Sternheimer-Malbin Staining to Detect Decoy Cells in Urine of 213 Kidney Transplant Patients

Yan, Lizhi; Guo, Hongbo; Han, Lizhong; Huang, Hualiang; Shen, Yan; He, Jing; Liu, Jinlin

doi:10.1016/j.transproceed.2020.01.044

Cited by 6 publications

(3 citation statements)

References 18 publications

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“…Testing urine sediment for decoy cells has, without debate, gained importance, and various methods, such as Sternberg-Malbin staining, have improved availability and show excellent discriminative potential from other cells of the urinary tract [41,42]. Combined confocal immunostaining for proteins specific to the early and late stages of infection will extend the specificity for counting polyomavirus-infected cells beyond simple decoy staining.…”

Section: Discussionmentioning

confidence: 99%

Lessons from Polyomavirus Immunofluorescence Staining of Urinary Decoy Cells

Pajenda

Hevesi

Eder

et al. 2023

Life

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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.

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Section: Discussionmentioning

confidence: 99%

Lessons from Polyomavirus Immunofluorescence Staining of Urinary Decoy Cells

Pajenda

Hevesi

Eder

et al. 2023

Life

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“…Urine decoy test also has poor test performance with reportedly low specificity and positive predictive value. 7,39 Newer methods to detect infected cells such as the SV40T immunoperoxidase staining of exfoliated urinary tubular cells have been developed for improved test specificity [112][113][114] but they are yet to be widely adopted in clinical practice.…”

Section: Examination Of Urine Cytology For Decoy Cells or Infected Cellsmentioning

confidence: 99%

Polyoma BK Virus in Kidney Transplant Recipients: Screening, Monitoring, and Management

et al. 2021

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Polyomavirus BK virus (BKPyV) infection is an important complication of kidney transplantation and allograft failure. The prevalence of viremia is 10%-15%, compared with BK-associated nephropathy (BKPyVAN) at 3%-5%. Given that there are no effective antiviral prophylaxis or treatment strategies for BKPyVAN, active screening to detect BKPyV viremia is recommended, particularly during the early posttransplant period. Immunosuppression reduction to allow viral clearance may avoid progression to severe and irreversible allograft damage. The frequency and duration of screening are highly variable between transplant centers because the evidence is reliant largely on observational data. While the primary treatment goals center on achieving viral clearance through immunosuppression reduction, prevention of subsequent acute rejection, premature graft loss, and return to dialysis remain as major challenges. Treatment strategies for BKPyV infection should be individualized to the recipient's underlying immunological risk and severity of the allograft infection. Efficacy data for adjuvant therapies including intravenous immunoglobulin and cidofovir are sparse. Future well-powered and high-quality randomized controlled trials are needed to inform evidence-based clinical practice for the management of BKPy infection.

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“…Recently, authors have tested other noninvasive diagnostic methods, such as the RTqPCR of urine sediment cells, to see if they are useful as biomarkers for BKPyVAN [15][16][17]. Immunostaining of decoy cells [18] or cytology using the Sternheim-Malbin method, which is quicker to perform, has also been shown to be effective [19]. It is assumed that the decoy cells are highly infected virus-replicating cells that originate from the tubular elements of the nephron and from further distally.…”

Section: Introductionmentioning

confidence: 99%

Acute Kidney Injury and BK Polyomavirus in Urine Sediment Cells

Pajenda,

Gerges,

Freire

et al. 2023

IJMS

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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.

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Sternheimer-Malbin Staining to Detect Decoy Cells in Urine of 213 Kidney Transplant Patients

Cited by 6 publications

References 18 publications

Lessons from Polyomavirus Immunofluorescence Staining of Urinary Decoy Cells

Lessons from Polyomavirus Immunofluorescence Staining of Urinary Decoy Cells

Polyoma BK Virus in Kidney Transplant Recipients: Screening, Monitoring, and Management

Acute Kidney Injury and BK Polyomavirus in Urine Sediment Cells

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