Journal: Acta Pharmaceutica Sinica. B

Article Title: Arsenic trioxide-based nanoparticles for enhanced chemotherapy by activating pyroptosis

doi: 10.1016/j.apsb.2025.08.003

Figure Lengend Snippet: Immunomodulatory effects of AsMn/Dz@BSA-FA. Analysis of the proportion of (A) CD11c + CD86 + cells, (B) CD3 + CD4 + cells, and (C) CD3 + CD8 + cells in tumor tissues of each group of mice using flow cytometry. (D) Immunofluorescence detection of changes in CD86, CD11c, CD8, and CD4 in tumor tissue after treatment with different formulations, Scale bar: 100 μm. (E) Expression levels of CD86, CD8, and CD4 in tumor tissue after treatment with different formulations. Changes in (F) TNF- α and (G)IL-6 levels in tumor tissue after treatment with different formulations. Data are presented as mean ± SD ( n = 5). ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

Article Snippet: APC Anti-Mouse CD11c Antibody, PE Anti-Mouse CD86 Antibody, FITC Anti-Mouse CD3 Antibody, APC Anti-Mouse CD4 Antibody, PE Anti-Mouse CD8a Antibody and Purified Anti-Mouse CD16/32 Antibody were from Elabscience (Wuhan, China).

Techniques: Flow Cytometry, Immunofluorescence, Expressing

Journal: Journal of Virology

Article Title: CD8+ T Cell Immunodominance in Lymphocytic Choriomeningitis Virus Infection Is Modified in the Presence of Toll-Like Receptor Agonists

doi: 10.1128/jvi.05996-11

Figure Lengend Snippet: FIG. 1. Immunodominance hierarchies of LCMV-specific CD8 T cells are altered by TLR23-L administration. Mice were injected with 500 PFU LCMV s.c. with pIC (100 g) and pam3cysk4 (20 g) individually or in combination before quantifying T cell responses in the spleen at days 8 (A) and 12 (B). LCMV-specific CD8 T cell responses were estimated for the NP396, NP205, GP33, or GP276 epitope (A and B) using ICS and measuring IFN- production after in vitro restimulation. Dot plots are representative of immunodominance profiles (i), and representative data of one experiment out of three independent trials standard deviations from triplicate animals in each condition are shown (ii). The controls

Article Snippet: Splenocytes then were stained with PE-Cy5-conju- gated, rat anti-mouse CD8 clone 53-6.7 (Cedarlane) at 4°C for 30 min.

Techniques: Injection, In Vitro

Journal: Journal of Virology

Article Title: CD8+ T Cell Immunodominance in Lymphocytic Choriomeningitis Virus Infection Is Modified in the Presence of Toll-Like Receptor Agonists

doi: 10.1128/jvi.05996-11

Figure Lengend Snippet: FIG. 2. Tetramer analysis of altered immunodominance between NP396 and GP276. Mice were injected with 500 PFU LCMV s.c. as described in Materials and Methods, with or without pIC and pam3cysk4 individually or in combination. (A to C) LCMV-specific CD8 T cell responses in the spleen were estimated at day 12 with tetramer staining for the NP396 or GP276 epitopes. (A) Dot plots show tetramer-positive CD8 cells from a representative mouse. (B) Graphs summarize the data from three experiments (n 3 mice in each trial). (C) Number of CD8 T cells that are tetramer positive for NP396 or GP276 were estimated. Controls (C) represent infected spleens with CD8 labeling to adjust for the compensation. Naïve splenocytes stained with the same tetramers gave similar background data (data not shown). For immunodominance analyses between TLR-L-treated and untreated mice, the condition where NP396 becomes subdominant is depicted by an asterisk.

Article Snippet: Splenocytes then were stained with PE-Cy5-conju- gated, rat anti-mouse CD8 clone 53-6.7 (Cedarlane) at 4°C for 30 min.

