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ici 118 551  (MedChemExpress)


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    MedChemExpress ici 118 551
    (A and B) Distinct expression of Adrb2 in neutrophils. (A) The average expression of adrenergic receptors was projected onto the UMAP plot of neutrophil subtypes defined in . (B) Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice 9 weeks after the onset of primary breast cancer. Expression levels of adrenergic receptors were determined by the qPCR analyses. n = 4. (C) The sympathetic neurotransmitter norepinephrine directly promotes the function of immunosuppressive neutrophils. Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice and then in vitro treated with norepinephrine (NE). mRNA levels of Cd274 and Il18 were examined by the qPCR analyses. n = 3, mean ± SEM, * p < 0.05 (Student’s t -test). (D to G) Ablation of sympathetic signals abolishes immunosuppressive neutrophils in lung tumors. (D and E) The wild-type mice were intranasally treated with 6-OHDA or control vehicle [PBS containing 0.01% vitamin C (Vc)] and then inoculated with LLC cancer cells. (D) Representative images of the lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (E) Numbers of immunosuppressive neutrophils in the lung tumors of Vc-treated or 6-OHDA-treated mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (F and G) MMTV-PyMT +/- female mice at 1 week after the onset of primary breast cancer were intranasally treated with 6-OHDA or control vehicle. (F) Representative images of the metastatic lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (G) Numbers of immunosuppressive neutrophils in the metastatic lung tumors of Vc-treated or 6-OHDA-treated MMTV-PyMT +/- mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (H and I) Blockage of the Adrb2 signal potentiates the anti-PD-L1 immunotherapy. MMTV-PyMT +/- female mice at 7 weeks after the onset of primary breast cancer were intravenously injected with the anti-PD-L1 antibody once per week without or with the daily treatment of the Adrb2 antagonist <t>ICI-118,551.</t> The intact lung tissues (left lobes) of the mice at 9 weeks after the onset of primary breast cancer were processed through the whole-tissue PyMT-immunolabeling. (H) Representative 3D-projection images at 1.26× magnification of the lightsheet imaging were shown. (I) Numbers and sizes of the metastatic lung tumors of MMTV-PyMT +/- mice under indicated conditions were counted. 10 mice per condition, * p < 0.05 (two-way ANOVA test).
    Ici 118 551, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Sympathetic Signals Promote Immunosuppressive Neutrophils in Lung Tumors"

    Article Title: Sympathetic Signals Promote Immunosuppressive Neutrophils in Lung Tumors

