Dendritic Cells Can Be Primed to Concurrently Activate Both Unconventional Gamma/delta and Conventional alpha/beta T Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3893-3893
Author(s):  
Francesca Fiore ◽  
Barbara Castella ◽  
Barbara Nuschak ◽  
Raffaello Bertieri ◽  
Sara Mariani ◽  
...  
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Immune Responses ◽  
Tumor Cells ◽  
Peripheral Blood ◽  
Adoptive Cell Therapy ◽  
Effector Memory ◽  
Gamma Delta T Cells ◽  
Gamma Delta ◽  
Alpha Beta

Abstract Vgamma9/Vdelta2 (gamma/delta) T cells represent the major subset of unconventional T cells circulating in the peripheral blood. Gamma/delta T cells play a major role in immune defenses against microbes, stressed cells and tumor cells. This property is based on their capability to naturally recognize phosphoantigens (pAgs), which are produced via the mevalonate (Mev) or the DOXP pathway in mammalian and nonmammalian cells, and induced self-ligands, which are de novo expressed or upregulated on the surface of stressed or tumor cells. Interestingly, gamma/delta T cells can also be activated by aminobisphosphonates (ABP)-treated monocytes. We have previously shown that ABP specifically target the Mev pathway of monocytes and induce the accumulation of phosphorylated Mev metabolites naturally recognized by gamma/delta T cells. The aim of this work was to determine whether ABP-treated dendritic cells (DC) can also activate gamma/delta T cells and whether this activation, if any, is detrimental or beneficial to the generation of antigen (Ag)-specific MHC-restricted immune responses mediated by conventional alpha/beta T cells. To this end, we have generated highly purified immature (iDC) and mature DC (mDC) from peripheral blood monocytes of healthy donors and incubated with zoledronic acid (Zol) for 24 hours. Zol is the most potent ABP currently available for clinical use. Zol treatment did not affect the phenotype and immunostimulatory properties of iDC and mDC. Zol-treated iDC and mDC induced a rapid and vigorous expansion of central memory and effector memory gamma/delta T cells. Zol-treated iDC were more potent inducers of gamma/delta T-cell activation than mDC and monocytes. Activated gamma/delta T cells displayed antitumor activity and expressed on the cell surface the appropriate antigen repertoire to target secondary lymphoid organs and exert costimulatory activity on conventional alpha/beta T cells. Indeed, an in vitro model showed that antigen-specific MHC-restricted immune responses againt the influenza matrix peptide were significantly improved by the concurrent activation of gamma/delta T cells. This is the first report showing that: 1) DC can simultaneously be primed to activate both gamma/delta and alpha/beta T cells; 2) the former act as cellular adjuvants for the development of adaptive immune responses. In conclusion, large numbers of gamma/delta T cells with effector and costimulatory activities can rapidly be generated by Zol-treated iDC/mDC. This strategy is worth of further investigation to improve adoptive cell therapy and vaccine interventions against tumors and infections.

Enhanced ability of dendritic cells to stimulate innate and adaptive immunity on short-term incubation with zoledronic acid

Blood ◽  
2007 ◽  
Vol 110 (3) ◽  
pp. 921-927 ◽  
Author(s):  
Francesca Fiore ◽  
Barbara Castella ◽  
Barbara Nuschak ◽  
Raffaello Bertieri ◽  
Sara Mariani ◽  
...  
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Zoledronic Acid ◽  
Immune Responses ◽  
T Cell Activation ◽  
Peripheral Blood ◽  
Adoptive Cell Therapy ◽  
Γδ T Cells ◽  
Effector Memory ◽  
Αβ T Cells

Abstract Vγ9/Vδ2 (γδ) T cells play a major role in innate immunity against microbes, stressed, and tumor cells. They represent less than 5% of peripheral blood lymphocytes but can be activated and expanded in vitro by aminobisphosphonates (ABP)–treated monocytes. The aim of this work was to determine whether ABP-treated dendritic cells (DCs) can also activate γδ T cells and regulate immune responses mediated by conventional αβ T cells. Highly purified immature (iDC) and mature DC (mDC) were generated from peripheral blood monocytes of healthy donors and incubated with zoledronic acid (Zol) for 24 hours. Zol-treated iDC and mDC retained their immunostimulatory properties and induced the vigorous expansion of central memory and effector memory γδ T cells. γδ T cells displayed antitumor activity and appropriate cell surface antigens to target secondary lymphoid organs and exert costimulatory activity. Antigen-specific MHC-restricted immune responses, mediated by conventional αβ T cells, were improved by the concurrent γδ T-cell activation. In conclusion, large numbers of γδ T cells with effector and costimulatory activities are rapidly generated by Zol-treated iDC/mDC. This strategy is worthy of further investigation to improve adoptive cell therapy and vaccine interventions against tumors and infections.

