 327 recipients (12%) developed post-transplant BK viremia over a median follow-up time of 5 years. The incidence rate of BK viremia was lowest in patients with PKD (1.46 per 100 person-years) compared to other causes of ESRD (DM = 2.06, HTN = 2.65, and GN = 2.01 per 100 person-years). A diagnosis of PKD was associated with a lower risk of post-transplant BK viremia (adjusted HR (95% CI) = 0.67 (0.48-0.95), P = 0.02). BK nephropathy was significantly less common in patients with PKD (0.21 per 100 person-years) compared to those with HTN (0.80 per 100 person-years, P ≤ 0.001). Among patients with PKD, the risk of BK viremia was lower in patients with nephrectomy, compared to those without nephrectomy (adjusted HR (95% CI) = 0.42 (0.19-0.92), P < 0.05). CONCLUSION: ESRD due to PKD is associated with a lower risk of post-transplant BK infection. The renal tubular epithelial cells in PKD are unique; they are in a proliferative but non-differentiated state. Whether this characteristic of renal tubular epithelial cells alters the BK viral reservoir or replication in PKD patients warrants further study.


Asunto(s)
Virus BK/aislamiento & purificación , Fallo Renal Crónico/cirugía , Trasplante de Riñón/efectos adversos , Enfermedades Renales Poliquísticas/cirugía , Infecciones por Polyomavirus/epidemiología , Infecciones Tumorales por Virus/epidemiología , Adulto , Células Epiteliales/virología , Femenino , Humanos , Fallo Renal Crónico/etiología , Túbulos Renales/citología , Túbulos Renales/virología , Masculino , Persona de Mediana Edad , Enfermedades Renales Poliquísticas/complicaciones , Infecciones por Polyomavirus/virología , Estudios Retrospectivos , Factores de Riesgo , Infecciones Tumorales por Virus/virología
12.
World J Gastroenterol ; 24(21): 2300-2310, 2018 Jun 07.
Artículo en Inglés | MEDLINE | ID: mdl-29881239

RESUMEN

AIM: To evaluate the differences in acute kidney injury (AKI) between acute-on-chronic liver failure (ACLF) and decompensated cirrhosis (DC) patients. METHODS: During the period from December 2015 to July 2017, 280 patients with hepatitis B virus (HBV)-related ACLF (HBV-ACLF) and 132 patients with HBV-related DC (HBV-DC) who were admitted to our center were recruited consecutively into an observational study. Urine specimens were collected from all subjects and the levels of five urinary tubular injury biomarkers were detected,including neutrophil gelatinase-associated lipocalin (NGAL), interleukin-18 (IL-18), liver-type fatty acid binding protein (L-FABP), cystatin C (CysC), and kidney injury molecule-1 (KIM-1). Simultaneously, the patient demographics, occurrence and progression of AKI, and response to terlipressin therapy were recorded. All patients were followed up for 3 mo or until death after enrollment. RESULTS: AKI occurred in 71 and 28 of HBV-ACLF and HBV-DC patients, respectively (25.4% vs 21.2%, P = 0.358). Among all patients, the levels of four urinary biomarkers (NGAL, CysC, L-FABP, IL-18) were significantly elevated in patients with HBV-ACLF and AKI (ACLF-AKI), compared with that in patients with HBV-DC and AKI (DC-AKI) or those without AKI. There was a higher proportion of patients with AKI progression in ACLF-AKI patients than in DC-AKI patients (49.3% vs 17.9%, P = 0.013). Forty-three patients with ACLF-AKI and 19 patients with DC-AKI were treated with terlipressin. The response rate of ACLF-AKI patients was significantly lower than that of patients with DC-AKI (32.6% vs 57.9%, P = 0.018). Furthermore, patients with ACLF-AKI had the lowest 90 d survival rates among all groups (P < 0.001). CONCLUSION: AKI in ACLF patients is more likely associated with structural kidney injury, and is more progressive, with a poorer response to terlipressin treatment and a worse prognosis than that in DC patients.


