Decoy Cells Research Articles

Shedding Light on Viral Shedding: Novel Insights into Nuclear Assembly, Cytoplasmic Transformation and Extracellular Vesicle Release of the BK Virus.

Despite the high prevalence of BK polyomavirus (BKPyV) and the associated risk for BKPyV-associated nephropathy (BKPyVAN) in kidney transplant (KTX) recipients, many details on viral processes such as replication, maturation, assembly and virion release from host cells have not been fully elucidated. VP1 is a polyomavirus-specific protein that is expressed in the late phase of its replicative cycle with important functions in virion assembly and infectious particle release. This study investigated the localization and time-dependent changes in the distribution of VP1-positive viral particles and their association within the spectrum of differing cell morphologies that are observed in the urine of KTX patients upon active BKPyV infection. We found highly differing recognition patterns of two anti-VP1 antibodies with respect to intracellular and extracellular VP1 localization, pointing towards independent binding sites that were seemingly associated with differing stages of virion maturation. Cells originating from single clones were stably cultured out of the urine sediment of KTX recipients with suspected BKPyVAN. The cell morphology, polyploidy, virus replication and protein production were investigated by confocal microscopy using both a monoclonal (mAb 4942) and a polyclonal rabbit anti-VP1-specific antibody (RantiVP1 Ab). Immunoblotting was performed to investigate changes in the VP1 protein. Both antibodies visualized VP1 and the mAb 4942 recognized VP1 in cytoplasmic vesicles exhibiting idiomorphic sizes when released from the cells. In contrast, the polyclonal antibody detected VP1 within the nucleus and in cytoplasm in colocalization with the endoplasmic reticulum marker CNX. At the nuclear rim, VP1 was recognized by both antibodies. Immunoblotting revealed two smaller versions of VP1 in urinary decoy cell extracts, potentially from different translation start sites as evaluated by in silico analysis. Oxford Nanopore sequencing showed integration of BKPyV DNA in chromosomes 3, 4 and 7 in one of the five tested primary cell lines which produced high viral copies throughout four passages before transcending into senescence. The different staining with two VP1-specific antibodies emphasizes the modification of VP1 during the process of virus maturation and cellular exit. The integration of BKPyV into the human genome leads to high virus production; however, this alone does not transform the cell line into a permanently cycling and indefinitely replicating one.

The miR-29-3p family suppresses inflammatory osteolysis.

Osteoclasts are the cells primarily responsible for inflammation-induced bone loss, as is particularly seen in rheumatoid arthritis. Increasing evidence suggests that osteoclasts formed under homeostatic versus inflammatory conditions may differ in phenotype. While microRNA-29-3p family members (miR-29a-3p, miR-29b-3p, miR-29c-3p) promote the function of RANKL-induced osteoclasts, the role of miR-29-3p during inflammatory TNF-α-induced osteoclastogenesis is unknown. We used bulk RNA-seq, histology, qRT-PCR, reporter assays, and western blot analysis to examine bone marrow monocytic cell cultures and tissue from male mice in which the function of miR-29-3p family members was decreased by expression of a miR-29-3p tough decoy (TuD) competitive inhibitor in the myeloid lineage (LysM-cre). We found that RANKL-treated monocytic cells expressing the miR-29-3p TuD developed a hypercytokinemia/proinflammatory gene expression profile in vitro, which is associated with macrophages. These data support the concept that miR-29-3p suppresses macrophage lineage commitment and may have anti-inflammatory effects. In correlation, when miR-29-3p activity was decreased, TNF-α-induced osteoclast formation was accentuated in an in vivo model of localized osteolysis and in a cell-autonomous manner in vitro. Further, miR-29-3p targets mouse TNF receptor 1 (TNFR1/Tnfrsf1a), an evolutionarily conserved regulatory mechanism, which likely contributes to the increased TNF-α signaling sensitivity observed in the miR-29-3p decoy cells. Whereas our previous studies demonstrated that the miR-29-3p family promotes RANKL-induced bone resorption, the present work shows that miR-29-3p dampens TNF-α-induced osteoclastogenesis, indicating that miR-29-3p has pleiotropic effects in bone homeostasis and inflammatory osteolysis. Our data supports the concept that the knockdown of miR-29-3p activity could prime myeloid cells to respond to an inflammatory challenge and potentially shift lineage commitment toward macrophage, making the miR-29-3p family a potential therapeutic target for modulating inflammatory response.

The Second International Consensus Guidelines on the Management of BK Polyomavirus in Kidney Transplantation.