scholarly journals 688 In vivo expansion of gamma delta T cells by a CD19-targeted butyrophilin heterodimer leads to elimination of peripheral B cells

2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A727-A727
Author(s):  
Suresh De Silva ◽  
George Fromm ◽  
Louis Gonzalez ◽  
Arpita Patel ◽  
Kyung Yoon ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Tumor Cells ◽  
Mechanism Of Action ◽  
Peripheral Blood ◽  
Fusion Proteins ◽  
Gamma Delta T Cells ◽  
Gamma Delta ◽  
Gamma Delta T Cell

BackgroundA primary mechanism of cancer immunotherapy resistance involves downregulation of specific antigens or major histocompatibility complex based antigen presentation, which renders tumor cells invisible to alpha-beta T cells, but not gamma-delta T cells. Recently, a two-step model of gamma-delta T cell activation has emerged, wherein one butyrophilin (BTN, ie. BTN2A1) directly binds the gamma-delta TCR but is only activated if certain molecular patterns (eg. phosphoantigens) facilitate recruitment of a second BTN (ie. BTN3A1) into a complex to form a BTN2A1/3A1 heterodimer. The BTN2A1/3A1 complex specifically activates the predominant gamma-delta T cell population in the peripheral blood, comprising the Vg9d2 T cell receptor (TCR), but does not activate the primary gamma-delta T cell population in mucosal tissues, comprising the Vg4 TCR. The unique mechanism of action and specificity of gamma-delta TCR/BTN interactions suggests that therapeutic proteins comprising specific BTN heterodimers could be used to target specific gamma-delta T cell populations, with a lower risk of off-target activation common with CD3-directed T cell engagers.MethodsHuman BTN2A1/3A1-Fc-CD19scFv and mouse BTNL1/6-Fc-CD19scFv heterodimeric fusion proteins were purified and binding to CD19 or the respective gamma-delta TCRs was assessed by ELISA, Octet and flow cytometry using gd T-cells isolated from human peripheral blood and mouse intestinal tissue. The functionality of the constructs to activate gamma-delta T cells and mediate killing of tumor cells was assessed using live cell imaging in vitro as well as a murine B-cell lymphoma model in vivo.ResultsThe CD19-targeting scFv domains of the BTN heterodimer fusion proteins bound to human and mouse CD19 with low nanomolar affinity. The BTN2A1/3A1-Fc-CD19scFv compound specifically bound to the Vg9d2 TCR on human gd T cells while the mouse BTNL1/6-Fc-CD19scFv bound to Vg7d4 TCR on mouse gd T cells. Both compounds were able to activate gd T cells in a co-culture assay resulting in degranulation and increased surface expression of CD107a and also increased apoptosis of CD19+ tumor cells. Intraperitoneal administration of the mouse BTNL1/6-Fc-CD19scFv led to anti-tumor effects in A20 tumor bearing BALB/c mice. Phenotyping from BTNL1/6-Fc-CD19scFv treated mice revealed profound and rapid expansion of the endogenous gamma-delta T cells in the circulation and tumor, with concomitant depletion of peripheral CD19+ B-cells, confirming the mechanism of action of the heterodimer as a gamma-delta T cell specific engager.ConclusionsThese results provide proof of mechanism for in vivo manipulation of gamma-delta T cells using antigen-targeted butyrophilin heterodimeric fusion proteins for the treatment of cancer.

PD-1 (CD279) Contributes to the Exhaustion of Gamma/Delta-T Cells in Tumor-Bearing Mice

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 839-839 ◽  
Author(s):  
Richard D Lopez ◽  
Shin Mineishi ◽  
Lawrence S. Lamb ◽  
Hyung-Gyoon Kim ◽  
Benjamin Beck
Keyword(s):  
T Cells ◽  
Peripheral Blood ◽  
Ex Vivo ◽  
Gamma Delta T Cells ◽  
Gamma Delta ◽  
Tumor Bearing ◽  
Functionally Impaired ◽  
Alpha Beta

