Monoculture-derived T lymphocytes specific for multiple viruses expand and produce clinically relevant effects in immunocompromised individuals

Abstract

Immunocompromised individuals are at high risk for life-threatening diseases, especially those caused by cytomegalovirus (CMV), Epstein-Barr virus (EBV) and adenovirus. Conventional therapeutics are primarily active only against CMV, and resistance is frequent. Adoptive transfer of polyclonal cytotoxic T lymphocytes (CTLs) specific for CMV or EBV seems promising, but it is unclear whether this strategy can be extended to adenovirus, which comprises many serotypes. In addition, the preparation of a specific CTL line for each virus in every eligible individual would be impractical. Here we describe genetic modification of antigen-presenting cell lines to facilitate the production of CD4(+) and CD8(+) T lymphocytes specific for CMV, EBV and several serotypes of adenovirus from a single cell culture. When administered to immunocompromised individuals, the single T lymphocyte line expands into multiple discrete virus-specific populations that supply clinically measurable antiviral activity. Monoculture-derived multispecific CTL infusion could provide a safe and efficient means to restore virus-specific immunity in the immunocompromised host.

Umbilical cord blood graft engineering: challenges and opportunities

“…In the adult donor setting, virus-specific T-cells (VSTs) can be generated from the donor, either by culture with modified APCs [82][83][84][85][86][87] or peptide multimers. 88,89 Rapid isolation strategies such as 'gamma catch' can be utilized when VSTs occur at high frequency in the donor's blood.…”

“…83,84 However, there are considerable barriers to successful use of adoptive immunotherapy with VSTs post-UCBT; specifically, there are small numbers of T-cells available for manipulation and their phenotype is naïve. [85][86][87]94,95 Hanley et al have shown that multi-VSTs against EBV, CMV and ADV can be generated from naïve cord blood cells using a complex method of transducing EBV-LCLs transduced with CMVpp65 via an adenoviral vector and these are highly active against virus, despite recognition of non-canonical CMV and EBV epitopes. These VSTs generated from UCB have been shown to be efficacious for treatment of viral infection post-allograft.…”

Polyomavirus JC-targeted T-cell therapy for progressive multiple leukoencephalopathy in a hematopoietic cell transplantation recipient

“…[10][11][12] In HSCT recipients, there is a rationale for enhancing/rebuilding JCV-specific immunity through adoptive cell therapy, as there is sound evidence that the transfer of donor-derived, Ag-specific T-lymphocytes can restore protective immunity and control other viral infections. [13][14][15][16] We report the case of a patient presenting with JCVassociated PML 5 years after HSCT, who improved after immunosuppression discontinuation and combined treatment, including donor-derived JCV-specific CTL.…”

HLA-restricted presentation of WT1 tumor antigen in B-lymphoblastoid cell lines established using a maxi-EBV system

“…11,12 A previously reported mini-EBV system is an attractive alternative, as one can convert PBMC to transgeneexpressing LCLs via just single infection of mini-EBV. 13 On the other hand, production of mini-EBVs requires EBV packaging cell lines, and, as a result, recombinational events between mini-EBVs and helper virus genomes can be problematic.…”

“…10 Alternatively, foreign antigens can be expressed in LCLs by infecting LCLs with recombinant retroviruses 11 or adenoviruses expressing foreign antigens. 12 To simplify the establishment of LCLs presenting foreign antigen, a single-step strategy was developed by utilizing so-called 'mini-EBVs'. 13 Mini-EBV (71 kb in size) contains minimal viral genes that are essential for B-cell immortalization, 14,15 whereas maxi-EBV (175 kb in size) contains an entire EBV genome.…”