BK polyomavirus (BKPyV) remains a significant challenge after kidney transplantation. International experts reviewed current evidence and updated recommendations according to Grading of Recommendations, Assessment, Development, and Evaluations (GRADE). Risk factors for BKPyV-DNAemia and biopsy-proven BKPyV-nephropathy include recipient older age, male sex, donor BKPyV-viruria, BKPyV-seropositive donor/-seronegative recipient, tacrolimus, acute rejection, and higher steroid exposure. To facilitate early intervention with limited allograft damage, all kidney transplant recipients should be screened monthly for plasma BKPyV-DNAemia loads until month 9, then every 3 mo until 2 y posttransplant (3 y for children). In resource-limited settings, urine cytology screening at similar time points can exclude BKPyV-nephropathy, and testing for plasma BKPyV-DNAemia when decoy cells are detectable. For patients with BKPyV-DNAemia loads persisting >1000 copies/mL, or exceeding 10 000 copies/mL (or equivalent), or with biopsy-proven BKPyV-nephropathy, immunosuppression should be reduced according to predefined steps targeting antiproliferative drugs, calcineurin inhibitors, or both. In adults without graft dysfunction, kidney allograft biopsy is not required unless the immunological risk is high. For children with persisting BKPyV-DNAemia, allograft biopsy may be considered even without graft dysfunction. Allograft biopsies should be interpreted in the context of all clinical and laboratory findings, including plasma BKPyV-DNAemia. Immunohistochemistry is preferred for diagnosing biopsy-proven BKPyV-nephropathy. Routine screening using the proposed strategies is cost-effective, improves clinical outcomes and quality of life. Kidney retransplantation subsequent to BKPyV-nephropathy is feasible in otherwise eligible recipients if BKPyV-DNAemia is undetectable; routine graft nephrectomy is not recommended. Current studies do not support the usage of leflunomide, cidofovir, quinolones, or IVIGs. Patients considered for experimental treatments (antivirals, vaccines, neutralizing antibodies, and adoptive T cells) should be enrolled in clinical trials.

JC Virus in Kidney Transplant Population: Are We Cautious Enough?

The John Cunningham virus (JCV) is a polyomavirus that usually infects people at a young age and does not cause any symptoms in immunocompetent individuals. However, in immunocompromised individuals, such as kidney transplant recipients, JCV can cause severe and potentially fatal disease. Unfortunately, JCV has not been researched as extensively as the BK virus and is not mentioned in relevant kidney transplant guidelines. This lack of attention to JCV can lead to less consideration in kidney transplant patients' care. Surveillance using locally available diagnostic methods is of the utmost importance. The presence of JCV can be diagnosed with urine decoy cells, viruria, or viremia verified by the PCR method. A low threshold for considering JCV as a possible cause of any neurological or renal dysfunction in kidney transplant recipients must be maintained. In such cases, kidney and brain biopsy are indicated. Maintaining the appropriate immunosuppression while avoiding over-immunosuppression to prevent JCV disease is crucial, and the approach should be individual, according to overall immunological risk. We hypothesize that the presence of the JCV can indicate overt immunosuppression and identify kidney transplant recipients more prone to opportunistic infections and diseases, including some malignancies. To explore that, future observational studies are needed.

Detection of Bk Polyomavirus Antigen in Patients with Acute Lymphoblastic Leukemia

Background: BK polyomavirus that is acquired in early childhood as BKPyV is near ubiquitous in adults with a seroprevalence of >80%, it has been found as the main cause of hemorrhagic cystitis in hematopoietic stem cell transplantation patients, due to immunosuppression regimen that lead to the activation of the virus from the latency status and lead to increased viral shedding in urine (viuria).
 Objective: To investigate the frequency of BK Polyomavirus antigen excretion in urine of children with acute lymphoblastic leukemia with and without chemotherapy and compare it with normal controls. 
 Patients and Methods: A case-control study conducted from December 2021 to May 2022 in Baghdad, Iraq on leukemia patients of Central Pediatrics Hospital (Al-Eskan ). Urine samples and urine sediment smears were collected from 60 acute lymphoblastic leukemia patients. And compared with 60 apparently healthy age and sex-matched children, BK polyomavirus antigen in urine was detected using Enzyme-linked Immunosorbent Assay and urine cytology were Pap stained for the detection of decoy cells (DCs).
 Results: Positive BKPyV antigen in urine was seen in 55 (91.7%) of Acute lymphoblastic leukemia patients and 39 (65.0%) in controls (p=<0.001) and all the patients were decoy negative. There was no significant effect of the positivity of antigenuria on neither leukocytes level nor on the occurance of relapse in leukemia patients. 
 Conclusion: The very high frequency of BKPyV in the urine signifies the importance of reactivation of this virus in ALL patients with and without chemotherapy.

Detection of Bk Polyomavirus Antigen in Patients with Acute Lymphoblastic Leukemia

Background: BK polyomavirus that is acquired in early childhood as BKPyV is near ubiquitous in adults with a seroprevalence of >80%, it has been found as the main cause of hemorrhagic cystitis in hematopoietic stem cell transplantation patients, due to immunosuppression regimen that lead to the activation of the virus from the latency status and lead to increased viral shedding in urine (viuria). Objective: To investigate the frequency of BK Polyomavirus antigen excretion in urine of children with acute lymphoblastic leukemia with and without chemotherapy and compare it with normal controls. Patients and Methods: A case-control study conducted from December 2021 to May 2022 in Baghdad, Iraq on leukemia patients of Central Pediatrics Hospital (Al-Eskan ). Urine samples and urine sediment smears were collected from 60 acute lymphoblastic leukemia patients. And compared with 60 apparently healthy age and sex-matched children, BK polyomavirus antigen in urine was detected using Enzyme-linked Immunosorbent Assay and urine cytology were Pap stained for the detection of decoy cells (DCs). Results: Positive BKPyV antigen in urine was seen in 55 (91.7%) of Acute lymphoblastic leukemia patients and 39 (65.0%) in controls (p=<0.001) and all the patients were decoy negative. There was no significant effect of the positivity of antigenuria on neither leukocytes level nor on the occurance of relapse in leukemia patients. Conclusion: The very high frequency of BKPyV in the urine signifies the importance of reactivation of this virus in ALL patients with and without chemotherapy. Keywords: Acute lymphoblastic leukemia, BK Polyomavirus, Antigenuria

Acute Kidney Injury and BK Polyomavirus in Urine Sediment Cells.

