Ex vivo generation of cytotoxic T lymphocytes specific for one or two distinct viruses for the prophylaxis of patients receiving an allogeneic bone marrow transplant

Abstract

Summary:Human adenovirus (AdV) infection and EBV-lymphoproliferative disease (LPD) are serious complications following allogeneic stem cell transplantation. In the healthy individual these viruses cause minor, self-limiting diseases but in the immunocompromised patient they are responsible for significant morbidity and mortality. The limitations of anti-viral drugs and a better understanding of the cellular immune response to viral pathogens have prompted interest in developing adoptive immunotherapy for transplant patients. Ex vivo expanded cytotoxic T lymphocytes (CTLs) specific for EBV have been used effectively both as prophylaxis against EBV-LPD and as treatment of established EBV + lymphoma. To generate CTLs specific for AdV, we infected immature dendritic cells with virus, in the presence of lipid, and subsequently used these cells to stimulate PBMNCs. Cytotoxicity assays showed that the resulting CTLs specifically lysed AdV-expressing targets and that this was mediated predominantly by CD4 + T cells. To generate CTLs specific for both AdV and EBV, we developed a CD40 ligand co-culture system to infect B-lymphoblastoid cell lines (LCLs) with high efficiency. PBMNCs from healthy AdV-seropositive donors were stimulated weekly with autologous AdV + -LCLs. Chromium release assays demonstrated that the resultant CTLs had specificity against both EBV and AdV and that this was mediated by both CD4 + and CD8 + T cells. Our findings have potential implications for post-transplant AdV and EBV immunotherapy in recipients of allogeneic stem cell transplants. Bone Marrow Transplantation (2001) 27, 53-64.

Mouse Adenovirus Type 1 Infection in SCID Mice: an Experimental Model for Antiviral Therapy of Systemic Adenovirus Infections

“…The reduction of immunosuppression, combined with antiviral therapy initiated as early as possible after the adenovirus infection, remains the mainstay of therapy (1,8,9,22). Alternatively, there may be a therapeutic benefit with adenovirus-specific T cells, similar to that described for Epstein-Barr virus and cytomegalovirus (46,50). Due to extensive cross-reactivity of adenovirus-specific T cells across serotypes, this approach may be feasible (20,51).…”

Adenovirus infection in children after allogeneic stem cell transplantation: diagnosis, treatment and immunity

“…Various research groups are currently investigating the role of CD4 þ and CD8 þ T cells in the cellular immune response to HAdV. 23,[25][26][27] Together with a more detailed analysis of the origin of the infectious HAdV in the setting of allogeneic SCT and of the HAdV type(s) infecting a patient, this research will certainly enhance our knowledge of the interaction between HAdV and the immune system. This will be of benefit not only for recipients of stem cell grafts but also for recipients of solid organ transplants and other immunocompromised patients in which HAdV infection may have serious complications.…”

Adenovirus infections following allogeneic stem cell transplantation: incidence and outcome in relation to graft manipulation, immunosuppression, and immune recovery

“…Flomenberg et al demonstrated adenovirus-specific cellular immunity in healthy donors, 25,26 and others have also reported the generation of adenovirus-specific CTLs. 27,28 Unlike neutralizing antibodies, adenovirus CTLs may not be serotype restricted, suggesting that serotype variation might not be a stumbling block for developing adoptive cellular therapy. 29 We are currently investigating the kinetics of adenovirusspecific T-cell recovery following an allograft and whether this is predictive of the acquisition and outcome of adenovirus infections.…”