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Comparative Study
. 2017 Mar 7;18(1):15.
doi: 10.1186/s12865-017-0194-z.

Validation of T-Track® CMV to Assess the Functionality of Cytomegalovirus-Reactive Cell-Mediated Immunity in Hemodialysis Patients

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Free PMC article
Comparative Study

Validation of T-Track® CMV to Assess the Functionality of Cytomegalovirus-Reactive Cell-Mediated Immunity in Hemodialysis Patients

Bernhard Banas et al. BMC Immunol. .
Free PMC article

Abstract

Background: Uncontrolled cytomegalovirus (CMV) replication in immunocompromised solid-organ transplant recipients is a clinically relevant issue and an indication of impaired CMV-specific cell-mediated immunity (CMI). Primary aim of this study was to assess the suitability of the immune monitoring tool T-Track® CMV to determine CMV-reactive CMI in a cohort of hemodialysis patients representative of patients eligible for renal transplantation. Positive and negative agreement of T-Track® CMV with CMV serology was examined in 124 hemodialysis patients, of whom 67 (54%) revealed a positive CMV serostatus. Secondary aim of the study was to evaluate T-Track® CMV performance against two unrelated CMV-specific CMI monitoring assays, QuantiFERON®-CMV and a cocktail of six class I iTAg™ MHC Tetramers.

Results: Positive T-Track® CMV results were obtained in 90% (60/67) of CMV-seropositive hemodialysis patients. In comparison, 73% (45/62) and 77% (40/52) positive agreement with CMV serology was achieved using QuantiFERON®-CMV and iTAg™ MHC Tetramer. Positive T-Track® CMV responses in CMV-seropositive patients were dominated by pp65-reactive cells (58/67 [87%]), while IE-1-responsive cells contributed to an improved (87% to 90%) positive agreement of T-Track® CMV with CMV serology. Interestingly, T-Track® CMV, QuantiFERON®-CMV and iTAg™ MHC Tetramers showed 79% (45/57), 87% (48/55) and 93% (42/45) negative agreement with serology, respectively, and a strong inter-assay variability. Notably, T-Track® CMV was able to detect IE-1-reactive cells in blood samples of patients with a negative CMV serology, suggesting either a previous exposure to CMV that yielded a cellular but no humoral immune response, or TCR cross-reactivity with foreign antigens, both suggesting a possible protective immunity against CMV in these patients.

Conclusion: T-Track® CMV is a highly sensitive assay, enabling the functional assessment of CMV-responsive cells in hemodialysis patients prior to renal transplantation. T-Track® CMV thus represents a valuable immune monitoring tool to identify candidate transplant recipients potentially at increased risk for CMV-related clinical complications.

Trial registration: ClinicalTrials.gov NCT02630537.

Keywords: CMV; Cell-mediated immunity; Cytomegalovirus; Hemodialysis; IE-1; IFN-γ ELISpot; QuantiFERON®-CMV; T-Track® CMV; iTAg™ MHC Tetramers; pp65.

Figures

Fig. 1
Fig. 1
CMV-specific immunity in hemodialysis patients measured with T-Track® CMV (a), QuantiFERON®-CMV (b) and iTAg™ MHC Tetramers (c). a Spot-forming cells (SFC) in IFN-γ ELISpot after in vitro stimulation of PBMC from CMV-seronegative (n = 57) and CMV-seropositive (n = 67) hemodialysis patients with T-activated® aIE-1 and app65 proteins, or with medium (unst.) as a negative control. SFC levels are presented as log10-transformed values in scatter plots, including median values (horizontal black lines). The horizontal grey dashed line indicates the positivity cut-off (10 SFC / 200,000 PBMC). b CD8+-secreted IFN-γ levels were measured by ELISA following the stimulation of whole blood from CMV-seronegative (n = 57) and CMV-seropositive (n = 66) hemodialysis patients with HLA class I-specific peptides. Test results were considered positive when IFN-γ levels ≥ 0.2 IU/mL (grey dashed line). Indeterminate results (4/66 seropositive and 2/57 seronegative patients) are not represented; therefore the scatter plots represent the results of 62 seropositive and 55 seronegative assays. *, values ≥ 10 IU/mL cannot be quantitatively evaluated; consequently, no median values were depicted. c PBMC of CMV-seronegative (n = 45) and CMV-seropositive (n = 52) hemodialysis patients were stained with a mixture of six iTAg™ MHC class I Tetramers, and CMV peptide-specific CD8+ T cells were quantified by flow cytometry. Test results were considered positive when ≥ 0.1% of total CD8+ T cells were tetramer-positive (grey dashed line). The scatter plots show median values (horizontal black lines)

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References

    1. Hanley PJ, Bollard CM. Controlling cytomegalovirus: helping the immune system take the lead. Viruses. 2014;6:2242–58. doi: 10.3390/v6062242. - DOI - PMC - PubMed
    1. Crough T, Khanna R. Immunobiology of human cytomegalovirus: from bench to bedside. Clin Microbiol Rev. 2009;22:76–98. doi: 10.1128/CMR.00034-08. - DOI - PMC - PubMed
    1. Schleiss MR. Cytomegalovirus in the neonate: immune correlates of infection and protection. Clin Dev Immunol. 2013;2013:501801. doi: 10.1155/2013/501801. - DOI - PMC - PubMed
    1. Rossini G, Cerboni C, Santoni A, Landini MP, Landolfo S, Gatti D, et al. Interplay between human cytomegalovirus and intrinsic/innate host responses: a complex bidirectional relationship. Mediators Inflamm. 2012;2012:607276. doi: 10.1155/2012/607276. - DOI - PMC - PubMed
    1. Jeitziner SM, Walton SM, Torti N, Oxenius A. Adoptive transfer of cytomegalovirus-specific effector CD4+ T cells provides antiviral protection from murine CMV infection. Eur J Immunol. 2013;43:2886–95. doi: 10.1002/eji.201343690. - DOI - PubMed

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