Polyomaviruses are widespread, with BK viruses being most common in humans who require immunosuppression due to allotransplantation. Infection with BK polyomavirus (BKV) may manifest as BK virus-associated nephropathy and hemorrhagic cystitis. Established diagnostic methods include the detection of polyomavirus in urine and blood by PCR and in tissue biopsies via immunohistochemistry. In this study, 79 patients with pathological renal retention parameters and acute kidney injury (AKI) were screened for BK polyomavirus replication by RNA extraction, reverse transcription, and virus-specific qPCR in urine sediment cells. A short fragment of the VP2 coding region was the target of qPCR amplification; patients with (n = 31) and without (n = 48) a history of renal transplantation were included. Urine sediment cell immunofluorescence staining for VP1 BK polyomavirus protein was performed using confocal microscopy. In 22 patients with acute renal injury, urinary sediment cells from 11 participants with kidney transplantation (KTX) and from 11 non-kidney transplanted patients (nonKTX) were positive for BK virus replication. BK virus copies were found more frequently in patients with AKI stage III (n = 14). Higher copy numbers were detected in KTX patients having experienced BK polyoma-nephropathy (BKPyVAN) in the past or diagnosed recently by histology (5.6 × 109-3.1 × 1010). One patient developed BK viremia following delayed graft function (DGF) with BK virus-positive urine sediment. In nonKTX patients with BK copies, decoy cells were absent; however, positive staining of cells was found with epithelial morphology. Decoy cells were only found in KTX patients with BKPyVAN. In AKI, damage to the tubular epithelium itself may render the epithelial cells more permissive for polyoma replication. This non-invasive diagnostic approach to assess BK polyomavirus replication in urine sediment cells has the potential to identify KTX patients at risk for viremia and BKPyVAN during AKI. This method might serve as a valuable screening tool for close monitoring and tailored immunosuppression decisions.

Human polyomaviruses JCPyV and MCPyV in urothelial cell carcinoma: a single institution experience.

Urothelial cell carcinoma (UCC) is the most common type of urinary bladder. JCPyV and BKPyV have been detected in the urine and tissue of urothelial cell carcinomas (UCC) in immunocompetent patients. Here, we investigated the presence of several HPyVs in UCC samples using diverse molecular techniques to study the prevalence of HPyVs in UCC. A large single-institution database of urine cytology specimens (UCS; n = 22.867 UCS) has previously been searched for decoy cells (n = 30), suggesting polyomavirus infection. The available urine sediments and formalin-fixed paraffin-embedded (FFPE) tissue samples of UCC patients were tested for the presence of JCPyV-LTAg expression by immunohistochemistry (IHC) labeled with SV40-LTAg antibody (clone: PAb416) and subsequent PCR followed by sequencing. In addition, the presence of the oncogenic Merkel cell polyomavirus (MCPyV) and the presence of human polyomavirus 6 (HPyV6) and 7 (HPyV7) DNA were tested with DNA PCR or IHC. Of the 30 patients harboring decoy cells, 14 were diagnosed with UCC of the urinary bladder (14/30; 46.6%) before presenting with decoy cells in the urine. The SV40-LTAg IHC was positive in all 14 UCC urine sediments and negative in the FFPE tissues. JCPyV-DNA was identified in all five available UCS and in three FFPE samples of UCC (three of 14; 21.4%). Two UCC cases were positive for MCPyV-DNA (two of 14; 14.3%), and one of them showed protein expression by IHC (one of 14; 7.1%). All specimens were HPyV6 and HPyV7 negative. Our findings show the presence of JCPyV in the urine and UCC of immunocompetent patients. Moreover, MCPyV was detected in two UCC cases. In total, five UCC cases showed the presence of either JCPyV or MCPyV. The evidence here supports the hypothesis that these viruses might sporadically be associated with UCC. Further studies are needed to confirm the relevance of JCPyV or MCPyV as a possible risk factor for UCC development.

