In vitro expansion of polyclonal T-cell subsets for adoptive immunotherapy by recombinant modified vaccinia Ankara.
OBJECTIVE
Adoptive cellular therapy of cytomegalovirus (CMV)-specific T cells in allogeneic hematopoietic stem cell transplantation (HSCT) patients is a promising approach for controlling CMV viremia and its morbidity. We sought to develop a clinically suitable strategy to dually expand infusible CD8(+) and CD4(+) T-cell subsets specific for CMV.


METHODS
Polyclonal CMV T-cell lines were generated using peripheral blood mononuclear cell (PBMCs) treated with synthetic single-stranded CpG motif-containing oligodeoxynucleotides (ODNs) and infected with recombinant (r) modified vaccinia Ankara (MVA) expressing CMV antigens. Cultures derived from 12 healthy CMV-positive donors were analyzed using chromium release and lymphoproliferation assays, intracellular staining for interferon-gamma (IFN-gamma), and HLA tetramers.


RESULTS
A 3-day incubation with a combination of ODN 2006 and 2216 was found to reproducibly generate a highly rMVA infectable population of PBMCs with concomitant high expression of CMV antigens. CpG ODN-treated autologous PBMCs infected with rMVA elicited a 30-fold average expansion of both CMV-specific CD4(+) and CD8(+) T cells in 10 days. The enriched T-cell populations showed minimal alloreactivity, high levels of CMV-specific HLA class I tetramer binding, cytotoxic activity, and IFN-gamma production from both CD8(+) and CD4(+) T cells.


CONCLUSIONS
The ability to quickly produce autologous professional antigen-presenting cells, capable of stimulating clinically useful amounts of CMV-specific CD4(+) and CD8(+) T-cell lines, enhances the attractiveness of using rMVA for immunotherapeutic interventions to manage HSCT-related CMV disease.
Laboratory of Vaccine Research, Beckman Research Institute of the City of Hope, CA 91010, USA. clarosa@coh.org
Corinna
C
La Rosa
Zhongde
Z
Wang
Simon F
SF
Lacey
Maria M
MM
Lalimarmo
Aparna
A
Krishnan
Jeff
J
Longmate
Don J
DJ
Diamond
D017951:Antigen Presentation / Q000187:drug effects / Q000276:immunology; D000938:Antigen-Presenting Cells / Q000276:immunology* / Q000821:virology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000637:transplantation; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000637:transplantation; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D018899:CpG Islands / Q000276:immunology; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology*; D003586:Cytomegalovirus Infections / Q000209:etiology / Q000276:immunology* / Q000628:therapy; D006680:HLA Antigens / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects / Q000379:methods; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D007371:Interferon-gamma / Q000276:immunology; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology*; D017710:Lymphocyte Transfusion / Q000379:methods; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology; D014184:Transplantation, Homologous; D014616:Vaccinia virus / Q000235:genetics / Q000276:immunology
D006680:HLA Antigens; D009838:Oligodeoxyribonucleotides; D007371:Interferon-gamma