anti arginase 1  (Boster Bio)

Journal: Nature

Article Title: Tumour–brain crosstalk restrains cancer immunity via a sensory–sympathetic axis

doi: 10.1038/s41586-025-10028-8

Figure Lengend Snippet: a-b , Experimental setup, representative IF images, and quantification showing the depletion efficiency of Npy2r + or P2ry1 + VSNs in Npy2r-ires-cre; lsl-DTR mice (n = 4 PBS, n = 6 DT) or P2ry1-ires-cre; lsl-DTR mice (n = 4 PBS, n = 8 DT) that were injected with PBS or DT in the VNG. Percentages of DTR+ neurons in total TUBB3 + cells in the VNG were calculated; p < 0.0001 for both (a) and (b). c , UMAP plots showing Trpv1, Npy2r and P2ry1 expression in lung-innervating VSNs (K1-K3, L1 and A3) from the single-cell RNA-seq dataset described in Zhao et al. 2022 . d , Quantification of anterogradely labelled Trpv1 + or Npy2r + vagal sensory fibers around tumour regions or adjacent healthy alveolar regions ( Trpv1-cre : n = 5 tumour regions, n = 7 healthy regions, p = 0.0205; Npy2r-ires-cre : n = 6 tumour regions, n = 6 healthy regions, p = 0.0122). e , Experimental setup, representative IF images, and quantification showing the depletion efficiency of Trpv1 + VSNs in Trpv1-cre; lsl-DTR mice that were injected with PBS or DT in the VNG (n = 4 PBS, n = 4 DT, p < 0.0001). f , Representative images and quantification of Npy2r (cyan) and Trpv1 (red) by RNA-scope in the VNG from Trpv1-cre, lsl-DTR mice vagally injected with PBS (n = 4) or DT (n = 4). g , Experimental scheme and tumour burden quantification indicated by total lung weight in lsl-DTR (n = 9) or Trpv1-cre;lsl-DTR (n = 8) mice both receiving identical vagal DT treatment. p = 0.0024. h , Flow-cytometric analysis of tumour-bearing lungs from lsl-DTR (n = 7) or Trpv1-cre;lsl-DTR (n = 5) mice both receiving identical vagal DT treatment. Left, percentage of alveolar macrophages (AMs) expressing ARG1, p = 0.0233; middle, percentage of CD8 + T cells co-expressing IFN-γ and TNF-α, p = 0.0449; right, percentage of CD4 + T cells expressing IFN-γ, p = 0.0213. Data are expressed as mean ± s.e.m. Unpaired, two-tailed Student’s T-test ( a , b , d , e , g , h ) was performed.

Article Snippet: After blocking, slides were stained with anti-ARG1 monoclonal antibody (93668, 1:250, Cell Signaling Technology) or anti-CD8a monoclonal antibody (4SM15, 1:250, Invitrogen).

Techniques: Injection, Expressing, Single Cell, RNA Sequencing, RNAscope, Two Tailed Test

Journal: Nature

Article Title: Tumour–brain crosstalk restrains cancer immunity via a sensory–sympathetic axis

