BK virus-induced tubulo-interstitial nephritis in a renal transplant recipient

We present a case of renal BK virus infection with renal allograft dysfunction. Renal allograft biopsy showed mononuclear infiltrates in the interstitium and viral inclusions in the tubular epithelial cells. Infected cells were stained with an anti-polyomavirus antibody. The polymerase chain reaction (PCR) performed on blood, urine, and on the DNA extracted from renal tissue showed the presence of the BK virus DNA sequence. The immunosuppressive therapy including tacrolimus, prednisone, and mycophenolate mofetil was reduced leading to an improvement of the renal function. BK virus infection is now recognized as a cause of renal allograft dysfunction, and has been observed with increasing frequency in recent years. Reactivation of the latent virus occurs in immunocompromised hosts such as organ recipients with immunosuppressive treatment. Histologically, renal BK virus infection is characterized by a lymphocytic interstitial infiltrate, and could mimic acute rejection. The pathologist should diagnose the viral infection and may be helped by urine cytology and immunohistochemistry. An accurate diagnosis is important because antirejection therapy favors the decline of the renal function. Enhanced new immunotherapy protocols seem to be the main risk factor for this infection. The response to reduced immunosuppression is variable with reports of an end stage renal failure in 70% of the patients after 18 months.     

Testing for polyomavirus type BK DNA in plasma to identify renal-allograft recipients with viral nephropathy

BACKGROUND: Reactivation of polyomavirus type BK (BK virus) is increasingly recognized as a cause of severe renal-allograft dysfunction. Currently, patients at risk for nephropathy due to infection with the BK virus are identified by the presence of cells containing viral inclusion bodies ("decoy cells") in the urine or by biopsy of allograft tissue. METHODS: In a retrospective analysis, we performed polymerase-chain-reaction assays for BK virus DNA in plasma samples from 9 renal-allograft recipients with BK virus nephropathy; 41 renal-allograft recipients who did not have signs of nephropathy, 16 of whom had decoy cells in the urine; and as immunocompromised controls, 17 patients who had human immunodeficiency virus type 1 (HIV-1) infection (stage C3 according to the classification of the Centers for Disease Control and Prevention) and who had not undergone transplantation. RESULTS: In all nine patients with BK virus nephropathy, BK virus DNA was detected in the plasma at the time of the initial histologic diagnosis (a mean [+/-SD] of 46+/-28 weeks after transplantation) and during the course of histologically diagnosed, persistent BK virus disease. In three of the six patients with nephropathy who were studied serially after transplantation, BK virus DNA was initially undetectable but was detected 16 to 33 weeks before nephropathy became clinically evident and was confirmed by biopsy. Tests for BK virus DNA in plasma became negative and the nephropathy resolved after the doses of immunosuppressive drugs were decreased in two patients and after removal of the renal allograft in three patients. BK virus DNA was found in the plasma of only 2 of the 41 renal-allograft recipients who had no signs of nephropathy and in none of the patients with HIV-1 infection. CONCLUSIONS: Testing for BK virus DNA in plasma from renal-allograft recipients with use of the polymerase chain reaction is a sensitive and specific method for identifying viral nephropathy.     

BK polyomavirus diversity—Why viral variation matters

BK polyomavirus (BKPyV or BKV) is a non‐enveloped, circular double‐stranded DNA virus that may exceed 80% seroprevalence in adults. BKV infection typically occurs during childhood, and the majority of adults are latently infected. While BKV infection is rarely associated with clinical disease in most individuals, in immunosuppressed individuals, reactivation may cause kidney (BK‐associated nephropathy) or bladder (hemorrhagic cystitis and ureteral stenosis) injury. No antiviral therapies have been approved for the treatment of BKV infection. Reducing immunosuppression is the most effective therapy, although this is not feasible in many patients. Thus, a robust understanding of viral pathogenesis and viral diversity remains important for the development of future therapeutic strategies. Studies of BKV diversity are quite sparse compared to other common viral infections; thus, much of our understanding of BVK variability and evolution relies heavily analogous studies of other viruses such as HIV or viral hepatitis. We provide a comprehensive review of BKV diversity at the population and individual level with careful consideration of how viral variability may impact viral replication, pathogenesis, tropism, and protein function. We also discuss a number of outstanding questions related to BK virus diversity that should be explored rigorously in future studies.     

