Activating and propagating polyclonal gamma delta T cells with broad specificity for malignancies.
OBJECTIVE
To activate and propagate populations of γδ T cells expressing polyclonal repertoire of γ and δ T-cell receptor (TCR) chains for adoptive immunotherapy of cancer, which has yet to be achieved.


METHODS
Clinical-grade artificial antigen-presenting cells (aAPC) derived from K562 tumor cells were used as irradiated feeders to activate and expand human γδ T cells to clinical scale. These cells were tested for proliferation, TCR expression, memory phenotype, cytokine secretion, and tumor killing.


RESULTS
γδ T-cell proliferation was dependent upon CD137L expression on aAPC and addition of exogenous IL2 and IL21. Propagated γδ T cells were polyclonal as they expressed TRDV1, TRDV2-2, TRDV3, TRDV5, TRDV7, and TRDV8 with TRGV2, TRGV3F, TRGV7, TRGV8, TRGV9*A1, TRGV10*A1, and TRGV11 TCR chains. IFNγ production by Vδ1, Vδ2, and Vδ1(neg)Vδ2(neg) subsets was inhibited by pan-TCRγδ antibody when added to cocultures of polyclonal γδ T cells and tumor cell lines. Polyclonal γδ T cells killed acute and chronic leukemia, colon, pancreatic, and ovarian cancer cell lines, but not healthy autologous or allogeneic normal B cells. Blocking antibodies demonstrated that polyclonal γδ T cells mediated tumor cell lysis through combination of DNAM1, NKG2D, and TCRγδ. The adoptive transfer of activated and propagated γδ T cells expressing polyclonal versus defined Vδ TCR chains imparted a hierarchy (polyclonal>Vδ1>Vδ1(neg)Vδ2(neg)>Vδ2) of survival of mice with ovarian cancer xenografts.


CONCLUSIONS
Polyclonal γδ T cells can be activated and propagated with clinical-grade aAPCs and demonstrate broad antitumor activities, which will facilitate the implementation of γδ T-cell cancer immunotherapies in humans.
Departments of Pediatrics, University of Texas Graduate School of Biomedical Sciences at Houston, Houston, Texas.
Drew C
DC
Deniger
Departments of Pediatrics.
Sourindra N
SN
Maiti
Departments of Pediatrics.
Tiejuan
T
Mi
Departments of Pediatrics.
Kirsten C
KC
Switzer
Cancer Biology.
Vijaya
V
Ramachandran
Departments of Pediatrics, University of Texas Graduate School of Biomedical Sciences at Houston, Houston, Texas.
Lenka V
LV
Hurton
Departments of Pediatrics.
Sonny
S
Ang
Departments of Pediatrics.
Simon
S
Olivares
Departments of Pediatrics.
Brian A
BA
Rabinovich
Departments of Pediatrics.
M Helen
MH
Huls
Departments of Pediatrics, University of Texas Graduate School of Biomedical Sciences at Houston, Houston, Texas.
Dean A
DA
Lee
Experimental Therapeutics, and.
Robert C
RC
Bast
Stem Cell Transplantation and Cellular Therapy, University of Texas MD Anderson Cancer Center; and.
Richard E
RE
Champlin
Departments of Pediatrics, University of Texas Graduate School of Biomedical Sciences at Houston, Houston, Texas ljncooper@mdanderson.org.
Laurence J N
LJ
Cooper
D019264:Adoptive Transfer; D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D016207:Cytokines / Q000096:biosynthesis / Q000494:pharmacology; D004195:Disease Models, Animal; D015870:Gene Expression; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D008213:Lymphocyte Activation / Q000276:immunology*; D051379:Mice; D008822:Mice, Transgenic; D055655:NK Cell Lectin-Like Receptor Subfamily K / Q000378:metabolism; D009369:Neoplasms / Q000276:immunology* / Q000378:metabolism* / Q000401:mortality / Q000628:therapy; D012333:RNA, Messenger; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics / Q000378:metabolism*; D037182:T-Cell Antigen Receptor Specificity / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*; D023041:Xenograft Model Antitumor Assays; D000098862:T Lineage-Specific Activation Antigen 1
D000945:Antigens, Differentiation, T-Lymphocyte; D000098862:T Lineage-Specific Activation Antigen 1; D016207:Cytokines; C527085:KLRK1 protein, human; D055655:NK Cell Lectin-Like Receptor Subfamily K; D012333:RNA, Messenger; D016692:Receptors, Antigen, T-Cell, gamma-delta; D007371:Interferon-gamma