doi: 10.1038/s41586-025-10028-8

Figure Lengend Snippet: a , Representative Ce3D images showing enrichment of immune cells in proximity to tumour (outlined by orange dashed line)-innervating nerve fibres ( n = 3 mice). Scale bar: 100 μm (left), 50 μm (right). b , Experimental setup (top), representative H&E-stained lung sections (bottom left) and tumour quantification (bottom right) from Trpv1-cre; LSL-DTR mice injected with PBS ( n = 4) or DT ( n = 4) in the VNG, orthotopically transplanted with KP tumour cells and treated with anti-CD4 plus anti-CD8 antibodies. P = 0.9810. Scale bars, 1 mm. c , Representative flow cytometry plots and quantification of IFNγ + TNF + CD8 T cells and IFNγ + CD4 T cells in tumour-bearing lungs from Trpv1-cre; LSL-DTR mice injected with PBS ( n = 4) or DT ( n = 5) in the VNG (CD8: P = 0.0002; CD4: P = 0.0073). d , Representative immunofluorescence images and quantification of CD8 T cells within grade-matched, size-matched lung tumours from Trpv1-cre; LSL-DTR mice injected with PBS or DT in the VNG. Each dot represents a tumour (PBS: n = 62 tumours from 4 mice, DT: n = 84 tumours from 4 mice, P < 0.0001). Scale bars, 100 μm. e , Representative flow cytometry histogram and quantification of ARG1 in alveolar macrophages (AMs) from Trpv1-cre; LSL-DTR mice injected with PBS ( n = 4) or DT ( n = 5) in the VNG. P = 0.0154. gMFI, geometric mean fluorescence intensity. f , Representative immunofluorescence images and quantification of ARG1 + cells within grade-matched, size-matched lung tumours from Trpv1-cre; LSL-DTR mice injected with PBS or DT in the VNG. Each dot represents a tumour (PBS: n = 62 tumours from 4 mice, DT: n = 84 tumours from 4 mice, P = 0.0087). Scale bars, 100 μm. g , Experimental setup (left), representative H&E-stained lung sections (middle) and tumour quantification (right) from Trpv1-cre; LSL-DTR mice that received VNG PBS (G1, n = 7), VNG PBS plus intratracheal clodronate (G2, n = 7, P = 0.0329 compared to G1) or VNG DT plus intratracheal clodronate (G3, n = 6, P = 0.9994 compared to G2). Results shown are from one experiment, representative of at least two independent experiments. Scale bars, 1 mm. Data are expressed as mean ± s.e.m. Unpaired, two-tailed Student’s t -test ( b – f ) or one-way ANOVA with Tukey’s multiple comparisons ( g ).

Article Snippet: After blocking, slides were stained with anti-ARG1 monoclonal antibody (93668, 1:250, Cell Signaling Technology) or anti-CD8a monoclonal antibody (4SM15, 1:250, Invitrogen).

Techniques: Staining, Injection, Flow Cytometry, Immunofluorescence, Fluorescence, Two Tailed Test

Journal: Nature

Article Title: Tumour–brain crosstalk restrains cancer immunity via a sensory–sympathetic axis

