Rapid expansion of cytomegalovirus-specific cytotoxic T lymphocytes by artificial antigen-presenting cells expressing a single HLA allele.
Cytomegalovirus (CMV) is a major threat in patients undergoing allogeneic bone marrow transplantation. The adoptive transfer of CMV-specific cytotoxic T lymphocytes (CTLs) expanded from the blood of CMV-seropositive donors has been shown to effectively control CMV infection. However, the requirement for safe and effective antigen-presenting cells (APCs) for each patient precludes broad applicability of this successful form of therapy. Here we analyze the ability of artificial APCs (AAPCs) to activate and expand CMV-specific CTLs from peripheral blood of seropositive HLA A2.1+ donors. We demonstrate that AAPCs expressing the CMV P495 peptide or the full-length pp65 protein stimulate P495-specific CTLs at least as effectively as autologous, peptide-pulsed, peripheral blood mononuclear cells or EBV-transformed B cells. Starting from 100 mL of blood, the AAPCs reliably yield clinically relevant CTL numbers after a single stimulation. CTLs activated on AAPCs effectively kill CMV-infected fibroblasts and have a Tc1 memory effector phenotype identical to that of CTLs generated with autologous APCs. AAPCs thus offer a rapid, controlled, convenient, and highly reproducible system for expanding CMV-specific CTLs. Furthermore, the CTL expansion obtained with AAPCs encoding full-length pp65 indicates that AAPCs may be used to present known as well as unknown CTL epitopes in the context of the AAPC's HLA.
Papanicolaou
Genovefa A
GA
Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
Latouche
Jean-Baptiste
JB
Tan
Cuiwen
C
Dupont
Jakob
J
Stiles
Jeffrey
J
Pamer
Eric G
EG
Sadelain
Michel
M
D000938:Antigen-Presenting Cells / Q000166:cytology / Q000276:immunology / Q000821:virology; D016026:Bone Marrow Transplantation; D002455:Cell Division / Q000276:immunology; D016207:Cytokines / Q000276:immunology; D003587:Cytomegalovirus / Q000276:immunology*; D003586:Cytomegalovirus Infections / Q000276:immunology / Q000517:prevention & control* / Q000628:therapy; D005434:Flow Cytometry; D015789:HLA-A2 Antigen / Q000276:immunology*; D006801:Humans; D007156:Immunologic Memory / Q000276:immunology; D016130:Immunophenotyping; D016219:Immunotherapy, Adoptive; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology* / Q000276:immunology / Q000821:virology*
D016207:Cytokines; D015789:HLA-A2 Antigen