rabbit anti mouse primary cd8  (Cell Signaling Technology Inc)

Journal: Pharmaceuticals

Article Title: AHR Agonist ITE Boosted PD1 Antibody’s Effects by Inhibiting Myeloid-Derived Cells Suppressive Cells in an Orthotopic Mouse Glioma Model

doi: 10.3390/ph18040471

Figure Lengend Snippet: Synergistic effect of ITE and PD1 antibody on cytotoxic immune cell infiltration. ( A,B ) Gating Strategy for myeloid cells. R2 represents single cells, R3 represents live cells, R4 represents CD45+ immune cells, R5 represents non-neutrophils, and R6 represents myeloid cells. R7 was composed of CD11c+ lymphocytes, NK cells, and monocytes. ( C – G ) Effects of DMSO, ITE, PD1 antibody, and ITE+PD1 antibody treatments on immune cells in brain tumors and spleen. ( C ) Percentage of CD45+ cells in brain tumors and spleen (* p < 0.05). ( D ) Percentage of CD4-CD8+T cells in brain tumors and spleen (* p < 0.05). ( E ) Immunofluorescence and immunohistochemical staining of frozen brain tumor sections to detect CD8 T-cell antigens; blue: cell nucleus, red: CD8. ( F ) Detection of IFN-γ in brain by ELISA. ( G ) Percentage of CD4+T cells in brain tumors and spleen (** p < 0.05). ( H ) Immunofluorescence and immunohistochemical staining of frozen brain tumor sections to detect CD4 T-cell antigens; blue: cell nucleus, green: CD3, red: CD4. ( I ) Percentage of CD4+CD8+T cells in brain tumors (* p < 0.05, ** p < 0.01). ( J , K ) Th17/Treg cells in brain tumor.

Article Snippet: The slices were treated in 3% hydrogen peroxide for 15 min at room temperature, washed with PBS for three times, incubated with normal goat serum at room temperature, and then incubated with rabbit anti-mouse CD4 antibody and rabbit anti-mouse CD8 primary (Bioss, Beijing, China; 1:100, 4 °C overnight) antibodies.

Techniques: Immunofluorescence, Immunohistochemical staining, Staining, Enzyme-linked Immunosorbent Assay

Journal: Pharmaceuticals

Article Title: AHR Agonist ITE Boosted PD1 Antibody’s Effects by Inhibiting Myeloid-Derived Cells Suppressive Cells in an Orthotopic Mouse Glioma Model

doi: 10.3390/ph18040471

Figure Lengend Snippet: Primary antibody list.

Article Snippet: The slices were treated in 3% hydrogen peroxide for 15 min at room temperature, washed with PBS for three times, incubated with normal goat serum at room temperature, and then incubated with rabbit anti-mouse CD4 antibody and rabbit anti-mouse CD8 primary (Bioss, Beijing, China; 1:100, 4 °C overnight) antibodies.

Techniques:

Journal: NPJ Breast Cancer

Article Title: Agonistic CD40 elicits CD8+ T-cell-dependent primary responses and CD4+ T-cell-dependent long-term immunity in breast cancer

doi: 10.1038/s41523-025-00889-7

Figure Lengend Snippet: A Proportions of CD3+, CD8 +, and CD4 + T cells by flow cytometry in control and aCD40 treated Brpkp110 tumors on day 13 post treatment ( n = 10). B FoxP3 + CD4+ regulatory T cells on day 7 post-treatment ( n = 7). C Proportions of Granzyme B + T cells in subpopulations by flow cytometry in control and aCD40 treated Brpkp110 tumors on day 7 post implantation ( n = 10). D Images of immunofluorescent staining for CD8 (red) and nuclei (blue) and CD8 staining quantification in untreated (control) and aCD40 treated Brpkp110 tumors on day 7 post treatment ( n = 4–6). E Quantification of CD8 immunofluorescent staining in outer, middle, and inner thirds of control and aCD40 treated tumors on day 7 post treatment ( n = 5–6). F After 7 days of treatment with aCD40, Brpkp110 tumors were minced and cultured ex vivo. Supernatant was collected and pooled for each treatment group after 48 h and cytokines were measured ( n = 2, with 3 tumors pooled per group). G T cell activation and proliferation markers measured by flow cytometry in CD4+ and CD8 + T cell populations of control and aCD40 treated Brpkp110 TDLN on day 7 post-treatment ( n = 10). H Growth curves of control and aCD40 treated Brpkp110 tumors implanted into WT hosts with or without T cell depletions ( n = 17–20). I Tumor growth curve of aCD40 (treatment on Day 8) +/− FTY720 (treatment started on Day 7) treated Brpkp110 tumors ( n = 14–16). Data: ( A – C , D (right), E – G ) median, ( H , I ) mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Tissue sections were blocked with 5% donkey serum diluted in PBS with 0.3% Triton X-100 and stained with rabbit anti-mouse primary CD8 (clone: D4W2Z, Cell Signaling Technology 98941, 1:100) followed by donkey anti-rabbit Alexa Fluor 594 (Thermo Scientific A-21207, 1:250) secondary antibody.