Abstract Abstract 839 Objectives: Programmed death-1 (PD-1)/CD279 is an immunoinhibitory receptor that can be physiologically expressed on activated antigen-specific alpha/beta T-cells and is thought to play a role in maintaining a balance between T-cell activation and tolerance. Recently, both in vitro and in vivo, it has been shown that disrupting the interaction between PD-1 and its ligands can improve antitumor effects in preclinical and clinical models, this suggesting an important role played by this pathway in escape from immune surveillance. In comparison to healthy donors, gamma/delta-T cells found in tumor-bearing hosts can be diminished in number, or can be functionally impaired in a variety of important ways. While the mechanisms accounting for these numeric or functional defects have remained unclear, here we examine the extent to which PD-1 expression on gamma/delta-T cells may play a role in this process. Findings: We first noted that peripheral blood gamma/delta-T cells are diminished in numbers in patients newly diagnosed with cancer. In addition, these gamma/delta-T cells expanded poorly when cultured ex vivo. Similar to humans, in tumor-bearing mice, we found that peripheral blood gamma/delta-T cells are diminished in number and likewise, expand poorly when cultured ex vivo. Using FACS analysis of mouse peripheral blood, we first determined that a substantial proportion of gamma/delta-T cells are actively undergoing apoptosis in tumor-bearing mice compared to healthy mice. Further analysis revealed that PD-1 is significantly upregulated on gamma/delta-T cells taken from tumor-bearing mice compared to gamma/delta-T cells taken from healthy mice. In contrast, no difference of PD-1 expression was seen when comparing alpha/beta-T cells taken from tumor-bearing and healthy mice. Using in vitro co-culture studies, we next determined that apoptosis in gamma/delta-T cells can be induced by direct contact with malignant cells, but not by contact with non-malignant cells. We then showed in these cultures that PD-1 is upregulated on gamma/delta-T cells co-cultured with tumor cell lines. Moreover, we were able to determine that the PD-1-positive gamma/delta-T cells in these cultures were undergoing apoptosis to a greater extent than PD-1-negative gamma/delta-T cells in these same cultures. Finally, in vitro using CFSE-based methods, we showed that while gamma/delta-T cells isolated from healthy mice readily proliferate upon mitogen stimulation, in contrast, gamma/delta-T cells from tumor-bearing mice proliferate poorly under the same conditions. However, in spleen cell cultures derived from tumor-bearing mice, upon addition of a monoclonal antibody directed against PD-L1 (B7-H1), a ligand for PD-1, substantial restoration of gamma/delta-T cell proliferation occurs. Conclusion: Until now, the role played by PD-1 in the exhaustion of tumor-reactive gamma/delta T-cells has not been explored. Using in vitro and in vivo models, we show that the PD-1 pathway is a potentially important mechanism by which gamma/delta T-cells are either functionally impaired or otherwise exhausted in tumor-bearing mice. These findings suggest that by disrupting the PD-1 pathway, it may be possible to “revive” or “rescue” gamma/delta T-cells in tumor-bearing hosts. Disclosures: No relevant conflicts of interest to declare.

Chronic Myeloid Leukemia Patients Undergoing Interferon Alpha Therapy Exhibit Normal Peripheral Blood Gamma Delta T Cells That May Be Expanded in Vitro to Generate Predominantly CD45RA-Positive Effector Memory Cells for Immunotherapy

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 3729-3729
Author(s):  
Gabrielle Melanie Siegers ◽  
Anna Kreutzman ◽  
Mette Matilda Ilander ◽  
Satu Mustjoki ◽  
Lynne Marie Postovit
Keyword(s):  
T Cells ◽  
Chronic Myeloid Leukemia ◽  
Peripheral Blood ◽  
Myeloid Leukemia ◽  
Healthy Donor ◽  
Effector Memory ◽  
Gamma Delta T Cells ◽  
Healthy Controls ◽  
Gamma Delta

