Derivation of human T lymphocytes from cord blood and peripheral blood with antiviral and antileukemic specificity from a single culture as protection against infection and relapse after stem cell transplantation

Abstract

IntroductionInfections, malignant relapse, and graft-versus-host disease (GVHD), continue to cause significant morbidity and mortality after hematopoietic stem cell (HSC) or cord blood (CB) transplantation. Virus infections such as cytomegalovirus (CMV), EpsteinBarr virus (EBV), and adenovirus (AdV) are particularly problematic and remain difficult to treat, especially after umbilical CB transplantation. [1][2][3][4][5] Although ganciclovir/foscarnet may help prevent or treat CMV 6 and CD20-specific antibody may control EBV-associated lymphoproliferation, 7 these drugs are expensive and are often toxic or ineffective due to primary or secondary resistance. 6 Moreover, AdV infections are increasingly common and effective treatments are not currently available. 8 The other major cause of morbidity and mortality is relapse, occurring in more than 30% of transplant recipients with B-cell acute lymphoblastic leukemia (B-ALL), [9][10][11][12] with few appealing therapeutic options and less than 10% long-term survival. 13,14 Although donor lymphocyte infusions can be used after HSC transplantation to treat both viral infections and leukemia relapse, these are associated with potentially life-threatening GVHD, 15 have a low success rate in relapsed B-ALL, 15,16 and are unavailable for CB transplant recipients. An alternative for viral infections is the adoptive transfer of cytotoxic T lymphocytes (CTLs) directed to CMV, 17,18 EBV,19,20 and, more recently, AdV, 21,22 which can rapidly reconstitute antiviral immunity after HSC transplantation without causing GVHD. Infusion of peripheral blood-derived T-lymphocyte lines enriched in cells simultaneously recognizing CMV, EBV, and AdV (multivirus-specific CTLs [MV-CTLs]) reproducibly controls infections due to all 3 viruses after allogeneic HSC transplantation. 21 Importantly, functional CMV-, EBV-, and AdV-specific CTLs can now also be generated from naive T cells isolated from CB units. 23 It is also possible to infuse leukemia-specific CTLs into patients after HSC transplantation 24,25 ; these can be generated by stimulating peripheral blood mononuclear cells with apoptotic leukemic blasts. 25,26 Unfortunately, however, the paucity of antigen-specific CTL precursors and the need to separate graft-versus-tumor from the graft-versus-host effect may require extensive culture to generate sufficient numbers of cells for adoptive T-cell therapy. 25,26 To overcome this difficulty, investigators have used T lymphocytes engineered to express chimeric antigen receptors (CARs) directed to self-antigens expressed by tumor cells. 27,28 For example, T cells expressing a CAR specific for the CD19 molecule 29,30 may be able to prevent or treat leukemia relapse in B-ALL patients as these cells almost invariably express CD19.It would be appealing to combine these approaches to prepare a single product containing CTLs that were virus specific (through The online version of this article contains a data supplement.The publication costs of this article were defrayed in part by page charge paym...

Haploidentical related-donor hematopoietic cell transplantation in children using megadoses of CliniMACs-selected CD34+ cells and a fixed CD3+ dose

“…30,31 The high numbers of cells that can be harvested from a haploidentical donor greatly facilitates this process, while attempts to do this with UCB may require utilization of additional units. 32 Two novel aspects of this trial were the decreased dose of ATG compared with prior trials of haploidentical HCT and the use of a fixed CD3 dose. Less ATG did not appear to affect engraftment rates, which were similar in this trial (90%) and the Perugia trial (93%).…”

T-cell and natural killer cell therapies for hematologic malignancies after hematopoietic stem cell transplantation: enhancing the graft-versus-leukemia effect

“…64 Addition of cytokine signals such as IL-21 65 and IL-15 66 has also been explored. Other attempts to improve in vivo persistence involve utilizing the endogenous signaling provided by latent viruses 67,68 which have been explored clinically in neuroblastoma and B-cell leukemias/lymphomas. 69,70 Most recently, improvements within the construct itself have also allowed improvements in CAR T-cell persistence and efficacy.…”

Umbilical cord blood graft engineering: challenges and opportunities

“…This bi-specific approach using virus and CD19 + tumor-specific CTLs is being translated to the UCBT setting. 107 The MD Anderson investigators have used the sleeping beauty transposase/transposon system to electroporate a CD19 construct into the UCB of four patients who received those cells following UCBT. 108 The CB-CD19-CAR + T-cells were well tolerated and accrual to the study continues.…”