Preclinical evaluation of ex vivo expanded/activated γδ T cells for immunotherapy of glioblastoma multiforme.
We have previously shown that expanded/activated γδ T cells from healthy donors are cytotoxic to GBM cell lines and primary GBM explants. In this report, we examined the therapeutic effect of intracranial infusion of expanded/activated γδ T cells on human minimal and established U251 tumor xenografts in athymic nude mice. Immunohistochemistry was used to determine the presence of NKG2D ligands on cell lines and tumors, and blocking studies were used to determine the effect of these ligands on γδ T cell recognition. Expanded/activated γδ T cells were prepared by 18-day culture in RPMI, human serum (HS), anti-CD2, IL-12, IFN-γ, and OKT-3. Anti-GBM activity of the cell product was assessed using in vitro cytotoxicity assays against the GBM cell line U251MG in suspension and in adherent culture. Ex vivo expanded/activated γδ T cells were of the effector/memory phenotype, expressed Th1 cytokines, and effectively killed U251 cells in vitro. Xenografts were prepared using a U251 cell line following transfection with a firefly luciferase gene to monitor tumor progression. Mice treated with γδ T cells showed slower progression of both new and established GBM xenografts versus mice that received vehicle only as determined by photon emission over time. Median survival was improved in all γδ T cell treated groups between 32 and 50 days by Kaplan-Meier analysis. U251 cells expressed ULBP-2 and ULBP-3, although blocking of these reduced in vitro cytotoxicity of γδ T cells to U251MG by only 33 and 25%, respectively. These studies show that expanded/activated γδ T cells can mediate killing of new or established GBM xenografts, reduce tumor progression, and constitute a potentially effective novel immunotherapeutic strategy against GBM.
Bryant
Nichole L
NL
Departments of Pediatrics, School of Medicine, University of Alabama at Birmingham, Suite 541 Tinsley Harrison Tower, 1530 Third Avenue South, 1900 University Blvd, Birmingham, AL 35294, USA.
Gillespie
G Yancey
GY
Lopez
Richard D
RD
Markert
James M
JM
Cloud
Gretchen A
GA
Langford
Catherine P
CP
Arnouk
Hilal
H
Su
Yun
Y
Haines
Hilary L
HL
Suarez-Cuervo
Catalina
C
Lamb
Lawrence S
LS
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D005434:Flow Cytometry / Q000379:methods; D058851:GPI-Linked Proteins / Q000378:metabolism; D005909:Glioblastoma / Q000276:immunology* / Q000401:mortality / Q000628:therapy*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000276:immunology*; D016019:Survival Analysis; D013601:T-Lymphocytes / Q000145:classification / Q000166:cytology / Q000276:immunology*; D013997:Time Factors; D014162:Transfection / Q000379:methods; D014409:Tumor Necrosis Factor-alpha; D023041:Xenograft Model Antitumor Assays
D015703:Antigens, CD; D058851:GPI-Linked Proteins; D036341:Intercellular Signaling Peptides and Proteins; D016692:Receptors, Antigen, T-Cell, gamma-delta; D014409:Tumor Necrosis Factor-alpha; C423230:ULBP2 protein, human; C423229:ULBP3 protein, human