Abstract Abstract 3729 Comprising <10% of T lymphocytes in human blood, gamma delta T cells (GDTc) are cytotoxic immune surveillance cells that are an attractive option for adoptive immunotherapy. We have previously shown that healthy donor GDTc expanded in vitro are cytotoxic to EM-2 and K562 Ph+ leukemia cell lines but not to autologous peripheral blood mononuclear cells (PBMCs). Additionally, we have noted an oligoclonal expansion of GDTc in interferon alpha (IFNα) monotherapy treated chronic myeloid leukemia (CML) patients. The current study is aimed at elucidating the role that these cells play in the remission of IFNα-treated CML patients. Immunophenotyping of thawed PBMC samples from CML patients undergoing (IFN-ON) or who had discontinued IFNα monotherapy (IFN-OFF) revealed similar central CD45RA-CD27+ (5.7% ± 2.7%, mean ± standard error, n=6) and CD45RA-CD27- effector memory (3.7% ± 2.0%, n=6) GDTc pools compared to those from healthy age-matched donors (4.4% ± 1.2% and 4.7% ± 1.4% respectively, n=3), suggesting that GDTc pools are normal in these patients. However, differences approaching significance were observed between IFN-ON (n=3) and healthy controls (n=3), with IFN-ON exhibiting less CD45RA+CD27+ naïve GDTc (24.6% ± 6.7% compared to 48.2% ± 7.7%, p=0.08) and more terminally differentiated CD45RA+CD27- RA+ effector memory GDTc (66.4% ± 11.8% compared to 42.7 ± 7.3%, p=0.16). The percentage of GDTc expressing the activating Natural Killer cell receptor NKG2D was more variable and significantly lower in freshly thawed IFN-OFF patient samples (53.1% ± 4.6%, p=0.03) as compared to age-matched healthy donor controls (68.4% ± 1.5%). A similar trend was also observed between IFN-ON patients and healthy controls (IFN-ON 54.5% ± 8.8%, p=0.08). Although GDTc are typically difficult to recover and expand from cryopreserved PBMCs, we have developed a new protocol with which we can expand patient GDTc, potentially enabling the generation of therapeutic doses on demand. We achieved variable expansions of 41- to 9097-fold from patient samples over 21 days, with four of six patient cultures achieving over 100-fold GDTc expansion. Of these four cultures, two were from each patient group, suggesting that IFN-ON or –OFF status did not correlate with the ability of these cells to expand in vitro. Importantly, expanded GDTc were mostly terminally differentiated CD45RA+CD27- RA+ effector memory GDTc (52% ± 11.7%, n=6). Of the total GDTc population, 65.9% ± 4.6% expressed NKG2D (n=6), suggesting that our protocol can generate terminally differentiated cytotoxic GDTc. We are now investigating whether the cells expanded in vitro are the clonal GDTc population previously identified in our patients and whether these expanded cells are able to kill Ph+ leukemia cells. Our findings will be important not only for understanding the mechanism of IFNα-induced cure of the patients, but also for developing GDTc therapies for CML and other malignancies. Disclosures: Mustjoki: Bristol-Myers Squibb: Honoraria, Research Funding; Novartis: Honoraria.

Anti-Tumor Actions of Trametes Versicolor Extract Polysaccharide K Include Activation of Dendritic Cells and Gamma Delta T Cells

Planta Medica ◽  
2013 ◽  
Vol 79 (10) ◽  
Author(s):  
CA Wenner ◽  
C Inatsuka ◽  
T Davis Smith ◽  
M Sasagawa ◽  
MR Martzen ◽  
...  
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Trametes Versicolor ◽  
Gamma Delta T Cells ◽  
Gamma Delta ◽  
Polysaccharide K

scholarly journals Transient increase in circulating gamma/delta T cells during Plasmodium vivax malarial paroxysms.

1994 ◽  
Vol 179 (1) ◽  
pp. 311-315 ◽  
Author(s):  
M K Perera ◽  
R Carter ◽  
R Goonewardene ◽  
K N Mendis
Keyword(s):  
T Cells ◽  
Plasmodium Vivax ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Clinical Symptoms ◽  
Gastrointestinal Symptoms ◽  
Gamma Delta T Cells ◽  
Gamma Delta ◽  
Endemic Region ◽  
Gastrointestinal Pathology

The percentage of peripheral blood mononuclear cells (PBMC) bearing the CD3+ phenotype and the alpha/beta and gamma/delta T cell receptors (TCR) in PBMC were examined in Plasmodium vivax malaria patients and convalescents. The cells were labeled with monoclonal antibodies, stained with either fluorescence or phycoerythrin, and examined by ultraviolet (UV) microscopy. A highly significant increase in both the proportion and the absolute numbers of gamma/delta T cells (p &lt; 0.005 and &lt; 0.001, respectively, Student's t test) was observed in nonimmune P. vivax patients during clinical paroxysms compared to nonmalarial controls. These T cells, which normally constitute not more than 3-5% of PBMC, constituted &lt; or = to 30% of PBMC during paroxysms in these nonimmune patients in whom the clinical symptoms were severe. A less significant increase of gamma/delta T cells were also observed in these nonimmune patients during infection, between paroxysms and during convalescence. In contrast, in an age-matched group of semi-immune patients resident in a malaria-endemic region of the country, in whom the clinical disease was comparatively mild, there was no increase in gamma/delta T cells either during infection, even during paroxysms, or convalescence. The severity of disease symptoms in patients as measured by a clinical score correlated positively with the proportion of gamma/delta T cells in peripheral blood (r = 0.53, p &lt; 0.01), the most significant correlation being found between the prevalence and severity of gastrointestinal symptoms, nausea, anorexia, and vomiting, and the proportion of gamma/delta T cells (r = 0.49, p = 0.002). These findings suggest that gamma/delta T cells have a role to play in the pathogenesis of malaria, possibly in the general constitutional disturbances and particularly in gastrointestinal pathology in malaria.