    Journal: bioRxiv

    doi: 10.1101/2024.02.15.580434

    (A and B) Distinct expression of Adrb2 in neutrophils. (A) The average expression of adrenergic receptors was projected onto the UMAP plot of neutrophil subtypes defined in . (B) Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice 9 weeks after the onset of primary breast cancer. Expression levels of adrenergic receptors were determined by the qPCR analyses. n = 4. (C) The sympathetic neurotransmitter norepinephrine directly promotes the function of immunosuppressive neutrophils. Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice and then in vitro treated with norepinephrine (NE). mRNA levels of Cd274 and Il18 were examined by the qPCR analyses. n = 3, mean ± SEM, * p < 0.05 (Student’s t -test). (D to G) Ablation of sympathetic signals abolishes immunosuppressive neutrophils in lung tumors. (D and E) The wild-type mice were intranasally treated with 6-OHDA or control vehicle [PBS containing 0.01% vitamin C (Vc)] and then inoculated with LLC cancer cells. (D) Representative images of the lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (E) Numbers of immunosuppressive neutrophils in the lung tumors of Vc-treated or 6-OHDA-treated mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (F and G) MMTV-PyMT +/- female mice at 1 week after the onset of primary breast cancer were intranasally treated with 6-OHDA or control vehicle. (F) Representative images of the metastatic lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (G) Numbers of immunosuppressive neutrophils in the metastatic lung tumors of Vc-treated or 6-OHDA-treated MMTV-PyMT +/- mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (H and I) Blockage of the Adrb2 signal potentiates the anti-PD-L1 immunotherapy. MMTV-PyMT +/- female mice at 7 weeks after the onset of primary breast cancer were intravenously injected with the anti-PD-L1 antibody once per week without or with the daily treatment of the Adrb2 antagonist ICI-118,551. The intact lung tissues (left lobes) of the mice at 9 weeks after the onset of primary breast cancer were processed through the whole-tissue PyMT-immunolabeling. (H) Representative 3D-projection images at 1.26× magnification of the lightsheet imaging were shown. (I) Numbers and sizes of the metastatic lung tumors of MMTV-PyMT +/- mice under indicated conditions were counted. 10 mice per condition, * p < 0.05 (two-way ANOVA test).
    Figure Legend Snippet: (A and B) Distinct expression of Adrb2 in neutrophils. (A) The average expression of adrenergic receptors was projected onto the UMAP plot of neutrophil subtypes defined in . (B) Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice 9 weeks after the onset of primary breast cancer. Expression levels of adrenergic receptors were determined by the qPCR analyses. n = 4. (C) The sympathetic neurotransmitter norepinephrine directly promotes the function of immunosuppressive neutrophils. Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice and then in vitro treated with norepinephrine (NE). mRNA levels of Cd274 and Il18 were examined by the qPCR analyses. n = 3, mean ± SEM, * p < 0.05 (Student’s t -test). (D to G) Ablation of sympathetic signals abolishes immunosuppressive neutrophils in lung tumors. (D and E) The wild-type mice were intranasally treated with 6-OHDA or control vehicle [PBS containing 0.01% vitamin C (Vc)] and then inoculated with LLC cancer cells. (D) Representative images of the lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (E) Numbers of immunosuppressive neutrophils in the lung tumors of Vc-treated or 6-OHDA-treated mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (F and G) MMTV-PyMT +/- female mice at 1 week after the onset of primary breast cancer were intranasally treated with 6-OHDA or control vehicle. (F) Representative images of the metastatic lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (G) Numbers of immunosuppressive neutrophils in the metastatic lung tumors of Vc-treated or 6-OHDA-treated MMTV-PyMT +/- mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (H and I) Blockage of the Adrb2 signal potentiates the anti-PD-L1 immunotherapy. MMTV-PyMT +/- female mice at 7 weeks after the onset of primary breast cancer were intravenously injected with the anti-PD-L1 antibody once per week without or with the daily treatment of the Adrb2 antagonist ICI-118,551. The intact lung tissues (left lobes) of the mice at 9 weeks after the onset of primary breast cancer were processed through the whole-tissue PyMT-immunolabeling. (H) Representative 3D-projection images at 1.26× magnification of the lightsheet imaging were shown. (I) Numbers and sizes of the metastatic lung tumors of MMTV-PyMT +/- mice under indicated conditions were counted. 10 mice per condition, * p < 0.05 (two-way ANOVA test).

    Techniques Used: Expressing, In Vitro, Control, Immunostaining, Injection, Immunolabeling, Imaging



