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• W2076139592 abstract "To the Editor: Hammer and colleagues report on BKV-specific T-cell responses in peripheral blood mononuclear cells (PBMC) of 15 viremic kidney transplant recipients (1Hammer MH Brestich G Andree H et al.HLA-independent method to monitor polyoma BK virus-specific CD4+ and CD8+ T-cell immunity..Am J Transplant. 2006; 6: 625-631Abstract Full Text Full Text PDF PubMed Scopus (56) Google Scholar). After stimulation with overlapping peptides spanning the viral capsid protein VP1, interferon-γ (IFNγ)-producing CD4+ T cells were observed in 7 patients (47%), including 2 patients (13%) with a CD8+ T-cell response. All patients with a detectable T-cell response had plasma BKV loads of <250 000 copies (c)/mL. Both patients with VP1-specific CD8+ T cells were the only ones to lose their grafts during follow-up. The authors concluded that high BKV load of <250 000 c/mL correlated with peripheral BKV-specific T-cell responses and that BKV-specific CD8+ responses indicated a risk for subsequent allograft loss (1Hammer MH Brestich G Andree H et al.HLA-independent method to monitor polyoma BK virus-specific CD4+ and CD8+ T-cell immunity..Am J Transplant. 2006; 6: 625-631Abstract Full Text Full Text PDF PubMed Scopus (56) Google Scholar). Using ELISpot and flow cytometry for intracellular cytokine staining, we detected BKV-specific PBMC responses in renal transplant patients with plasma BKV loads clearly below 250 000 c/mL. In patient 1 (51 100 c/mL), both large T-antigen (LT)-VP1-specific IFNγ-producing PBMC were detectable by ELISpot (542 and 392 SFU/106PBMC). By flow cytometry, this response included LT-and VP1-specific CD4+ T cells (0.16% and 0.11%) and CD8+ T cells (0.04% and 0.06%), respectively. A similar BKV-specific cellular immune profile was found in patient 2 (300 c/mL). Patient 3 (944 c/mL) is remarkable for clear responses by ELISpot analysis which could not readily be attributed to CD4+ or CD8+ T cells. This discordance may reflect differences in sensitivity and/or an early phase of cellular immune effectors including NK-cells (2Comoli P Azzi A Maccario R et al.Polyomavirus BK-specific immunity after kidney transplantation..Transplantation. 2004; 78: 1229-1232Crossref PubMed Scopus (98) Google Scholar). In patient 4 (9600 c/mL), LT-and VP1-specific CD4+ T-cell, but no CD8+ T-cell responses were detectable.In patients 5 and 6 with persisting BKV viremia (38 400 and 98 000 c/mL), only very low responses were detectable by ELISpot, but none by flow cytometry. Overall, we found no correlation of BKV-specific cellular immune responses and the level of plasma BKV load. However, all patients with detectable cellular responses had a decline of BKV loads in the preceding 4–12 weeks. Interestingly, LT-and VP1-specific responses were not always concordant. Similarly, LT-specific T-cell responses were seen in patient 7 with stable allograft function after clearing BKV viremia and polyomavirus-associated nephropathy (PVAN) more than 12 months ago, and in patient 8 being on hemodialysis after allograft loss due to PVAN 15 months ago (Table 1).Table 1Plasma BKV load and large T-and VP1-specific interferon-γ production in peripheral blood mononuclear cellsPatientAge/SexSerum creatinine (μmol/L)ImmunosuppressionBKV load plasma (copies/mL)BKV replication dynamicsΔ log10PBMCCD4+ T cellsCD8+ T cellsLTVP1LTVP1LTVP1ELISpotFlow cytometryFlow cytometry(IFNγ SFU/106cells)IFNγ (%)IFNγ (%)136/F113Sir/MMF ↓51 100Decreasing-35423920.160.110.040.06261/F141Sir/MMF ↓300Decreasing-3973730.040.100.020.07326/M104CyA/Aza944Decreasing-267444negnegnegneg453/M115CyA/MMF/Pre9600Decreasing–0.5492270.030.02negneg550/M344CyA/Tac/Lef38 400Stable+0.31013neg0.01negneg645/M155CyA/MMF/Pre98 000Stable-0.237156neg0.01negneg750/F203CyA/MMF/Pre0Cleared PVAN752760.020.070.090.07865/MHemodialysisNo0Graft