Dataset Information

Natural killer T cell activation increases iNOS⁺CD206^- M1 macrophage and controls the growth of solid tumor.

ABSTRACT:

Background

NKT cells play an important role in anti-tumor immunity. Alpha-galactosylceramide (?-GalCer), a synthetic glycolipid is presented to natural killer T (NKT) cells by most antigen-presenting cells through CD1d molecules leading to activation of NKT cells. However, the precise mechanisms of how ?-GalCer-activated NKT regulate the polarization of the macrophages and effector T cells in the solid tumor are not studied adequately.

Methods

We induced solid tumor in C57BL/6 mice by subcutaneous injection of B16F10 cell line (1 X 10⁶ cells) and monitored the tumor growth. Animals were given an intraperitoneal injection of ?-GalCer (2??g/injection) in 200??l PBS on day +?1, +?5, +?10, +?15, and?+?20 (with respect to tumor cell injection). Immune cells were characterized using flow cytometry and immunofluorescence staining. NK cells, Gr1⁺ cells, and F4/80⁺ macrophages in the mice were depleted by intravenous injection of cell-specific antibodies. Statistical analysis was performed using Student's t-test or one-way ANOVA.

Results

Our results showed that intratumoral NKT cells have a lower frequency of CD69, CD25, CD122, and IFN-?R expression; produced less inflammatory cytokines such as IFN-?, TNF-?, and GM-CSF; higher frequency CD62L⁺ NKT cells; and also showed reduced proliferation as compared to the splenic NKT cells. Mice treated with ?-GalCer showed a significantly increased frequency of IFN-?-producing NKT cells, CD8⁺ T cells, and effector Th1 cells. Depletion of NK cells in ?-GalCer-treated mice showed a lower frequency of IFN-?-producing CD4⁺ and CD8⁺ T cells in the tumor and prevented the ?-GalCer-induced tumor growth. NKT cell activation with ?-GalCer treatment significantly increased the iNOS⁺CD206^- M1-macrophages and reduced the iNOS^-CD206⁺ M2-macrophages in the spleen and tumor, and depletion of F4/80⁺ macrophages prevented the ?-GalCer-induced reduction in the tumor growth.

Conclusions

We showed that activation of NKT cell with ?-GalCer modulates the frequency of M1-macrophages and effector Th1 cells in the secondary lymphoid tissues and tumor microenvironment and inhibit tumor growth. The finding suggests that activation of NKT cells with ?-GalCer may provide an effective anti-cancer outcome.

SUBMITTER: Paul S

PROVIDER: S-EPMC6685184 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature

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Publications

Natural killer T cell activation increases iNOS<sup>+</sup>CD206<sup>-</sup> M1 macrophage and controls the growth of solid tumor.

Paul Sourav S Chhatar Sushanta S Mishra Amrita A Lal Girdhari G

Journal for immunotherapy of cancer 20190806 1

<h4>Background</h4>NKT cells play an important role in anti-tumor immunity. Alpha-galactosylceramide (α-GalCer), a synthetic glycolipid is presented to natural killer T (NKT) cells by most antigen-presenting cells through CD1d molecules leading to activation of NKT cells. However, the precise mechanisms of how α-GalCer-activated NKT regulate the polarization of the macrophages and effector T cells in the solid tumor are not studied adequately.<h4>Methods</h4>We induced solid tumor in C57BL/6 mic ...[more]