Techniques: Flow Cytometry, Control, Staining, Cell Culture, Ex Vivo, Activation Assay

Journal: NPJ Breast Cancer

Article Title: Agonistic CD40 elicits CD8+ T-cell-dependent primary responses and CD4+ T-cell-dependent long-term immunity in breast cancer

doi: 10.1038/s41523-025-00889-7

Figure Lengend Snippet: A Tumor growth curves (left) and tumor volume changes compared to pretreatment on day 26 post implantation (right) of Brpkp110 tumors. Indicated treatments initiated on day 7 post implantation ( n = 18–20). B Tumor growth curves (left) and tumor volume changes compared to pretreatment on day 26 post implantation (right) of E0771 tumors. Indicated treatments initiated on day 8 post implantation ( n = 12–15). C Tumor growth curves (left) and tumor volume changes compared to pretreatment on day 31 post implantation (right) of AT3 tumors. Indicated treatments initiated on day 9 post implantation ( n = 14). D Tumor growth curves (left) and tumor volume changes compared to pretreatment on day 34 post implantation (right) of EpH4 1424 tumors. Indicated treatments initiated on day 5 post implantation ( n = 14–16). E Brpkp110 tumor growth curves (left) and volume changes compared to pretreatment (right) on day 27 post implantation in control and aCD40+ICB treated hosts with and without CD8 + T cell depletions. Indicated treatments initiated on day 7 post implantation ( n = 12–15). F Brpkp110 tumor growth curves (left) and volume changes compared to pretreatment (right) on day 25 post implantation in control and aCD40+ICB treated hosts with and without CD4 + T cell depletions. Indicated treatments initiated on day 8 post implantation ( n = 14–18). G Brpkp110 tumor growth curves (left) and volume changes compared to pretreatment (right) on day 27 post implantation in control and aCD40+ICB treated hosts with and without CD4+ and CD8 + T cell depletions. Indicated treatments initiated on day 7 post implantation ( n = 12–18). H Tumor growth curves (left) and volume changes compared to pretreatment (right) on day 26 post Brpkp110 tumor implantation into WT and BATF3 KO hosts. Indicated treatments initiated on day 7 post implantation ( n = 14–18). Data: ( A – H left) mean ± SEM, ( A – H right) each column represents individual tumor. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Tissue sections were blocked with 5% donkey serum diluted in PBS with 0.3% Triton X-100 and stained with rabbit anti-mouse primary CD8 (clone: D4W2Z, Cell Signaling Technology 98941, 1:100) followed by donkey anti-rabbit Alexa Fluor 594 (Thermo Scientific A-21207, 1:250) secondary antibody.

Techniques: Control, Tumor Implantation

Journal: NPJ Breast Cancer

Article Title: Agonistic CD40 elicits CD8+ T-cell-dependent primary responses and CD4+ T-cell-dependent long-term immunity in breast cancer