Is There Still a Role for Urine Cytology in Kidney Transplantation? Experience Report in a Teaching Hospital

Introduction: Urinary cytology has been used as a monitoring strategy for kidney transplant patients, live under chronic immunosuppression, as it is a non-invasive and low-cost method. The early detection of infections can prevent possible dysfunction and graft loss. However, its use has been decreased. Methods: We conducted a prospective study on voided urine samples from 29 consecutive patients undergoing kidney transplantation over a continuous period of 17 months, collected during their first-year post-transplantation. One technician prepared the samples by cytocentrifugation, Papanicolaou staining, and one pathologist analyzed them. Decoy cells (DCs) positive samples were prepared, by other two technicians, for contrast phase analysis, ultrastructural study, and immunofluorescence for SV40 T antigen. Results: We evaluated 252 urine samples from 26 patients. Two consecutive samples (0.8%) out of five, from the same patient (3.8%), referring to the fourth- and fifth-week post-transplant, showed low-grade squamous intraepithelial lesions; only rare epithelial cells with atypia of undetermined significance were detected (0.4%) in the sample corresponding to the sixth month post-transplant. The patient’s subsequent gynecological examination and Pap smear revealed no changes. Pseudohyphae and fungus yeasts were found in five samples (2.0%) from four patients (15.4%). Trichosporon sp. was identified in the urine of one of these patients. Decoy cells were detected in 25 samples (9.9 %) from six patients (23.1%). Two patients had sustained DCs shedding; one of them, presenting dirty background smears, developed Polyomavirus-associated nephropathy. Ultrastructural study of DCs showed icosahedral viral particles. Immunofluorescence (SV40 T antigen) was positive in DCs nuclei. Analysis by contrast phase was successful in samples with numerous DCs. Conclusion: Systematic urinary cytology after transplantation helps detect some infection signs. Patients with sustained DCs shedding and dirty background smears deserve special clinical attention. Electron microscopy and immunofluorescence (SV40 T antigen) are alternative techniques to detect polyomavirus reactivation. The findings suggest that urinary cytology still plays a role in kidney transplantation.

A case of acute kidney injury due to native kidney BK polyomavirus-associated nephropathy in a human T-lymphotropic virus type 1 carrier

BackgroundBK polyomavirus-associated nephropathy (BKPyVAN) has become a major cause of kidney dysfunction and graft loss in kidney transplant recipients. On rare occasion, polyomavirus has also been known to affect native kidneys of immunocompromised individuals. Only a small number of opportunistic infections have been reported in the carrier phase of human T-lymphotropic virus type 1 (HTLV-1). This is the first reported case of BKPyVAN in native kidneys of an HTLV-1 carrier.Case presentationA 61-year-old man was referred to our hospital from a primary care physician for work-up and treatment of pneumonia. He was diagnosed with Pneumocystis pneumonia and identified as a HTLV-1 carrier who had not yet developed adult T-cell leukemia (ATL). The pneumonia was successfully treated with sulfamethoxazole-trimethoprim. He had never been diagnosed with any kind of kidney dysfunction. Laboratory investigations showed a serum creatinine of 5.3 mg/dL, and urinary sediment showed cells with nuclear enlargement and inclusion bodies suggesting viral infection. The urinary Papanicolaou stain showed inclusions in swollen, ground-glass nuclei, typical of “decoy cells”. Renal biopsy showed degeneration of tubules with epithelial enlargement, vacuolar degeneration, nuclear inclusion bodies, and detachment from the tubular basement membrane. Tubular nuclei showed positive staining positive for simian virus 40 large-T antigen. Polymerase chain reaction tests for BK polyomavirus DNA of both urine and plasma were positive. These findings confirmed a diagnosis of BKPyVAN. Intravenous immunoglobulin therapy did not improve renal function, necessitating maintenance hemodialysis therapy.ConclusionsBKPyVAN should be considered when acute kidney injury occurs with opportunistic infection. HTLV-1 carriers can develop opportunistic infections even before the onset of ATL.

Modelling BK Polyomavirus dissemination and cytopathology using polarized human renal tubule epithelial cells.

Most humans have a lifelong imperceptible BK Polyomavirus (BKPyV) infection in epithelial cells lining the reno-urinary tract. In kidney transplant recipients, unrestricted high-level replication of donor-derived BKPyV in the allograft underlies polyomavirus-associated nephropathy, a condition with massive epithelial cell loss and inflammation causing premature allograft failure. There is limited understanding on how BKPyV disseminates throughout the reno-urinary tract and sometimes causes kidney damage. Tubule epithelial cells are tightly connected and have unique apical and basolateral membrane domains with highly specialized functions but all in vitro BKPyV studies have been performed in non-polarized cells. We therefore generated a polarized cell model of primary renal proximal tubule epithelial cells (RPTECs) and characterized BKPyV entry and release. After 8 days on permeable inserts, RPTECs demonstrated apico-basal polarity. BKPyV entry was most efficient via the apical membrane, that in vivo faces the tubular lumen, and depended on sialic acids. Progeny release started between 48 and 58 hours post-infection (hpi), and was exclusively detected in the apical compartment. From 72 hpi, cell lysis and detachment gradually increased but cells were mainly shed by extrusion and the barrier function was therefore maintained. The decoy-like cells were BKPyV infected and could transmit BKPyV to uninfected cells. By 120 hpi, the epithelial barrier was disrupted by severe cytopathic effects, and BKPyV entered the basolateral compartment mimicking the interstitial space. Addition of BKPyV-specific neutralizing antibodies to this compartment inhibited new infections. Taken together, we propose that during in vivo low-level BKPyV replication, BKPyV disseminates inside the tubular system, thereby causing minimal damage and delaying immune detection. However, in kidney transplant recipients lacking a well-functioning immune system, replication in the allograft will progress and eventually cause denudation of the basement membrane, leading to an increased number of decoy cells, high-level BKPyV-DNAuria and DNAemia, the latter a marker of allograft damage.