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    (A and B) Distinct expression of Adrb2 in neutrophils. (A) The average expression of adrenergic receptors was projected onto the UMAP plot of neutrophil subtypes defined in . (B) Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice 9 weeks after the onset of primary breast cancer. Expression levels of adrenergic receptors were determined by the qPCR analyses. n = 4. (C) The sympathetic neurotransmitter norepinephrine directly promotes the function of immunosuppressive neutrophils. Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice and then in vitro treated with norepinephrine (NE). mRNA levels of Cd274 and Il18 were examined by the qPCR analyses. n = 3, mean ± SEM, * p < 0.05 (Student’s t -test). (D to G) Ablation of sympathetic signals abolishes immunosuppressive neutrophils in lung tumors. (D and E) The wild-type mice were intranasally treated with 6-OHDA or control vehicle [PBS containing 0.01% vitamin C (Vc)] and then inoculated with LLC cancer cells. (D) Representative images of the lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (E) Numbers of immunosuppressive neutrophils in the lung tumors of Vc-treated or 6-OHDA-treated mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (F and G) MMTV-PyMT +/- female mice at 1 week after the onset of primary breast cancer were intranasally treated with 6-OHDA or control vehicle. (F) Representative images of the metastatic lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (G) Numbers of immunosuppressive neutrophils in the metastatic lung tumors of Vc-treated or 6-OHDA-treated MMTV-PyMT +/- mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (H and I) Blockage of the Adrb2 signal potentiates the anti-PD-L1 immunotherapy. MMTV-PyMT +/- female mice at 7 weeks after the onset of primary breast cancer were intravenously injected with the anti-PD-L1 antibody once per week without or with the daily treatment of the Adrb2 antagonist <t>ICI-118,551.</t> The intact lung tissues (left lobes) of the mice at 9 weeks after the onset of primary breast cancer were processed through the whole-tissue PyMT-immunolabeling. (H) Representative 3D-projection images at 1.26× magnification of the lightsheet imaging were shown. (I) Numbers and sizes of the metastatic lung tumors of MMTV-PyMT +/- mice under indicated conditions were counted. 10 mice per condition, * p < 0.05 (two-way ANOVA test).
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    (A and B) Distinct expression of Adrb2 in neutrophils. (A) The average expression of adrenergic receptors was projected onto the UMAP plot of neutrophil subtypes defined in . (B) Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice 9 weeks after the onset of primary breast cancer. Expression levels of adrenergic receptors were determined by the qPCR analyses. n = 4. (C) The sympathetic neurotransmitter norepinephrine directly promotes the function of immunosuppressive neutrophils. Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice and then in vitro treated with norepinephrine (NE). mRNA levels of Cd274 and Il18 were examined by the qPCR analyses. n = 3, mean ± SEM, * p < 0.05 (Student’s t -test). (D to G) Ablation of sympathetic signals abolishes immunosuppressive neutrophils in lung tumors. (D and E) The wild-type mice were intranasally treated with 6-OHDA or control vehicle [PBS containing 0.01% vitamin C (Vc)] and then inoculated with LLC cancer cells. (D) Representative images of the lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (E) Numbers of immunosuppressive neutrophils in the lung tumors of Vc-treated or 6-OHDA-treated mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (F and G) MMTV-PyMT +/- female mice at 1 week after the onset of primary breast cancer were intranasally treated with 6-OHDA or control vehicle. (F) Representative images of the metastatic lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (G) Numbers of immunosuppressive neutrophils in the metastatic lung tumors of Vc-treated or 6-OHDA-treated MMTV-PyMT +/- mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (H and I) Blockage of the Adrb2 signal potentiates the anti-PD-L1 immunotherapy. MMTV-PyMT +/- female mice at 7 weeks after the onset of primary breast cancer were intravenously injected with the anti-PD-L1 antibody once per week without or with the daily treatment of the Adrb2 antagonist <t>ICI-118,551.</t> The intact lung tissues (left lobes) of the mice at 9 weeks after the onset of primary breast cancer were processed through the whole-tissue PyMT-immunolabeling. (H) Representative 3D-projection images at 1.26× magnification of the lightsheet imaging were shown. (I) Numbers and sizes of the metastatic lung tumors of MMTV-PyMT +/- mice under indicated conditions were counted. 10 mice per condition, * p < 0.05 (two-way ANOVA test).