doi: 10.1038/s41523-025-00889-7

Figure Lengend Snippet: A Proportions of circulating effector memory (CD44 + CD62L-), central memory (CD44 + CD62L+), and naïve (CD44-CD62L-) CD4+ (left) and CD8+ (right) in blood, 3 months post treatment induced tumor clearance ( n = 5–6, data representative of 2 experiments with similar results). B Secondary Brpkp110 tumor rechallenge of naïve and previously Brpkp110 tumor-bearing mice cured after aCD40 + ICB, at least 2 months post primary tumor clearance ( n = 12–14, data representative of 3 experiments with similar results). C Control and rechallenge tumor growth in T cell sufficient ( n = 6–12) and T cell depleted hosts ( n = 12–14, data representative of 2 experiments with similar results). D Brpkp110 tumor growth curves in intra-tumoral (IT) vehicle (control) and IT aCD40 treated hosts. aCD40 administered tumors denoted as aCD40 IT and contralateral untreated tumors denoted as CD40 IT Distant ( n = 9–12, data representative of 2 experiments with similar results). E Brpkp110 tumor growth curves in intra-tumoral (IT) vehicle (control) and IT or intraperitoneal (IP) aCD40 or ICB received hosts ( n = 4–9, data representative of 2 experiments with similar results). Data: ( A ) median, ( C – E ) mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Tissue sections were blocked with 5% donkey serum diluted in PBS with 0.3% Triton X-100 and stained with rabbit anti-mouse primary CD8 (clone: D4W2Z, Cell Signaling Technology 98941, 1:100) followed by donkey anti-rabbit Alexa Fluor 594 (Thermo Scientific A-21207, 1:250) secondary antibody.

Techniques: Control

Journal: Nature Neuroscience

Article Title: T cell-mediated microglial activation triggers myelin pathology in a mouse model of amyloidosis

doi: 10.1038/s41593-024-01682-8

Figure Lengend Snippet: a , Representative image showing CD8 + T-cells (green) in the cortex and corpus callosum of 5xFAD mice aged 10 months. Plaques are stained with Thiazine Red (ThR, red). Scale bar, 10 µm. Quantifications indicate the number of CD8 + T cells. Statistical significance was determined for n = 3 animals using unpaired two-sided Student’s t-test (GM, WT 10 m, 5xFAD 10 m, *p = 0.017; WM, WT 4 m, 5XFAD 4 m, *p = 0.011; WM, WT 10 m, 5xFAD 10 m, ***p = 0.0006). b , Representative image showing STAT1 + (grey) CC1 + oligodendrocytes (green) in the cortex (upper panel) and corpus callosum (lower panel) of WT and 5xFAD mice aged 4 and10 months. Plaques are stained with Thiazine Red (ThR, red). Scale bar, 10 µm. Quantifications indicate the percentage of CC1 + oligodendrocytes also positive for STAT1. Statistical significance was determined for n = 3-4 animals using unpaired two-sided Student’s t-test (GM, WT 10 m, 5xFAD 10 m, *p = 0.037; WM, WT 10 m, 5xFAD 10 m, *p = 0.040). c , Representative image showing Serpina3n + (grey) CC1 + oligodendrocytes (green) in the cortex (upper panel) and corpus callosum (lower panel) of WT and 5xFAD mice aged 4 and10 months. Plaques are stained with Thiazine Red (ThR, red). Scale bar 10 µm. Quantifications indicate the percentage of CC1 + oligodendrocytes also positive for Serpina3n. Statistical significance was determined for n = 3-4 animals using unpaired two-sided Student’s t-test (GM, WT 10 m, 5xFAD 10 m, *p = 0.029; WM, WT 4 m, 5XFAD 4 m, *p = 0.028; WM, WT 10 m, 5xFAD 10 m, *p = 0.001). Each point on the graph represents one animal. Data is presented as mean ± SEM.

Article Snippet: The primary mouse anti-CD8 antibody (1:50 dilution, clone C8/144B, catalog no. M7103, Dako) was incubated overnight and developed with 3,3′-diaminobenzidine (Dako REAL EnVision, catalog no. K500711-2, Dako) until saturation of the chromogenic reaction.

Techniques: Staining

Journal: Nature Neuroscience

Article Title: T cell-mediated microglial activation triggers myelin pathology in a mouse model of amyloidosis