doi: 10.1038/s41586-025-10028-8

Figure Lengend Snippet: a , Quantification of neutrophils, cDC1s, cDC2s, or monocytes in Trpv1-cre; lsl-DTR mice injected with PBS (n = 4) or DT (n = 5) in the VNG. b , Quantification of the absolute numbers of IFNγ + TNFα + CD8 T cells and IFNγ + CD4 T cells in tumour-bearing lungs from Trpv1-cre; lsl-DTR mice injected with PBS (n = 9) or DT (n = 8) in the VNG (CD8: p = 0.0192; CD4: p = 0.0131). c , Quantification of IFNγ + TNFα + CD8 T cells and IFNγ + CD4 T cells in the tumour-draining lymph nodes (LNs) from Trpv1-cre; lsl-DTR mice injected with PBS (n = 4) or DT (n = 5) in the VNG (CD8 p = 0.0010, CD4 p = 0.0970). d-e , Quantification of IFNγ + TNFα + CD8 T cells and IFNγ + CD4 T cells in tumour-bearing lungs (d) or tumour-draining LNs (e) from Npy2r-ires-cre; lsl-DTR mice injected with PBS (n = 8) or DT (n = 10) in the VNG. CD8 p = 0.0357, CD4 p = 0.0878 in d . CD8 p = 0.0001, CD4 p = 0.0008 in e . f , Quantification of IFNγ + TNFα + CD8 T cells and IFNγ + CD4 T cells in the spleens from Trpv1-cre; lsl-DTR mice injected with PBS (n = 4) or DT (n = 5) in the VNG. CD8 p = 0.2305, CD4 p = 0.8389. g , Quantification of IFNγ + TNFα + CD8 T cells and IFNγ + CD4 T cells in the spleens from Npy2r-ires-cre; lsl-DTR mice injected with PBS (n = 8) or DT (n = 10) in the VNG. CD8 p = 0.4003, CD4 p = 0.8316. h , Quantification of surface MHC-II expression in AMs from Trpv1-cre; lsl-DTR mice that received VNG injection of PBS (n = 4) or DT (n = 5). p = 0.0029. i , Quantification of ARG1 and surface MHC-II expression in AMs from Npy2r-ires-cre; lsl-DTR mice that received VNG injection of PBS (n = 4) or DT (n = 5). ARG1 p = 0.0222, MHC-II p = 0.1213. j-k , Quantification of IFNγ + TNFα + CD8 T cells, IFNγ + CD4 T cells, and AM ARG1 expression in tumour-bearing lungs ( j ), or quantification of IFNγ + TNFα + CD8 T cells and IFNγ + CD4 T cells in the tumour-draining lymph nodes ( k ) from P2ry1-ires-cre; lsl-DTR mice that received VNG injection of PBS (n = 4) or DT (n = 5). l-m , Quantification of AM frequency or absolute number ( l ), or neutrophils, cDC1s, cDC2s, or monocytes (m) in tumour-bearing lungs from Trpv1-cre; lsl-DTR mice intratracheally injected with vehicle control (n = 6) or clodronate liposome (n = 11). Left, p < 0.0001; right, p = 0.0002 in l . n , Experimental setup and cDC1 quantification in tumour-bearing lungs from chimeric Trpv1-cre; lsl-DTR mice reconstituted with WT (n = 10) or Batf3 − / − (n = 17) bone marrow (BM) cells. p < 0.0001. o , Quantification of lung tumour burden in chimeric Trpv1-cre; lsl-DTR mice reconstituted with WT BM and injected with PBS (n = 6) or DT (n = 4) in the VNG ( p = 0.1148), or those reconstituted with Batf3 − / − BM and injected with PBS (n = 9) or DT (n = 8) in the VNG ( p = 0.0046). All experiments were independently repeated at least twice with similar results. Data are expressed as mean ± s.e.m. Unpaired, two-tailed Student’s T-test ( a-n ) or one-way ANOVA with Tukey’s multiple comparisons ( o ) was performed.

Article Snippet: After blocking, slides were stained with anti-ARG1 monoclonal antibody (93668, 1:250, Cell Signaling Technology) or anti-CD8a monoclonal antibody (4SM15, 1:250, Invitrogen).

Techniques: Injection, Expressing, Control, Two Tailed Test

Journal: Nature

Article Title: Tumour–brain crosstalk restrains cancer immunity via a sensory–sympathetic axis

doi: 10.1038/s41586-025-10028-8

Figure Lengend Snippet: a-b , Heatmap (a) and violin plots (b) showing the expression of genes encoding candidate neuropeptides in lung-innervating Npy2r + or P2ry1 + VSNs based on the single-cell RNA-seq dataset described in Zhao et al. 2022 . Npy2r + and P2ry1 + VSNs were defined by Npy2r or P2ry1 > 0.1, respectively. P values in b were generated by two-sided Wilcoxon rank-sum test. c-e , Experimental setup and quantification of lung tumour burden by total lung weight (c), quantification of AM ARG1 expression (d), quantification of IFNγ + TNFα + CD8 T cells and IFNγ + CD4 T cells (e) in tumour-bearing lungs from WT mice that were intraperitoneally (i.p.) injected daily with vehicle control (Ctrl, n = 11) or BIBN4096 (n = 7) starting from Day 7 (7 days post tumour inoculation). f-h , Experimental setup and quantification of lung tumour burden by total lung weight (f), quantification of AM ARG1 expression (g), quantification of IFNγ + TNFα + CD8 T cells and IFNγ + CD4 T cells (h) in tumour-bearing lungs from WT mice that were intratracheally (i.t.) treated daily with vehicle control (Ctrl, n = 6) or BIBN4096 (n = 6) starting from Day 7 (7 days post tumour inoculation). i-k , Experimental setup and quantification of lung tumour burden by total lung weight (i), quantification of AM ARG1 expression (j), quantification of IFNγ + TNFα + CD8 T cells and IFNγ + CD4 T cells (k) in tumour-bearing lungs from WT mice that were i.p. injected daily with BIBN4096 (n = 7) or vehicle (Ctrl, n = 7) starting from Day -2 (2 days before tumour inoculation). Data are expressed as mean ± s.e.m. n.s. = not significant ( p > 0.05) by unpaired, two-tailed Student’s T-test ( c-k ).

