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anti mouse cd153 antibody (R&D Systems)

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R&D Systems anti mouse cd153 antibody
Anti Mouse Cd153 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+mouse+cd153+antibody/Mouse+CD30+Ligand%2FTNFSF8+Antibody/pmc07235045__41467_2020_16347_MOESM3_ESM-53-2-9
Average 85 stars, based on 1 article reviews

anti mouse cd153 antibody - by Bioz Stars, 2026-09

85/100 stars

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Western Blot:

Article Title: The CD153 vaccine is a senotherapeutic option for preventing the accumulation of senescent T cells in mice
Article Snippet: For the analysis of the immunohistochemical staining images, the image files were renamed and assigned a random numerical code so that the investigators were blinded during all image analysis. re research | rep o rtin g su m m ary O cto ber2018 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology Animals and other organisms Human research participants Clinical data Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Validation ELISA: horseradish peroxidase (HRP)-conjugated antibodies specific for mouse IgG (1:1000; GE Healthcare, UK) and anti-mouse IgG subclass-specific HRP-conjugated antibodies (1:1000; IgG1, IgG2b, IgG2c and IgG3, Abcam). .. Western blot: anti-mouse CD153 antibody (0.05 #g/ml, Cat# AF732; R&D Systems) and horseradish peroxidase (HRP)-conjugated antibodies specific for mouse IgG (1:2000; GE Healthcare, UK). .. CDC assay: anti-mouse CD153 antibody (functional grade, RM153, Cat# 14-1531-85; eBioscience) and anti-mouse major histocompatibility complex (MHC) class I antibody (H-2Kd/H-2Dd, functional grade, 34-1-2S, Cat# 16-5998-82 Lot# 2003128; eBioscience).

Image Search Results

Figure 3. CD153 engagement restores TCR signaling in CD153+ SA-T cells (A and B) CD153+ SA-T cells and PD-1– CD25– CD44hi cells were enriched from aged WT B6 mice, and stimulated with anti-CD3 and/or anti-CD28 Abs in the presence or absence of RM153 for 3 days. (A) Representative ﬂow cytometry histograms of Ki67 expression and the percentages of Ki67+ cells in the presence of the indicated combinations of Abs (n = 3). (B) Secretion of cytokines and chemokines in the culture supernatants (n = 3). UD, undetected.

Journal: Cell reports

Article Title: cis interaction of CD153 with TCR/CD3 is crucial for the pathogenic activation of senescence-associated T cells.

doi: 10.1016/j.celrep.2022.111373

Figure Lengend Snippet: Figure 3. CD153 engagement restores TCR signaling in CD153+ SA-T cells (A and B) CD153+ SA-T cells and PD-1– CD25– CD44hi cells were enriched from aged WT B6 mice, and stimulated with anti-CD3 and/or anti-CD28 Abs in the presence or absence of RM153 for 3 days. (A) Representative ﬂow cytometry histograms of Ki67 expression and the percentages of Ki67+ cells in the presence of the indicated combinations of Abs (n = 3). (B) Secretion of cytokines and chemokines in the culture supernatants (n = 3). UD, undetected.

Article Snippet: Stable transfectants expressing CD153-Clover (CD153Clov), CD153DCD or CD153 proteins were obtained by incubation with puromycin (EL4; 2 mg/mL) for at least 2 weeks and subsequent FACS, or by magnetic cell separation (MACS system; Miltenyi Biotec, Gaithersburg, MD, USA) using PE-anti-CD153 Ab (RM153) and anti-PE Microbeads (Miltenyi).

Techniques: Cytometry, Expressing

B cells and senescence-associated T cells (SA-Ts) accumulate in salivary glands of aged mice, particularly in female mice. ( A ) Proportion of lymphocytes in the spleen (Sp) and salivary glands (SG) of aged C57BL/6N mice (CD4 + : n = 6–7 mice per group, CD8 + : n = 4–5 mice per group, CD19 + : n = 4–6 mice per group). ( B ) CD44/CD62L expression in CD4 + T cells and PD-1/CD153 expression in TEM cells in the spleens of young and aged mice as detected by a flow cytometric analysis. Results are representative of those from each group of mice. ( C ) Proportions of SA-Ts in the spleens of young (Y) and aged (A) mice ( n = 5–6 mice per group). ( D ) PD-1/CD153 expression in TEM cells in the salivary glands of male and female aged mice as detected by a flow cytometric analysis. Results are representative of those from each group of mice. ( E ) Proportions of SA-Ts in the salivary glands of young (Y) and aged (A) mice ( n = 5–6 mice per group). ( F ) Numbers of CD19 + B cells and SA-Ts in the salivary glands of young (Y) and aged (A) mice ( n = 4–6 mice per group). ( G ) Proportions of CD19 + B cells and SA-Ts in the salivary glands of female young (Y), middle-aged (M), and aged (A) mice ( n = 4–5 mice per group). Values are shown as means ± SEM. ( A , C , E , F ) * p < 0.05, ** p < 0.01, NS, not significant (the Student’s unpaired t -test). ( G ) ** p < 0.01 versus young mice (Dunnett’s multiple-comparison test).