Efficient migration of dendritic cells toward lymph node chemokines and induction of TH1 responses require maturation stimulus and apoptotic cell interaction

Blood ◽  
2005 ◽  
Vol 106 (5) ◽  
pp. 1734-1741 ◽  
Author(s):  
Nicolas Bertho ◽  
Henri Adamski ◽  
Louis Toujas ◽  
Martine Debove ◽  
Jean Davoust ◽  
...  
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Lymph Node ◽  
Immune Responses ◽  
Tumor Cells ◽  
Apoptotic Cell ◽  
Apoptotic Cells ◽  
Naive T Cells ◽  
Naïve T Cells ◽  
Tnf Α

Abstract Dendritic cells (DCs) have the unique ability to initiate primary immune responses, and they can be conditioned for vaccinal purposes to present antigens after the engulfment of apoptotic cells. To recruit the rare antigen-specific naive T cells, DCs require a maturation step and subsequent transport toward lymph node (LN). To date, prostaglandin E2 (PGE2) is the best-characterized compound inducing this LN-directed migration in vitro, but PGE2 may skew the immune responses in a TH2 direction. We demonstrate here that on incubation with apoptotic tumor cells and tumor necrosis factor-α (TNF-α) or lipopolysaccharide (LPS), human monocyte-derived DCs become fully mature and acquire high migratory capacities toward LN-directing chemokines. The migration of TNF-α-treated DCs occurs only after cotreatment with apoptotic cells but not with necrotic cells. DC migration requires CD36 expression and incubation with apoptotic cells in the presence of heat-labile serum components. Moreover, on treatment with apoptotic cells and LPS, the migrating DCs are able to recruit naive T cells to generate TH1 immune responses. Our results show that the cotreatment of DCs with apoptotic tumor cells and inflammatory signals is promising for the design of an antitumoral DC-based vaccine. (Blood. 2005;106:1734-1741)

scholarly journals Germinal center formation, immunoglobulin class switching, and autoantibody production driven by "non alpha/beta" T cells.

1996 ◽  
Vol 183 (5) ◽  
pp. 2271-2282 ◽  
Author(s):  
L Wen ◽  
W Pao ◽  
F S Wong ◽  
Q Peng ◽  
J Craft ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Lupus Erythematosus ◽  
Cell Receptor ◽  
Gamma Delta T Cells ◽  
Gamma Delta ◽  
Autoantibody Production ◽  
Ige Synthesis ◽  
Alpha Beta ◽  
T Cell Help

The production of class-switched antibodies, particularly immunoglobulin (Ig) G1 and IgE, occurs efficiently in T cell receptor (TCR) alpha-/- mice that are congenitally devoid of alpha/beta T cells. This finding runs counter to a wealth of data indicating that IgG1 and IgE synthesis are largely dependent on the collaboration between B and alpha/beta T cells. Furthermore, many of the antibodies synthesized in TCR alpha-/- mice are reactive to a similar spectrum of self-antigens as that targeted by autoantibodies characterizing human systemic lupus erythematosus (SLE). SLE, too, is most commonly regarded as an alpha/beta T cell-mediated condition. To distinguish whether the development of autoantibodies in TCR alpha-/- mice is due to an intrinsic de-regulation of B cells, or to a heretofore poorly characterized collaboration between B and "non-alpha/beta T" cells, the phenotype has been reconstituted by transfer of various populations of B and non-alpha/beta T cells including cloned gamma/delta T cells derived from TCR alpha-/- mice, to severe combined immunodeficient (SCID) mice. The results establish that the reproducible production of IgG1 (including autoantibodies) is a product of non-alpha/beta T cell help that can be provided by gamma/delta T cells. This type of B-T collaboration sustains the production of germinal centers, lymphoid follicles that ordinarily are anatomical signatures of alpha/beta T-B cell collaboration. Thus, non-alpha/beta T cell help may drive Ig synthesis and autoreactivity under various circumstances, especially in cases of alpha/beta T cell immunodeficiency.

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