Electron microscopy in the diagnosis of BK-polyoma virus infection in the transplanted kidney

BK polyomavirus has become an important etiologic agent responsible for significant morbidity in renal transplant recipients. This virus can be detected in transitional cells in the urine (decoy cells) using cytology, but correlation with allograft function status and histologic evidence of renal involvement is poor. Accurate diagnosis of BK polyomavirus infection requires a high index of suspicion and utilization of ancillary diagnostic techniques in many cases. Electron microscopy is very sensitive in depicting the presence of BK virions, but the finding of viral particles is not by itself diagnostic of BK interstitial nephritis. Management of patients with polyoma virus nephropathy is difficult since there is no specific antiviral therapy available at this time.     

The use of brincidofovir for the treatment of mixed dsDNA viral infection

Double-stranded DNA (dsDNA) viral infections constitute a major complication following solid organ and stem cell transplantation. Few therapeutic options are currently available for the treatment of such infections in highly immunocompromised hosts. Brincidofovir is an oral investigational drug with broad antiviral activity against dsDNA viruses in vitro, but clinical experience is limited. Here we report a young female who developed a mixed infection with adenovirus, cytomegalovirus, Epstein–Barr virus and BK polyomavirus after an allogeneic stem cell transplant, and was successfully treated with brincidofovir.     

BK Polyomavirus Interstitial Nephritis in a Renal Allograft Recipient

Human polyomavirus (PV) interstitial nephritis has recently been recognized as a cause of severe renal allograft dysfunction. It occurs in immunosuppressed patients after reactivation of the latent virus PV type BK (BK virus) in the renal epithelium. BK disease is defined as a morphologically manifest renal infection with cytopathic signs accompanied by varying degrees of interstitial inflammatory cell infiltrates and functional impairment. It is also identified by the presence of cells containing viral inclusion bodies (decoy cells) in the urine. The authors report a case of BK PV interstitial nephritis in a 36-year-old renal allograft recipient. Under light microscopy the chief diagnostic indicator was detection of intranuclear viral inclusions, which were found exclusively in tubular epithelial cells. Cells with viral changes were often enlarged with nuclear atypia and chromatin basophilia. Widespread interstitial plasma cell infiltrates associated with tubulitis were present. Intranuclear paracrystalline arrays of virus particles 35-38 nm in diameter were present as characteristic ultrastructural indicators. Urine samples revealed decoy cells with ground-glass-type intranuclear inclusions positive for BK virus by electron microcopy.     

Diagnosis of childhood BK virus cystitis by electron microscopy and PCR.

A case of BK virus cystitis in a 5 year old boy is reported. This patient, who was not immunocompromised, had had acute cystitis for two weeks. Many intracytoplasmic inclusions were observed in urinary sediment smears stained by the Papanicolaou method. Electron microscopic examination showed virus particles, presumed to be human polyomavirus, in the nuclei of the degenerated urothelial cells. A DNA sequence of the BK virus was detected in 200-300 urothelial cells in Papanicolaou stained smears by the polymerase chain reaction. BK virus is an unusual cause of symptomatic cystitis in a healthy child.     

BK polyomavirus interstitial nephritis in a renal allograft recipient

Human polyomavirus (PV) interstitial nephritis has recently been recognized as a cause of severe renal allograft dysfunction. It occurs in immunosuppressed patients after reactivation of the latent virus PV type BK (BK virus) in the renal epithelium. BK disease is defined as a morphologically manifest renal infection with cytopathic signs accompanied by varying degrees of interstitial inflammatory cell infiltrates and functional impairment. It is also identified by the presence of cells containing viral inclusion bodies (decoy cells) in the urine. The authors report a case of BK PV interstitial nephritis in a 36-year-old renal allograft recipient. Under light microscopy the chief diagnostic indicator was detection of intranuclear viral inclusions, which were found exclusively in tubular epithelial cells. Cells with viral changes were often enlarged with nuclear atypia and chromatin basophilia. Widespread interstitial plasma cell infiltrates associated with tubulitis were present. Intranuclear paracrystalline arrays of virus particles 35-38 nm in diameter were present as characteristic ultrastructural indicators. Urine samples revealed decoy cells with ground-glass-type intranuclear inclusions positive for BK virus by electron microcopy.     