Journal: International Journal of Molecular Sciences

Article Title: Chemokines Up-Regulated in Epithelial Cells Control Senescence-Associated T Cell Accumulation in Salivary Glands of Aged and Sjögren’s Syndrome Model Mice

doi: 10.3390/ijms22052302

Figure Lengend Snippet: B cells and senescence-associated T cells (SA-Ts) accumulate in salivary glands of aged mice, particularly in female mice. ( A ) Proportion of lymphocytes in the spleen (Sp) and salivary glands (SG) of aged C57BL/6N mice (CD4 + : n = 6–7 mice per group, CD8 + : n = 4–5 mice per group, CD19 + : n = 4–6 mice per group). ( B ) CD44/CD62L expression in CD4 + T cells and PD-1/CD153 expression in TEM cells in the spleens of young and aged mice as detected by a flow cytometric analysis. Results are representative of those from each group of mice. ( C ) Proportions of SA-Ts in the spleens of young (Y) and aged (A) mice ( n = 5–6 mice per group). ( D ) PD-1/CD153 expression in TEM cells in the salivary glands of male and female aged mice as detected by a flow cytometric analysis. Results are representative of those from each group of mice. ( E ) Proportions of SA-Ts in the salivary glands of young (Y) and aged (A) mice ( n = 5–6 mice per group). ( F ) Numbers of CD19 + B cells and SA-Ts in the salivary glands of young (Y) and aged (A) mice ( n = 4–6 mice per group). ( G ) Proportions of CD19 + B cells and SA-Ts in the salivary glands of female young (Y), middle-aged (M), and aged (A) mice ( n = 4–5 mice per group). Values are shown as means ± SEM. ( A , C , E , F ) * p < 0.05, ** p < 0.01, NS, not significant (the Student’s unpaired t -test). ( G ) ** p < 0.01 versus young mice (Dunnett’s multiple-comparison test).

Article Snippet: Frozen sections of salivary gland tissue were fixed with methanol/acetone (1:1), blocked using 5% normal goat serum (WAKO)/0.3% Triton X-100 (Sigma) in phosphate-buffered saline (PBS), and stained with FITC-conjugated anti-mouse CD326 (EpCAM) mAb (118207, BioLegend), Alexa Fluor 647-conjugated anti-mouse CD4 mAb (100426, BioLegend), anti-mouse CD153 mAb (14-1531-85, Thermo Fisher Scientific), and an anti-mouse CXCL13 polyclonal antibody (PA1-29046, Thermo Fisher Scientific).

Techniques: Expressing, Comparison

Enhanced migratory rate of CD4 + T cells toward CXCL13 isolated from aged mice. ( A ) The cell surface expression of CXCR5 on naïve and TEM cells from the spleens of young (Y) and aged (A) female C57BL/6N mice was detected by flow cytometry ( n = 5–6 mice per group). Left histograms show a representative of those from each group of mice. Values are shown as means ± SEM. * p < 0.05 (the Student’s unpaired t -test). ( B – D ) Dose-dependent migratory response of TEM cells ( B ), naïve cells ( C ), and PD-1 + CD153 + and PD-1 + CD153 − TEM cells ( D ) to increasing concentrations of CXCL13 in aged female mice. Values are shown as the means ± SEM of tetraplicate measurements. These results are two independent experiments. * p < 0.05 and ** p < 0.01 versus the control group (the CXCL13 concentration is 0 μg/mL) (Dunnett’s multiple comparison test). ( E ) CD153 + CD4 + T cell localization in the SG of aged female mice was detected by an immunofluorescence analysis. Lower images show high magnification (×100) of the square area of the upper image. CD4 (green), EpCAM (red), CD153 (purple), CD4 and CD153 (white), and nuclei (blue). The scale bars of upper and lower images are 20 and 10 μm, respectively.

