Cytomegalovirus-Specific Cytotoxic T Lymphocytes Can Be Efficiently Expanded from Granulocyte Colony-Stimulating Factor–Mobilized Hemopoietic Progenitor Cell Products Ex Vivo and Safely Transferred to Stem Cell Transplantation Recipients to Facilitate Immune Reconstitution

Abstract

Uncontrolled cytomegalovirus (CMV) reactivation after allogeneic hematopoietic stem cell transplantation causes significant morbidity and mortality. Adoptive transfer of CMV-specific cytotoxic T lymphocytes (CTLs) is a promising therapy to treat reactivation and prevent viral disease. In this article, we describe the generation of clinical-grade CMV-specific CTLs directly from granulocyte colony-stimulating factor-mobilized hemopoietic progenitor cell (G-HPC) products collected for transplantation. This method requires less than 2.5% of a typical G-HPC product to reproducibly expand CMV-specific CTLs ex vivo. Comparison of 11 CMV CTL lines generated from G-HPC products with 52 CMV CTL lines generated from nonmobilized peripheral blood revealed similar expansion kinetics and phenotype. G-HPC-derived CTLs produced IFN-γ after reexposure to CMVpp65 antigen and exhibited CMV-directed cytotoxicity but no alloreactivity against transplantation recipient-derived cells. Seven patients received CMV-specific CTL lines expanded from G-HPC products in a prophylactic adoptive immunotherapy phase I/II clinical trial. Use of G-HPC products will facilitate integration of CTL generation into established quality systems of transplantation centers and more rapid inclusion of T cell therapies into routine clinical care.

CMV-specific immune reconstitution following allogeneic stem cell transplantation

“…[34][35][36] Reconstitution of CMV cell mediated immunity via adoptive transfer of CMV specific immune effectors has been shown to control CMV infection in both animal models 37,38 and human trials. [39][40][41][42][43][44][45][46][47] In otherwise healthy individuals, both CD8…”

“…102 CMV specific immune reconstitution via adoptive transfer of ex vivo isolated or expanded donor derived virus specific T cells (CMVVSTs) has now been performed successfully in a number of clinical trials. [39][40][41][42][43][44][45][46][47]58,59 CMV-VSTs can be manufactured in a variety of ways. Time consuming ex vivo culture methods using limiting dilution cloning or EBV transformed lymphocytes as antigen presenting cells (APCs) have largely been replaced with more rapid techniques using alternative APCs (activated monocytes, rapidly matured monocyte derived DCs or artificial APCs), various antigen sources and shorter culture times.…”

Humanes Cytomegalievirus (HCMV)

“…B. bei haploidenter Transplantation) beinhalten. Einige Studien konnten die Effizienz in der Therapie und die Vermeidung des Auftretens von HCMV-Erkrankungen nachweisen [91,92,96,100,[104][105][106][107]. Die Gabe von virusspezifischen Spender-T-Lymphozyten einige Wochen nach Transplantation senkt zusätzlich das Risiko des Auftretens einer GVHD, ohne dass die Ursachen dafür bisher bekannt sind [58,108].…”

“…Unklar ist derzeit, ob antigenspezifische CD4-oder CD8-positive T-Zellen einzeln oder gemeinsam appliziert werden sollten [105]. Erwähnenswert ist der Ansatz, die Expansion aus einem kleinen Aliquot des Stammzelltransplantats vorzunehmen und damit die logistisch oft problematische Wiedereinbestellung des Spenders zur Gewinnung virusspezifischer T-Zellen zu vermeiden [106,107]. Die Fortschritte auf diesem Gebiet sind vielversprechend [109].…”

The immune response to cytomegalovirus in allogeneic hematopoietic stem cell transplant recipients

“…Studies of Feuchtinger et al and Einsele et al have indicated the significance of antiviral effector functions of T helper cells in maintaining CTL responses after adoptive transfer [133,141]. There are few studies in Phase I/II that have demonstrated the feasibility of transferring CMV-specific T cells [133,134,[142][143][144]. They have indicated that restoration of CMV-specific immunity can be accelerated without serious immediate infusion-related toxicities, although some patients can develop GvHD after infusion, which is correlated with clinical protection.…”