Asunto(s)
Lesión Renal Aguda/etiología , Insuficiencia Hepática Crónica Agudizada/complicaciones , Cirrosis Hepática/complicaciones , Lipresina/análogos & derivados , Vasoconstrictores/uso terapéutico , Lesión Renal Aguda/tratamiento farmacológico , Lesión Renal Aguda/epidemiología , Lesión Renal Aguda/orina , Insuficiencia Hepática Crónica Agudizada/virología , Adulto , Biomarcadores/orina , Progresión de la Enfermedad , Femenino , Virus de la Hepatitis B/aislamiento & purificación , Humanos , Túbulos Renales/patología , Túbulos Renales/virología , Cirrosis Hepática/virología , Lipresina/uso terapéutico , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Factores de Riesgo , Terlipresina , Resultado del Tratamiento
13.
J Virol ; 92(2)2018 01 15.
Artículo en Inglés | MEDLINE | ID: mdl-29093088

RESUMEN

Prior studies have found that HIV, through the Vpr protein, promotes genome reduplication (polyploidy) in infection-surviving epithelial cells within renal tissue. However, the temporal progression and molecular regulation through which Vpr promotes polyploidy have remained unclear. Here we define a sequential progression to Vpr-mediated polyploidy in human renal tubule epithelial cells (RTECs). We found that as in many cell types, Vpr first initiates G2 cell cycle arrest in RTECs. We then identified a previously unreported cascade of Vpr-dependent events that lead to renal cell survival and polyploidy. Specifically, we found that a fraction of G2-arrested RTECs reenter the cell cycle. Following this cell cycle reentry, two distinct outcomes occur. Cells that enter complete mitosis undergo mitotic cell death due to extra centrosomes and aberrant division. Conversely, cells that abort mitosis undergo endoreplication to become polyploid. We further show that multiple small-molecule inhibitors of the phosphatidylinositol 3-kinase-related kinase (PIKK) family, including those that target ATR, ATM, and mTOR, indirectly prevent Vpr-mediated polyploidy by preventing G2 arrest. In contrast, an inhibitor that targets DNA-dependent protein kinase (DNA-PK) specifically blocks the Vpr-mediated transition from G2 arrest to polyploidy. These findings outline a temporal, molecularly regulated path to polyploidy in HIV-positive renal cells.IMPORTANCE Current cure-focused efforts in HIV research aim to elucidate the mechanisms of long-term persistence of HIV in compartments. The kidney is recognized as one such compartment, since viral DNA and mRNA persist in the renal tissues of HIV-positive patients. Further, renal disease is a long-term comorbidity in the setting of HIV. Thus, understanding the regulation and impact of HIV infection on renal cell biology will provide important insights into this unique HIV compartment. Our work identifies mechanisms that distinguish between HIV-positive cell survival and death in a known HIV compartment, as well as pharmacological agents that alter these outcomes.


Asunto(s)
Células Epiteliales/metabolismo , Células Epiteliales/virología , VIH-1/fisiología , Interacciones Huésped-Patógeno , Mitosis , Poliploidía , Productos del Gen vpr del Virus de la Inmunodeficiencia Humana/metabolismo , Muerte Celular , Línea Celular , Supervivencia Celular , Proteína Quinasa Activada por ADN/antagonistas & inhibidores , Técnica del Anticuerpo Fluorescente , Puntos de Control de la Fase G2 del Ciclo Celular , Humanos , Túbulos Renales/citología , Túbulos Renales/virología , Microscopía Fluorescente , Modelos Biológicos , Inhibidores de Proteínas Quinasas/farmacología , Transducción de Señal
14.
Transplantation ; 101(8): 1820-1829, 2017 08.
Artículo en Inglés | MEDLINE | ID: mdl-27755502