Correlates of quantitative measurement of BK polyomavirus (BKV) DNA with clinical course of BKV infection in renal transplant patients

BK virus-allograft nephropathy (BKVAN) is an increasingly recognized complication after kidney transplantation. Quantitative tests have been advocated to monitor patients, but data demonstrating their efficacy are relatively limited. We developed a real-time PCR assay to quantitate BK virus loads in the setting of renal transplantation, and we correlated the BK virus load with clinical course and with the presence of BK virus in renal biopsy specimens. BK virus loads were measured in urine, plasma, and kidney biopsy samples in three clinical settings: (i) patients with asymptomatic BK viruria, (ii) patients with active BKVAN, and (iii) patients with resolved BKVAN. Active BKVAN was associated with BK viremia greater than 5 x 103 copies/ml and with BK viruria greater than 107 copies/ml in all cases. Resolution of nephropathy led to resolution of viremia, decreased viruria levels, and disappearance of viral inclusions, but low-level viral DNA persisted in biopsy specimens even for patients whose viruria was cleared. All but one patient in the resolved BKVAN group carried a urinary viral load below 107 copies/ml. Viral loads in patients with asymptomatic viruria were generally lower but in some cases overlapped with levels more typical of BKVAN. One patient with asymptomatic viruria and with a viral load overlapping values seen in BKVAN had developed nephropathy by the time of follow-up. In conclusion, serial measurement of viral loads by quantitative PCR is a useful tool in monitoring the course of BK virus infection. The results should be interpreted in conjunction with the clinical picture and biopsy findings.     

BK Polyomavirus in Renal Transplants: Role of Electron Microscopy and Immunostaining in Detecting Early Infection

Reactivation of BK polyomavirus (BKV) is increasingly recognized as a cause of failure of renal allografts. Since no specific treatment is available for this infection, early diagnosis is important, as it allows for early intervention and possible recovery of renal function. Forty-four consecutive renal transplant biopsies performed over a 2-year period were included in the study. In addition to evaluation of renal biopsy tissue sections using routine histochemical stains, CD3, CD20, BK virus immunostains using the specific BK virus and the SV40 antibodies and electron microscopy studies were performed. None of the transplant cases but one exhibited classical histologic viral changes. Viral particles were seen by EM in 19%, and BK-virus positivity was identified in only 43% of these cases. CD20-rich inflammatory infiltrates predominated in cases in which either positive BK stain and/or viral particles were identified ultrastructurally. A combined approach using electron microscopic and immunohistochemical evaluation can be utilized effectively to identify BK virus-associated nephropathy at an early phase facilitating early clinical intervention.     

BK polyomavirus in renal transplants: role of electron microscopy and immunostaining in detecting early infection

Reactivation of BK polyomavirus (BKV) is increasingly recognized as a cause of failure of renal allografts. Since no specific treatment is available for this infection, early diagnosis is important, as it allows for early intervention and possible recovery of renal function. Forty-four consecutive renal transplant biopsies performed over a 2-year period were included in the study. In addition to evaluation of renal biopsy tissue sections using routine histochemical stains, CD3, CD20, BK virus immunostains using the specific BK virus and the SV40 antibodies and electron microscopy studies were performed. None of the transplant cases but one exhibited classical histologic viral changes. Viral particles were seen by EM in 19%, and BK-virus positivity was identified in only 43% of these cases. CD20-rich inflammatory infiltrates predominated in cases in which either positive BK stain and/or viral particles were identified ultrastructurally. A combined approach using electron microscopic and immunohistochemical evaluation can be utilized effectively to identify BK virus-associated nephropathy at an early phase facilitating early clinical intervention.     

New commercially available PCR and microplate hybridization assay for detection and differentiation of human polyomaviruses JC and BK in cerebrospinal fluid, serum, and urine samples