loss post-PVAN20470.030.110.020.05Aza = azathioprine; CyA = cyclosporine; Lef = leflunomide; MMF = mycophenolate mofetil; Tac = tacrolimus; Pre = prednisone; Sir = sirolimus; PVAN = polyomavirus-associated nephropathy; LT = BKV large T-antigen; VP1 = BKV viral capsid protein-1; SFU = mean spot-forming units; IFNγ= interferon-γ; neg = negative (<0.01%).BKV load was determined by real-time PCR as described with a cut-off of 300 c/mL plasma. ELISpots were performed in triplicate for interferon-γ detection after overnight stimulation of 2.5 × 105PBMC with 2 µg/mL of 15-mer peptide pools of 11 amino acid overlap spanning LT or VP1 versus negative control medium. Intracellular cytokine staining for IFN-γ detection and flow cytometry was performed after 6 h stimulation with LT or VP1 peptide pools and addition of brefeldin A. Open table in a new tab Aza = azathioprine; CyA = cyclosporine; Lef = leflunomide; MMF = mycophenolate mofetil; Tac = tacrolimus; Pre = prednisone; Sir = sirolimus; PVAN = polyomavirus-associated nephropathy; LT = BKV large T-antigen; VP1 = BKV viral capsid protein-1; SFU = mean spot-forming units; IFNγ= interferon-γ; neg = negative (<0.01%). BKV load was determined by real-time PCR as described with a cut-off of 300 c/mL plasma. ELISpots were performed in triplicate for interferon-γ detection after overnight stimulation of 2.5 × 105PBMC with 2 µg/mL of 15-mer peptide pools of 11 amino acid overlap spanning LT or VP1 versus negative control medium. Intracellular cytokine staining for IFN-γ detection and flow cytometry was performed after 6 h stimulation with LT or VP1 peptide pools and addition of brefeldin A. In our series, VP1-specific CD8+ T-cell responses were not associated with poor allograft function. This discrepancy may be due to lower immune effector frequencies which, apart from technical differences, could reflect the stage of PVAN, immune reconstitution dynamics after reducing immunosuppression or other factors. Indeed, Hammer et al. were able to raise VP1-specific CD8+ T cells from allograft biopsies from patients without PBMC responses, all of whom maintained allograft function (1Hammer MH Brestich G Andree H et al.HLA-independent method to monitor polyoma BK virus-specific CD4+ and CD8+ T-cell immunity..Am J Transplant. 2006; 6: 625-631Abstract Full Text Full Text PDF PubMed Scopus (56) Google Scholar). It is unclear why the authors concluded that these cells originated in the donor rather than from efficient homing of recipient T-cells to the site of replication. It is well recognized that duration and levels of plasma BKV loads are important surrogates of viral tissue damage (3Funk G Steiger J Hirsch HH Rapid dynamics of polyomavirus type BK in renal transplant recipients..J Infect Dis. 2006; 193: 80-87Crossref PubMed Scopus (79) Google Scholar, 4Hirsch HH Knowles W Dickenmann M et al.Prospective study of polyomavirus BK replication and nephropathy in renal transplant recipients..N Engl J Med. 2002; 347: 488-496Crossref PubMed Scopus (983) Google Scholar, 5Drachenberg CB Papadimitriou JC Hirsch HH et al.Histological patterns of polyomavirus nephropathy: Correlation with outcome and viral load..Am J Transplant. 2004; 4: 2082-2092Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar), which might also determine specific immune-mediated and possibly allospecific collateral damage. Thus, careful investigation of the dynamics of BKV replication, together with the qualitative and quantitative patterns of BKV-specific cellular immune responses, seems to be needed to elucidate the balance of virus and immune response and to identify the most informative tests." @default.
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• W2076139592 title "BKV Replication and Cellular Immune Responses in Renal Transplant Recipients" @default.
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