Journal: International Journal of Molecular Sciences

Article Title: Chemokines Up-Regulated in Epithelial Cells Control Senescence-Associated T Cell Accumulation in Salivary Glands of Aged and Sjögren’s Syndrome Model Mice

doi: 10.3390/ijms22052302

Figure Lengend Snippet: Enhanced migratory rate of CD4 + T cells toward CXCL13 isolated from aged mice. ( A ) The cell surface expression of CXCR5 on naïve and TEM cells from the spleens of young (Y) and aged (A) female C57BL/6N mice was detected by flow cytometry ( n = 5–6 mice per group). Left histograms show a representative of those from each group of mice. Values are shown as means ± SEM. * p < 0.05 (the Student’s unpaired t -test). ( B – D ) Dose-dependent migratory response of TEM cells ( B ), naïve cells ( C ), and PD-1 + CD153 + and PD-1 + CD153 − TEM cells ( D ) to increasing concentrations of CXCL13 in aged female mice. Values are shown as the means ± SEM of tetraplicate measurements. These results are two independent experiments. * p < 0.05 and ** p < 0.01 versus the control group (the CXCL13 concentration is 0 μg/mL) (Dunnett’s multiple comparison test). ( E ) CD153 + CD4 + T cell localization in the SG of aged female mice was detected by an immunofluorescence analysis. Lower images show high magnification (×100) of the square area of the upper image. CD4 (green), EpCAM (red), CD153 (purple), CD4 and CD153 (white), and nuclei (blue). The scale bars of upper and lower images are 20 and 10 μm, respectively.

Techniques: Isolation, Expressing, Flow Cytometry, Control, Concentration Assay, Comparison, Immunofluorescence

Increases in SA-Ts, but not B cells, in salivary glands of aly/aly mice. ( A ) PD-1 + CD153 + expression in TEM cells in the spleens of aly/+ and aly/aly female mice (age 16 weeks) as detected by a flow cytometric analysis. Results are representative of those from each group of mice. ( B ) Numbers and proportions of SA-Ts in the spleens of aly/+ and aly/aly female mice ( n = 4–5 mice per group). ( C ) PD-1 + CD153 + expression in TEM cells in the salivary glands of aly/+ and aly/aly female mice (age 16 weeks) as detected by a flow cytometric analysis. Results are representative of those from each group of mice. ( D ) Numbers and proportions of SA-Ts in the salivary glands of aly/+ and aly/aly female mice ( n = 3–4 mice per group). ( E ) The frequencies of CD19 + B cells gated on lymphocytes in the salivary glands of aly/+ and aly/aly female mice (age 16 weeks) were assessed by flow cytometry. Results are representative of those from each group of mice. ( F ) Proportions of CD19 + B cells in the SG of aly/+ and aly/aly female mice (age 16 weeks) ( n = 3–5 mice per group). ( G ) The cell surface expression of CXCR4 and CXCR5 on SA-Ts from the spleens of aged and aly/aly female mice was detected by flow cytometry. Histograms show a representative of two independent experiments. Values are shown as means ± SEM. * p < 0.05 and ** p < 0.01 (the Student’s unpaired t -test). ( H ) Senescence-associated staining for SPiDER-βGal in frozen sections of salivary glands harvested from aly/+ and aly/aly female mice (age 12 weeks). Nuclei were stained with DAPI. Bars = 50 μm.

Journal: International Journal of Molecular Sciences

Article Title: Chemokines Up-Regulated in Epithelial Cells Control Senescence-Associated T Cell Accumulation in Salivary Glands of Aged and Sjögren’s Syndrome Model Mice

doi: 10.3390/ijms22052302

Figure Lengend Snippet: Increases in SA-Ts, but not B cells, in salivary glands of aly/aly mice. ( A ) PD-1 + CD153 + expression in TEM cells in the spleens of aly/+ and aly/aly female mice (age 16 weeks) as detected by a flow cytometric analysis. Results are representative of those from each group of mice. ( B ) Numbers and proportions of SA-Ts in the spleens of aly/+ and aly/aly female mice ( n = 4–5 mice per group). ( C ) PD-1 + CD153 + expression in TEM cells in the salivary glands of aly/+ and aly/aly female mice (age 16 weeks) as detected by a flow cytometric analysis. Results are representative of those from each group of mice. ( D ) Numbers and proportions of SA-Ts in the salivary glands of aly/+ and aly/aly female mice ( n = 3–4 mice per group). ( E ) The frequencies of CD19 + B cells gated on lymphocytes in the salivary glands of aly/+ and aly/aly female mice (age 16 weeks) were assessed by flow cytometry. Results are representative of those from each group of mice. ( F ) Proportions of CD19 + B cells in the SG of aly/+ and aly/aly female mice (age 16 weeks) ( n = 3–5 mice per group). ( G ) The cell surface expression of CXCR4 and CXCR5 on SA-Ts from the spleens of aged and aly/aly female mice was detected by flow cytometry. Histograms show a representative of two independent experiments. Values are shown as means ± SEM. * p < 0.05 and ** p < 0.01 (the Student’s unpaired t -test). ( H ) Senescence-associated staining for SPiDER-βGal in frozen sections of salivary glands harvested from aly/+ and aly/aly female mice (age 12 weeks). Nuclei were stained with DAPI. Bars = 50 μm.