Article Snippet: After blocking, slides were stained with anti-ARG1 monoclonal antibody (93668, 1:250, Cell Signaling Technology) or anti-CD8a monoclonal antibody (4SM15, 1:250, Invitrogen).

Techniques: Expressing, Single Cell, RNA Sequencing, Generated, Injection, Control, Two Tailed Test

Journal: Nature

Article Title: Tumour–brain crosstalk restrains cancer immunity via a sensory–sympathetic axis

doi: 10.1038/s41586-025-10028-8

Figure Lengend Snippet: a , Experimental setup (left), representative H&E-stained lung sections (middle) and tumour quantification (right) from Trpv1-cre; LSL-DTR mice that received VNG PBS injection (G1, n = 12), VNG DT injection (G2, n = 13) or VNG DT injection plus daily aerosolized salbutamol treatment (G3, n = 12). Lung tumour burden was assessed by tumour/tissue area (G1 versus G2: P = 0.0133; G2 versus G3: P = 0.0131) and total lung mass (G1 versus G2: P = 0.0101; G2 versus G3: P = 0.0005). Scale bars, 1 mm. b , Representative H&E-stained lung sections and tumour quantification from wild-type (WT, n = 7) and Adrb2 − / − mice ( n = 7). Lung tumour burden was assessed as tumour/tissue area ( P = 0.0398) and total lung mass ( P < 0.0001). Scale bars, 1 mm. c , d , Quantification of ARG1 + alveolar macrophages ( c ), IFNγ + TNF + CD8 T cells and TNF + CD4 T cells ( d ) in tumour-bearing lungs from wild-type ( n = 7) and Adrb2 − / − mice ( n = 5–6). c , P = 0.0004. d , CD8: P = 0.0037; CD4: P = 0.0213. e , Experimental setup (left), representative H&E-stained lung sections (middle) and tumour quantification (right) from irradiated Trpv1-cre; LSL-DTR mice that were reconstituted with wild-type or Adrb2 − / − bone marrow (BM) and injected with PBS or DT in the VNG. WT BM: n = 8 PBS (G1), n = 8 DT (G2); Adrb2 − / − BM: n = 10 PBS (G3), n = 10 DT (G4); pooled from 2 independent experiments. Scale bars, 1 mm. 5.1× and 1.3× denote fold change of tumour/tissue area between indicated groups. f , g , Quantification of ARG1 + alveolar macrophages ( f ), IFNγ + TNF + CD8 T cells and TNF + CD4 T cells ( g ) in tumour-bearing lungs from bone marrow chimeric mice described in e . h , i , Arg1 expression in cultured alveolar macrophages measured by quantitative PCR with reverse transcription (RT–qPCR). h , Wild-type alveolar macrophages were untreated or treated with 10 μM noradrenaline (NA) ( n = 4 per group, P < 0.0001). i , Wild-type or Adrb2 − / − alveolar macrophages were pretreated with 10 μM noradrenaline and then either left untreated ( n = 4 per group, P = 0.0001) or treated with TES ( n = 4 per group , P = 0.0073). Data are presented as fold change relative to untreated wild-type alveolar macrophages ( h , i ). Results shown are representative of at least two independent experiments. Data are expressed as mean ± s.e.m. Unpaired, two-tailed Student’s t -test ( b – d , h ) or one-way ANOVA with Tukey’s multiple comparisons ( a , e – g , i ). Exact P values for e – g are provided in the .

Article Snippet: After blocking, slides were stained with anti-ARG1 monoclonal antibody (93668, 1:250, Cell Signaling Technology) or anti-CD8a monoclonal antibody (4SM15, 1:250, Invitrogen).