Techniques: Injection, Staining, Infection, Labeling

Journal: Journal of Virology

Article Title: CD8+ T Cell Immunodominance in Lymphocytic Choriomeningitis Virus Infection Is Modified in the Presence of Toll-Like Receptor Agonists

doi: 10.1128/jvi.05996-11

Figure Lengend Snippet: FIG. 7. Analyses of the conditions that favor changing immunodominance hierarchies during viral infection. Mice were injected with 500 PFU LCMV-WE and TLR23L with TLR4-L (10 g) simultaneously or TLR23L 3 days prior to virus infection. For different flank conditions, mice were injected with virus in one flank and TLR23L in another flank. Eight days p.i., LCMV-specific CD8 T cell responses were estimated using ICS. The data are representative of three experiments. For immunodominance analyses of TLR-L-treated and untreated mice, the change in the profile where NP396 becomes subdominant was depicted by an asterisk.

Article Snippet: Splenocytes then were stained with PE-Cy5-conju- gated, rat anti-mouse CD8 clone 53-6.7 (Cedarlane) at 4°C for 30 min.

Techniques: Infection, Injection, Virus

Journal: Frontiers in Immunology

Article Title: Response gene to complement 32 promotes tumorigenesis by mediating DNA damage repair and inhibits CD8+ T cells infiltration in diffuse large B-cell lymphoma

doi: 10.3389/fimmu.2025.1591615

Figure Lengend Snippet: RGC32 knockdown promoted DNA damage and CD8+ T cells. infiltration in DLBCL TME. (A, B) Western blot was conducted to assess the protein levels of phosphorylated (p) ATM/ATR/CHK1, p-H2AX, IRF1 and PD-L1 in cells transfected with shControl and shRGC32. (C, D) In vivo , IHC and flow cytometry showed knockdown of RGC32 increased CD8 expression. (E) IHC images represented the association between expressions of CD8 and RGC32 in DLBCL tissues. Bar = 5μm.

Article Snippet: Cell surface CD8a and CD3 of TILs was stained with CD8a Flow antibody (Proteintech, PE-65069, 0.3μl) and CD3 Flow antibody (Proteintech, APC-65077, 0.5μl), and tested by FACS- 240 Navios Flow Cytometer (Beckman Coulter Inc. USA).

Techniques: Knockdown, Western Blot, Transfection, In Vivo, Flow Cytometry, Expressing

Journal: Frontiers in Immunology

Article Title: Response gene to complement 32 promotes tumorigenesis by mediating DNA damage repair and inhibits CD8+ T cells infiltration in diffuse large B-cell lymphoma

doi: 10.3389/fimmu.2025.1591615

Figure Lengend Snippet: Schematic diagram of pathways regulated by RGC32 in DLBCL. RGC32 promotes DNA damage repair in DLBCL cells by activating the ATM/ATR/CHK1 pathway. The reduction of DNA damage leads to a decrease in tumor neoantigen load, and the ATM/ATR/CHK1 pathway can activate IRF1 to promote PD-L1 expression. Both of these aspects could inhibit the infiltration of CD8+ T lymphocytes in the tumor microenvironment, thereby promoting the occurrence and development of DLBCL.

Article Snippet: Cell surface CD8a and CD3 of TILs was stained with CD8a Flow antibody (Proteintech, PE-65069, 0.3μl) and CD3 Flow antibody (Proteintech, APC-65077, 0.5μl), and tested by FACS- 240 Navios Flow Cytometer (Beckman Coulter Inc. USA).

Techniques: Expressing

Journal: JCI Insight

Article Title: Combined single-cell transcriptome and immune repertoire analysis reveals hepatic and renal immune injury by heat stroke

doi: 10.1172/jci.insight.189825

Figure Lengend Snippet: ( A ) The subclustering of CD8 + T cells identified 6 cell subclusters. ( B ) Dot plot illustrating the scaled expression of representative marker genes across the 6 CD8 + T cell subclusters. ( C ) Pie diagrams displaying the proportion of sample group contributions per annotated CD8 + T cell subclusters. ( D ) Relative contribution of the 6 CD8 + cell subclusters in different sample groups. P values were obtained using the 1-way Kruskal-Wallis test with post hoc Dunn’s test. ( E ) Frequencies of HS-associated CD8 + T cells as determined by flow cytometry. P values were obtained using the 1-way Kruskal-Wallis test with post hoc Dunn’s test. ( F ) Dot plot showing the expression of cytotoxic effector features in CD8 + T cell subtypes derived from the HC, HE, and HS groups. ( G ) Bar plot showing the percentage distribution of TCR clonality types among CD8 + T cell subsets. ( H ) Heatmap illustrating the overlap of TCR clonality types among CD8 + T cell subsets. ( I ) Schematic showing the differentiation trajectory among each CD8 + T cell subset. ( J ) Heatmap displaying dynamic changes in gene expression along the pseudotime of CD8 + T cell differentiation trajectory.