RESUMEN

BACKGROUND: BK polyomavirus (BKV)-associated nephropathy is a threat to kidney allograft survival affecting up to 15% of renal transplant patients. Previous studies revealed that tubular epithelial cells (TEC) show a limited response towards BKV infection. Here we investigated the interplay between BKV and TEC in more detail. In particular, we questioned whether BKV suppresses and/or evades antiviral responses. METHODS: Human primary TEC and peripheral blood mononuclear cells were infected with BKV Dunlop strain or other viruses. Moreover, TEC were stimulated with genomic double-stranded (ds)DNA or IFN. Viral replication and cellular responses were measured using quantitative real time PCR and multiplex assay. RESULTS: BKV infection of primary human TEC did not induce an antiviral response, whereas infection with influenza A virus, herpes simplex virus 1, or cytomegalovirus induced a strong antiviral response measured by upregulation of interferon-stimulated genes, such as CXCL10 and DAI. In addition, intracellular delivery of dsDNA or stimulation with IFN did elicit a rapid and pronounced response. However, BKV infection did not affect dsDNA-induced gene expression, indicating BKV did not modulate the antiviral response. Prestimulation of primary TEC with IFNα or dsDNA did not hamper replication of BKV, whereas influenza and herpes simplex virus 1 replication were clearly reduced. In contrast, BKV infection of leukocytes did elicit an antiviral response. CONCLUSIONS: BKV specifically evades innate immunity in TEC and is not susceptible to an intrinsic interferon response, which may facilitate latent presence of the virus in this cell type.


Asunto(s)
Virus BK/genética , Inmunidad Innata , Enfermedades Renales/virología , Trasplante de Riñón , Túbulos Renales/patología , Infecciones por Polyomavirus/virología , ARN Viral/genética , Células Cultivadas , Citometría de Flujo , Rechazo de Injerto/inmunología , Rechazo de Injerto/patología , Rechazo de Injerto/virología , Humanos , Enfermedades Renales/metabolismo , Enfermedades Renales/patología , Túbulos Renales/metabolismo , Túbulos Renales/virología , Leucocitos Mononucleares , Infecciones por Polyomavirus/metabolismo , Infecciones por Polyomavirus/patología , Reacción en Cadena en Tiempo Real de la Polimerasa , Replicación Viral
15.
J Virol ; 90(14): 6379-6386, 2016 07 15.
Artículo en Inglés | MEDLINE | ID: mdl-27147740

RESUMEN

UNLABELLED: The APOBEC3 family of DNA cytosine deaminases has important roles in innate immunity and cancer. It is unclear how DNA tumor viruses regulate these enzymes and how these interactions, in turn, impact the integrity of both the viral and cellular genomes. Polyomavirus (PyVs) are small DNA pathogens that contain oncogenic potentials. In this study, we examined the effects of PyV infection on APOBEC3 expression and activity. We demonstrate that APOBEC3B is specifically upregulated by BK polyomavirus (BKPyV) infection in primary kidney cells and that the upregulated enzyme is active. We further show that the BKPyV large T antigen, as well as large T antigens from related polyomaviruses, is alone capable of upregulating APOBEC3B expression and activity. Furthermore, we assessed the impact of A3B on productive BKPyV infection and viral genome evolution. Although the specific knockdown of APOBEC3B has little short-term effect on productive BKPyV infection, our informatics analyses indicate that the preferred target sequences of APOBEC3B are depleted in BKPyV genomes and that this motif underrepresentation is enriched on the nontranscribed stand of the viral genome, which is also the lagging strand during viral DNA replication. Our results suggest that PyV infection upregulates APOBEC3B activity to influence virus sequence composition over longer evolutionary periods. These findings also imply that the increased activity of APOBEC3B may contribute to PyV-mediated tumorigenesis. IMPORTANCE: Polyomaviruses (PyVs) are a group of emerging pathogens that can cause severe diseases, including cancers in immunosuppressed individuals. Here we describe the finding that PyV infection specifically induces the innate immune DNA cytosine deaminase APOBEC3B. The induced APOBEC3B enzyme is fully functional and therefore may exert mutational effects on both viral and host cell DNA. We provide bioinformatic evidence that, consistent with this idea, BK polyomavirus genomes are depleted of APOBEC3B-preferred target motifs and enriched for the corresponding predicted reaction products. These data imply that the interplay between PyV infection and APOBEC proteins may have significant impact on both viral evolution and virus-induced tumorigenesis.


