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Clinical applications of gamma delta T cells with multivalent immunity.

Deniger DC, Moyes JS, Cooper LJ - Front Immunol (2014)

Bottom Line: However, these cells represent a small fraction (1-5%) of the peripheral T-cell pool and require activation and propagation to achieve clinical benefit.Recent advances using immobilized antigens, agonistic monoclonal antibodies (mAbs), tumor-derived artificial antigen presenting cells (aAPC), or combinations of activating mAbs and aAPC have been successful in expanding gamma delta T cells with oligoclonal or polyclonal TCR repertoires.Gamma delta T cells are also amenable to genetic modification as evidenced by introduction of αβ TCRs, chimeric antigen receptors, and drug-resistance genes.

View Article: PubMed Central - PubMed

Affiliation: Surgery Branch, National Cancer Institute , Bethesda, MD , USA.

ABSTRACT
γδ T cells hold promise for adoptive immunotherapy because of their reactivity to bacteria, viruses, and tumors. However, these cells represent a small fraction (1-5%) of the peripheral T-cell pool and require activation and propagation to achieve clinical benefit. Aminobisphosphonates specifically expand the Vγ9Vδ2 subset of γδ T cells and have been used in clinical trials of cancer where objective responses were detected. The Vγ9Vδ2 T cell receptor (TCR) heterodimer binds multiple ligands and results in a multivalent attack by a monoclonal T cell population. Alternatively, populations of γδ T cells with oligoclonal or polyclonal TCR repertoire could be infused for broad-range specificity. However, this goal has been restricted by a lack of applicable expansion protocols for non-Vγ9Vδ2 cells. Recent advances using immobilized antigens, agonistic monoclonal antibodies (mAbs), tumor-derived artificial antigen presenting cells (aAPC), or combinations of activating mAbs and aAPC have been successful in expanding gamma delta T cells with oligoclonal or polyclonal TCR repertoires. Immobilized major histocompatibility complex Class-I chain-related A was a stimulus for γδ T cells expressing TCRδ1 isotypes, and plate-bound activating antibodies have expanded Vδ1 and Vδ2 cells ex vivo. Clinically sufficient quantities of TCRδ1, TCRδ2, and TCRδ1(neg)TCRδ2(neg) have been produced following co-culture on aAPC, and these subsets displayed differences in memory phenotype and reactivity to tumors in vitro and in vivo. Gamma delta T cells are also amenable to genetic modification as evidenced by introduction of αβ TCRs, chimeric antigen receptors, and drug-resistance genes. This represents a promising future for the clinical application of oligoclonal or polyclonal γδ T cells in autologous and allogeneic settings that builds on current trials testing the safety and efficacy of Vγ9Vδ2 T cells.

No MeSH data available.


Related in: MedlinePlus

Methodologies for expanding γδ T cells ex vivo. (A) A generalized schematic for the use of aminobisphosphonates (Zol, zoledronic acid) or synthetic phosphoantigens (BrHPP, bromohydrin pyrophosphate; 2M3B1PP, 2-methyl-3-butenyl-1-pyrophosphate) and interleukin-2 (IL-2) to expand γδ T cells from peripheral blood mononuclear cells (PBMC). (B) Plate-bound MHC class-I chain-related (MICA) and IL-2 were used to expand γδ T cells from colon and ovarian tumor tissues. (C) Immobilized antibodies (Ab) were used to expand γδ T cells from PBMC in three scenarios: (top) PBMC directly stimulated with anti-pan-TCRγδ Ab and IL-2, (middle) PBMC depleted of CD4 and CD8 T cells followed by two rounds of stimulus with anti-CD3 Ab (OKT3), IL-2, and IL-4, and (bottom) PBMC were depleted of non-adherent cells, stimulated with anti-CD2 Ab (S5.2), interferon-γ (IFNγ), and IL-12, then stimulated with OKT3 and IL-2. (D) Schematic for the use of artificial antigen presenting cells (aAPC) to expand γδ T cells from PBMC in two scenarios: (top) PBMC was depleted of CD56+ NK cells then of other non-γδ T cells (TCRγ/δ+ magnetic bead kit) so that γδ T cell were isolated by “negative selection” and co-cultured recursively with aAPC, IL-2, and IL-21 for 2–3 rounds of stimulation; (bottom) PBMC was depleted of CD14+ monocytes and “positively selected” with TCRγδ magnetic beads then co-cultured recursively with anti-TCRγδ Ab-loaded aAPC, IL-2, and IL-21 for 2–3 rounds of stimulation.
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Figure 1: Methodologies for expanding γδ T cells ex vivo. (A) A generalized schematic for the use of aminobisphosphonates (Zol, zoledronic acid) or synthetic phosphoantigens (BrHPP, bromohydrin pyrophosphate; 2M3B1PP, 2-methyl-3-butenyl-1-pyrophosphate) and interleukin-2 (IL-2) to expand γδ T cells from peripheral blood mononuclear cells (PBMC). (B) Plate-bound MHC class-I chain-related (MICA) and IL-2 were used to expand γδ T cells from colon and ovarian tumor tissues. (C) Immobilized antibodies (Ab) were used to expand γδ T cells from PBMC in three scenarios: (top) PBMC directly stimulated with anti-pan-TCRγδ Ab and IL-2, (middle) PBMC depleted of CD4 and CD8 T cells followed by two rounds of stimulus with anti-CD3 Ab (OKT3), IL-2, and IL-4, and (bottom) PBMC were depleted of non-adherent cells, stimulated with anti-CD2 Ab (S5.2), interferon-γ (IFNγ), and IL-12, then stimulated with OKT3 and IL-2. (D) Schematic for the use of artificial antigen presenting cells (aAPC) to expand γδ T cells from PBMC in two scenarios: (top) PBMC was depleted of CD56+ NK cells then of other non-γδ T cells (TCRγ/δ+ magnetic bead kit) so that γδ T cell were isolated by “negative selection” and co-cultured recursively with aAPC, IL-2, and IL-21 for 2–3 rounds of stimulation; (bottom) PBMC was depleted of CD14+ monocytes and “positively selected” with TCRγδ magnetic beads then co-cultured recursively with anti-TCRγδ Ab-loaded aAPC, IL-2, and IL-21 for 2–3 rounds of stimulation.