Techniques: Expressing, Flow Cytometry, Staining

Journal: Current protocols in cytometry

Article Title: Acquisition of high-quality spectral flow cytometry data

doi: 10.1002/cpcy.74

Figure Lengend Snippet: T cell surface antibody cocktail

Article Snippet: T cell surface antibody cocktail Materials (See for antibody specifics) table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Fluorophore Marker Clone Effective Concentration BB515 Ly-6A/E (Sca-1) D7 0.2 μg/mL Alexa Fluor 532 CD3 17A2 4 μg/mL PE Dazzle 594 CD62L MEL-14 0.4 μg/mL PE Cy5 CD122 TM- β 1 2 μg/mL PerCP Cy5.5 CD28 37.51 4 μg/mL PerCP e710 PD-1 RMP1–30 2 μg/mL APC R700 CD103 M290 1 μg/mL APC Fire750 CD44 IM7 0.2 μg/mL BV480 CD4 RM4–5 2 μg/mL BV570 CD8 53–6.7 1 μg/mL BV605 CD152 (CTLA-4) UC10–4B9 4 μg/mL BV650 CD27 LG.3A10 2 μg/mL BV711 CD153 RM153 2 μg/mL BV786 KLRG-1 2F1 2 μg/mL - CD16/32 93 2.5 μg/mL Open in a separate window T cell surface antibody cocktail list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 FACS Staining Buffer (see recipe) Brilliant Stain Buffer (BD Biosciences, cat# 566349) Rat Anti Ly-6A/E, BB515 (BD Biosciences Cat# 565397, RRID: AB_2739218) Anti-mouse CD3, Alexa Fluor 532 (Thermo Fisher Scientific Cat# 58-0032-82, RRID: AB_11217479) Anti-mouse CD62L, PE/Dazzle 594 (BioLegend Cat# 104448, RRID:AB_2566163) Anti-mouse CD122, PE/Cy5 (BioLegend Cat# 123220, RRID: AB_2715962) Anti-mouse CD28, PerCP/Cyanine 5.5 (BioLegend Cat# 102114, RRID: AB_2073850) Anti-mouse PD-1, PerCP-eFluor 710 (Thermo Fisher Scientific Cat# 46-9981-82, RRID: AB_11151142) Anti-mouse CD103, APC-R700 (BD Biosciences Cat# 565529, RRID: AB_2739282) Anti-mouse CD44, APC/Fire 750 (BioLegend Cat# 103062, RRID: AB_2616727) Anti-mouse CD4, Brilliant Violet 480 (BD Biosciences Cat# 565634, RRID: AB_2739312) Anti-mouse CD8, Brilliant Violet 570 (BioLegend Cat# 100739, RRID: AB_10897645) Anti-mouse CD152, Brilliant Violet 605 (BioLegend Cat# 106323, RRID: AB_2566467) Anti-mouse CD27, Brilliant Violet 650 (BioLegend Cat# 124233, RRID: AB_2687192) Anti-mouse CD153, Brilliant Violet 711 (BD Biosciences Cat# 740751, RRID: AB_2740419) Anti-mouse KLRG-1, Brilliant Violet 786 (BD Biosciences Cat# 565477, RRID: AB_2739256) Anti-mouse CD16/32 (Biolegend, cat# 101330) list-behavior=enumerated prefix-word= mark-type=lower-alpha max-label-size=0 Prepare surface antibody cocktail in FACS Staining Buffer with a 1:10 dilution of Brilliant Violet Buffer.

