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

Abstract

The online version of this article has a Supplementary Appendix. BackgroundHuman 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 MethodsThe 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. ResultsClearance 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-γ-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. ConclusionsThis 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.Key words: allogeneic stem cell transplantation, adoptive immunotherapy, adenovirus infection, adenovirus-specific T cells.Citation: Zandvliet ML, Falkenburg JH, van Liempt E, Lankester AC, Kalpoe JS, Kester MG, van der Steen DM, van Tol MJ, Willemze R, Guchelaar HJ, Schilham MW, and Meij P. Combined CD8 + and CD4 + adenovirus hexon-specific T cells associated with viral clearance after stem cell transplantation as treatment for adenovirus infection. Haematologica 2010;95(11):1943-1951. doi:10.3324/haematol.2010

Generation of a multipathogen-specific T-cell product for adoptive immunotherapy based on activation-dependent expression of CD154

“…Immunity to AdV is atypical, because this virus shows a predominance of memory CD4 ϩ T cells in the peripheral blood (supplemental Figure 1), 38 and the ratio of the enriched CD4 ϩ and CD8 ϩ T-cell population is therefore comparable to the ratio of the precursor cells. This is probably the case because of MHC class I down-regulation in the target cells after infection with AdV to evade immune recognition and the development of a classic, CD8 ϩ -mediated cytotoxic adaptive immune response.…”

“…23 These data illustrate that the use of synthetic peptides that can be readily produced under GMP regulations and that have previously been shown to be safe may simplify the generation of clinical-grade, multipathogen-specific T-cell lines. 38 To ensure in vivo expansion, persistence, and optimal activity of adoptively transferred pathogen-specific T cells, both CD4 ϩ and CD8 ϩ effector T cells need to be present for efficient protection against viruses, 38,39 and CD4 ϩ T H 1 cells for protection against fungi. 40 The CD154 isolation method favors enrichment of CD4 ϩ T cells because of its predominant expression on activated CD4 ϩ T cells; therefore, it is especially beneficial for MHC class II antigens such as fungal antigens, which primarily induce T H 1 immunity.…”

Adenovirus Infections in Immunocompetent and Immunocompromised Patients

“…More specifically, the most prominent risk factors include allogeneic transplantations with in vivo and/or ex vivo T-cell depletion, grafts from unrelated donors or cord blood, treatment with the anti-CD52 antibody alemtuzumab (Campath) or anti-thymocyte globulin (ATG), and the presence of graft-versus-host disease (GvHD) grades III and IV associated with the use of immunosuppressive agents (129)(130)(131)(132)(133)(134)(135)(136). Additionally, severe lymphopenia, with CD3 ϩ cell counts of Ͻ300 per l peripheral blood (PB), and an absence of HAdV-specific T cells play an important role in the development of viral disease (122,124,(137)(138)(139)(140)(141). In contrast to the case with allo-HSCT recipients, a donor-positive and recipient-negative HAdV serostatus appears to be a risk factor for a severe course of infection in patients undergoing solid organ transplantation (43,142,143).…”

“…Healthy individuals usually carry HAdV-specific T cells, which can be identified by various methods, such as gamma interferon secretion assays, cytokine flow cytometry, or detection of MHC class I multimers (118,122,123). The absence of HAdVspecific T cells has a negative impact on the course of HAdV infections, and conversely, reconstitution of the HAdV-specific Tcell response correlates with viral clearance (122,124). The finding that many CD4-or CD8-restricted hexon epitopes are shared among different HAdV species and types suggests that T cells with such specificities can be protective against most, if not all, human adenonoviruses, and this fact can be exploited for vaccine-based or adoptive T-cell transfer immunotherapy for treating infections by these viruses, as outlined below.…”

The Effect of Cidofovir on Adenovirus Plasma DNA Levels in Stem Cell Transplantation Recipients without T Cell Reconstitution

“…In pediatric hematopoietic stem cell transplantation (HSCT) recipients, HAdV reactivations or primary infections can progress to viremia and disseminated disease. It is broadly accepted that T cells are essential for the protection from and clearance of HAdV viremia [2][3][4]. However, in the absence of T cell surveillance, the mortality of HAdV viremia is high because of progression to HAdV-related multiorgan failure [4][5][6][7][8][9].…”