Mentions: Immunotherapy with γδ T cells requires their activation and expansion as they comprise only a small percentage of circulating T cells. Interleukin-2 (IL-2) and activating CD3 antibody (OKT3), commonly used for the propagation of αβ T cells directly from peripheral blood mononuclear cells (PBMC), do not reliably expand γδ T cells without further manipulation and so alternative approaches are needed. Aminobisphosphonates, e.g., Zoledronic Acid (Zol), used in the treatment of bone-related diseases, e.g., osteoporosis, resulted in in vivo propagation of γδ T cells, and the use of aminobisphosphonates has been subsequently translated into laboratory practice to grow γδ T cells ex vivo (Figure 1A) (42, 43). Aminobisphosphonates inhibit cholesterol synthesis and result in the accumulation of phosphoantigen intermediates in the mevalonate–CoA pathway, including IPP, a ligand for Vγ9Vδ2 (44). However, only the Vγ9Vδ2 T-cell subset is reactive to cells treated with phosphoantigens (45, 46). Synthetic phosphoantigens, e.g., bromohydrin pyrophosphate (BrHPP) (47) and 2-methyl-3-butenyl-1-pyrophosphate (2M3B1PP) (48), can mimic aminobisphosphonates and stimulate Vγ9Vδ2 T cells for proliferation.


Clinical applications of gamma delta T cells with multivalent immunity.

Deniger DC, Moyes JS, Cooper LJ - Front Immunol (2014)

Methodologies for expanding γδ T cells ex vivo. (A) A generalized schematic for the use of aminobisphosphonates (Zol, zoledronic acid) or synthetic phosphoantigens (BrHPP, bromohydrin pyrophosphate; 2M3B1PP, 2-methyl-3-butenyl-1-pyrophosphate) and interleukin-2 (IL-2) to expand γδ T cells from peripheral blood mononuclear cells (PBMC). (B) Plate-bound MHC class-I chain-related (MICA) and IL-2 were used to expand γδ T cells from colon and ovarian tumor tissues. (C) Immobilized antibodies (Ab) were used to expand γδ T cells from PBMC in three scenarios: (top) PBMC directly stimulated with anti-pan-TCRγδ Ab and IL-2, (middle) PBMC depleted of CD4 and CD8 T cells followed by two rounds of stimulus with anti-CD3 Ab (OKT3), IL-2, and IL-4, and (bottom) PBMC were depleted of non-adherent cells, stimulated with anti-CD2 Ab (S5.2), interferon-γ (IFNγ), and IL-12, then stimulated with OKT3 and IL-2. (D) Schematic for the use of artificial antigen presenting cells (aAPC) to expand γδ T cells from PBMC in two scenarios: (top) PBMC was depleted of CD56+ NK cells then of other non-γδ T cells (TCRγ/δ+ magnetic bead kit) so that γδ T cell were isolated by “negative selection” and co-cultured recursively with aAPC, IL-2, and IL-21 for 2–3 rounds of stimulation; (bottom) PBMC was depleted of CD14+ monocytes and “positively selected” with TCRγδ magnetic beads then co-cultured recursively with anti-TCRγδ Ab-loaded aAPC, IL-2, and IL-21 for 2–3 rounds of stimulation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263175&req=5