doi: 10.1038/s41593-024-01682-8

Figure Lengend Snippet: a , SEM images showing myelin abnormalities (green) in WT and 5xFAD mice aged 10 months. The asterisk shows an axonal swelling with thin myelin. b , c , Images ( b ) and quantification ( c ) of different myelin abnormalities in 5xFAD brain sections. Unpaired two-sided Student’s t -test for n = 3–4 animals (GM, WT 10 months, 5xFAD 10 months, * P = 0.018; WM, WT 4 months, 5xFAD 4 months, * P = 0.041; WM, WT 10 months, 5xFAD 10 months, ** P = 0.002). d , Image of BCAS1 + (green) swellings (shown with white arrowheads) lacking a nucleus (stained with DAPI) in 5xFAD mice aged 10 months. Thiazine red (ThR) indicates plaques. Quantifications of BCAS1 + swellings in 4-month old and 10-month old WT and 5xFAD mice. Unpaired two-sided Student’s t -test for n = 3 animals (WT 10 m, 5xFAD 10 m, * P = 0.019). e , Image and quantification showing the percentage of EdU + (gray) OLIG2 + (green) cells in the cortex of control or CD8 + T cell-depleted 5xFAD mice. ThR (red) indicates plaques; unpaired two-sided Student’s t -test for n = 5 animals (control, anti-CD8, * P = 0.027). f , SEM images and quantification of myelinated axons with myelin abnormalities (green) in the cortex of control or CD8 + T cell-depleted 5xFAD mice; unpaired Student’s t -test for n = 3 animals (control, anti-CD8, ** P = 0.002). g , Image and quantification showing the percentage of Serpina3n + (gray) CC1 + (green) oligodendrocytes in the cortex of control or CD8 + T cell-depleted 5xFAD mice. ThR (red) indicates plaques. Unpaired two-sided Student’s t -test for n = 4 animals (control, anti-CD8, * P = 0.049). h , Behavior of control or CD8 + T cell-depleted WT and 5xFAD mice in the Barnes maze. Left: Tracing graphs in platform trials in the Barnes maze. Right: Quantification of the target quadrant time (%) on the platform during the training sessions and relative escape latency to the target hole in the probe trial. One-way analysis of variance (ANOVA) with Tukey’s post hoc analysis for n = 9–10 animals (WT control, 5xFAD control, *** P = 0.0000000069; 5xFAD control, 5xFAD anti-CD8, ** P = 0.0019). Each point on the graph represents one animal. Data are presented as the mean ± s.e.m. a , f , Scale bar, 1 µm. b , Scale bar, 0.4 µm. d , e , g , Scale bar, 10 µm.

Article Snippet: The primary mouse anti-CD8 antibody (1:50 dilution, clone C8/144B, catalog no. M7103, Dako) was incubated overnight and developed with 3,3′-diaminobenzidine (Dako REAL EnVision, catalog no. K500711-2, Dako) until saturation of the chromogenic reaction.

Techniques: Staining, Control

Journal: Nature Neuroscience

Article Title: T cell-mediated microglial activation triggers myelin pathology in a mouse model of amyloidosis

doi: 10.1038/s41593-024-01682-8

Figure Lengend Snippet: a , Representative image and quantification showing number of CD8 + T cells in mice (7.5 month-old) treated with anti-CD8 and isotype control antibodies. Each dot (red) represents one CD8 + T cell. Scale bar, 500 µm. Statistical significance was determined for n = 9–10 animals using unpaired two-sided Student’s t-test (control, anti-CD8, ***p = 0.001). b , Representative image showing Aβ plaque load (green) in mice (7.5 month-old) treated with anti-CD8 and isotype control antibodies. Scale bar, 500 µm. Quantifications indicate proportion of Aβ positive area normalized to total area in the cortex. Statistical significance was determined for n = 9 animals using unpaired Student’s t-test. c , Representative image and quantification indicating number of BCAS1 + (green) swellings lacking a nucleus (stained with DAPI) in the cortex of 7.5 month-old 5xFAD mice treated with anti-CD8 and isotype control antibodies. Scale bar, 10 µm. Statistical significance was determined for n = 5 animals using unpaired two-sided Student’s t-test (control, anti-CD8, *p = 0.038). d , Behavior of 7.5 month-old WT and 5xFAD mice following 6-week treatment with anti-CD8 and isotype control antibodies in an open-field test. Quantifications indicate fecal boli count, total distance travelled, frequency to enter the center and cumulative time spent in the center. Statistical significance was determined for n = 9-10 animals by one-way ANOVA with Tukey’s post hoc test. (Fecal boli count, WT, 5xFAD +control, **p = 0.0017; Distance traveled, WT, 5xFAD +control,*p = 0.03; Center enter frequency, 5xFAD +control, 5xFAD +anti-CD8, *p = 0.02; Cumulative duration in center, 5xFAD +control, 5xFAD +anti-CD8, *p = 0.01). e , Behavior of 7.5 month-old mice following 6-week treatment with anti-CD8 and isotype control antibodies in an elevated plus maze test. Quantifications indicate fecal boli count, total distance travelled, total time spent in the open arms and total time spent in the closed arms. Statistical significance was determined for n = 9-10 animals by one-way ANOVA with Tukey’s post hoc test. (Fecal boli count, WT, 5xFAD +control, *p = 0.03; Open arm duration, WT, 5xFAD +control, *p = 0.03; Open arm duration, WT, 5xFAD +anti-CD8, ***p = 0.0001; Close arm duration, WT, 5xFAD +anti-CD8, *p = 0.02). Each point on the graph represents one animal. Data is presented as mean ± SEM.

