Performance of gastrointestinal pathologists within a national digital review panel for Barrett's oesophagus in the Netherlands: Results of 80 prospective biopsy reviews

Aims: The histopathological diagnosis of low-grade dysplasia (LGD) in Barrett's oesophagus (BO) is associated with poor interobserver agreement and guidelines dictate expert review. To facilitate nationwide expert review in the Netherlands, a web-based digital review panel has been set up, which currently consists of eight 'core' pathologists. The aim of this study was to evaluate if other pathologists from the Dutch BO expert centres qualify for the expert panel by assessing their performance in 80 consecutive LGD reviews submitted to the panel. Methods: Pathologists independently assessed digital slides in two phases. Both phases consisted of 40 cases, with a group discussion after phase I. For all cases, a previous consensus diagnosis made by five core pathologists was available, which was used as reference. The following criteria were used: (1) percentage of 'indefinite for dysplasia' diagnoses, (2) percentage agreement with consensus diagnosis and (3) proportion of cases with a consensus diagnosis of dysplasia underdiagnosed as non-dysplastic. Benchmarks were based on scores of the core pathologists. Results: After phase I, 1/7 pathologists met the benchmark scor

Adherence to pre-set benchmark quality criteria to qualify as expert assessor of dysplasia in Barrett’s esophagus biopsies – towards digital review of Barrett’s esophagus

Background: Dysplasia assessment of Barrett’s esophagus biopsies is associated with low observer agreement; guidelines advise expert review. We have developed a web-based review panel for dysplastic Barrett’s esophagus biopsies. Objective: The purpose of this study was to test if 10 gastrointestinal pathologists working at Dutch Barrett’s esophagus expert centres met pre-set benchmark scores for quality criteria. Methods: Ten gastrointestinal pathologists twice assessed 60 digitalized Barrett’s esophagus cases, enriched for dysplasia; then

IMPLEMENTATION ROUTES OF ADVANCED THERAPY MEDICINAL PRODUCTS DEVELOPED IN ACADEMIA IN CLINICAL PRACTICE IN THE NETHERLANDS

Clinical Pharmacy and Toxicolog

Co-ordinated isolation of CD8(+) and CD4(+) T cells recognizing a broad repertoire of cytomegalovirus pp65 and IE1 epitopes for highly specific adoptive immunotherapy

Background aims. Adoptive transfer of cytomegalovirus (CMV)-specific memory T cells can be used for treatment of CMV reactivation after allogeneic stem cell transplantation. As co-ordinated CD8(+) and CD4(+) T cells specific for a broad repertoire of CMV epitopes may be most effective for adoptive immunotherapy, the aim of this study was to isolate these cells from peripheral blood of CMV seropositive donors, irrespective of their HLA type. Methods. Activation of CMV-specific CD8(+) and CD4(+) T cells was compared after stimulation of donor peripheral blood with minimal epitope peptides, pools of overlapping 15-mer peptides or full-length protein. Furthermore, the kinetics of interferon (IFN)-gamma production after stimulation was analyzed to determine the optimal time-point for IFN-gamma-based isolation of CMV-specific T cells. The specificity, phenotype and functionality of generated T-cell lines were analyzed. Results. CMV protein-spanning 15-mer peptide pools induced simultaneous activation of both CD8(+) and CD4(+) CMV-specific T cells, while full-length CMV protein only efficiently activated CD4(+) CMV-specific T cells. Isolation of IFN-gamma-secreting cells at the peak of the IFN-gamma response after 4-h stimulation with CMV pp65 and IE1 peptide pools resulted in efficient enrichment of CMV-specific T cells. The T-cell lines contained high frequencies of CD8(+) and CD4(+) T cells recognizing multiple CMV pp65 and IE1 epitopes, and produced IFN-gamma and tumor necrosis factor (TNF)-alpha upon specific restimulation. Conclusions. This study provides a feasible strategy for the rapid generation of clinical-grade CD8(+) and CD4(+) T-cell lines with high specificity for multiple CMV pp65 and IE1 epitopes, which may be used for effective adoptive immunotherapy