JC and BK human polyomaviruses (family Polyomaviridae) may cause severe neurological or urinary tract pathologies in immunocompromised hosts. In the present study, we evaluated a new commercially available PCR and microplate colorimetric hybridization assay for the standardized differential detection of JC virus (JCV) and BK virus (BKV) genomes in clinical samples. This JC/BK Consensus test was first evaluated by testing serial dilutions of JCV or BKV plasmid DNA standards and was then compared with an in-house reference PCR assay for the detection of JC and BK virus genomes in 70 cerebrospinal fluid (CSF) samples of patients with neurological disorders and in 75 serum or plasma samples and 125 urine samples of renal graft recipients. This new test allowed a limit of detection of 10 copies and 1 copy of JC and BK virus genomes, respectively, and was able to differentiate various levels of JCV, BKV, and mixed JCV and BKV DNA genomes in a single reaction tube. Our results showed 100% specificity and sensitivity for the JC/BK Consensus test with CSF samples. With serum or plasma samples, this test had a sensitivity and a specificity of 100% for both JCV and mixed JCV and BKV DNA detection and a sensitivity and a specificity of 100 and 97.8% for BKV DNA detection, respectively. With urine samples, the sensitivity and specificity were 100 and 96.6%, respectively, for JCV DNA detection; 100 and 89.4%, respectively, for BKV DNA detection; and 44.4 and 100%, respectively, for mixed JCV and BKV DNA detection. In conclusion, our data indicate that this new test, the JC/BK Consensus test, is valuable for the sensitive and specific differential detection of single JCV and BKV infections in CSF, serum or plasma, and urine samples. The use of this reliable PCR assay would improve the routine virological diagnosis as well as the clinical care of immunocompromised patients with polyomavirus-related pathologies.     

Renal failure due to BK virus infection in an immunodeficient child

We report a child with cartilage-hair hypoplasia and Hodgkin's disease who developed progressive renal failure and died following infection with a polyomavirus, BK virus. Renal biopsy showed interstitial inflammation, tubular atrophy, and intranuclear inclusions in tubular epithelium, with large numbers of papovavirus particles within the cells. BK virus infection was confirmed by polymerase chain reaction on renal biopsy material and in urine and the demonstration of a high titre of haemagglutination inhibition (HI) and IgM antibody to the virus in the patient's serum. This case emphasises the need to study in depth any unusual clinical manifestation in immunocompromised patients to delineate better the full clinical impact of less well-established pathogens such as BK virus. © 1995 Wiley-Liss, Inc.     

Antibodies to dsDNA are Produced During Primary BK Virus Infection in Man,Indicating that Anti-dsDNA Antibodies may be Related to Virus Replication In Vivo

Experimental immunizations with both the Polyomavirus BK and with the isolated viral genomic dsDNA regularly induce antibodies with a relative affinity for BK virus dsDNA. In the present study we demonstrate that the anti-dsDNA responses to BK virus in experimental animals also appear during natural BK virus infection in man. Fifty-nine children were examined over time for serological signs of primary BK virus infection. Of eight children found to undergo primary infection with BK virus, anti-BK dsDNA antibodies appeared in all. In 4 of the 8 patients the antibodies cross-reacted significantly with mammalian dsDNA, and weak cross-reactions were also noted in at least three other patients. The antibodies resembled those induced in the experimental model with regard to their relative affinity for BK dsDNA. In contrast, most, but not all, anti-dsDNA antibodies from 10 SLE patients cross-reacted extensively with dsDNA from viral and mammalian origin. Thus, a dsDNA virus like BK virus may provoke immunological intolerance to dsDNA, but, with qualities different from those produced during SLE. The present observations demonstrate that induction of anti-dsDNA antibodies is not restricted to experimental immunization of animals, but does also take place in humans during naturally acquired BK virus infection. The relevance of this model for the spontaneous production of anti-dsDNA antibodies is discussed.     

Human parainfluenza virus giant cell pneumonia following cord blood transplant associated with pulmonary alveolar proteinosis

Giant cell pneumonia secondary to human parainfluenza virus 3 has been reported only rarely in immunocompromised hosts. The few cases documented after bone marrow transplant have resulted in significant morbidity and mortality. To our knowledge, this entity has not been described following umbilical cord blood transplant. Pulmonary alveolar proteinosis, a rare condition that has been reported with increasing frequency in association with immunocompromise and infections, has not been documented in the setting of either umbilical cord blood transplant or human parainfluenza viral infection. We report what we believe is the first documented case of giant cell pneumonia caused by human parainfluenza virus 3 in an umbilical cord blood transplant recipient. To our knowledge, a unique associated feature of this case, a pulmonary alveolar proteinosis-like reaction, has not been reported previously in association with human parainfluenza virus pneumonia.     