Techniques: Marker, Concentration Assay

a – c Seven-week-old C57BL/6J mice were immunized with the CD153-Alum vaccine (# a – e ) or the KLH-Alum vaccine on days 0 and 14. a The titer against recombinant mouse CD153 is expressed as the OD at 450 nm at 28 days postimmunization. # a – e , n = 4, 6, 6, 3, 3, respectively; KLH, n = 3. b The antibody recognizes the rmCD153 protein by western blot analysis. As a negative control, 100 ng of rmOPN was applied to lane 1, while 30 and 100 ng of rmCD153 were applied to lanes 2 and 3, respectively. c The levels of IgG1 (Th2 response) and IgG2b, IgG2c, and IgG3 (Th1 response) in mice immunized with the CD153#D-Alum vaccine ( n = 3) or the CD153#D-CpG vaccine ( n = 6) on days 0, 14, and 28. Sera were collected on day 42. IgG1, p = 0.011; IgG2b, p = 0.0067; IgG2c, p = 0.0027; IgG3, p = 0.0029. Left; The titer against CD153-BSA is expressed as the half-maximal binding (OD 50%). Right; ratio of the titer against CD153-BSA-specific IgG2b:IgG1 ( p = 0.022), IgG2c:IgG1 ( p = 0.0091) and IgG3:IgG1 ( p = 0.21). d The proportions of CD153 + senescent T cells in splenic tissues and VAT of male C57BL/6J mice at the ages of 20 and 3 months (normal diet). Senescent T cells were defined as PD-1 + CD153 + cells in CD4 + , CD44 hi , CD62L lo cells. e The proportion of senescent T cells induced by R848 administration in splenic tissues of male C57BL/6J mice ( n = 6 in each group) at the ages of 18 weeks with or without the CD153-CpG vaccine or the KLH-CpG vaccine. PBS vs. R848, p = 0.0086; PBS vs. KLH-CpG, p = 0.0052; PBS vs. CD153-CpG, p = 0.98; R848 vs. KLH-CpG, p > 0.99; R848 vs. CD153-CpG, p = 0.020; KLH-CpG vs. CD153-CpG, p = 0.012. All the data are expressed as the mean ± SEM. Statistical evaluation was performed by one sided ( a ) and by two sided ( c , e ); analysis of variance (ANOVA) test ( a ); unpaired two-tailed t-test ( c ), natural logarithmic transformation; Tukey’s multiple comparison test e ; * p < 0.05; ** p < 0.01; **** p < 0.0001; ND, not detected.

Journal: Nature Communications

Article Title: The CD153 vaccine is a senotherapeutic option for preventing the accumulation of senescent T cells in mice

doi: 10.1038/s41467-020-16347-w

Figure Lengend Snippet: a – c Seven-week-old C57BL/6J mice were immunized with the CD153-Alum vaccine (# a – e ) or the KLH-Alum vaccine on days 0 and 14. a The titer against recombinant mouse CD153 is expressed as the OD at 450 nm at 28 days postimmunization. # a – e , n = 4, 6, 6, 3, 3, respectively; KLH, n = 3. b The antibody recognizes the rmCD153 protein by western blot analysis. As a negative control, 100 ng of rmOPN was applied to lane 1, while 30 and 100 ng of rmCD153 were applied to lanes 2 and 3, respectively. c The levels of IgG1 (Th2 response) and IgG2b, IgG2c, and IgG3 (Th1 response) in mice immunized with the CD153#D-Alum vaccine ( n = 3) or the CD153#D-CpG vaccine ( n = 6) on days 0, 14, and 28. Sera were collected on day 42. IgG1, p = 0.011; IgG2b, p = 0.0067; IgG2c, p = 0.0027; IgG3, p = 0.0029. Left; The titer against CD153-BSA is expressed as the half-maximal binding (OD 50%). Right; ratio of the titer against CD153-BSA-specific IgG2b:IgG1 ( p = 0.022), IgG2c:IgG1 ( p = 0.0091) and IgG3:IgG1 ( p = 0.21). d The proportions of CD153 + senescent T cells in splenic tissues and VAT of male C57BL/6J mice at the ages of 20 and 3 months (normal diet). Senescent T cells were defined as PD-1 + CD153 + cells in CD4 + , CD44 hi , CD62L lo cells. e The proportion of senescent T cells induced by R848 administration in splenic tissues of male C57BL/6J mice ( n = 6 in each group) at the ages of 18 weeks with or without the CD153-CpG vaccine or the KLH-CpG vaccine. PBS vs. R848, p = 0.0086; PBS vs. KLH-CpG, p = 0.0052; PBS vs. CD153-CpG, p = 0.98; R848 vs. KLH-CpG, p > 0.99; R848 vs. CD153-CpG, p = 0.020; KLH-CpG vs. CD153-CpG, p = 0.012. All the data are expressed as the mean ± SEM. Statistical evaluation was performed by one sided ( a ) and by two sided ( c , e ); analysis of variance (ANOVA) test ( a ); unpaired two-tailed t-test ( c ), natural logarithmic transformation; Tukey’s multiple comparison test e ; * p < 0.05; ** p < 0.01; **** p < 0.0001; ND, not detected.

Article Snippet: For the CDC assay, 2 × 10 4 LPS-stimulated RAW 264.7 cells suspended in DMEM supplemented with 1% FBS were mixed with purified IgG antibodies at various concentrations (30, 100, 300 μg/ml) or positive control antibodies and incubated at 4 °C for 1 h. The positive control antibodies included an anti-mouse CD153 antibody (functional grade, RM153; eBioscience) and an anti-mouse major histocompatibility complex (MHC) class I antibody (H-2Kd/H-2Dd, functional grade, 34-1-2S; eBioscience).