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    (A and B) Distinct expression of Adrb2 in neutrophils. (A) The average expression of adrenergic receptors was projected onto the UMAP plot of neutrophil subtypes defined in . (B) Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice 9 weeks after the onset of primary breast cancer. Expression levels of adrenergic receptors were determined by the qPCR analyses. n = 4. (C) The sympathetic neurotransmitter norepinephrine directly promotes the function of immunosuppressive neutrophils. Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice and then in vitro treated with norepinephrine (NE). mRNA levels of Cd274 and Il18 were examined by the qPCR analyses. n = 3, mean ± SEM, * p < 0.05 (Student’s t -test). (D to G) Ablation of sympathetic signals abolishes immunosuppressive neutrophils in lung tumors. (D and E) The wild-type mice were intranasally treated with 6-OHDA or control vehicle [PBS containing 0.01% vitamin C (Vc)] and then inoculated with LLC cancer cells. (D) Representative images of the lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (E) Numbers of immunosuppressive neutrophils in the lung tumors of Vc-treated or 6-OHDA-treated mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (F and G) MMTV-PyMT +/- female mice at 1 week after the onset of primary breast cancer were intranasally treated with 6-OHDA or control vehicle. (F) Representative images of the metastatic lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (G) Numbers of immunosuppressive neutrophils in the metastatic lung tumors of Vc-treated or 6-OHDA-treated MMTV-PyMT +/- mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (H and I) Blockage of the Adrb2 signal potentiates the anti-PD-L1 immunotherapy. MMTV-PyMT +/- female mice at 7 weeks after the onset of primary breast cancer were intravenously injected with the anti-PD-L1 antibody once per week without or with the daily treatment of the Adrb2 antagonist <t>ICI-118,551.</t> The intact lung tissues (left lobes) of the mice at 9 weeks after the onset of primary breast cancer were processed through the whole-tissue PyMT-immunolabeling. (H) Representative 3D-projection images at 1.26× magnification of the lightsheet imaging were shown. (I) Numbers and sizes of the metastatic lung tumors of MMTV-PyMT +/- mice under indicated conditions were counted. 10 mice per condition, * p < 0.05 (two-way ANOVA test).
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    (A and B) Distinct expression of Adrb2 in neutrophils. (A) The average expression of adrenergic receptors was projected onto the UMAP plot of neutrophil subtypes defined in . (B) Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice 9 weeks after the onset of primary breast cancer. Expression levels of adrenergic receptors were determined by the qPCR analyses. n = 4. (C) The sympathetic neurotransmitter norepinephrine directly promotes the function of immunosuppressive neutrophils. Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice and then in vitro treated with norepinephrine (NE). mRNA levels of Cd274 and Il18 were examined by the qPCR analyses. n = 3, mean ± SEM, * p < 0.05 (Student’s t -test). (D to G) Ablation of sympathetic signals abolishes immunosuppressive neutrophils in lung tumors. (D and E) The wild-type mice were intranasally treated with 6-OHDA or control vehicle [PBS containing 0.01% vitamin C (Vc)] and then inoculated with LLC cancer cells. (D) Representative images of the lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (E) Numbers of immunosuppressive neutrophils in the lung tumors of Vc-treated or 6-OHDA-treated mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (F and G) MMTV-PyMT +/- female mice at 1 week after the onset of primary breast cancer were intranasally treated with 6-OHDA or control vehicle. (F) Representative images of the metastatic lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (G) Numbers of immunosuppressive neutrophils in the metastatic lung tumors of Vc-treated or 6-OHDA-treated MMTV-PyMT +/- mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (H and I) Blockage of the Adrb2 signal potentiates the anti-PD-L1 immunotherapy. MMTV-PyMT +/- female mice at 7 weeks after the onset of primary breast cancer were intravenously injected with the anti-PD-L1 antibody once per week without or with the daily treatment of the Adrb2 antagonist <t>ICI-118,551.</t> The intact lung tissues (left lobes) of the mice at 9 weeks after the onset of primary breast cancer were processed through the whole-tissue PyMT-immunolabeling. (H) Representative 3D-projection images at 1.26× magnification of the lightsheet imaging were shown. (I) Numbers and sizes of the metastatic lung tumors of MMTV-PyMT +/- mice under indicated conditions were counted. 10 mice per condition, * p < 0.05 (two-way ANOVA test).
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    (A and B) Distinct expression of Adrb2 in neutrophils. (A) The average expression of adrenergic receptors was projected onto the UMAP plot of neutrophil subtypes defined in . (B) Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice 9 weeks after the onset of primary breast cancer. Expression levels of adrenergic receptors were determined by the qPCR analyses. n = 4. (C) The sympathetic neurotransmitter norepinephrine directly promotes the function of immunosuppressive neutrophils. Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice and then in vitro treated with norepinephrine (NE). mRNA levels of Cd274 and Il18 were examined by the qPCR analyses. n = 3, mean ± SEM, * p < 0.05 (Student’s t -test). (D to G) Ablation of sympathetic signals abolishes immunosuppressive neutrophils in lung tumors. (D and E) The wild-type mice were intranasally treated with 6-OHDA or control vehicle [PBS containing 0.01% vitamin C (Vc)] and then inoculated with LLC cancer cells. (D) Representative images of the lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (E) Numbers of immunosuppressive neutrophils in the lung tumors of Vc-treated or 6-OHDA-treated mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (F and G) MMTV-PyMT +/- female mice at 1 week after the onset of primary breast cancer were intranasally treated with 6-OHDA or control vehicle. (F) Representative images of the metastatic lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (G) Numbers of immunosuppressive neutrophils in the metastatic lung tumors of Vc-treated or 6-OHDA-treated MMTV-PyMT +/- mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (H and I) Blockage of the Adrb2 signal potentiates the anti-PD-L1 immunotherapy. MMTV-PyMT +/- female mice at 7 weeks after the onset of primary breast cancer were intravenously injected with the anti-PD-L1 antibody once per week without or with the daily treatment of the Adrb2 antagonist <t>ICI-118,551.</t> The intact lung tissues (left lobes) of the mice at 9 weeks after the onset of primary breast cancer were processed through the whole-tissue PyMT-immunolabeling. (H) Representative 3D-projection images at 1.26× magnification of the lightsheet imaging were shown. (I) Numbers and sizes of the metastatic lung tumors of MMTV-PyMT +/- mice under indicated conditions were counted. 10 mice per condition, * p < 0.05 (two-way ANOVA test).