BK virus histopathologic disease severity does not predict allograft outcome in renal transplant recipients

AimsBK polyomavirus nephropathy (BKPyVN) is an important cause of allograft failure after renal transplantation. Despite early screening for the virus, allograft loss from BKPyVN is still experienced in up to 14% of all renal transplant recipients. The aim of this study was to investigate the association between BKPyVN histopathologic disease severity and allograft outcome at our center.MethodsKidney transplant recipients who had undergone transplantation between 2002 and 2014 with biopsy proven BKPyVN were eligible for this retrospective study. Each biopsy was re-evaluated by a single pathologist blinded to the clinical data and scored according to the Banff criteria for rejection and BKPyVN. Serum creatinine and BK viral load at the time of biopsy diagnosis as well as allograft outcomes to include allograft survival and serum BK viremia resolution were collected for each recipient to determine if BK virus histopathologic disease severity could predict allograft outcome.ResultsTwenty cases of BKPyVN were identified from 1031 total renal transplants performed. There was no statistical association between allograft loss and BKPyVN histopathology (p = 0.49). There was also no statistical association between BKPyVN histopathology and BK viral load at the time of biopsy diagnosis (p = 0.38) or serum BK viremia resolution (p = 0.16).ConclusionsBKPyVN histopathology does not appear to be useful in predicting renal allograft outcome in those recipients diagnosed with BKPyVN which is in contrast to some previously published data.     

The human polyomavirus BK (BKPyV): virological background and clinical implications

Polyomavirus BK (BKPyV) infects most people subclinically during childhood and establishes a lifelong infection in the renourinary tract. In most immunocompetent individuals, the infection is completely asymptomatic, despite frequent episodes of viral reactivation with shedding into the urine. In immunocompromised patients, reactivation followed by high‐level viral replication can lead to severe disease: 1–10% of kidney transplant patients develop polyomavirus‐associated nephropathy (PyVAN) and 5–15% of allogenic hematopoietic stem cell transplant patients develop polyomavirus‐associated haemorrhagic cystitis (PyVHC). Other conditions such as ureteric stenosis, encephalitis, meningoencephalitis, pneumonia and vasculopathy have also been associated with BKPyV infection in immunocompromised individuals. Although BKPyV has been associated with cancer development, especially in the bladder, definitive evidence of a role in human malignancy is lacking. Diagnosis of PyVAN and PyVHC is mainly achieved by quantitative PCR of urine and plasma, but also by cytology, immunohistology and electron microscopy. Despite more than 40 years of research on BKPyV, there is still no effective antiviral therapy. The current treatment strategy for PyVAN is to allow reconstitution of immune function by reducing or changing the immunosuppressive medication. For PyVHC, treatment is purely supportive. Here, we present a summary of the accrued knowledge regarding BKPyV.     

Structural proteins of a human papovavirus (BK virus): a comparison with the structural proteins of simian virus 40.

The protein composition of SV40 and BK virus was analyzed by polyacrylamide gel electrophoresis. The data indicate that the electrophoretic mobility of the virion polypeptides coded by the viral genome is different in SV40 and BK virus, whereas the electrophoretic mobility of the virion polypeptides originated from the host cell is the same in the two viruses. One more polypeptide was detected in BK virus as compared to SV40.     

On the biological origin of anti-double-stranded (ds)DNA antibodies: systemic lupus erythematosus-related anti-dsDNA antibodies are induced by polyomavirus BK in lupus-prone (NZBxNZW) F1 hybrids,but not in normal mice

We have recently demonstrated that polyomavirus BK and isolated BK double-stranded (ds)DNA have a strong potential for induction of anti-dsDNA antibodies. Here, data are presented that demonstrate that normal mice (a term used in this report for mice not predisposed to a lupus-like syndrome) of four different strains responded to both BK virus and BK dsDNA by producing transient antibodies binding preferentially to the viral dsDNA itself. These antibodies did not bind in the Crithidia luciliae assay, and did not seem to be of pathogenic significance, as neither signs of proteinuria nor immunochemical signs of glomerulonephritis developed in these mice. In contrast, 5-week-old (NZBxNZW)F₁ mice developed strong and persistent anti-dsDNA antibodies in response to BK virus and BK dsDNA, with similar features to those of anti-dsDNA antibodies from individuals with systemic lupus erythematosus: they reacted strongly in the Crithidia luciliae assay and cross-reacted with viral as well as with mammalian dsDNA. Furthermore, persistent proteinuria and glomerulonephritis, with demonstrable heavy mesangial deposits of immune complexes containing IgG anti-dsDNA antibodies, developed 2--3 months earlier than in spontaneously autoimmune control mice. The relevance of these observations to a viral origin of anti-dsDNA antibodies in lupus is discussed.