Techniques: Recombinant, Western Blot, Negative Control, Binding Assay, Two Tailed Test, Transformation Assay, Comparison

a Time course of HFD loading and injection of vaccines. Mice immunized with the CD153-Alum vaccine or the KLH-Alum vaccine ( n = 3 in each group) were vaccinated at the ages of 7, 9, 11, 13, and 15 weeks and fed a HFD from the age of 15 weeks. Mice immunized with the CD153-CpG vaccine or the KLH-CpG vaccine ( n = 6 in each group) were vaccinated at the ages of 7, 9, 11, and 16 weeks and fed a HFD from the age of 11 weeks. Mice in the ND (normal diet) control group ( n = 5) were unvaccinated and fed a ND, while mice in the HFD control group ( n = 6) were unvaccinated and fed a HFD from the age of 11 weeks. b Titers against CD153-BSA in mice immunized with the CD153-Alum vaccine ( n = 3) or the CD153-CpG vaccine ( n = 6) during HFD loading. Titers are expressed as the half-maximal binding (OD 50%). c The body weights of mice in the ND control group ( n = 5), mice in the HFD control group ( n = 6), and mice immunized with the CD153-CpG vaccine ( n = 6) or the KLH-CpG vaccine ( n = 6) before metabolic measurement. Asterisk indicates the CD153-CpG group vs. the KLH-CpG group. d The weekly average amount of HFD intake per mouse (g/week/mouse) in mice immunized with the CD153-CpG vaccine or the KLH-CpG vaccine during HFD loading. Data were collected from 11 to 19 weeks of age ( n = 8 in each group). e The average VO 2 values during the light (8:00–20:00) and dark (20:00–8:00) periods in mice in the HFD control group ( n = 6) and in mice immunized with the CD153-CpG vaccine ( n = 6) or the KLH-CpG vaccine ( n = 6) at the age of 21–22 weeks. All the data are expressed as the mean ± SEM. Statistical evaluation was performed by two sided; Tukey’s multiple comparison test ( c , d ); Bonferroni correction, HFD vs. KLH-CpG vs. CD153-CpG ( e ); * p < 0.05; ** p < 0.01; *** p < 0.001.

Journal: Nature Communications

Article Title: The CD153 vaccine is a senotherapeutic option for preventing the accumulation of senescent T cells in mice

doi: 10.1038/s41467-020-16347-w

Figure Lengend Snippet: a Time course of HFD loading and injection of vaccines. Mice immunized with the CD153-Alum vaccine or the KLH-Alum vaccine ( n = 3 in each group) were vaccinated at the ages of 7, 9, 11, 13, and 15 weeks and fed a HFD from the age of 15 weeks. Mice immunized with the CD153-CpG vaccine or the KLH-CpG vaccine ( n = 6 in each group) were vaccinated at the ages of 7, 9, 11, and 16 weeks and fed a HFD from the age of 11 weeks. Mice in the ND (normal diet) control group ( n = 5) were unvaccinated and fed a ND, while mice in the HFD control group ( n = 6) were unvaccinated and fed a HFD from the age of 11 weeks. b Titers against CD153-BSA in mice immunized with the CD153-Alum vaccine ( n = 3) or the CD153-CpG vaccine ( n = 6) during HFD loading. Titers are expressed as the half-maximal binding (OD 50%). c The body weights of mice in the ND control group ( n = 5), mice in the HFD control group ( n = 6), and mice immunized with the CD153-CpG vaccine ( n = 6) or the KLH-CpG vaccine ( n = 6) before metabolic measurement. Asterisk indicates the CD153-CpG group vs. the KLH-CpG group. d The weekly average amount of HFD intake per mouse (g/week/mouse) in mice immunized with the CD153-CpG vaccine or the KLH-CpG vaccine during HFD loading. Data were collected from 11 to 19 weeks of age ( n = 8 in each group). e The average VO 2 values during the light (8:00–20:00) and dark (20:00–8:00) periods in mice in the HFD control group ( n = 6) and in mice immunized with the CD153-CpG vaccine ( n = 6) or the KLH-CpG vaccine ( n = 6) at the age of 21–22 weeks. All the data are expressed as the mean ± SEM. Statistical evaluation was performed by two sided; Tukey’s multiple comparison test ( c , d ); Bonferroni correction, HFD vs. KLH-CpG vs. CD153-CpG ( e ); * p < 0.05; ** p < 0.01; *** p < 0.001.

