Functionally active virus-specific T cells that target CMV, adenovirus, and EBV can be expanded from naive T-cell populations in cord blood and will target a range of viral epitopes.
The naive phenotype of cord blood (CB) T cells may reduce graft-versus-host disease after umbilical cord blood transplantation, but this naivety and their low absolute numbers also delays immune reconstitution, producing higher infection-related mortality that is predominantly related to CMV, adenovirus (Adv), and EBV. Adoptive immunotherapy with peripheral blood-derived virus-specific cytotoxic T lymphocytes (CTLs) can effectively prevent viral disease after conventional stem cell transplantation, and we now describe the generation of single cultures of CTLs from CB that are specific for multiple viruses. Using EBV-infected B cells transduced with a clinical-grade Ad5f35CMVpp65 adenoviral vector as sources of EBV, Adv, and CMV antigens, we expanded virus-specific T cells even from CB T cells with a naive phenotype. After expansion, each CTL culture contained both CD8(+) and CD4(+) T-cell subsets, predominantly of effector memory phenotype. Each CTL culture also had HLA-restricted virus-specific cytotoxic effector function against EBV, CMV, and Adv targets. The CB CTLs recognized multiple viral epitopes, including CD4-restricted Adv-hexon epitopes and immunosubdominant CD4- and CD8-restricted CMVpp65 epitopes. Notwithstanding their naive phenotype, it is therefore possible to generate trivirus-specific CTLs in a single culture of CB, which may be of value to prevent or treat viral disease in CB transplant recipients. This study is registered at www.clinicaltrials.gov as NCT00078533.
Hanley
Patrick J
PJ
Department of Immunology, Baylor College of Medicine, Texas Children's Hospital and The Methodist Hospital, Center for Cell and Gene Therapy, Houston, TX 77030, USA.
Cruz
Conrad Russell Young
CR
Savoldo
Barbara
B
Leen
Ann M
AM
Stanojevic
Maja
M
Khalil
Mariam
M
Decker
William
W
Molldrem
Jeffrey J
JJ
Liu
Hao
H
Gee
Adrian P
AP
Rooney
Cliona M
CM
Heslop
Helen E
HE
Dotti
Gianpietro
G
Brenner
Malcolm K
MK
Shpall
Elizabeth J
EJ
Bollard
Catherine M
CM
D000256:Adenoviridae / Q000378:metabolism*; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry*; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism / Q000821:virology; D004854:Herpesvirus 4, Human / Q000378:metabolism*; D006801:Humans; D016130:Immunophenotyping; D008954:Models, Biological; D009035:Mothers; D010455:Peptides / Q000737:chemistry; D010641:Phenotype; D013601:T-Lymphocytes / Q000166:cytology* / Q000378:metabolism / Q000821:virology
D000939:Epitopes; D010455:Peptides