Techniques: Staining, Injection, Irradiation, Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Reverse Transcription, Quantitative RT-PCR, Two Tailed Test

Journal: Nature

Article Title: Tumour–brain crosstalk restrains cancer immunity via a sensory–sympathetic axis

doi: 10.1038/s41586-025-10028-8

Figure Lengend Snippet: a , Quantification of the number of KP tumour cells that were untreated (Ctrl) or treated with 10 μM norepinephrine (NE) for three days (triplicates for each condition). b , Experimental setup, representative H&E-stained lung sections, and quantification of lung tumour burden from Trpv1-cre; lsl-DTR ; Adrβ2 − / − mice injected with PBS (n = 9) or DT (n = 6) in the VNG. c , Quantification of ARG1 + AMs, IFNγ + TNFα + CD8 T cells, and TNFα + CD4 T cells in tumour-bearing lungs from the same mice as b. d-e , Quantification of ARG1 and surface MHC-II expression in AMs ( d ), and quantification of IFNγ + TNFα + CD8 T cells and TNFα + CD4 T cells ( e ) in tumour-bearing lungs from Trpv1-cre; lsl-DTR mice that received VNG PBS injection (n = 12, G1), VNG DT injection (n = 13, G2), or VNG DT injection plus daily aerosolized salbutamol treatment (n = 12, G3). d , ARG1: p = 0.1276 (G1 vs G2), p = 0.2619 (G2 vs G3); MHC-II: p < 0.0001 (G1 vs G2, G2 vs G3). e , CD8: p = 0.0797 (G1 vs G2), p = 0.0276 (G2 vs G3); CD4: p = 0.0456 (G1 vs G2), p = 0.0271 (G2 vs G3). f , Expressions of genes encoding adrenergic receptors in AMs sorted from healthy (n = 3) or tumour-bearing (n = 6) lungs as determined by bulk RNA-sequencing. g , Adrβ1 and Adrβ2 expression in AMs and CD8 T cells sorted from tumour-bearing lungs (n = 2) as determined by RT-qPCR. p = 0.0015. h , Tumour cell viability from in vitro T cell-killing assay. OT-I CD8 + T cells were co-cultured with OVA-expressing KP tumour cells in the presence or absence of norepinephrine (NE, 10 µM). Percentages of live tumour cells were determined by flow cytometry 24 h after co-culture. i , Experimental scheme, tumour quantification by total lung weight and representative H&E lung sections of WT or Adrβ2 − / − mice that were inoculated with KP tumour cells and intratracheally treated with PBS or clodronate liposomes. WT + PBS (n = 6, G1), Adrβ2 − / − + PBS (n = 7, G2), WT + clodronate (n = 6, G3), Adrβ2 − / − + clodronate (n = 8, G4). G1 vs G2: p = 0.0047, G3 vs G4: p = 0.5043, G1 vs G3: p = 0.0587, G2 vs G4: p = 0.9799. j , Quantification of IFNγ + TNFα + CD8 T cells and TNFα + CD4 T cells in tumour-bearing lungs from the same cohort in i . CD8: p = 0.0361 (G1 vs G2), p = 0.2254 (G3 vs G4), p = 0.0357 (G1 vs G3), p = 0.1563 (G2 vs G4); CD4: p = 0.0053 (G1 vs G2), p = 0.9741 (G3 vs G4), p = 0.1673 (G1 vs G3), p = 0.6366 (G2 vs G4). k , Quantification of IFNγ + TNFα + CD8 T cells and TNFα + CD4 T cells in tumour-bearing lungs from Trpv1-cre; lsl-DTR mice that were injected with PBS or DT in the VNG (n = 6 PBS, n = 5 DT), and intratracheally treated with clodronate liposome. l , Experimental setup of the mixed bone marrow chimera experiment in which irradiated Npy2r-ires-cre; lsl-DTR mice were reconstituted with Adrβ2 − / − and CD11c-cre; Pparg fl/fl bone marrow mixed at 1:1 ratio and injected with PBS or DT in the VNG (n = 12 PBS, n = 11 DT). CD11c-cre; Pparg fl/fl mice are deficient in AMs . m-o , Quantification of AM ARG1 and surface MHC-II expression ( m ), IFNγ + TNFα + CD8 T cells and TNFα + CD4 T cells ( n ) in the tumour-bearing lungs, and quantification of lung tumour burden ( o , n = 6 PBS, n = 7 DT) from the same chimeric cohort in l . p , Representative flow cytometry plots showing the gating strategy for exhausted CD8 T cells (Tex, defined as PD1 + TIM3 + TCF1 − CD44 + CD8 T cells). q-t , Quantification of Tex populations in tumour-bearing lungs from Npy2r-ires-cre; lsl-DTR mice injected with PBS or DT in the VNG ( q , n = 4 PBS, n = 6 DT), from Trpv1-cre; lsl-DTR mice that were injected with PBS or DT in the VNG ( r , n = 4 PBS, n = 5 DT), from WT ( s , n = 7) versus Adrβ2 − / − mice ( s , n = 6), and from vagal DT-injected Trpv1-cre; lsl-DTR mice that were either left untreated or treated daily with aerosolized salbutamol ( t , n = 13 DT, n = 12 DT+ salbutamol). Data are expressed as mean ± s.e.m. and representative of at least two independent experiments. Unpaired, two-tailed Student’s T-test ( a-c, g, k, m-o, q-t ) or one-way ANOVA with Tukey’s multiple comparisons ( d-e, h-j ) was performed.