Techniques: Injection, Vaccines, Control, Binding Assay, Comparison

a – c Blood glucose concentrations and percent basal glucose in mice immunized with the Alum-conjugated vaccine (CD153-Alum or KLH-Alum) ( a ; n = 3 in each group) or the CpG-conjugated vaccine (CD153-CpG or KLH-CpG) ( b ; n = 6 in each group) as determined by the intraperitoneal insulin tolerance test (ipITT). Mice in both the ND (normal diet, n = 5) and HFD (high-fat diet, n = 6) control groups were treated in a manner similar to the vaccinated group. Asterisk indicates the CD153-CpG group vs. the KLH-CpG group. c Area under the curve (AUC) of blood glucose levels as determined by the ITT. The AUC was estimated using the trapezoidal rule. d – f Blood glucose concentrations in mice immunized with the Alum-conjugated vaccine ( d ; n = 3 in each group) or the CpG-conjugated vaccine ( e ; n = 6 in each group) as determined by the oral glucose tolerance test (OGTT). Mice in both the ND (normal diet, n = 5) and HFD (high-fat diet, n = 6) control groups were treated in a manner similar to the vaccinated group. Asterisk indicates the CD153-CpG group vs. the KLH-CpG group. Dagger indicates the CD153-CpG group vs. the HFD control group. f AUC of blood glucose levels as determined by the OGTT. The AUC was estimated using the trapezoidal rule. g HOMA-IR index after 6 h of fasting. Alum-conjugated vaccine group, n = 3 in each group; CpG-conjugated vaccine group, n = 6 in each group; ND control group, n = 5; HFD control group, n = 6. All the data are expressed as the mean ± SEM. Statistical evaluation was performed by two sided; unpaired two-tailed t-test, KLH-Alum vs. CD153-Alum ( c , f , g ); Bonferroni correction, HFD vs. KLH-CpG vs. CD153-CpG (C, F and G); Tukey’s multiple comparison test ( a , b , d , e ); * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; †††† p < 0.0001.

Journal: Nature Communications

Article Title: The CD153 vaccine is a senotherapeutic option for preventing the accumulation of senescent T cells in mice

doi: 10.1038/s41467-020-16347-w

Figure Lengend Snippet: a – c Blood glucose concentrations and percent basal glucose in mice immunized with the Alum-conjugated vaccine (CD153-Alum or KLH-Alum) ( a ; n = 3 in each group) or the CpG-conjugated vaccine (CD153-CpG or KLH-CpG) ( b ; n = 6 in each group) as determined by the intraperitoneal insulin tolerance test (ipITT). Mice in both the ND (normal diet, n = 5) and HFD (high-fat diet, n = 6) control groups were treated in a manner similar to the vaccinated group. Asterisk indicates the CD153-CpG group vs. the KLH-CpG group. c Area under the curve (AUC) of blood glucose levels as determined by the ITT. The AUC was estimated using the trapezoidal rule. d – f Blood glucose concentrations in mice immunized with the Alum-conjugated vaccine ( d ; n = 3 in each group) or the CpG-conjugated vaccine ( e ; n = 6 in each group) as determined by the oral glucose tolerance test (OGTT). Mice in both the ND (normal diet, n = 5) and HFD (high-fat diet, n = 6) control groups were treated in a manner similar to the vaccinated group. Asterisk indicates the CD153-CpG group vs. the KLH-CpG group. Dagger indicates the CD153-CpG group vs. the HFD control group. f AUC of blood glucose levels as determined by the OGTT. The AUC was estimated using the trapezoidal rule. g HOMA-IR index after 6 h of fasting. Alum-conjugated vaccine group, n = 3 in each group; CpG-conjugated vaccine group, n = 6 in each group; ND control group, n = 5; HFD control group, n = 6. All the data are expressed as the mean ± SEM. Statistical evaluation was performed by two sided; unpaired two-tailed t-test, KLH-Alum vs. CD153-Alum ( c , f , g ); Bonferroni correction, HFD vs. KLH-CpG vs. CD153-CpG (C, F and G); Tukey’s multiple comparison test ( a , b , d , e ); * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; †††† p < 0.0001.