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    (A and B) Distinct expression of Adrb2 in neutrophils. (A) The average expression of adrenergic receptors was projected onto the UMAP plot of neutrophil subtypes defined in . (B) Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice 9 weeks after the onset of primary breast cancer. Expression levels of adrenergic receptors were determined by the qPCR analyses. n = 4. (C) The sympathetic neurotransmitter norepinephrine directly promotes the function of immunosuppressive neutrophils. Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice and then in vitro treated with norepinephrine (NE). mRNA levels of Cd274 and Il18 were examined by the qPCR analyses. n = 3, mean ± SEM, * p < 0.05 (Student’s t -test). (D to G) Ablation of sympathetic signals abolishes immunosuppressive neutrophils in lung tumors. (D and E) The wild-type mice were intranasally treated with 6-OHDA or control vehicle [PBS containing 0.01% vitamin C (Vc)] and then inoculated with LLC cancer cells. (D) Representative images of the lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (E) Numbers of immunosuppressive neutrophils in the lung tumors of Vc-treated or 6-OHDA-treated mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (F and G) MMTV-PyMT +/- female mice at 1 week after the onset of primary breast cancer were intranasally treated with 6-OHDA or control vehicle. (F) Representative images of the metastatic lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (G) Numbers of immunosuppressive neutrophils in the metastatic lung tumors of Vc-treated or 6-OHDA-treated MMTV-PyMT +/- mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (H and I) Blockage of the Adrb2 signal potentiates the anti-PD-L1 immunotherapy. MMTV-PyMT +/- female mice at 7 weeks after the onset of primary breast cancer were intravenously injected with the anti-PD-L1 antibody once per week without or with the daily treatment of the Adrb2 antagonist <t>ICI-118,551.</t> The intact lung tissues (left lobes) of the mice at 9 weeks after the onset of primary breast cancer were processed through the whole-tissue PyMT-immunolabeling. (H) Representative 3D-projection images at 1.26× magnification of the lightsheet imaging were shown. (I) Numbers and sizes of the metastatic lung tumors of MMTV-PyMT +/- mice under indicated conditions were counted. 10 mice per condition, * p < 0.05 (two-way ANOVA test).
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    (A and B) Distinct expression of Adrb2 in neutrophils. (A) The average expression of adrenergic receptors was projected onto the UMAP plot of neutrophil subtypes defined in . (B) Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice 9 weeks after the onset of primary breast cancer. Expression levels of adrenergic receptors were determined by the qPCR analyses. n = 4. (C) The sympathetic neurotransmitter norepinephrine directly promotes the function of immunosuppressive neutrophils. Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice and then in vitro treated with norepinephrine (NE). mRNA levels of Cd274 and Il18 were examined by the qPCR analyses. n = 3, mean ± SEM, * p < 0.05 (Student’s t -test). (D to G) Ablation of sympathetic signals abolishes immunosuppressive neutrophils in lung tumors. (D and E) The wild-type mice were intranasally treated with 6-OHDA or control vehicle [PBS containing 0.01% vitamin C (Vc)] and then inoculated with LLC cancer cells. (D) Representative images of the lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (E) Numbers of immunosuppressive neutrophils in the lung tumors of Vc-treated or 6-OHDA-treated mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (F and G) MMTV-PyMT +/- female mice at 1 week after the onset of primary breast cancer were intranasally treated with 6-OHDA or control vehicle. (F) Representative images of the metastatic lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (G) Numbers of immunosuppressive neutrophils in the metastatic lung tumors of Vc-treated or 6-OHDA-treated MMTV-PyMT +/- mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (H and I) Blockage of the Adrb2 signal potentiates the anti-PD-L1 immunotherapy. MMTV-PyMT +/- female mice at 7 weeks after the onset of primary breast cancer were intravenously injected with the anti-PD-L1 antibody once per week without or with the daily treatment of the Adrb2 antagonist ICI-118,551. The intact lung tissues (left lobes) of the mice at 9 weeks after the onset of primary breast cancer were processed through the whole-tissue PyMT-immunolabeling. (H) Representative 3D-projection images at 1.26× magnification of the lightsheet imaging were shown. (I) Numbers and sizes of the metastatic lung tumors of MMTV-PyMT +/- mice under indicated conditions were counted. 10 mice per condition, * p < 0.05 (two-way ANOVA test).