Asunto(s)
Citidina Desaminasa/metabolismo , Regulación de la Expresión Génica , Genoma Viral , Túbulos Renales/enzimología , Antígenos de Histocompatibilidad Menor/metabolismo , Infecciones por Polyomavirus/virología , Poliomavirus/patogenicidad , Replicación Viral , Células Cultivadas , Citidina Desaminasa/antagonistas & inhibidores , Citidina Desaminasa/genética , Humanos , Túbulos Renales/virología , Antígenos de Histocompatibilidad Menor/genética , Poliomavirus/genética , Infecciones por Polyomavirus/patología , ARN Interferente Pequeño/genética , Activación Transcripcional , Regulación hacia Arriba
16.
Am J Transplant ; 16(3): 821-32, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26639422

RESUMEN

BK polyomavirus (BKPyV) replication causes nephropathy and premature kidney transplant failure. Insufficient BKPyV-specific T cell control is regarded as a key mechanism, but direct effects of immunosuppressive drugs on BKPyV replication might play an additional role. We compared the effects of mammalian target of rapamycin (mTOR)- and calcineurin-inhibitors on BKPyV replication in primary human renal tubular epithelial cells. Sirolimus impaired BKPyV replication with a 90% inhibitory concentration of 4 ng/mL by interfering with mTOR-SP6-kinase activation. Sirolimus inhibition was rapid and effective up to 24 h postinfection during viral early gene expression, but not thereafter, during viral late gene expression. The mTORC-1 kinase inhibitor torin-1 showed a similar inhibition profile, supporting the notion that early steps of BKPyV replication depend on mTOR activity. Cyclosporine A also inhibited BKPyV replication, while tacrolimus activated BKPyV replication and reversed sirolimus inhibition. FK binding protein 12kda (FKBP-12) siRNA knockdown abrogated sirolimus inhibition and increased BKPyV replication similar to adding tacrolimus. Thus, sirolimus and tacrolimus exert opposite effects on BKPyV replication in renal tubular epithelial cells by a mechanism involving FKBP-12 as common target. Immunosuppressive drugs may therefore contribute directly to the risk of BKPyV replication and nephropathy besides suppressing T cell functions. The data provide rationales for clinical trials aiming at reducing the risk of BKPyV replication and disease in kidney transplantation.


Asunto(s)
Virus BK/fisiología , Células Epiteliales/virología , Túbulos Renales/virología , Sirolimus/farmacología , Proteína 1A de Unión a Tacrolimus/metabolismo , Tacrolimus/farmacología , Replicación Viral/efectos de los fármacos , Western Blotting , Células Cultivadas , Células Epiteliales/metabolismo , Técnica del Anticuerpo Fluorescente , Humanos , Inmunosupresores/uso terapéutico , Lactante , Túbulos Renales/metabolismo , Infecciones por Polyomavirus/tratamiento farmacológico , Infecciones por Polyomavirus/metabolismo , Infecciones por Polyomavirus/virología , ARN Mensajero/genética , ARN Interferente Pequeño/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteína 1A de Unión a Tacrolimus/antagonistas & inhibidores , Proteína 1A de Unión a Tacrolimus/genética , Infecciones Tumorales por Virus/tratamiento farmacológico , Infecciones Tumorales por Virus/metabolismo , Infecciones Tumorales por Virus/virología
17.
Cancer Lett ; 356(2 Pt B): 953-61, 2015 Jan 28.
Artículo en Inglés | MEDLINE | ID: mdl-25444910

