Measuring Antiviral Capacity of T Cell Responses to Adenovirus

Abstract

Adenoviruses are a major cause of infectious mortality in children following allogeneic hematopoietic stem cell transplantation, with adoptive transfer of adenovirus-specific T cells being an effective therapeutic approach. We have previously shown that T cells specific for the peptide epitope LTDLGQNLLY were protective. In this study, we aimed to establish a viral dissemination assay to measure the antiviral capacity of T cells specific for this and other peptide epitopes in an infectious setting. We used replication-competent adenovirus 11 (Ad11pGFP) and adenovirus 5 containing adenovirus 35 fiber (Ad5F35GFP) viruses and T cells specific for HLA-A*01-restricted LTDLGQNLLY, HLA-B*07-restricted KPYSGTAYNAL, and HLA-A*02-restricted LLDQLIEEV peptide epitopes. T cells in PBMC from healthy donors were expanded with peptide and IL-2 or treated with IL-2 alone to serve as nonstimulated control cells, and then these expanded or nonstimulated CD8 + cells were purified and cocultured with autologous monocytes infected with adenovirus at low multiplicity of infection. After 3 d, the number of infected GFP + monocytes and, hence, viral dissemination was quantified by flow cytometry. T cells expanded with LTDLGQNLLY peptide from multiple HLA-A*01 + donors prevented adenovirus dissemination, and nonstimulated T cells did not prevent dissemination, thus, indicating that LTDLGQNLLY-specific T cells have high antiviral capacity. Similarly, expanded KPYSGTAYNAL-and LLDQLIEEV-specific T cells could prevent viral dissemination. However, the frequency of expanded T cells specific for these last two epitopes was variable between donors with consequent variable prevention of adenoviral dissemination. Taken together, we demonstrate that T cells specific for three peptide epitopes, from both structural and nonstructural proteins, can prevent adenoviral dissemination and provide a novel method to measure the antiviral capacity of adenovirus-specific T cell responses.

Targeted in-vitro-stimulation reveals highly proliferative multi-virus-specific human central memory T cells as candidates for prophylactic T cell therapy

“…Beside the verification of the high proliferative capacity of virus-specific T CM in the GMP-product, the PSPA helped us to identify additional antiviral T cell specificities within the T CM compartment. Two AdV epitope-specific CD8 + T cell populations [40,41,63] that had been undetectable in the pre-PSPA T cell screening became visible after stimulation. The frequency of AdV-specific T cells in the peripheral blood is generally low [63] and is dominated by CD4 + T cells as depicted by ICS before PSPA.…”

“…Two AdV epitope-specific CD8 + T cell populations [40,41,63] that had been undetectable in the pre-PSPA T cell screening became visible after stimulation. The frequency of AdV-specific T cells in the peripheral blood is generally low [63] and is dominated by CD4 + T cells as depicted by ICS before PSPA. In this regard, the PSPA could become a valuable diagnostic tool to amplify the sensitivity for rare specificities within the T CM compartment.…”

Characterization of the conserved regions of E1A protein from human adenovirus for reinforcement of cytotoxic T lymphocytes responses to the all genogroups causes ocular manifestation through an in silico approach

“…The data obtained by these researchers indicate that 32 percent of T-cells specific to the LLDQLIEEV epitope amino acid sequence of the E1A inhibited adenoviral propagation. Their results showed that even low levels of specific T cells can accelerate to control the infection and recommend that the performance of the T-cell response is critical to prevention ( 22 ).…”

Changes of Host Immunity Mediated by IFN-γ+ CD8+ T Cells in Children with Adenovirus Pneumonia in Different Severity of Illness

“…Although few studies focus on the relationship between adenovirus pathogenesis and T cells, some comparative studies still show that T cell depletion is related to the mortality of adenovirus-infected hematopoietic cell transplant patients [27,28]. Furthermore, some studies have manifested that adenovirus-specific T cells have a specific antiviral ability against adenovirus [29]. In addition, previous studies have explored the effect of CD8 + T cells on the pathogenesis of mouse adenovirus-1 (MAV-1) by establishing an animal model of lung inflammation induced by MAV-1 [30].…”