Article Snippet: The primary mouse anti-CD8 antibody (1:50 dilution, clone C8/144B, catalog no. M7103, Dako) was incubated overnight and developed with 3,3′-diaminobenzidine (Dako REAL EnVision, catalog no. K500711-2, Dako) until saturation of the chromogenic reaction.

Techniques: Control, Staining

Journal: Nature Neuroscience

Article Title: T cell-mediated microglial activation triggers myelin pathology in a mouse model of amyloidosis

doi: 10.1038/s41593-024-01682-8

Figure Lengend Snippet: a , Image and quantification showing number of CD8 + T cells (green) in the cortex of control or anti-PD1 and CTLA4 (ICB) treated 5xFAD mice. Plaques are stained with Thiazine Red (ThR, red). Scale bar, 10 µm. Statistical significance was determined for n = 4-5 animals using unpaired two-sided Student’s t-test (control, ICB, *p = 0.048). b , Image and quantification showing Aβ plaque load (red) normalized to total area in the cortex for control or anti-PD1 and CTLA4 (ICB) treated 5xFAD mice. Scale bar, 500 µm. Statistical significance was determined for n = 4-5 animals using unpaired Student’s t-test. c , Representative image and quantification showing percentage of Serpina3n + (grey) CC1 + (green) oligodendrocytes in the cortex of control or anti-PD1 and CTLA4 (ICB) treated 5xFAD mice. Plaques are stained with Thiazine Red (ThR, red). Scale bar, 10 µm. Statistical significance was determined for n = 3-4 animals using unpaired two-sided Student’s t-test (control, ICB, *p = 0.036). d , SEM images and quantification of myelinated axons with myelin abnormalities (green) in the cortex of control or anti-PD1 and CTLA4 (ICB) treated 5xFAD mice. Statistical significance was determined for n = 3 animals using unpaired two-sided Student’s t-test (control, ICB, *p = 0.003). e , Image and quantification showing percentage of EdU + (grey) OLIG2 + (green) cells in the cortex of control or anti-PD1 and CTLA4 (ICB) treated 5xFAD mice. Plaques are stained with Thiazine Red (ThR, red). Scale bar, 10 µm. Statistical significance was determined for n = 3-5 animals using unpaired two-sided Student’s t-test (control, ICB, *p = 0.019). f , Image and quantification indicating percentage of MHCII + (green) IBA1 + (grey) cells in the cortex of control or anti-PD1 and CTLA4 (ICB) treated 5xFAD mice. Plaques are stained with Thiazine Red (ThR, red). Scale bar, 10 µm. Statistical significance was determined for n = 4-6 animals using unpaired two-sided Student’s t-test (control, ICB, *p = 0.015). g , Behavior of control or anti-PD1 and CTLA4 (ICB) treated 5xFAD mice in an open-field test. Quantifications indicate fecal boli count, total distance travelled, frequency to enter the center and cumulative time spent in the center. Statistical significance was determined for n = 4-5 animals using unpaired Student’s t-test (Distance traveled, *p = 0.037; Center enter frequency, *p = 0.041; Cumulative duration in center, *p = 0.014). h , Behavior of control or anti-PD1 and CTLA4 (ICB) treated 5xFAD mice in an elevated plus maze test. Quantifications indicate fecal boli count, total distance travelled, total time spent in the open arms and closed arms. Statistical significance was determined for n = 4-5 animals using unpaired Student’s t-test (Fecal boli count, *p = 0.036; Distance traveled, *p = 0.037). Each point on the graph represents one animal. Data is presented as mean ± SEM.