RESUMEN

Human papillomavirus (HPV) DNA integrations may affect therapeutic responses in cancers through ATM network-related DNA damage response (DDR). We studied whether cisplatin-induced DDR was altered in human HK-2 renal tubular cells immortalized by HPV16 E6/E7 genes. Cytotoxicity assays utilized thiazolyl blue dye and DDR was identified by gene expression differences, double-strand DNA breaks, ATM promoter activity, and analysis of cell cycling and side population cells. After cisplatin, HK-2 cells showed greater ATM promoter activity indicating activation of this network, but DDR was muted, since little γH2AX was expressed, DNA strand breaks were absent and cells continued cycling. When HK-2 cells were treated with the MDM2 antagonist inducing p53, nutlin-3, or p53 transcriptional activator, tenovin-1, cell growth decreased but cisplatin toxicity was unaffected. By contrast, arsenic trioxide, which by inhibiting wild-type p53-induced phosphatase-1 that serves responses downstream of p53, and by depolymerizing tubulin, synergistically enhanced cisplatin cytotoxicity including loss of SP cells. Our findings demonstrated that HPV16 E6/E7 altered DDR through p53-mediated cell growth controls, which may be overcome by targeting of WIP1 and other processes, and thus should be relevant for treating renal cell carcinoma.


Asunto(s)
Apoptosis/efectos de los fármacos , Arsenicales/farmacología , Carcinoma de Células Renales/tratamiento farmacológico , Transformación Celular Viral/efectos de los fármacos , Cisplatino/farmacología , Túbulos Renales/efectos de los fármacos , Proteínas Oncogénicas Virales/metabolismo , Óxidos/farmacología , Proteínas E7 de Papillomavirus/metabolismo , Proteínas Represoras/metabolismo , Antineoplásicos/farmacología , Trióxido de Arsénico , Proteínas de la Ataxia Telangiectasia Mutada/genética , Western Blotting , Carcinoma de Células Renales/genética , Carcinoma de Células Renales/virología , Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Transformación Celular Viral/genética , Ensayo Cometa , Roturas del ADN de Doble Cadena/efectos de los fármacos , Sinergismo Farmacológico , Histonas/genética , Histonas/metabolismo , Papillomavirus Humano 16/genética , Humanos , Neoplasias Renales/tratamiento farmacológico , Neoplasias Renales/genética , Neoplasias Renales/virología , Túbulos Renales/metabolismo , Túbulos Renales/virología , Proteínas Oncogénicas Virales/genética , Proteínas E7 de Papillomavirus/genética , Infecciones por Papillomavirus/genética , Infecciones por Papillomavirus/virología , Regiones Promotoras Genéticas/genética , Proteínas Represoras/genética , Células Tumorales Cultivadas , Proteína p53 Supresora de Tumor/genética , Proteína p53 Supresora de Tumor/metabolismo
18.
J Gen Virol ; 95(Pt 11): 2346-2355, 2014 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-25146006

RESUMEN

A major research priority for HIV eradication is the elucidation of the events involved in HIV reservoir establishment and persistence. Cell-to-cell transmission of HIV represents an important area of study as it allows for the infection of cell types which are not easily infected by HIV, leading to the establishment of long-lived viral reservoirs. This phenomenon enables HIV to escape elimination by the immune system. This process may also enable HIV to escape suppressive effects of anti-retroviral drugs. During cell-to-cell transmission of HIV, a dynamic series of events ensues at the virological synapse that promotes viral dissemination. Cell-to-cell transmission involves various types of cells of the immune system and this mode of transmission has been shown to have an important role in sexual and mother-to-child transmission of HIV and spread of HIV within the central nervous system and gut-associated lymphoid tissues. There is also evidence that cell-to-cell transmission of HIV occurs between thymocytes and renal tubular cells. Herein, following a brief review of the processes involved at the virological synapse, evidence supporting the role for cell-to-cell transmission of HIV in the maintenance of the HIV reservoir will be highlighted. Therapeutic considerations and future directions for this area of research will also be discussed.


Asunto(s)
Infecciones por VIH/virología , Linfocitos T CD4-Positivos/virología , Sistema Nervioso Central/virología , Células Dendríticas/virología , Reservorios de Enfermedades/virología , Femenino , Tracto Gastrointestinal/virología , VIH/inmunología , VIH/patogenicidad , Infecciones por VIH/inmunología , Infecciones por VIH/transmisión , Humanos , Sinapsis Inmunológicas/virología , Transmisión Vertical de Enfermedad Infecciosa , Túbulos Renales/virología , Macrófagos/virología , Masculino , Embarazo
19.
Kidney Int ; 86(2): 266-74, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24827777