Article Snippet: The following antibodies were used: anti-human CD4 (APC/cyanine, 7317417, BioLegend), anti-human CD3 (APC, APC-65151, Proteintech), anti-human GZMA (PE/Cyanine, 7507221, BioLegend), anti-human CD8a (PE, PE-65144, Proteintech), anti-human GZMB (Pacific Blue, 515407, BioLegend), anti-human CD161 (FITC, FITC-65115, Proteintech), anti-human CD127 (BV510, 563086, BD Bioscience), anti-human CD56 (PE, PE-65067, Proteintech), and anti-human CD16 (PerCP-Cy 5.5, 565421, BD Biosciences).

Techniques: Expressing, Marker, Flow Cytometry, Derivative Assay, Gene Expression, Cell Differentiation

Journal: International Immunology

Article Title: Cancer-derived cholesterol sulfate is a key mediator to prevent tumor infiltration by effector T cells

doi: 10.1093/intimm/dxac002

Figure Lengend Snippet: CS production and CD8 + T-cell infiltration are inversely correlated in colon cancer tissue samples. (A) Comparison of the SULT2B1 expression in 17 human cancer types. Data are from the Human Protein Atlas ( https://www.proteinatlas.org ) and are indicated as medians of the fragments per kilobase million (FPKM) values. (B) Kaplan–Meier curves showing survival of colon cancer patients with high ( n = 404) or low ( n = 193) SULT2B1 expression. The cut-off value was set at 5.68. P = 0.0077 (log-rank test). (C) Comparison of CS production between normal colon tissues and colon cancer tissues. Data are presented as the mean ± SD. ** P < 0.01 (two-tailed Mann–Whitney test). (D) Representative images for CS production and CD8 + T-cell infiltration in colon cancer tissue samples. Scale bars indicate 1 mm (the second column) and 250 µm (the third and fourth columns). The graph shows the comparison of the number of infiltrating CD8 + T cells between the CS-high and CS-low regions ( n = 10). Data are presented as the mean ± SD. ** P < 0.01 (two-tailed unpaired Student’s t -test).

Article Snippet: The following antibodies and reagents were used: anti-Fcg III/II receptor (2.4G2, 0.5 μg ml −1 ; TONBO Biosciences, San Diego, CA, USA, for blocking), APC-conjugated anti-mouse CD274 (PD-L1; 10F.9G2, 2 μg ml −1 ; Biolegend, San Diego, CA, USA), APC-conjugated Rat IgG2b (RTK4530, 2 μg ml −1 ; Biolegend), PE-conjugated anti-MHC II (M5/114.15.2, 0.2 μg ml −1 ; Invitrogen), PE-conjugated Rat IgG2b (A95-1; 0.2 μg ml −1 ; BD Pharmingen), FITC-conjugated anti-mouse CD3ε (145-2C11, 5 μg ml −1 ; BD Pharmingen), FITC-conjugated anti-mouse CD4 (RM4-5, 5 μg ml −1 ; BD Pharmingen), PE-conjugated anti-mouse CD4 (RM4-5, 2 μg ml −1 ; BD Pharmingen), PE-conjugated anti-mouse CD8a (53-6.7, 2 μg ml −1 ; TONBO Biosciences), PerCP-Cy5.5-conjugated anti-mouse CD8a (2.43, 2 μg ml −1 ; TONBO Biosciences), PerCP-Cy5.5-conjugated anti-mouse Vα2 (B20.1, 2 μg ml −1 ; BD Pharmingen), biotin-conjugated anti-mouse Vβ5 (MR9-4, 5 μg ml −1 ; BD Pharmingen), APC-streptavidin (0.4 μg ml −1 ; BD Pharmingen), PE-conjugated anti-mouse CD366 (TIM-3; 5D12, 2 μg ml −1 ; BD Pharmingen), PerCP-Cy5.5-conjugated anti-mouse CD45 (30-F11, 2 μg ml −1 ; Biolegend), APC-conjugated anti-mouse CD279 [programmed cell death 1 (PD-1); RMP1-30, 2 μg ml −1 ; Biolegend], biotin-conjugated anti-mouse F4/80 (BM8.1, 5 μg ml −1 ; TONBO Biosciences), PE-Cy7-conjugated anti-mouse CD3ε (145-2C11, 2 μg ml −1 ; BD Pharmingen) and PE-Cy7-conjugated anti-mouse CD45 (30-F11, 2 μg ml −1 ; Biolegend).