PMID: 31387637

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Project description:Macrophages play an important role throughout the body. Antiinflammatory macrophages expressing the macrophage mannose receptor (MMR, CD206) are involved in disease development, ranging from oncology to atherosclerosis and rheumatoid arthritis. [68Ga]Ga-NOTA-anti-CD206 single-domain antibody (sdAb) is a PET tracer targeting CD206. This first-in-human study, as its primary objective, evaluated the safety, biodistribution, and dosimetry of this tracer. The secondary objective was to assess its tumor uptake. Methods: Seven patients with a solid tumor of at least 10 mm, an Eastern Cooperative Oncology Group score of 0 or 1, and good renal and hepatic function were included. Safety was evaluated using clinical examination and blood sampling before and after injection. For biodistribution and dosimetry, PET/CT was performed at 11, 90, and 150 min after injection; organs showing tracer uptake were delineated, and dosimetry was evaluated. Blood samples were obtained at selected time points for blood clearance. Metabolites in blood and urine were assessed. Results: Seven patients were injected with, on average, 191 MBq of [68Ga]Ga-NOTA-anti-CD206-sdAb. Only 1 transient adverse event of mild severity was considered to be possibly, although unlikely, related to the study drug (headache, Common Terminology Criteria for Adverse Events grade 1). The blood clearance was fast, with less than 20% of the injected activity remaining after 80 min. There was uptake in the liver, kidneys, spleen, adrenals, and red bone marrow. The average effective dose from the radiopharmaceutical was 4.2 mSv for males and 5.2 mSv for females. No metabolites were detected. Preliminary data of tumor uptake in cancer lesions showed higher uptake in the 3 patients who subsequently progressed than in the 3 patients without progression. One patient could not be evaluated because of technical failure. Conclusion: [68Ga]Ga-NOTA-anti-CD206-sdAb is safe and well tolerated. It shows rapid blood clearance and renal excretion, enabling high contrast-to-noise imaging at 90 min after injection. The radiation dose is comparable to that of routinely used PET tracers. These findings and the preliminary results in cancer patients warrant further investigation of this tracer in phase II clinical trials.

Project description:BackgroundThe anti-tumor immunity of natural killer (NK) cells can be paralyzed by the CD73-induced generation of immunosuppressive adenosine from precursor ATP within the hypoxic microenvironment of solid tumors. In an effort to redirect purinergic immunosuppression of NK cell anti-tumor function, we showed, for the first time, that immunometabolic combination treatment with NKG2D-engineered CAR-NK cells alongside blockade of CD73 ectonucleotidase activity can result in significant anti-tumor responses in vivo.MethodsNK cells were engineered non-virally with NKG2D.CAR-presenting vectors based on the piggyBac transposon system with DAP10 and CD3ζ co-signaling domains. The anti-tumor immunity of NKG2D.CAR.NK cells in combination with CD73 targeting was evaluated against multiple solid tumor targets in vitro and humanized mouse xenografts in immunodeficient tumor-bearing mice in vivo. Intratumoral migration was evaluated via immunohistochemical staining, while degranulation capacity and IFN-γ production of NK cells were measured in response to solid tumor targets.ResultsOur results showed that CD73 blockade can mediate effective purinergic reprogramming and enhance anti-tumor cytotoxicity both in vitro and in vivo by enhancing the killing ability of CAR-engineered NK cells against CD73+ solid tumor targets via mechanisms that might imply alleviation from adenosinergic immunometabolic suppression. CD73 blockade improved the intratumoral homing of CD56+ CAR-NK cells in vivo. These engineered NK cells showed synergistic therapeutic efficacy in combination with CD73 targeting against CD73+ human lung cancer xenograft models. Interestingly, CD73 blockade could inhibit tumor growth in vivo independently of adaptive immune cells, innate immunity or NK cell-mediated ADCC.ConclusionsImmunotherapies targeting the adenosinergic signaling cascade, which act by neutralizing CD73 ectoenzymatic activity, had thus far not been evaluated in humanized tumor models, nor had the implication of innate immunity been investigated. Taken together, our pre-clinical efficacy data demonstrate, for the first time, the potential of targeting CD73 to modulate purinergic signaling and enhance adoptive NK cell immunotherapy via mechanisms that could implicate autocrine tumor control as well as by mediating adenosinergic signaling.

Dataset Information

Natural killer T cell activation increases iNOS⁺CD206^- M1 macrophage and controls the growth of solid tumor.

Background

Methods

Results

Conclusions

Publications

Natural killer T cell activation increases iNOS<sup>+</sup>CD206<sup>-</sup> M1 macrophage and controls the growth of solid tumor.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

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Dataset Information

Natural killer T cell activation increases iNOS+CD206- M1 macrophage and controls the growth of solid tumor.

Background

Methods

Results

Conclusions

Publications

Natural killer T cell activation increases iNOS<sup>+</sup>CD206<sup>-</sup> M1 macrophage and controls the growth of solid tumor.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Natural killer T cell activation increases iNOS⁺CD206^- M1 macrophage and controls the growth of solid tumor.