Role of cytomegalovirus (CMV)-specific polyfunctional CD8+ T-cells and antibodies neutralizing virus epithelial infection in the control of CMV infection in an allogeneic stem-cell transplantation setting

Abstract

The role of cytomegalovirus (CMV)-specific polyfunctional CD8 + T-cells and that of antibodies neutralizing virus epithelial infection (AbNEI) in the control of CMV DNAemia were investigated in 39 CMV-seropositive allogeneic stem-cell transplant (Allo-SCT) recipients with (n524) or without (n515) CMV DNAemia. AbNEI levels were monitored prospectively by means of a neutralization assay employing retinal epithelial cells (ARPE-19) and the recombinant CMV strain BADrUL131-Y4. Quantification of CMV-specific polyfunctional CD8 + T-cells (expressing two or three of the following markers: IFN-cc, TNF-a and CD107a) in whole blood was performed by flow cytometry for intracellular cytokine staining. We found no differences in the dynamic pattern of AbNEI in patients with or without subsequent CMV DNAemia. Baseline and peak AbNEI titres were not predictive of the dynamics of CMV replication within episodes. No correlation was found between CMV DNA loads and AbNEI levels during episodes of CMV DNAemia (r50.09; 95 % confidence interval 20.52 to 0.64; P50.78). The detection of pp65/IE-1 CMV-specific polyfunctional CD8 + T-cells was associated with low-level virus replication within subsequent episodes of CMV DNAemia. Interestingly, the presence of AbNEI titres (inverse) .4.7 log 2 was predictive of the occurrence of CMV DNAemia (sensitivity, 83 %; specificity, 80 %). Our findings provide an insight to the role of humoral and cellular immunity in the control of CMV infection in an Allo-SCT setting.

Peptide vaccination in the presence of adjuvants in patients after hematopoietic stem cell transplantation with CD4+ T cell reconstitution elicits consistent CD8+ T cell responses

“…This is in line with reports from Spanish colleagues 35 who described a synergy of humoral and cellular immune responses against the virus.…”

Treatment of CMV infection after allogeneic hematopoietic stem cell transplantation

“…The observation that patients with high baseline and peak AbNEIs levels were more likely to develop CMV-DNAemia was of interest. Possible explanations include a major role played by memory B cells of donor and recipient origins [122]. The gH/gL-pentamer complex represents the platform for prophylactic Pentamer-based vaccines of absolute interest [123,124].…”

IL28B genetic variation and cytomegalovirus‐specific T‐cell immunity in allogeneic stem cell transplant recipients

“…The inclusion criteria were the following: (i) availability of data on CMV‐specific CD8 + T‐cell immunity obtained within the episode of active CMV infection; (ii) availability of plasma specimens for IL‐28B quantitation obtained prior to the time of immunological analyses; and (iii) availability of data on the donor IL28B genotype. The patients in cohorts A and B had been included in different studies previously published by our group [Solano et al, ; Tormo et al, ,, ; Giménez et al, ,]. In all these studies, CMV‐specific IFN‐γ‐producing CD8 + T cells, but not CD4 + T cells, were planned to be enumerated.…”

“…On the day of testing, stimulated blood was thawed at 37°C, washed, permeabilized, and stained with a combination of labeled monoclonal antibodies (anti‐IFN‐γ‐FITC, anti‐CD69‐PE, anti‐CD4, or CD8‐PerCP‐Cy5.5, and anti‐CD3‐APC, when the IFN‐γ CD8 + kit was used) for 30 min at room temperature. In some experiments, anti‐CD3–APCCy7 (BD, Biosciences) was used instead of anti‐CD3‐APC [Giménez et al, ,]. Appropriate isotype controls were used (BD FastImmune TMc2a/c1/CD8/CD3 reacting with keyhole limpet hemocyanin; BD Biosciences).…”