Figure 1: Methodologies for expanding γδ T cells ex vivo. (A) A generalized schematic for the use of aminobisphosphonates (Zol, zoledronic acid) or synthetic phosphoantigens (BrHPP, bromohydrin pyrophosphate; 2M3B1PP, 2-methyl-3-butenyl-1-pyrophosphate) and interleukin-2 (IL-2) to expand γδ T cells from peripheral blood mononuclear cells (PBMC). (B) Plate-bound MHC class-I chain-related (MICA) and IL-2 were used to expand γδ T cells from colon and ovarian tumor tissues. (C) Immobilized antibodies (Ab) were used to expand γδ T cells from PBMC in three scenarios: (top) PBMC directly stimulated with anti-pan-TCRγδ Ab and IL-2, (middle) PBMC depleted of CD4 and CD8 T cells followed by two rounds of stimulus with anti-CD3 Ab (OKT3), IL-2, and IL-4, and (bottom) PBMC were depleted of non-adherent cells, stimulated with anti-CD2 Ab (S5.2), interferon-γ (IFNγ), and IL-12, then stimulated with OKT3 and IL-2. (D) Schematic for the use of artificial antigen presenting cells (aAPC) to expand γδ T cells from PBMC in two scenarios: (top) PBMC was depleted of CD56+ NK cells then of other non-γδ T cells (TCRγ/δ+ magnetic bead kit) so that γδ T cell were isolated by “negative selection” and co-cultured recursively with aAPC, IL-2, and IL-21 for 2–3 rounds of stimulation; (bottom) PBMC was depleted of CD14+ monocytes and “positively selected” with TCRγδ magnetic beads then co-cultured recursively with anti-TCRγδ Ab-loaded aAPC, IL-2, and IL-21 for 2–3 rounds of stimulation.
Mentions: Immunotherapy with γδ T cells requires their activation and expansion as they comprise only a small percentage of circulating T cells. Interleukin-2 (IL-2) and activating CD3 antibody (OKT3), commonly used for the propagation of αβ T cells directly from peripheral blood mononuclear cells (PBMC), do not reliably expand γδ T cells without further manipulation and so alternative approaches are needed. Aminobisphosphonates, e.g., Zoledronic Acid (Zol), used in the treatment of bone-related diseases, e.g., osteoporosis, resulted in in vivo propagation of γδ T cells, and the use of aminobisphosphonates has been subsequently translated into laboratory practice to grow γδ T cells ex vivo (Figure 1A) (42, 43). Aminobisphosphonates inhibit cholesterol synthesis and result in the accumulation of phosphoantigen intermediates in the mevalonate–CoA pathway, including IPP, a ligand for Vγ9Vδ2 (44). However, only the Vγ9Vδ2 T-cell subset is reactive to cells treated with phosphoantigens (45, 46). Synthetic phosphoantigens, e.g., bromohydrin pyrophosphate (BrHPP) (47) and 2-methyl-3-butenyl-1-pyrophosphate (2M3B1PP) (48), can mimic aminobisphosphonates and stimulate Vγ9Vδ2 T cells for proliferation.

Bottom Line: However, these cells represent a small fraction (1-5%) of the peripheral T-cell pool and require activation and propagation to achieve clinical benefit.Recent advances using immobilized antigens, agonistic monoclonal antibodies (mAbs), tumor-derived artificial antigen presenting cells (aAPC), or combinations of activating mAbs and aAPC have been successful in expanding gamma delta T cells with oligoclonal or polyclonal TCR repertoires.Gamma delta T cells are also amenable to genetic modification as evidenced by introduction of αβ TCRs, chimeric antigen receptors, and drug-resistance genes.

View Article: PubMed Central - PubMed

Affiliation: Surgery Branch, National Cancer Institute , Bethesda, MD , USA.

ABSTRACT
γδ T cells hold promise for adoptive immunotherapy because of their reactivity to bacteria, viruses, and tumors. However, these cells represent a small fraction (1-5%) of the peripheral T-cell pool and require activation and propagation to achieve clinical benefit. Aminobisphosphonates specifically expand the Vγ9Vδ2 subset of γδ T cells and have been used in clinical trials of cancer where objective responses were detected. The Vγ9Vδ2 T cell receptor (TCR) heterodimer binds multiple ligands and results in a multivalent attack by a monoclonal T cell population. Alternatively, populations of γδ T cells with oligoclonal or polyclonal TCR repertoire could be infused for broad-range specificity. However, this goal has been restricted by a lack of applicable expansion protocols for non-Vγ9Vδ2 cells. Recent advances using immobilized antigens, agonistic monoclonal antibodies (mAbs), tumor-derived artificial antigen presenting cells (aAPC), or combinations of activating mAbs and aAPC have been successful in expanding gamma delta T cells with oligoclonal or polyclonal TCR repertoires. Immobilized major histocompatibility complex Class-I chain-related A was a stimulus for γδ T cells expressing TCRδ1 isotypes, and plate-bound activating antibodies have expanded Vδ1 and Vδ2 cells ex vivo. Clinically sufficient quantities of TCRδ1, TCRδ2, and TCRδ1(neg)TCRδ2(neg) have been produced following co-culture on aAPC, and these subsets displayed differences in memory phenotype and reactivity to tumors in vitro and in vivo. Gamma delta T cells are also amenable to genetic modification as evidenced by introduction of αβ TCRs, chimeric antigen receptors, and drug-resistance genes. This represents a promising future for the clinical application of oligoclonal or polyclonal γδ T cells in autologous and allogeneic settings that builds on current trials testing the safety and efficacy of Vγ9Vδ2 T cells.

No MeSH data available.


Related in: MedlinePlus