Article Snippet: The primary mouse anti-CD8 antibody (1:50 dilution, clone C8/144B, catalog no. M7103, Dako) was incubated overnight and developed with 3,3′-diaminobenzidine (Dako REAL EnVision, catalog no. K500711-2, Dako) until saturation of the chromogenic reaction.

Techniques: Control, Staining

Journal: Nature Neuroscience

Article Title: T cell-mediated microglial activation triggers myelin pathology in a mouse model of amyloidosis

doi: 10.1038/s41593-024-01682-8

Figure Lengend Snippet: a , Representative image showing CD8 + T cells (green) in the cortex of APP-NLGF mice. Scale bar, 10 µm. Quantifications indicate the number of CD8 + T cells in the cortex of WT, APP-NLGF and APP-NLGF X RAG1KO mice aged 12 months. Statistical significance was determined for n = 3-4 animals by one-way ANOVA with Tukey’s post hoc test (WT, APP-NLGF , *p = 0.0326; APP-NLGF , APP-NLGF X RAG1KO , *p = 0.032). b , Representative scanning electron microscopy images of the cortex of WT, APP-NLGF and APP-NLGF X RAG1KO mice aged 12 months. Myelin abnormalities are shown in green. Scale bar, 1 µm. Quantifications indicate total percentage of myelinated axons with myelin abnormalities. Statistical significance was determined for n = 3 animals by one-way ANOVA with Tukey’s post hoc test (WT, APP-NLGF , **p = 0.001; APP-NLGF , APP-NLGF X RAG1KO , **p = 0.004). c , Representative image of BCAS1 + (green) swellings lacking a nucleus (stained with DAPI) in the cortex of WT, APP-NLGF and APP-NLGF X RAG1KO mice aged 12 months. Scale bar, 10 µm. d , Representative image showing EdU + (grey) OLIG2 + (green) cells in the cortex of WT, APP-NLGF and APP-NLGF X RAG1KO mice aged 12 months. Plaques are stained with Thiazine Red (ThR, red). Scale bar, 10 µm. e , Representative image showing MHCII + (green) IBA1 + (grey) cells in the cortex of WT, APP-NLGF and APP-NLGF X RAG1KO mice aged 12 months. Scale bar, 10 µm. f , Quantifications indicate the number of BCAS1 swellings (WT, APP-NLGF , ***p = 0.0008; APP-NLGF , APP-NLGF X RAG1KO , *p = 0.0154), percentage of IBA1 + cells positive for MHCII (WT, APP-NLGF , ***p = 0.0004; APP-NLGF , APP-NLGF X RAG1KO , ***p = 0.0008), percentage of OLIG2 + cells also positive for EdU (WT, APP-NLGF , **p = 0.0015; APP-NLGF , APP-NLGF X RAG1KO , *p = 0.0230). Statistical significance was determined for n = 3 animals by one-way ANOVA with Tukey’s post hoc test. Each point on the graph represents one animal. Data is presented as mean ± SEM.

Article Snippet: The primary mouse anti-CD8 antibody (1:50 dilution, clone C8/144B, catalog no. M7103, Dako) was incubated overnight and developed with 3,3′-diaminobenzidine (Dako REAL EnVision, catalog no. K500711-2, Dako) until saturation of the chromogenic reaction.

Techniques: Electron Microscopy, Staining

Journal: Nature Neuroscience

Article Title: T cell-mediated microglial activation triggers myelin pathology in a mouse model of amyloidosis