Lessons from Polyomavirus Immunofluorescence Staining of Urinary Decoy Cells.

Decoy cells that can be detected in the urine sediment of immunosuppressed patients are often caused by the uncontrolled replication of polyomaviruses, such as BK-Virus (BKV) and John Cunningham (JC)-Virus (JCV), within the upper urinary tract. Due to the wide availability of highly sensitive BKV and JCV PCR, the diagnostic utility of screening for decoy cells in urine as an indicator of polyomavirus-associated nephropathy (PyVAN) has been questioned by some institutions. We hypothesize that specific staining of different infection time-dependent BKV-specific antigens in urine sediment could allow cell-specific mapping of antigen expression during decoy cell development. Urine sediment cells from six kidney transplant recipients (five males, one female) were stained for the presence of the early BKV gene transcript lTag and the major viral capsid protein VP1 using monospecific antibodies, monoclonal antibodies and confocal microscopy. For this purpose, cyto-preparations were prepared and the BK polyoma genotype was determined by sequencing the PCR-amplified coding region of the VP1 protein. lTag staining began at specific sites in the nucleus and spread across the nucleus in a cobweb-like pattern as the size of the nucleus increased. It spread into the cytosol as soon as the nuclear membrane was fragmented or dissolved, as in apoptosis or in the metaphase of the cell cycle. In comparison, we observed that VP1 staining started in the nuclear region and accumulated at the nuclear edge in 6-32% of VP1+ cells. The staining traveled through the cytosol of the proximal tubule cell and reached high intensities at the cytosol before spreading to the surrounding area in the form of exosome-like particles. The spreading virus-containing particles adhered to surrounding cells, including erythrocytes. VP1-positive proximal tubule cells contain apoptotic bodies, with 68-94% of them losing parts of their DNA and exhibiting membrane damage, appearing as "ghost cells" but still VP1+. Specific polyoma staining of urine sediment cells can help determine and enumerate exfoliation of BKV-positive cells based on VP1 staining, which exceeds single-face decoy staining in terms of accuracy. Furthermore, our staining approaches might serve as an early readout in primary diagnostics and for the evaluation of treatment responses in the setting of reduced immunosuppression.

Urine CXCL10 to Assess BK Polyomavirus Replication After Kidney Transplantation.

Urine CXCL10 is a biomarker for renal allograft inflammation induced by rejection, urinary tract infection, or BK polyomavirus (BKPyV) replication. This study aimed to compare urine CXCL10 levels in different stages of BKPyV reactivation and to investigate urine CXCL10 as a biomarker for BKPyV replication. We included 763 urine samples (235 patients) from an interventional, randomized trial obtained in the context of regular screening for urine CXCL10 levels. All urine samples had a complete urine sediment analysis, no rejection episode noted within 30 d before urine collection, and a urine decoy cell analysis was conducted within ±3 d. Urine CXCL10 levels were 2.31 ng/mmol in samples without BKPyV viruria, slightly rose to 4.35 ng/mmol with BKPyV viruria, and then markedly increased to 16.42 ng/mmol when decoy cells were detectable, but still in the absence of BKPyV DNAemia ( P < 0.001). The highest urine CXCL10 values were observed in samples with BKPyV DNAemia (median 42.59 ng/mmol). The area under the curve of urine CXCL10 levels to detect ≥3 decoy cells was 0.816. At a CXCL10 cutoff of 3 ng/mmol, the negative predictive value was 97%. The area under the curve of urine CXCL10 levels to detect BKPyV DNAemia was 0.882, with a negative predictive value of 99% at a CXCL10 cutoff of 3 ng/mmol. Urine CXCL10 levels are already significantly elevated in BKPyV viruria (especially with decoy cell shedding) and further increase with BKPyV DNAemia. Low urine CXCL10 values can rule out the presence of ≥3 decoy cells and BKPyV DNAemia with high certainty.

Böbrek nakli yapılan hastalarda nakil öncesi ve nakil sonrası enfeksiyon etkenlerinin sıklığı

Amaç: Böbrek nakli, böbrek yetmezliğinin en önemli ve başarılı tedavi yöntemidir. Bu çalışmada, böbrek nakli alıcılarında nakil öncesi ve nakil sonrası ilk altı ayda Cytomegalovirus (CMV), BK virüs (BKV) ve bakteriyel etkenlerin sıklığının araştırılması amaçlanmıştır. &#x0D; Gereç ve Yöntem: Fakültemiz Organ Nakli Merkezinde böbrek nakli yapılan hastalardan, nakilden bir hafta önce ve nakilden sonra birinci, üçüncü ve altıncı aylarda alınan kan örneklerinde Real-time PCR ile CMV ve BKV araştırıldı. Kan, idrar, balgam/yara (gerekiyor ise) kültürleri yapıldı. İdrar sitolojisinde Decoy hücreleri değerlendirildi. &#x0D; Bulgular: Böbrek nakli alıcılarının yaş ortalaması 32,60±11,71 28 (% 62,2)’i erkek ve 39 (% 86,7)’u akraba olan canlı vericili, altısı kadavra vericili idi. Nakil sonrası 11/38 (% 28,9) hastada BKV, 25/41 (% 60,9) hastada CMV, 11/31 (% 35,4) hastada Decoy hücre pozitiflikleri saptandı. Real-time PCR pozitifliği en yüksek oran BKV için üçüncü ve altıncı aylarda iken, CMV için birinci ay olup altıncı aya doğru giderek azalma gösterdi. İdrar kültüründe Escherichia coli, Klebsiella pneumoniae, albicans-dışı Candida, Enterococcus faecalis, kan kültüründe Staphylococcus hominis, Streptecoccus epidermidis, solunum yolu örneklerinin kültüründe Acinetobacter baumannii, Klebsiella pneumoniae, Aspergillus fumigatus ve Candida albicans üredi..&#x0D; Sonuç: Böbrek nakli alıcılarımızda bakteriyel enfeksiyonlar erken dönemde gelişti. Real-time PCR pozitiflik oranı en yüksek BKV için üçüncü ve altıncı aylarda iken CMV için birinci ay olup altıncı aya doğru giderek azalma gösterdi. Böbrek nakil hastalarında Decoy hücre pozitifliği de BKV enfeksiyonu tanısı için önemli olabilir.