RESUMEN

Despite improved outcomes among persons living with HIV who are treated with antiretroviral therapy, they remain at increased risk for acute and chronic kidney diseases. Moreover, since HIV can infect renal epithelial cells, the kidney might serve as a viral reservoir that would need to be eradicated when attempting to achieve full virologic cure. In recent years, much progress has been made in elucidating the mechanism by which HIV infects renal epithelial cells and the viral and host factors that promote development of kidney disease. Polymorphisms in APOL1 confer markedly increased risk of HIV-associated nephropathy; however, the mechanism by which ApoL1 variants may promote kidney disease remains unclear. HIV-positive persons are at increased risk of acute kidney injury, which may be a result of a high burden of subclinical kidney disease and/or viral factors and frequent exposure to nephrotoxins. Despite the beneficial effect of antiretroviral therapy in preventing and treating HIVAN, and possibly other forms of kidney disease in persons living with HIV, some of these medications, including tenofovir, indinavir, and atazanavir can induce acute and/or chronic kidney injury via mitochondrial toxicity or intratubular crystallization. Further research is needed to better understand factors that contribute to acute and chronic kidney injury in HIV-positive patients and to develop more effective strategies to prevent and treat kidney disease in this vulnerable population.


Asunto(s)
Nefropatía Asociada a SIDA/etiología , Lesión Renal Aguda/etiología , Fármacos Anti-VIH/efectos adversos , Apolipoproteína L1 , Apolipoproteínas/genética , Genes Virales , Predisposición Genética a la Enfermedad , VIH-1/genética , VIH-1/patogenicidad , Humanos , Enfermedades del Complejo Inmune/etiología , Túbulos Renales/lesiones , Túbulos Renales/virología , Lipoproteínas HDL/genética , Modelos Biológicos , Podocitos/patología , Podocitos/virología , Polimorfismo Genético , Factores de Riesgo , Microangiopatías Trombóticas/etiología
20.
J Am Soc Nephrol ; 25(2): 407-19, 2014 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-24309185

RESUMEN

Since the recent publication of data showing favorable outcomes for patients with HIV-1 and ESRD, kidney transplantation has become a therapeutic option in this population. However, reports have documented unexplained reduced allograft survival in these patients. We hypothesized that the unrecognized infection of the transplanted kidney by HIV-1 can compromise long-term allograft function. Using electron microscopy and molecular biology, we examined protocol renal transplant biopsies from 19 recipients with HIV-1 who did not have detectable levels of plasma HIV-1 RNA at transplantation. We found that HIV-1 infected the kidney allograft in 68% of these patients. Notably, HIV-1 infection was detected in either podocytes predominately (38% of recipients) or tubular cells only (62% of recipients). Podocyte infection associated with podocyte apoptosis and loss of differentiation markers as well as a faster decline in allograft function compared with tubular cell infection. In allografts with tubular cell infection, epithelial cells of the proximal convoluted tubules frequently contained abnormal mitochondria, and both patients who developed features of subclinical acute cellular rejection had allografts with tubular cell infection. Finally, we provide a novel noninvasive test for determining HIV-1 infection of the kidney allograft by measuring HIV-1 DNA and RNA levels in patients' urine. In conclusion, HIV-1 can infect kidney allografts after transplantation despite undetectable viremia, and this infection might influence graft outcome.


Asunto(s)
Infecciones por VIH/virología , VIH-1/aislamiento & purificación , Fallo Renal Crónico/cirugía , Trasplante de Riñón , Riñón/virología , Trasplantes/virología , Adulto , Aloinjertos , Apoptosis , Biopsia , ADN Viral/orina , Femenino , Supervivencia de Injerto , Infecciones por VIH/complicaciones , Infecciones por VIH/orina , Hepatitis C Crónica/complicaciones , Humanos , Hibridación in Situ , Riñón/patología , Fallo Renal Crónico/complicaciones , Túbulos Renales/virología , Masculino , Microscopía Electrónica , Persona de Mediana Edad , Podocitos/virología , Reacción en Cadena de la Polimerasa , Proteinuria/etiología , ARN Viral/orina , Trasplantes/patología , Carga Viral
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