Techniques: Control, Two Tailed Test, Comparison

a The proportions of senescent T cells induced by HFD loading in the VAT of mice immunized with the Alum-conjugated vaccine or the CpG-conjugated vaccine. Senescent T cells were defined as PD-1 + CD153 + cells in CD4 + , CD44 hi , CD62L lo cells. Alum-conjugated vaccine group, n = 3 in each group; CpG-conjugated vaccine group, n = 6 in each group; ND control group, n = 5; HFD control group, n = 6. b The CDC activity of the CD153 vaccine-induced antibody was assessed ( n = 3 each sample) using 2 × 10 4 LPS-stimulated RAW 264.7 cells. The vaccine-induced IgG antibodies used were purified from the sera of HFD-loaded mice at the age of 22–25 weeks immunized with the CD153-Alum vaccine, CD153-CpG vaccine, KLH-Alum vaccine or KLH-CpG vaccine. N, no stimulation with antibody; P, 100 µg/ml anti-mouse MHC class I antibody (H-2Kd/H-2Dd); closed right triangle, the concentrations of purified IgG antibodies; opened right triangle, the concentrations of anti-mouse CD153 antibody (10, 30, 100 µg/ml, respectively). c , d VAT were collected from the four groups of mice (ND control group, HFD control group, KLH-CpG vaccine group, and CD153-CpG vaccine group; n = 3 in each group) according to the time course of HFD loading and injection of vaccines. c Quantification of macrophages in the crown-like structures among the four groups. HFD vs. KLH-CpG, p = 0.11; HFD vs. CD153-CpG, p = 0.0041; KLH-CpG vs. CD153-CpG, p = 0.15. 40–50 fields per sample, F4/80 (brown); scale bars: 100 µm. d Quantification of CD153-positive cells in the crown-like structures among the four groups. HFD vs. KLH-CpG, p = 0.024; HFD vs. CD153-CpG, p = 0.0015; KLH-CpG vs. CD153-CpG, p = 0.20. 40–50 fields per sample, CD153 (brown); scale bars: 100 µm. All the data are expressed as the mean ± SEM. Statistical evaluation was performed by two-sided; unpaired two-tailed t-test, KLH-Alum vs. CD153-Alum ( a ); Bonferroni correction, ND vs. HFD ( a ), HFD vs. KLH-CpG vs. CD153-CpG ( a , c , d ); * p < 0.05; ** p < 0.01; n.s. not significant.

Journal: Nature Communications

Article Title: The CD153 vaccine is a senotherapeutic option for preventing the accumulation of senescent T cells in mice

doi: 10.1038/s41467-020-16347-w

Figure Lengend Snippet: a The proportions of senescent T cells induced by HFD loading in the VAT of mice immunized with the Alum-conjugated vaccine or the CpG-conjugated vaccine. Senescent T cells were defined as PD-1 + CD153 + cells in CD4 + , CD44 hi , CD62L lo cells. Alum-conjugated vaccine group, n = 3 in each group; CpG-conjugated vaccine group, n = 6 in each group; ND control group, n = 5; HFD control group, n = 6. b The CDC activity of the CD153 vaccine-induced antibody was assessed ( n = 3 each sample) using 2 × 10 4 LPS-stimulated RAW 264.7 cells. The vaccine-induced IgG antibodies used were purified from the sera of HFD-loaded mice at the age of 22–25 weeks immunized with the CD153-Alum vaccine, CD153-CpG vaccine, KLH-Alum vaccine or KLH-CpG vaccine. N, no stimulation with antibody; P, 100 µg/ml anti-mouse MHC class I antibody (H-2Kd/H-2Dd); closed right triangle, the concentrations of purified IgG antibodies; opened right triangle, the concentrations of anti-mouse CD153 antibody (10, 30, 100 µg/ml, respectively). c , d VAT were collected from the four groups of mice (ND control group, HFD control group, KLH-CpG vaccine group, and CD153-CpG vaccine group; n = 3 in each group) according to the time course of HFD loading and injection of vaccines. c Quantification of macrophages in the crown-like structures among the four groups. HFD vs. KLH-CpG, p = 0.11; HFD vs. CD153-CpG, p = 0.0041; KLH-CpG vs. CD153-CpG, p = 0.15. 40–50 fields per sample, F4/80 (brown); scale bars: 100 µm. d Quantification of CD153-positive cells in the crown-like structures among the four groups. HFD vs. KLH-CpG, p = 0.024; HFD vs. CD153-CpG, p = 0.0015; KLH-CpG vs. CD153-CpG, p = 0.20. 40–50 fields per sample, CD153 (brown); scale bars: 100 µm. All the data are expressed as the mean ± SEM. Statistical evaluation was performed by two-sided; unpaired two-tailed t-test, KLH-Alum vs. CD153-Alum ( a ); Bonferroni correction, ND vs. HFD ( a ), HFD vs. KLH-CpG vs. CD153-CpG ( a , c , d ); * p < 0.05; ** p < 0.01; n.s. not significant.

Techniques: Control, Activity Assay, Purification, Injection, Vaccines, Two Tailed Test

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