Cytological Evaluation of Urinary Samples among Vesicovaginal Fistula Patients in National Obstetrics Fistula Centre, Southeastern Nigeria

Vesicovaginal fistula (VVF) is a non-physiological epithelial fistulous tract between the bladder and the vagina, resulting in continuous urinary incontinence. VVF occurs most commonly after obstetrical and gynecological injury. This study is a descriptive cross-sectional study designed to carry out cytological evaluation of urine samples of vesicovaginal fistula patients in National Obstetrics Fistula Centre, Southeastern Nigeria. The voided and catheterized urine samples collected were stained using Papanicolaou and Diff-Quick staining techniques. A total number of 123 patients were involved in this study. The mean age of the patients was 36.66 (± 6.21) years. The patients were grouped according to age into 4 groups: 1 (0.8%) was ≤ 24 years, 59 (48.0%) were 25 to 34 years, 43 (35.0%) were 35 to 44 years and 20 (16.2%) were ≥ 45 years of age. The parity status of these cases was grand multipara 29 (23.6%), multipara 32 (37.4%), primipara 46 (37.4%) and nullipara 16 (13.0%). Majority of the patients 111 (90.2%) were from rural areas, 8 (6.5%) from urban areas and 4 (3.3%) were from semi-urban areas. According to marital status, 91 (74.0%) were married, 16 (13.0%) were widowed, 5 (4.1%) were single and 11 (8.9%) were divorced. According to the period the patients had lived with fistula, majority of the patients 94 (76.4%) had lived less than one year with this problem, while 29 (23.6%) had suffered from VVF for 1-5 years. Cytological evaluation of the urine smears showed that a total of 92 (74.8%) urine samples had normal findings. The rest of the 31 (25.2%) urine samples showed a variety of findings. Out of these 31 samples, 16 (51.6%) showed inflammatory aggregates suggestive of urinary tract inflammation, 3 (9.7%) showed decoy cells suggestive of Polyoma virus infection, 7 (22.6%) cases showed perinuclei halo suggestive of Trichomonas vaginalis infection, 3 (9.7%) showed oval fat bodies suggestive of possible nephrotic syndrome, 8(25.8%) cases showed koilocytes, indicative of Human papilloma virus infections. Cases that showed presence of haematuria were 7 (22.6%). There was no case of tumor cells detected in all the urine samples examined. From the outcome of this study, it is important to take special note of the presence of the viral carcinogens: Polyoma virus and Human papilloma virus, since in theory and as reported in few case reports, malignancy of the urinary tract can lead to formation of vesicovaginal fistula.

Two Cases of BK Polyoma Virus Nephropathy in Patients with Isolated Heart Transplantation: Clinical Usefulness of Urinary Cytology

<h3>Introduction</h3> BK polyoma virus (BKpV) nephropathy (BKpVN) has been often reported in patients with kidney transplantation, whereas reports regarding BKpVN in patients with heart transplantation (HTx) is still sparse. <h3>Case Report</h3> We recently encountered two patients who developed BKpVN after HTx. Both patients (Case 1 and 2) coincidentally received HTx at the age of 56. A standard triple-drug regimen consisting of tacrolimus, mycophenolate mofetil (MMF), and a corticosteroid was started after surgery and the regimen was converted to everolimus (EVL) with reduced tacrolimus within 6 months after HTx because of donor transmitted atherosclerosis in both patients. Case 1 further underwent percutaneous coronary intervention 3.5 years after HTx. Despite renal-sparing immunosuppresion, values of creatinine begun to increase after HTx in both patients and they were diagnosed with BKpVN through high viral DNA copy number in the urine and blood. Regarding case 1, pathological analysis of kidney biopsy specimens disclosed interstitial infiltrate with lymphocytes associated with tubular injury which also lead to the diagnosis of BKpVN. Retrospectively, both patients were positive for decoy cell within 1 year after HTx therefore, these patients should have been recognized as high risk for BKpVN. After the diagnosis of BKpVN, immunosuppressive regimen of them was further changed to minimum dose of EVL (a trough concentration of 6-7 ng/ml) and MMF (500 mg daily) respectively under the guidance of myocardial biopsy and they have been uneventful for over a year after the diagnosis of BKpVN despite severe chronic kidney disease. <h3>Summary</h3> We presented two rare cases of BKpVN after HTx. In our institution, urinary cytology is regularly examined if there are decoy cells which we think are the sign of over immunosuppression. However, decoy cells detected by urinary cytology also suggest that there is subclinical BKpV infection. Urinary cytology, a noninvasive and cost-effective examination, should be routinely performed for the risk assessment of BKpVN even in HTx patients, since detecting decoy cell would be helpful for the early diagnosis of BKpVN.

