Show simple item record

dc.contributor.authorSundarasetty, Bala Sai
dc.contributor.authorSingh, Vijay Kumar
dc.contributor.authorSalguero, Gustavo
dc.contributor.authorGeffers, Robert
dc.contributor.authorRickmann, Mareike
dc.contributor.authorMacke, Laura
dc.contributor.authorBorchers, Sylvia
dc.contributor.authorFigueiredo, Constanca
dc.contributor.authorSchambach, Axel
dc.contributor.authorGullberg, Urban
dc.contributor.authorProvasi, Elena
dc.contributor.authorBonini, Chiara
dc.contributor.authorGanser, Arnold
dc.contributor.authorWoelfel, Thomas
dc.contributor.authorStripecke, Renata
dc.date.accessioned2013-05-02T11:03:32Zen
dc.date.available2013-05-02T11:03:32Zen
dc.date.issued2013-02en
dc.identifier.citationLentivirus-induced dendritic cells for immunization against high-risk WT1(+) acute myeloid leukemia. 2013, 24 (2):220-37 Hum. Gene Ther.en_GB
dc.identifier.issn1557-7422en
dc.identifier.pmid23311414en
dc.identifier.doi10.1089/hum.2012.128en
dc.identifier.urihttp://hdl.handle.net/10033/288407en
dc.description.abstractWilms' tumor 1 antigen (WT1) is overexpressed in acute myeloid leukemia (AML), a high-risk neoplasm warranting development of novel immunotherapeutic approaches. Unfortunately, clinical immunotherapeutic use of WT1 peptides against AML has been inconclusive. With the rationale of stimulating multiantigenic responses against WT1, we genetically programmed long-lasting dendritic cells capable of producing and processing endogenous WT1 epitopes. A tricistronic lentiviral vector co-expressing a truncated form of WT1 (lacking the DNA-binding domain), granulocyte-macrophage colony-stimulating factor (GM-CSF), and interleukin-4 (IL-4) was used to transduce human monocytes ex vivo. Overnight transduction induced self-differentiation of monocytes into immunophenotypically stable "SmartDC/tWT1" (GM-CSF(+), IL-4(+), tWT1(+), IL-6(+), IL-8(+), TNF-α(+), MCP-1(+), HLA-DR(+), CD86(+), CCR2(+), CCR5(+)) that were viable for 3 weeks in vitro. SmartDC/tWT1 were produced with peripheral blood mononuclear cells (PBMC) obtained from an FLT3-ITD(+) AML patient and surplus material from a donor lymphocyte infusion (DLI) and used to expand CD8(+) T cells in vitro. Expanded cytotoxic T lymphocytes (CTLs) showed antigen-specific reactivity against WT1 and against WT1(+) leukemia cells. SmartDC/tWT1 injected s.c. into Nod.Rag1(-/-).IL2rγc(-/-) mice were viable in vivo for more than three weeks. Migration of human T cells (huCTLs) to the immunization site was demonstrated following adoptive transfer of huCTLs into mice immunized with SmartDC/tWT1. Furthermore, SmartDC/tWT1 immunization plus adoptive transfer of T cells reactive against WT1 into mice resulted in growth arrest of a WT1(+) tumor. Gene array analyses of SmartDC/tWT1 demonstrated upregulation of several genes related to innate immunity. Thus, SmartDC/tWT1 can be produced in a single day of ex vivo gene transfer, are highly viable in vivo, and have great potential for use as immunotherapy against malignant transformation overexpressing WT1.
dc.language.isoenen
dc.rightsArchived with thanks to Human gene therapyen_GB
dc.titleLentivirus-induced dendritic cells for immunization against high-risk WT1(+) acute myeloid leukemia.en
dc.typeArticleen
dc.contributor.departmentDepartment of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, 30625 Hannover, Germany.en_GB
dc.identifier.journalHuman gene therapyen_GB
refterms.dateFOA2018-06-13T00:57:45Z
html.description.abstractWilms' tumor 1 antigen (WT1) is overexpressed in acute myeloid leukemia (AML), a high-risk neoplasm warranting development of novel immunotherapeutic approaches. Unfortunately, clinical immunotherapeutic use of WT1 peptides against AML has been inconclusive. With the rationale of stimulating multiantigenic responses against WT1, we genetically programmed long-lasting dendritic cells capable of producing and processing endogenous WT1 epitopes. A tricistronic lentiviral vector co-expressing a truncated form of WT1 (lacking the DNA-binding domain), granulocyte-macrophage colony-stimulating factor (GM-CSF), and interleukin-4 (IL-4) was used to transduce human monocytes ex vivo. Overnight transduction induced self-differentiation of monocytes into immunophenotypically stable "SmartDC/tWT1" (GM-CSF(+), IL-4(+), tWT1(+), IL-6(+), IL-8(+), TNF-α(+), MCP-1(+), HLA-DR(+), CD86(+), CCR2(+), CCR5(+)) that were viable for 3 weeks in vitro. SmartDC/tWT1 were produced with peripheral blood mononuclear cells (PBMC) obtained from an FLT3-ITD(+) AML patient and surplus material from a donor lymphocyte infusion (DLI) and used to expand CD8(+) T cells in vitro. Expanded cytotoxic T lymphocytes (CTLs) showed antigen-specific reactivity against WT1 and against WT1(+) leukemia cells. SmartDC/tWT1 injected s.c. into Nod.Rag1(-/-).IL2rγc(-/-) mice were viable in vivo for more than three weeks. Migration of human T cells (huCTLs) to the immunization site was demonstrated following adoptive transfer of huCTLs into mice immunized with SmartDC/tWT1. Furthermore, SmartDC/tWT1 immunization plus adoptive transfer of T cells reactive against WT1 into mice resulted in growth arrest of a WT1(+) tumor. Gene array analyses of SmartDC/tWT1 demonstrated upregulation of several genes related to innate immunity. Thus, SmartDC/tWT1 can be produced in a single day of ex vivo gene transfer, are highly viable in vivo, and have great potential for use as immunotherapy against malignant transformation overexpressing WT1.


Files in this item

Thumbnail
Name:
Sundarasetty et al_final.pdf
Size:
1.040Mb
Format:
PDF
Description:
allowed oublishers' PDF

This item appears in the following Collection(s)

Show simple item record