Article Snippet: After blocking, slides were stained with anti-ARG1 monoclonal antibody (93668, 1:250, Cell Signaling Technology) or anti-CD8a monoclonal antibody (4SM15, 1:250, Invitrogen).

Techniques: Staining, Injection, Expressing, RNA Sequencing, Quantitative RT-PCR, In Vitro, Cell Culture, Flow Cytometry, Co-Culture Assay, Liposomes, Irradiation, Two Tailed Test

Journal: Neuroscience

Article Title: Nrdp1-mediated Macrophage Phenotypic Regulation Promotes Functional Recovery in Mice with Mild Neurological Impairment after Intracerebral Hemorrhage.

doi: 10.1016/j.neuroscience.2024.02.028

Figure Lengend Snippet: Fig. 5. Nrdp1 overexpression promote Arg1, C/EBP-b, and IL-10 expression while inhibiting iNOS, IL-6, and TNF-a expression. (A-C) Representative western blotting images and quantitative analyses of Nrdp1(A), Arg1(B), iNOS(C) at different time points in S/ICH and Ad-N/ICH groups after ICH by western blot. (D-F) Representative western blotting images and quantitative analyses of Nrdp1(D), Arg1(E), iNOS(F) compared with baseline levels in S/ICH and Ad-N/ICH groups after ICH by western blot. (G) Representative western blotting images and quantitative analyses of C/EBP-b at different time points in S/ICH and Ad-N/ICH groups after ICH by western blot. (H-I) The correlation among the expression levels curves of Nrdp1, C/EBP-b, and Arg1 was approximately positive. (J-L) The expression levels of IL-6(J), IL-10(K), and TNF-a(L) at different time points after ICH by elisa. (Brain tissue samples were cubes with a radius of 1.5 mm centered on the blood injection site; N = 4 per experimental group per time point; ns: p > 0.05; *: p < 0.05; **: p < 0.01; ***: p < 0.001).

Article Snippet: The primary antibodies used were rabbit polyclonal antimouse iNOS (1:100, ab15323, Abcam, Cambridge, UK); rabbit monoclonal anti-mouse Arg1 (1:200, 93668S, Cell Signaling Technology, Boston, USA); rabbit monoclonal anti-mouse Iba1 (1:2000, ab178846, Abcam, Cambridge, UK);The secondary antibodies used were: HRP-conjugated goat anti-rabbit (1:1000, ab6721, Abcam, Cambridge, UK).

Techniques: Over Expression, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Injection