    Journal: bioRxiv

    Article Title: Sympathetic Signals Promote Immunosuppressive Neutrophils in Lung Tumors

    doi: 10.1101/2024.02.15.580434

    Figure Lengend Snippet: (A and B) Distinct expression of Adrb2 in neutrophils. (A) The average expression of adrenergic receptors was projected onto the UMAP plot of neutrophil subtypes defined in . (B) Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice 9 weeks after the onset of primary breast cancer. Expression levels of adrenergic receptors were determined by the qPCR analyses. n = 4. (C) The sympathetic neurotransmitter norepinephrine directly promotes the function of immunosuppressive neutrophils. Immunosuppressive neutrophils were FACS-sorted from the metastatic lung tumors of MMTV-PyMT +/- mice and then in vitro treated with norepinephrine (NE). mRNA levels of Cd274 and Il18 were examined by the qPCR analyses. n = 3, mean ± SEM, * p < 0.05 (Student’s t -test). (D to G) Ablation of sympathetic signals abolishes immunosuppressive neutrophils in lung tumors. (D and E) The wild-type mice were intranasally treated with 6-OHDA or control vehicle [PBS containing 0.01% vitamin C (Vc)] and then inoculated with LLC cancer cells. (D) Representative images of the lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (E) Numbers of immunosuppressive neutrophils in the lung tumors of Vc-treated or 6-OHDA-treated mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (F and G) MMTV-PyMT +/- female mice at 1 week after the onset of primary breast cancer were intranasally treated with 6-OHDA or control vehicle. (F) Representative images of the metastatic lung tumors processed by conventional tissue sectioning and CD274/Ly6G co-immunostaining were shown. (G) Numbers of immunosuppressive neutrophils in the metastatic lung tumors of Vc-treated or 6-OHDA-treated MMTV-PyMT +/- mice were quantified. n = 10, mean ± SEM, * p < 0.05 (Student’s t -test). (H and I) Blockage of the Adrb2 signal potentiates the anti-PD-L1 immunotherapy. MMTV-PyMT +/- female mice at 7 weeks after the onset of primary breast cancer were intravenously injected with the anti-PD-L1 antibody once per week without or with the daily treatment of the Adrb2 antagonist ICI-118,551. The intact lung tissues (left lobes) of the mice at 9 weeks after the onset of primary breast cancer were processed through the whole-tissue PyMT-immunolabeling. (H) Representative 3D-projection images at 1.26× magnification of the lightsheet imaging were shown. (I) Numbers and sizes of the metastatic lung tumors of MMTV-PyMT +/- mice under indicated conditions were counted. 10 mice per condition, * p < 0.05 (two-way ANOVA test).

    Article Snippet: For the administration of the Adrb2 antagonist, MMTV-PyMT +/- female mice at 7 weeks after the tumor onset were daily treated with ICI-118,551 (MedChemExpress, diluted in sterile PBS at 1 mg/ml) at 5 mg/kg of body weight via intraperitoneal injection.

    Techniques: Expressing, In Vitro, Control, Immunostaining, Injection, Immunolabeling, Imaging