Mesenchymal Stromal Cell Therapy Is Associated With Increased Adenovirus-Associated but Not Cytomegalovirus-Associated Mortality in Children With Severe Acute Graft-Versus-Host Disease

Transplantation and immunomodulatio

High Titers of Pre-existing Adenovirus Serotype-Specific Neutralizing Antibodies in the Host Predict Viral Reactivation After Allogeneic Stem Cell Transplantation in Children

Background. Human adenovirus (HAdV) infections are frequent in children after allogeneic stem cell transplantation (SCT) and may become fatal. Whether these infections occur through reactivation of endogenous virus or transmission via the graft remains a matter of debate. Methods. In a cohort of 24 pediatric patients who received SCT, infections with 1 or more of 5 serotypes of HAdV (1, 2, 5, 6, and 31) were detected by culture. Neutralizing antibody (NAb) titers were measured in vitro by means of a virus neutralization assay. Results. In 11 patients the infection was restricted to 1 site as demonstrated by culture only, and in 13 patients the HAdV infection was disseminated because plasma samples contained HAdV DNA. The 5 most commonly encountered HAdV serotypes caused 35 infectious episodes after SCT. Serum titers of NAb against these 5 serotypes of HAdV were measured before and after transplantation. High titers of NAb against a certain serotype in the recipient prior to SCT, reflecting previous infection, appeared to predispose for infection with the same serotype after SCT. In only 1 case of 41 independent samples of graft material, a very low level of HAdV DNA was detected. Antibody responses after SCT were detected in 21 of 35 infectious episodes. Conclusions. Together, these data suggest that adenoviral complications after SCT are caused by reactivation of endogenous persistent HAdV rather than by de novo infection from the donor or environment. This finding may offer a strategy of prophylactic treatment of high-risk patients before SCT to prevent infectious complications after allogeneic SCT.Transplantation and immunomodulatio

High titres of pre-existing adenovirus serotype-specific neutralizing antibodies predict viral reactivation after allogeneic stem cell transplantation in children

Transplantation and immunomodulatio

Combined CD8(+) and CD4(+) adenovirus hexon-specific T cells associated with viral clearance after stem cell transplantation as treatment for adenovirus infection

Background Human adenovirus can cause morbidity and mortality in immunocompromised patients after allogeneic stem cell transplantation. Reconstitution of adenovirus-specific CD4(+) T cells has been reported to be associated with sustained protection from adenovirus disease, but epitope specificity of these responses has not been characterized. Since mainly CD4(+) T cells and no CD8(+) T cells specific for adenovirus have been detected after allogeneic stem cell transplantation, the relative contribution of adenovirus-specific CD4(+) and CD8(+) T cells in protection from adenovirus disease remains to be elucidated. Design and Methods The presence of human adenovirus hexon-specific T cells was investigated in peripheral blood of pediatric and adult allogeneic stem cell transplant recipients, who showed spontaneous resolution of disseminated adenovirus infection. Subsequently, a clinical grade method was developed for rapid generation of adenovirus-specific T-cell lines for adoptive immunotherapy. Results Clearance of human adenovirus viremia coincided with emergence of a coordinated CD8(+) and CD4(+) T-cell response against adenovirus hexon epitopes in patients after allogeneic stem cell transplantation. Activation of adenovirus hexon-specific CD8(+) and CD4(+) T cells with a hexon protein-spanning peptide pool followed by interferon-gamma-based isolation allowed rapid expansion of highly specific T-cell lines from healthy adults, including donors with very low frequencies of adenovirus hexon-specific T cells. Adenovirus-specific T-cell lines recognized multiple MHC class I and II restricted epitopes, including known and novel epitopes, and efficiently lysed human adenovirus-infected target cells. Conclusions This study provides a rationale and strategy for the adoptive transfer of donor-derived human adenovirus hexon-specific CD8(+) and CD4(+) T cells for the treatment of disseminated adenovirus infection after allogeneic stem cell transplantation.Transplantation and immunomodulatio