Techniques: Comparison, Expressing, Two Tailed Test, MANN-WHITNEY

Journal: International Immunology

Article Title: Cancer-derived cholesterol sulfate is a key mediator to prevent tumor infiltration by effector T cells

doi: 10.1093/intimm/dxac002

Figure Lengend Snippet: CS production in tumors inhibits infiltration of effector T cells. (A) CS production in cells ( n = 6) and culture supernatant ( n = 5) were compared between E0771-SULT and E0771-MOCK. Data are presented as the mean ± SD. ** P < 0.01 (two-tailed unpaired Student’s t- test). (B) Trans-cancer migration assays showing that the presence of E0771-SULT (SULT+) and CS-treated E0771-MOCK (SULT–) in Matrigel reduces CCL21-induced T-cell migration ( n = 5–7). Data are presented as the mean ± SD. * P < 0.05, ** P < 0.01 (one-way ANOVA followed by Dunnett’s post hoc test). (C) In vitro growth was compared between E0771-MOCK and E0771-SULT ( n = 5). (D and E) After transplantation, tumor growth in wild-type (WT) or Sult2b1 –/– C57BL/6 mice was compared between E0771-MOCK and E0771-SULT ( n = 8–9). Data are presented as the mean + SD. * P < 0.05, ** P < 0.01 (two-tailed unpaired Student’s t -test). (F) viSNE plots highlighting the distribution of tumor-infiltrating lymphocytes in E0771-SULT and E0771-MOCK. (G) The percentages of CD8 + T cells and NK cells in infiltrating leukocytes and their expression of granzyme B were compared between E0771-SULT and E0771-MOCK ( n = 9). Data are presented as the mean ± SD. * P < 0.05 (two-tailed unpaired Student’s t -test). (H and I) After transplantation, tumor growth in BALB/c nude mice (H; n = 7) or DOCK2 –/– mice (I; n = 6) was compared between E0771-SULT and E0771-MOCK. (J) After transplantation, tumor growth in C57BL/6 mice was compared between Pan02-control and Pan02-ΔSULT ( n = 10, 8). Data are presented as the mean + SD. ** P < 0.01 (two-tailed Mann–Whitney test). (K) The percentages of CD8 + T cells in infiltrating leukocytes were compared between Pan02-control and Pan02-ΔSULT ( n = 9). Data are presented as the mean ± SD. ** P < 0.01 (two-tailed unpaired Student’s t -test).