MP37-15 DETECTION OF DECOY CELLS IN SIX MONTHS OR MORE ON URINARY CYTOLOGY EFFICIENTLY AND COST-EFFECTIVELY PREDICTS BK VIRUS NEPHROPATHY IN RENAL TRANSPLANT RECIPIENTS

You have accessJournal of UrologyRenal Transplantation & Vascular Surgery I (MP37)1 Sep 2021MP37-15 DETECTION OF DECOY CELLS IN SIX MONTHS OR MORE ON URINARY CYTOLOGY EFFICIENTLY AND COST-EFFECTIVELY PREDICTS BK VIRUS NEPHROPATHY IN RENAL TRANSPLANT RECIPIENTS Takanori Sekito, Motoo Araki, Kasumi Yoshinaga, Syogo Watari, Yuki Maruyama, Takuya Sadahira, Shingo Nishimura, Koichiro Wada, Tomoko Sako, Kohei Edamura, Yasuyuki Kobayashi, Masami Watanabe, Toyohiko Watanabe, Hiroyuki Yanai, and Yasutomo Nasu Takanori SekitoTakanori Sekito More articles by this author , Motoo ArakiMotoo Araki More articles by this author , Kasumi YoshinagaKasumi Yoshinaga More articles by this author , Syogo WatariSyogo Watari More articles by this author , Yuki MaruyamaYuki Maruyama More articles by this author , Takuya SadahiraTakuya Sadahira More articles by this author , Shingo NishimuraShingo Nishimura More articles by this author , Koichiro WadaKoichiro Wada More articles by this author , Tomoko SakoTomoko Sako More articles by this author , Kohei EdamuraKohei Edamura More articles by this author , Yasuyuki KobayashiYasuyuki Kobayashi More articles by this author , Masami WatanabeMasami Watanabe More articles by this author , Toyohiko WatanabeToyohiko Watanabe More articles by this author , Hiroyuki YanaiHiroyuki Yanai More articles by this author , and Yasutomo NasuYasutomo Nasu More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002046.15AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: BK virus nephropathy (BKVN) is one of the important causes of graft loss after kidney transplantation (KT). This study analyzed the association between the duration of positive decoy cells in urine cytology and BKVN. METHODS: Totally, 121 kidney transplant recipients were retrospectively evaluated. Urine cytology was routinely monitored every month during first 6 months after KT, every 1 to 2 months during second 6 months, every 1 to 3 months afterward. The best duration of consecutive presence of decoy cells to predict BKVN was analyzed by calculating the area under the curve (AUC) among each duration, and recipients were divided into two groups based on the best predictive performance. The effectiveness of SV40 immunostaining on urinary cytology was also analyzed. RESULTS: In total, 2,534 urine specimens, and 2,241 SV40 immunostaining for urine specimens were analyzed. The six consecutive months of decoy cells had best predictive performance in BKVN (AUC = 0.819). Recipients were divided into two groups based on the six or more consecutive months of decoy cells (decoy ≥ 6M, n=47) or less than six months (decoy < 6M, n=74). The incidence of BKVN in decoy ≥ 6M had significantly higher than those in decoy < 6M (5 vs 0, p = 0.008). The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) to predict BKVN by detecting decoy cells was 100%, 64%, 11% and 100% respectively. SV40 immunostaining provided slightly better specificity (68%) and PPV (12%) with equivalent sensitivity (100%) and NPV (100%) (Table 1). The monthly screening by decoy cells can annually save U.S. $256,520 in 121 patients (U.S. $2,120 per patient) compared to the monthly screening by plasma quantitative PCR, and U.S. $55,660 in 121 patients (U.S. $460 per patient) compared to the monthly screening by urinary SV40 immunostaining. CONCLUSIONS: The detection of decoy cells in six months or more on urine cytology can predict BKVN efficiently and cost-effectively in KT recipients. SV40 immunostaining on urine cytology adds minimal effectiveness. Source of Funding: None © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e652-e652 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Takanori Sekito More articles by this author Motoo Araki More articles by this author Kasumi Yoshinaga More articles by this author Syogo Watari More articles by this author Yuki Maruyama More articles by this author Takuya Sadahira More articles by this author Shingo Nishimura More articles by this author Koichiro Wada More articles by this author Tomoko Sako More articles by this author Kohei Edamura More articles by this author Yasuyuki Kobayashi More articles by this author Masami Watanabe More articles by this author Toyohiko Watanabe More articles by this author Hiroyuki Yanai More articles by this author Yasutomo Nasu More articles by this author Expand All Advertisement Loading ...

Presence of decoy cells for 6months on urine cytology efficiently predicts BK virus nephropathy in renal transplant recipients.