doi: 10.1038/s41593-024-01682-8

Figure Lengend Snippet: a , UMAP plot of microglia, colored according to the identified populations. b , Heatmaps of scaled average expression of selected marker genes of microglial populations. c , Left: UMAP plots of microglia. Right: Proportions of different microglial populations in control (gray) or CD8 + T cell-depleted (green) 5xFAD mice. The central line represents median, the boxes the upper and lower quartiles of the proportions, and the whiskers show the full range of all populations. Asterisks indicate statistical significance for n = 3 animals (using scCODA). Statistical significance was determined for n = 3 animals using scCODA. d , UMAP plots of microglia colored according to the expression of selected gene markers. e , Image and quantification showing the percentage of MHC-II + (green) IBA1 + (gray) cells in the cortex of control or CD8 + T cell-depleted 5xFAD mice. ThR (red) indicates plaques. Unpaired two-sided Student’s t -test for n = 5 animals (control, anti-CD8, * P = 0.010). Data are presented as the mean ± s.e.m. f , UMAP embedding and spatial plots of single-cell transcriptional profiles measured using MERFISH. Cells are colored according to the major cell type. n = 306,537 segmented single cells from five brain sections from n = 3, 9.5-month-old 5xFAD animals. g , Left: MERFISH spatial plot showing the values of ISG (top) and the MHC-II (bottom) expression score. The arrows in the zoom-in show the positions of T cells. Right: Plots show the cell type identities and expression of selected ISG and MHC-II genes. h , Volcano plot of differential gene expression between 50 nearest neighbors of T cells and remaining cells. Bonferroni-adjusted P values from a Wilcoxon rank-sum test are shown. i , Quantification of Cd74 + neighboring cells in proximity to control and T cells. Unpaired two-sided Student’s t -test (control, T cell, * P = 0.011). Data are presented as the mean ± s.e.m. Fifty neighboring cells each were analyzed for a total of 93 CD8 + T cells and 5,000 random cells. j , IHC of human hippocampal sections from 14 patients with AD showed CD8 + T cells (brown) in contact with MHC-II + myeloid cells (blue). The box plot shows the percentage of CD8 + T cells in contact (<50 µm) with MHC-II + cells in patients with AD (ABC score > 1). The central line represents the median, the boxes the upper and lower quartiles and the whiskers show the full range of the data. e , Scale bar, 10 µm. j , Scale bar, 50 µm.

Article Snippet: The primary mouse anti-CD8 antibody (1:50 dilution, clone C8/144B, catalog no. M7103, Dako) was incubated overnight and developed with 3,3′-diaminobenzidine (Dako REAL EnVision, catalog no. K500711-2, Dako) until saturation of the chromogenic reaction.

Techniques: Expressing, Marker, Control

Journal: Nature Neuroscience

Article Title: T cell-mediated microglial activation triggers myelin pathology in a mouse model of amyloidosis

doi: 10.1038/s41593-024-01682-8

Figure Lengend Snippet: a , Representative image showing Aβ plaque load (red) in 9 months old control and baricitinib treated mice. Quantifications indicate proportion of Aβ positive area normalised to total area in the cortex. Statistical significance was determined for n = 4-5 animals using unpaired Student’s t-test. b , Representative image showing CD8 + T cells (green) in the cortex of control and baricitinib treated mice (9 month-old). Scale bar, 10 µm. Quantifications indicate the number of CD8 + T cells in the cortex. Statistical significance was determined for n = 4-5 animals using unpaired Student’s t-test. c , Representative image showing STAT1 + (green) IBA1 + (grey) cells in control and baricitinib treated mice (7.5 months old). Plaques are stained with Methoxy-X04 (MX04, red). Scale bar, 10 µm. Quantifications indicate percentage of IBA1 + cells also positive for STAT1. Statistical significance was determined for n = 5 animals using unpaired two-sided Student’s t-test (control,baricitinib, **p = 0.002). d , Representative image showing P2RY12 + (red) IBA1 + (green) cells in the cortex of control and baricitinib treated mice (7.5 month-old). Scale bar, 10 µm. Quantifications indicate percentage of IBA1 + cells also positive for P2RY12 . Statistical significance was determined for n = 4 animals using unpaired two-sided Student’s t-test (Control, Baricitinib, *p = 0.04). e , Representative image showing STAT1 + (green) CAII + (red) oligodendrocytes in the cortex of control and baricitinib treated mice (7.5 month-old). Scale bar, 10 µm. Quantifications indicate percentage of CAII + cells also positive for STAT1. Statistical significance was determined for n = 5 animals using unpaired two-sided Student’s t-test (control, baricitinib, *p = 0.0296). Each point on the graph represents one animal. Data is presented as mean ± SEM.

Article Snippet: The primary mouse anti-CD8 antibody (1:50 dilution, clone C8/144B, catalog no. M7103, Dako) was incubated overnight and developed with 3,3′-diaminobenzidine (Dako REAL EnVision, catalog no. K500711-2, Dako) until saturation of the chromogenic reaction.

Techniques: Control, Staining