Article Snippet: The following antibodies and reagents were used: anti-Fcg III/II receptor (2.4G2, 0.5 μg ml −1 ; TONBO Biosciences, San Diego, CA, USA, for blocking), APC-conjugated anti-mouse CD274 (PD-L1; 10F.9G2, 2 μg ml −1 ; Biolegend, San Diego, CA, USA), APC-conjugated Rat IgG2b (RTK4530, 2 μg ml −1 ; Biolegend), PE-conjugated anti-MHC II (M5/114.15.2, 0.2 μg ml −1 ; Invitrogen), PE-conjugated Rat IgG2b (A95-1; 0.2 μg ml −1 ; BD Pharmingen), FITC-conjugated anti-mouse CD3ε (145-2C11, 5 μg ml −1 ; BD Pharmingen), FITC-conjugated anti-mouse CD4 (RM4-5, 5 μg ml −1 ; BD Pharmingen), PE-conjugated anti-mouse CD4 (RM4-5, 2 μg ml −1 ; BD Pharmingen), PE-conjugated anti-mouse CD8a (53-6.7, 2 μg ml −1 ; TONBO Biosciences), PerCP-Cy5.5-conjugated anti-mouse CD8a (2.43, 2 μg ml −1 ; TONBO Biosciences), PerCP-Cy5.5-conjugated anti-mouse Vα2 (B20.1, 2 μg ml −1 ; BD Pharmingen), biotin-conjugated anti-mouse Vβ5 (MR9-4, 5 μg ml −1 ; BD Pharmingen), APC-streptavidin (0.4 μg ml −1 ; BD Pharmingen), PE-conjugated anti-mouse CD366 (TIM-3; 5D12, 2 μg ml −1 ; BD Pharmingen), PerCP-Cy5.5-conjugated anti-mouse CD45 (30-F11, 2 μg ml −1 ; Biolegend), APC-conjugated anti-mouse CD279 [programmed cell death 1 (PD-1); RMP1-30, 2 μg ml −1 ; Biolegend], biotin-conjugated anti-mouse F4/80 (BM8.1, 5 μg ml −1 ; TONBO Biosciences), PE-Cy7-conjugated anti-mouse CD3ε (145-2C11, 2 μg ml −1 ; BD Pharmingen) and PE-Cy7-conjugated anti-mouse CD45 (30-F11, 2 μg ml −1 ; Biolegend).

Techniques: Two Tailed Test, Migration, In Vitro, Transplantation Assay, Expressing, Control, MANN-WHITNEY

Journal: International Immunology

Article Title: Cancer-derived cholesterol sulfate is a key mediator to prevent tumor infiltration by effector T cells

doi: 10.1093/intimm/dxac002

Figure Lengend Snippet: SULT2B1b-mediated CS production in tumors dampens anti-tumor T-cell responses. (A) The effect of OTI CD8 + T-cell transfer on tumor growth in C57BL/6 mice was compared between E.G7-OVA-MOCK ( n = 5) and E.G7-OVA-SULT ( n = 6). After the intravenous injection of activated Vα2 + Vβ5 + OTI T cells on day 10, the infiltration of OTI T cells was analyzed on day 15 ( n = 7). Data are presented as the mean + SD or ± SD. * P < 0.05, ** P < 0.01 (two-tailed Mann–Whitney test for both tumor growth and infiltration of OTI T cells). (B) The effect of OTI CD8 + T-cell transfer on tumor growth in C57BL/6 mice was compared between E0771-OVA-control ( n = 8) and E0771-OVA-SULT ( n = 8). After the intravenous injection of activated Vα2 + Vβ5 + OTI T cells on day 10, the infiltration of OTI T cells was analyzed on day 15 ( n = 7 or 8). Data are presented as the mean + SD or ± SD. * P < 0.05, ** P < 0.01 (two-tailed Mann–Whitney test for tumor growth and two-tailed unpaired Student’s t- test for infiltration of OTI T cells). (C) Schematic representation of the interaction between OTII CD4 + T cells and tumor cells expressing I-A b molecule covalently bound to OVA peptide. (D and E) The effect of OTII CD4 + T-cell transfer on tumor growth in C57BL/6 mice was compared between (D) E0771-control-I-A b /OVA ( n = 8) and E0771-SULT-I-A b /OVA ( n = 7) or (E) 3LL-control-I-A b /OVA ( n = 8) and 3LL-SULT-I-A b /OVA ( n = 8). After the intravenous injection of activated Vα2 + Vβ5 + OTII T cells on day 9, infiltration of OTII T cells was analyzed on day 14 (D, n = 9; E, n = 7). Data are presented as the mean + SD or ± SD. * P < 0.05, ** P < 0.01 (two-tailed unpaired Student’s t- test for tumor growth and two-tailed Mann–Whitney test for infiltration of OTII T cells). (F) Comparison of the effect of DOCK2 +/+ OTII CD4 + T cells and DOCK2 –/– OTII CD4 + T cells ( n = 7) on the growth of E0771-control-I-A b /OVA transplanted into C57BL/6 mice. Data are presented as the mean + SD. * P < 0.05 (two-tailed unpaired Student’s t- test).