To investigate the association between duration of consecutive presence of decoy cells on urine cytology and BK virus nephropathy after kidney transplantation. In total, 121 kidney transplant recipients were retrospectively evaluated. The best duration of consecutive presence of decoy cells that could be used to predict BK virus nephropathy was analyzed using the area under the curve for each duration, and recipients were divided into two groups based on the best predictive performance. The effectiveness of SV40 immunostaining on urinary cytology was also analyzed. In total, 2534 urine specimens as well as SV40 immunostaining in 2241 urine specimens were analyzed. Six consecutive months of decoy cell positivity had the best predictive performance for BK virus nephropathy (area under the curve=0.832). The incidence of BK virus nephropathy in recipients with positive decoy cells for 6months or more consecutive months (5/44) was significantly higher than in those who had positive decoy cells for less than 6months (0/77; P=0.005). Decoy cell positivity had a sensitivity, specificity, positive predictive value, and negative predictive value for BK virus nephropathy of 100%, 66%, 11%, and 100% respectively. SV40 immunostaining provided slightly better specificity (68%) and positive predictive value (12%). The detection of decoy cells at 6months or more on urine cytology had high predictive value for BK virus nephropathy in kidney transplant recipients. SV40 immunostaining on urine cytology added minimal diagnostic accuracy.

Early Detection Strategy of BK Polyomavirus Nephropathy in Patients undergoing Renal Transplant: A Single-Center Retrospective Study

Introduction: BK polyoma virus nephropathy represents one of the non-immunological causes of renal graft loss with a cumulative incidence between 5 and 10 percent, and a graft loss rate on BK virus nephropathy patients that ranges from 30 to 90 percent depending on the nephropathy status at the time of diagnosis (1). Objective: To determine the outcomes regarding the kidney graft survival and kidney function of an early detection strategy for BK Polyomavirus nephropathy which is drawn upon urinary cytology in order to look for Decoy cells. Methodology: Descriptive, retrospective study type. Adult renal transplant recipients/patients from the Interdisciplinary Renal Transplant program of the Colombia University Clinic were included in a time period from 2012 to 2018, in whom monthly post-transplant monitoring with positive urinary cytology was performed. Results: 303 patients were transplanted in the program, 107 patients with positive urinary cytology were included, wherein, men predominated with 72% and 45 years old being the average age (IQR: 18–72). The cumulative incidence of polyomavirus nephropathy, under this early detection strategy, was 9.2%, thus preserving the graft survival at 24 months in a 100% of the patients with the management strategies employed. Conclusions: This early detection and monitoring study utilizing post-transplant urinary cytology proved to be useful for the early diagnosis of BK Polyomavirus nephropathy with a favorable impact on the graft livelihood.

POS-739 AN INTERESTING CASE OF CHRONIC ALLOGRAFT DYSFUNCTION

Among renal transplants, chronic allograft dysfunction is defined as a condition in which irreversible damage to the kidney allograft occurs over a period of months. Here, we report a case of late presentation of graft dysfunction due to BK Virus with negative urine decoy cells & Negative BK Virus DNA PCR. A 18 year old young male underwent renal transplantation following which renal functions recovered to creatinine of 0.8 mg/dl with mother being donor. Patient was compliant & on regular follow up. After 1 year of renal transplantation, patient's creatinine was found to be 2.1 mg/dl with no symptoms and signs. Patient was admitted and was evaluated. Urine showed no proteinuria, blood & urine cultures were negative with normal counts. Ultrasound of transplant kidney showed no issues. Tacrolimus levels were within limits. All reversible factors contributing were ruled out. Viral etiology were sought for. CMV IgM was negative. Urinary decoy cells were not seen. BK virus DNA PCR also turned out to be negative. BK virus was ruled out of differentials. During the course of hospital stay, creatinine worsened to 2.7. At this stage notwithstanding , Renal biopsy of transplant kidney was done. Biopsy showed features of BK Virus nephropathy with viral cytopathic changes of the tubular cells. Among alloantigen independent causes of chronic allograft dysfunction; and among infections, CMV is the most common. Infections do appear in the early post transplant period when the patients are most immunosupressed. BK Virus is a polyoma virus hominis 1 which causes PVAN(polyoma virus associated nephropathy) in humans is seen in ∼5% of kidney transplant recipients and has emerged as an important cause of allograft dysfunction over the last 20 years with 5 year graft failure rate of 55% presenting with asymptomatic increase in serum creatinine.BK virus is diagnosed by Decoy cells in urine seen in over 90% of the infected patients. BK Virus DNA PCR is very sensitive . The gold standard being Renal biopsy of transplant kidney with tubulointerstitial nephritis , inclusions in tubular cells & Immunohistochemistry confirming BK Virus.In our case, interestingly, Decoy cells & DNA PCR was negative with positive Biopsy for BK Virus. In our case, Mycophenolate mofetil was stopped . Patient was treated with leflunomide, following which patients creatinine came down and stabilised at 2.5 mg/dl. BK Virus nephropathy is a great masqurader which may often camouflage itself & misguide the clinician with negative urine decoy cells & PCR. Prompt workup is necessary including renal biopsy if clinically suspicious. This is one such case report where BK Virus nephropathy was diagnosed.