Journal: Immunity & Ageing : I & A

Article Title: Ascorbic acid attenuates immunosenescence and cognitive decline via MYH9-Mediated CD8⁺ T cell differentiation

doi: 10.1186/s12979-025-00538-4

Figure Lengend Snippet: Ascorbic acid improves the cognitive level of aged mice and increases the number of CD8 + cells in aged mice. A Trajectory tracking map of mouse movement in the open field test, illustrating time spent in the central zone (seconds), average speed during movement episodes (cm/s), and total distance traveled within the central area. B Statistical chart of New Object Cognition Index. C Flow cytometry was utilized to analyze immune cell populations, including CD3 + T, CD4 + T, CD8 + T, CD11b + , and B cells, in the whole blood of middle-aged mice assigned to either the control group or the AA group ( D ). E Statistical analysis of the flow cytometry data was presented in a histogram format, with significance levels denoted as follows: ns ( P > 0.05), * ( P < 0.05), ** ( P < 0.01), and *** ( P < 0.001)

Article Snippet: The Rat IgG2b isotype and Anti-mouse CD8α-InVivo antibodies (Selleckchem) were each diluted to a final volume of 1 ml using sterile PBS and injected intraperitoneally at a dosage of 200 μg per mouse.

Techniques: Flow Cytometry, Control

Journal: Immunity & Ageing : I & A

Article Title: Ascorbic acid attenuates immunosenescence and cognitive decline via MYH9-Mediated CD8⁺ T cell differentiation

doi: 10.1186/s12979-025-00538-4

Figure Lengend Snippet: AA facilitates the early differentiation of T cells while concurrently suppressing myeloid differentiation. A A schematic depicting the early and long-term differentiation of T cells in an OP9-DL1 and lin-CD117 + HSC co-culture system is presented. B The diagram illustrates the progression of T cell maturation from CD44 and CD25 negative to positive selection. C Flow cytometry analysis and statistical bar graphs are utilized to demonstrate the AA-promoted changes in T cell DN stages, including the DN1 phase (CD44 + CD25 − ), DN2 phase (CD44 + CD25 + ), DN3 phase (CD44 − CD25 + ), and DN4 phase (CD44 − CD25 − ). D Flow cytometry analysis is utilized to examine the inhibition of myeloid differentiation, as well as to assess the long-term T cell differentiation promoted by AA in vitro ( E ). F and G Statistical bar graphs are generated to represent the flow cytometry results of CD8 + and CD4 + T cells, with significance levels denoted as ns: not significant ( P > 0.05), * ( P < 0.05), and ** ( P < 0.01), ( n = 3 for each group)

Article Snippet: The Rat IgG2b isotype and Anti-mouse CD8α-InVivo antibodies (Selleckchem) were each diluted to a final volume of 1 ml using sterile PBS and injected intraperitoneally at a dosage of 200 μg per mouse.

Techniques: Co-Culture Assay, Selection, Flow Cytometry, Inhibition, Cell Differentiation, In Vitro, Generated

Journal: Immunity & Ageing : I & A

Article Title: Ascorbic acid attenuates immunosenescence and cognitive decline via MYH9-Mediated CD8⁺ T cell differentiation

doi: 10.1186/s12979-025-00538-4

Figure Lengend Snippet: The Myh9 protein exhibits binding affinity towards AA and affects the proliferation of CD8 + T cells. A MetPro protein-metabolite interaction experimental workflow diagram. B Predicted binding differential protein KEGG pathway enrichment diagram. C Mass spectrometry predicted binding differential protein heatmap. D Protein interaction average degree analysis diagram. E Protein interaction network analysis diagram. F Concentration gradient binding curves of AA with Rhoa and Myh9 highlight the binding dynamics between these proteins. G Flow cytometry diagrams of CD8 + T cells for (Ga) control group, (Gb) AA group, (Gc) blebbistatin group, and (Gd) blebbistatin + AA group with statistical results ( H ) ( n = 3 for each group. ** p < 0.01, * p < 0.05). I Flow cytometry diagrams of CD11b + cells for (Ia) control group, (Ib) AA group, (Ic) blebbistatin group, and (Id) blebbistatin + AA group with statistical results ( J ) ( n = 3 for each group. **** p < 0.0001, *** p < 0.001, ** p < 0.01)

Article Snippet: The Rat IgG2b isotype and Anti-mouse CD8α-InVivo antibodies (Selleckchem) were each diluted to a final volume of 1 ml using sterile PBS and injected intraperitoneally at a dosage of 200 μg per mouse.

Techniques: Binding Assay, Mass Spectrometry, Concentration Assay, Flow Cytometry, Control

Journal: Immunity & Ageing : I & A

Article Title: Ascorbic acid attenuates immunosenescence and cognitive decline via MYH9-Mediated CD8⁺ T cell differentiation

doi: 10.1186/s12979-025-00538-4

Figure Lengend Snippet: Anti-CD8α antibody injection reduced the cognitive level of young mice. A Flow cytometry analysis of CD8 + T cells versus CD4 + T cells in the peripheral blood of mice after injection of IgG2b antibody and Anti-CD8α antibody ( B ). C Statistical histogram of CD8 + T flow cytometry results. D CD4 + T flow cytometry results statistical histogram. E Flow cytometry analysis of NK cells in mouse peripheral blood after IgG2b and Anti-CD8α antibody injection. F Statistical histogram of flow cytometry results of FNK cells. G Tracking diagram of mouse movement trajectory in open field test after antibody injection. H Time (s), speed, and distance traveled by the mouse in the central region during the open field test. I Statistical chart of new object cognition index ( n = 5 for each group, ns P > 0.05, * p < 0.05, *** P < 0.001)

Article Snippet: The Rat IgG2b isotype and Anti-mouse CD8α-InVivo antibodies (Selleckchem) were each diluted to a final volume of 1 ml using sterile PBS and injected intraperitoneally at a dosage of 200 μg per mouse.

Techniques: Injection, Flow Cytometry

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Dual inhibition of HERs and PD-1 counteract resistance in KRAS G12C -mutant head and neck cancer

doi: 10.1186/s13046-024-03227-0

Figure Lengend Snippet: Additive effect of the MRTX849/lapatinib combination on 4NQO-L sensitive tumors is modulated by the presence of CD8 + T-cells. A Growth curve (left) and tumor weight (right) of 4NQO-L tumors in WT mice treated with vehicle ( n = 5 mice, 10 tumors), MRTX849 ( n = 5 mice, 10 tumors), lapatinib ( n = 5 mice, 10 tumors), or the MRTX849/lapatinib combination ( n = 6 mice, 12 tumors). Error bars indicate SEM. Statistical significance was calculated using one-way ANOVA (**** p < 0.0001), ns denotes not significant. B IHC images showing the expression of Ki67 in tissue sections of 4NQO-L tumors treated with vehicle MRTX849, lapatinib, or the MRTX849/lapatinib combination (scale bars: 100 µm; inset 10 µm). n = 4 tumors and n = 27 analysis fields. Error bars indicate SD. Statistical significance was calculated using one-way ANOVA (** p < 0.01, **** p < 0.0001). C IHC images showing the infiltration of CD8 + T-cells in the tissue sections of 4NQO-L tumors treated with vehicle, MRTX849, lapatinib or the MRTX849/lapatinib combination (scale bar: 100 µm; inset: 10 µm). n = 4 tumors and n = 27 analysis fields. Error bars indicate SD. Statistical significance was calculated using one-way ANOVA (**** p < 0.0001). D Top: Tumor volume of the orthotopic 4NQO-L tumors in WT mice (left; n = 8 mice, 8 tumors) or NSG mice (right; n = 6 mice, 6 tumors) treated with vehicle or the MRTX849/lapatinib combination. Statistical significance was calculated using one-way ANOVA (**** p < 0.0001) Error bars indicate SEM. Bottom: Change in tumor volume from first to last day of treatment. E Top: Scheme of the experiment investigating the effect of CD8 + T-cell depletion on MRTX849/lapatinib efficacy. Bottom: Growth of 4NQO-L tumors in WT mice treated with IgG ( n = 6 mice, 11 tumors), αCD8 T cells depletion ( n = 6 mice, 12 tumors), IgG/MRTX849/lapatinib ( n = 5 mice, 10 tumors) or αCD8/MRTX849/lapatinib ( n = 6 mice, 12 tumors). Statistical significance was calculated using one-way ANOVA (**** p < 0.0001), ns denotes not significant

Article Snippet: In vivo PlusTM anti-mouse CD8α or IgG (In vivo PlusTM rat IgG2b) isotype controls, both from Bio X Cell, were used for CD8 depletion experiments.

Techniques: Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Dual inhibition of HERs and PD-1 counteract resistance in KRAS G12C -mutant head and neck cancer

doi: 10.1186/s13046-024-03227-0

Figure Lengend Snippet: MRTX849/lapatinib/αPD-1 treatment is ineffective in MRTX849-resistant tumors. A Growth curve of 4NQO-L KRAS G12C i acquired resistant tumors in WT mice treated with vehicle, MRTX849, lapatinib or MRTX849/lapatinib combination ( n = 6 mice, 12 tumors). B IHC images showing the infiltration of CD8 + T cells in the tissue sections of 4NQO-L KRAS G12C i acquired-resistance tumors treated with vehicle MRTX849, lapatinib, or MRTX849/lapatinib (scale bars: 100 µm; inset 10 µm). n = 4 tumors and n = 40 analysis fields. Error bars indicate SD. Statistical significance was calculated using one-way ANOVA (** p < 0.01, *** p < 0.001, **** p < 0.0001). C IHC images showing expression of PD-L1 in the tissue sections of 4NQO-L KRAS G12C i acquired resistant tumors treated with vehicle, MRTX849, lapatinib or MRTX849/lapatinib (scale bars: 100 µm; inset 10 µm). n = 4 tumors and n = 38 analysis fields. Error bars indicate SD. Statistical significance was calculated using one-way ANOVA (* p < 0.05, *** p < 0.001, **** p < 0.0001), ns denotes not significant. D IHC images showing the expression of αSMA in the tissue sections of 4NQO-L KRAS G12C i acquired resistant tumors treated with vehicle, MRTX849, lapatinib or MRTX849/lapatinib (scale bars: 100 µm; inset 10 µm). n = 4 tumors and n = 40 analysis fields. Error bars indicate SD. Statistical significance was calculated using one-way ANOVA (**** p < 0.0001), ns denotes not significant. E IHC images showing the infiltration of CD8 + T-cells in the tissue sections of 4NQO-L KRAS G12C i-acquired-resistance tumors (scale bars: 100 µm; inset 10 µm). n = 4 tumors and n = 41 analysis fields. Error bars indicate SD. Statistical significance was calculated using one-way ANOVA (**** p < 0.0001). F Growth curves of 4NQO-L KRAS G12C i-acquired-resistance tumors treated with IgG, IgG/MRTX849/lapatinib, MRTX849/αPD-1, or the combination of MRTX849/ lapatinib and αPD-1 ( n = 6 mice, 12 tumors). Statistical significance was calculated using one-way ANOVA, ns denotes not significant

Article Snippet: In vivo PlusTM anti-mouse CD8α or IgG (In vivo PlusTM rat IgG2b) isotype controls, both from Bio X Cell, were used for CD8 depletion experiments.

Techniques: Expressing

Journal: Cell

Article Title: Anti-NKG2A mAb Is a Checkpoint Inhibitor that Promotes Anti-tumor Immunity by Unleashing Both T and NK Cells

doi: 10.1016/j.cell.2018.10.014

Figure Lengend Snippet: NKG2A Is an Inhibitory Receptor that Blocks the Anti-tumor Efficacy of NK and CD8 + T Cells (A) Qa-1 b -sufficient or -deficient A20 tumor cells were engrafted subcutaneously (s.c.) in BALB/c mice. (B) BALB/c mice were treated with an anti-aGM1 pAbs or with control rabbit serum, an anti-CD8α mAb, or rat IgG2b isotype control and then subcutaneously engrafted with A20 tumor cells. Graphs show tumor growth in each individual mouse and combined survival curves. Complete regressions are indicated. log rank test, ∗∗ p = 0.0020; ns, no significant. (C) Experiment similar to that in (B), but with Qa-1 b KO A20 tumor cells. Complete regressions are indicated. log rank test, ∗∗∗ p = 0.0002 (NK cell depletion) and ∗∗∗ p = 0.0006 (CD8 + T cell depletion). See also Figure S1 .

Article Snippet: Anti-mouse CD8α V500 (53-6.7) , BD Biosciences , Cat#560776; RRID: AB_1937317.

Techniques:

Journal: Cell

Article Title: Anti-NKG2A mAb Is a Checkpoint Inhibitor that Promotes Anti-tumor Immunity by Unleashing Both T and NK Cells

doi: 10.1016/j.cell.2018.10.014

Figure Lengend Snippet: Combined Blockade of NKG2A and PD-1/PD-L1 Promotes Anti-tumor Immunity in A20 Tumor-Bearing BALB/c Mice (A) Flow cytometry characterization of NK and CD8 + TILs 19 days after A20 tumor cells engraftment. The spleen was used as control. Upper panels: representative fluorescence-activated cell sorting (FACS) profiles of PD-1 and NKG2A expression on NK and CD8 + T cells in the spleen and the tumor bed. Lower panels: pie chart analysis (mean ± SD). The data presented are the pooled results of three independent experiments (n = 12). (B) A20 tumor cells were engrafted in BALB/c mice. Tumor-bearing mice were then treated at 3- to 4-day intervals with an isotype control (IC), anti-NKG2A, anti-PD-L1, or a combination of these last two mAbs. Graphs show tumor growth in each individual mouse and combined survival curves. The data presented are the pooled results of three independent experiments. Complete regression are indicated. log rank test, ∗∗ p = 0.0087; ∗∗∗ p = 0.0001; ∗∗∗∗ p < 0.0001. (C) Experiment similar to that described in (B) but with treatment of the mice with an anti-asialo-GM1 pAbs or an anti-CD8α mAb 1 day before the initiation of immunotherapy with the combination of anti-NKG2A and anti-PD-L1 mAbs. Graphs show tumor growth in each individual and combined survival curves. Complete regression are indicated. log rank test, ∗ p < 0.0016; ∗∗ p < 0.01; ∗∗∗ p = 0.0001. See also Figure S2 .

Article Snippet: Anti-mouse CD8α V500 (53-6.7) , BD Biosciences , Cat#560776; RRID: AB_1937317.

Techniques: Flow Cytometry, Fluorescence, FACS, Expressing

Journal: Cell

Article Title: Anti-NKG2A mAb Is a Checkpoint Inhibitor that Promotes Anti-tumor Immunity by Unleashing Both T and NK Cells

doi: 10.1016/j.cell.2018.10.014

Figure Lengend Snippet: The Combined Blockade of NKG2A and PD-1/PD-L1 Promotes Anti-tumor Immunity in A20 Tumor-Bearing BALB/c Mice, Related to Figure 2 (A) A20 tumor cells were engrafted in BALB/c mice. Tumor-bearing mice were then treated at three- to four-day intervals with isotype control (IC) antibody, anti-NKG2A antibody, anti-PD-1 antibody or a combination of these last two antibodies. Graphs show tumor growth in each individual and combined survival curves. The data presented are the pooled results of two independent experiments. Log-rank test, ∗∗ p = 0.0087, ∗∗∗ p = 0.0001, ∗∗∗∗ p < 0.0001. (B) Experiment similar to that described in (A) but with treatment of the mice with an anti-asialo-GM1 pAbs or an anti-CD8α mAb one day before the initiation of immunotherapy. Graphs show tumor growth in each individual and combined survival curves. Log Rank test, ∗ p < 0.0016, ∗∗ p < 0.01, ∗∗∗ p = 0.0001.

Article Snippet: Anti-mouse CD8α V500 (53-6.7) , BD Biosciences , Cat#560776; RRID: AB_1937317.

Techniques:

Journal: Cell

Article Title: Anti-NKG2A mAb Is a Checkpoint Inhibitor that Promotes Anti-tumor Immunity by Unleashing Both T and NK Cells

doi: 10.1016/j.cell.2018.10.014

Figure Lengend Snippet:

Article Snippet: Anti-mouse CD8α V500 (53-6.7) , BD Biosciences , Cat#560776; RRID: AB_1937317.

Techniques: Purification, Recombinant, Selection, Staining, Software