Article Snippet: The following antibodies and reagents were used: anti-Fcg III/II receptor (2.4G2, 0.5 μg ml −1 ; TONBO Biosciences, San Diego, CA, USA, for blocking), APC-conjugated anti-mouse CD274 (PD-L1; 10F.9G2, 2 μg ml −1 ; Biolegend, San Diego, CA, USA), APC-conjugated Rat IgG2b (RTK4530, 2 μg ml −1 ; Biolegend), PE-conjugated anti-MHC II (M5/114.15.2, 0.2 μg ml −1 ; Invitrogen), PE-conjugated Rat IgG2b (A95-1; 0.2 μg ml −1 ; BD Pharmingen), FITC-conjugated anti-mouse CD3ε (145-2C11, 5 μg ml −1 ; BD Pharmingen), FITC-conjugated anti-mouse CD4 (RM4-5, 5 μg ml −1 ; BD Pharmingen), PE-conjugated anti-mouse CD4 (RM4-5, 2 μg ml −1 ; BD Pharmingen), PE-conjugated anti-mouse CD8a (53-6.7, 2 μg ml −1 ; TONBO Biosciences), PerCP-Cy5.5-conjugated anti-mouse CD8a (2.43, 2 μg ml −1 ; TONBO Biosciences), PerCP-Cy5.5-conjugated anti-mouse Vα2 (B20.1, 2 μg ml −1 ; BD Pharmingen), biotin-conjugated anti-mouse Vβ5 (MR9-4, 5 μg ml −1 ; BD Pharmingen), APC-streptavidin (0.4 μg ml −1 ; BD Pharmingen), PE-conjugated anti-mouse CD366 (TIM-3; 5D12, 2 μg ml −1 ; BD Pharmingen), PerCP-Cy5.5-conjugated anti-mouse CD45 (30-F11, 2 μg ml −1 ; Biolegend), APC-conjugated anti-mouse CD279 [programmed cell death 1 (PD-1); RMP1-30, 2 μg ml −1 ; Biolegend], biotin-conjugated anti-mouse F4/80 (BM8.1, 5 μg ml −1 ; TONBO Biosciences), PE-Cy7-conjugated anti-mouse CD3ε (145-2C11, 2 μg ml −1 ; BD Pharmingen) and PE-Cy7-conjugated anti-mouse CD45 (30-F11, 2 μg ml −1 ; Biolegend).

Techniques: Injection, Two Tailed Test, MANN-WHITNEY, Control, Expressing, Comparison

Journal: Frontiers in Medicine

Article Title: Analysis and prediction of immune cell infiltration characteristics in COPD: Folium isatidis and its active ingredients are able to combat lung lesions caused by COPD by correcting immune cell infiltration

doi: 10.3389/fmed.2025.1584411

Figure Lengend Snippet: IDR and FI modulated changes in immune cell infiltration caused by COPD. Flow cytometric analysis of (A) CD4 + T lymphocytes, (B) CD8 + T lymphocytes, (C) Treg cells, (D) MDSCs, (E) B lymphocytes, (F) NK cells, and (G) Eos. Proportion of (H) CD4 + T lymphocytes, (I) CD8 + T lymphocytes. (J) CD4/CD8 index. Proportion of (K) Treg cells, (L) MDSCs, (M) B lymphocytes, (N) NK cells, and (O) Eos. Data were expressed as mean ± SD, ### p < 0.001 vs. CON group, * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. MOD group ( n = 3).

Article Snippet: The Anti-Mouse antibodies CD3-APC (E-AB-F1013E), CD4-FITC (E-AB-F1097C), CD8a-PC5.5 (E-AB-F1104I), CD25-PE (E-AB-F1102D), Foxp3-APC (E-AB-F1238E), CD11b-FITC (E-AB-F1081C), Ly-6G/Ly-6C-APC (E-AB-F1120E), CD19-PE (E-AB-F0986D), NK1.1-PE (E-AB-F0987D) and F4/80-APC (E-AB-F0995E) were purchased from Elabscience (Wuhan, China).

Techniques: