Journal: Frontiers in Immunology

Article Title: Double negative T cells (CD4 - /CD8 - ) are associated with Trypanosoma cruzi persistence in the mouse colon during chronic Chagas disease

doi: 10.3389/fimmu.2026.1761769

Figure Lengend Snippet: Multiparameter spectral flow cytometry analysis of immune cells of the colonic lamina propria during acute and chronic T. cruzi infection. C57BL/6 mice were infected with 10 4 T. cruzi (TcCol-Nluc). Colonic lamina propria cells were isolated from uninfected (Control), acutely infected (Acute, 30 dpi) and chronically infected (Chronic, 90 dpi) mice and analysed by flow cytometry using the gating strategy shown in Supplementary Figure 1 . (A) Automated T-distributed stochastic neighbour embedding (t-SNE) 2D map of the flow cytometry data acquired from control and infected mice colon. (B) Upper panel; tSNE 2D map showing scaled expression of CD11b for myeloid cells, B220 for B cells and CD3 for T cells. Lower panel; tSNE 2D map showing the location of CD11b + myeloid cells, B220 + B cells and CD3 + T cells. (C) Heat maps for (left) cell surface markers expression (CD45, CD11b, B220, CD3, CD4 and CD8) and (right) groups (control, acute and chronic colons) for 22 cell clusters identified. (D) Percentage of each cluster for each group. Panel (D) shows auto-scaled cluster frequencies optimized for visualization within each group. For standardized quantitative comparison across groups, refer to the heat map in panel (C) . Arrows in (C) and (D) indicate immune cell clusters of myeloid cells (blue), B cells (green) or T cells (red).

Article Snippet: Antibodies were polyclonal goat IgG anti-mouse CD4 antibody (R&D Systems, MN), polyclonal rabbit IgG anti-mouse CD8 antibody (Novus Biologicals, CO), monoclonal Rat IgG2b Alexa Fluor ® 647 anti-mouse CD3 Antibody (BioLegend, CA), Donkey anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa FluorTM 488 (Thermo Fisher Scientific, CA), Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa FluorTM 568 (Thermo Fisher Scientific, CA) ( ).

Techniques: Flow Cytometry, Infection, Isolation, Control, Expressing, Comparison

Journal: Frontiers in Immunology

Article Title: Double negative T cells (CD4 - /CD8 - ) are associated with Trypanosoma cruzi persistence in the mouse colon during chronic Chagas disease

doi: 10.3389/fimmu.2026.1761769

Figure Lengend Snippet: Phenotypic analysis of T cells via multiparameter spectral flow cytometry in the colonic lamina propria during acute and chronic T. cruzi infection. C57BL/6 mice were infected with 10 4 T. cruzi (TcCol-Nluc). Colonic lamina propria cells were isolated from uninfected (Control), acutely infected (Acute, 30 dpi) and chronically infected (Chronic, 90 dpi) mice and analysed by flow cytometry using the gating strategy shown in Supplementary Figure 1 . (A) T cells composition by flow cytometry, (B) tSNE 2D map showing scaled expression of CD3, CD4, and CD8 cell makers (C) Heat map of T cells (CD45 + , CD3 + ) (highlighted clusters 1, 7, 9, 16, 20, 22). Arrows indicate immune cell clusters of inflammatory (purple) and regulatory (pink) double-negative (DN) T cells. (D) tSNE 2D map of DN T cells and percentage of T cells cluster for each group. Arrows indicate DN T cell clusters of inflammatory (purple) and regulatory (pink) cells. (E) Percentage of DN T cells with inflammatory/regulatory phenotypes in total immune cells (CD45 + cells). (F) Relative percentage of DN T cells with inflammatory/regulatory phenotypes in total DN T cells. Panel (D) shows auto-scaled cluster frequencies optimized for visualization within each group. For standardized quantitative comparison across groups, refer to the heat map in panel (C) . Bars in A represent mean ± SD, and individual symbols denote values from single mice (n = 15; 5 per set, 3 individual sets). Statistical comparisons were made by an unpaired t test: *p < 0.05, ****p < 0.0001.

Article Snippet: Antibodies were polyclonal goat IgG anti-mouse CD4 antibody (R&D Systems, MN), polyclonal rabbit IgG anti-mouse CD8 antibody (Novus Biologicals, CO), monoclonal Rat IgG2b Alexa Fluor ® 647 anti-mouse CD3 Antibody (BioLegend, CA), Donkey anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa FluorTM 488 (Thermo Fisher Scientific, CA), Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa FluorTM 568 (Thermo Fisher Scientific, CA) ( ).

Techniques: Flow Cytometry, Infection, Isolation, Control, Expressing, Comparison

Journal: Frontiers in Immunology

Article Title: Double negative T cells (CD4 - /CD8 - ) are associated with Trypanosoma cruzi persistence in the mouse colon during chronic Chagas disease

doi: 10.3389/fimmu.2026.1761769

Figure Lengend Snippet: Double-negative T cells phenotypes in the colonic lamina propria of C57BL/6 mice during T. cruzi infection. C57BL/6 mice were infected with 10 4 T. cruzi (TcCol-Nluc). Mice were euthanized during the acute (30 dpi) and chronic (90 dpi) phases. Colonic lamina propria cells were isolated from uninfected (Control; blue), acutely infected (Acute; pink) and chronically infected (Chronic; green) mice. Cells were gated on single cells, live, CD45 + , CD3 + , CD4 - , CD8 - events and subsequently on (A) inflammatory immune cell markers including CCR5, CXCR3 or Granzyme B, and (B) regulatory immune cells markers including CCR4, IL10Rα or IL10. Bars represent mean ± SD, and individual symbols denote values from single mice (n = 8-12; 4 per set, 2–3 individual sets). Statistical comparisons were made by an unpaired t test: *p < 0.05, **p < 0.01, ***p < 0.001, **** p ≤ 0.0001.

Article Snippet: Antibodies were polyclonal goat IgG anti-mouse CD4 antibody (R&D Systems, MN), polyclonal rabbit IgG anti-mouse CD8 antibody (Novus Biologicals, CO), monoclonal Rat IgG2b Alexa Fluor ® 647 anti-mouse CD3 Antibody (BioLegend, CA), Donkey anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa FluorTM 488 (Thermo Fisher Scientific, CA), Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa FluorTM 568 (Thermo Fisher Scientific, CA) ( ).

Techniques: Infection, Isolation, Control

Journal: Frontiers in Immunology

Article Title: Double negative T cells (CD4 - /CD8 - ) are associated with Trypanosoma cruzi persistence in the mouse colon during chronic Chagas disease

doi: 10.3389/fimmu.2026.1761769

Figure Lengend Snippet: Microscopy analysis of T. cruzi infected colons. C57BL/6 mice were infected with 10 4 T. cruzi (TcCol-Nluc-RFP). Colons were isolated from control and chronically infected mice (90 dpi), processed for microscopy and stained with CD3 (Blue), CD4 (Green) and CD8 (Red) specific antibodies ( Supplementary Table 1 , Antibody Panel 4) as described in the materials and methods. Immunofluorescence images of cross sections of colons obtained from non-infected mice (Control) and chronically infected mice (Chronic) are shown in panels (A, B) , respectively. In (A, B) , a low magnification image of the entire colon is shown in the upper left, a region of interest (ROI) 1 is shown at mid-magnification in the lower left, and a high magnification image of ROI 2 is shown in shown on the right. For ROI 2, an overlay image of the 3 channels CD3 (Blue), CD4 (Green) and CD8 (Red) is shown on the top right, and the individual channels are represented below as indicated. A representative result of 3 controls and 3 infected colons is shown in this figure. LP, Lamina Propria; solid white arrow heads, double-negative T cells (CD3 + , CD4 - , CD8 - ); open arrowheads, CD3 - , CD4 - , CD8 + cells; solid pink arrowheads, CD3 - , CD4 + , CD8 - cells; solid yellow arrowheads, CD3 - , CD4 + , CD8 + cells.

Article Snippet: Antibodies were polyclonal goat IgG anti-mouse CD4 antibody (R&D Systems, MN), polyclonal rabbit IgG anti-mouse CD8 antibody (Novus Biologicals, CO), monoclonal Rat IgG2b Alexa Fluor ® 647 anti-mouse CD3 Antibody (BioLegend, CA), Donkey anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa FluorTM 488 (Thermo Fisher Scientific, CA), Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa FluorTM 568 (Thermo Fisher Scientific, CA) ( ).

Techniques: Microscopy, Infection, Isolation, Control, Staining, Immunofluorescence

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Association Between Intra-Tumoral Immune Response and Programmed Death Ligand 1 (PD-L1) in Gastric Cancer

doi: 10.12659/MSM.916432

Figure Lengend Snippet: Expression patterns of PD-L1 and CD4/CD8 TILs in gastric cancer tissues. ( A1–A3, B1–B3 ) represents negative expression patterns of PD-L1 and CD4/CD8 in the same tumor tissues. ( A1–A3 ) Staining with 4× (bar=500 um). ( B1–B3 ) Staining with 20× (bar=100 um). ( C1–C3, D1–D3 ) represents positive expression patterns of PD-L1 and CD4/CD8 in the same tumor tissues: ( C1–C3 ) staining with 4× (bar=500 um); ( D1–D3 ) staining with 20× (bar=100 um). TILs, tumor infiltrating leucocytes.

Article Snippet: PD-L1 (28-8) (Roche Diagnostics, Basel, Switzerland), CD8 (OTI7C10; OriGene Technologies, Inc., Beijing, China) and CD4 (OTI1D6; OriGene Technologies, Inc.) monoclonal antibodies were used to stain the TMAs.

Techniques: Expressing, Staining

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Association Between Intra-Tumoral Immune Response and Programmed Death Ligand 1 (PD-L1) in Gastric Cancer

doi: 10.12659/MSM.916432

Figure Lengend Snippet: Association between tPD-L1 expression and CD8/CD4 TILs densities. ( A ) Gastric cancer non-metastasis patient group and ( B ) gastric cancer metastasis patient group. Bars represent means ± standard error. Four types of bars represent cell numbers of CD8, CD4 positive TILs expression in intra-tumor cell and intra-stromal respectively. The “+” represent positive expression patterns of PD-L1 in intra-tumor or intra-stromal cells; the “−” represent negative expression patterns of PD-L1 in tumor or stromal cells. M0=non-metastasis of gastric cancer; M1=metastasis of gastric cancer; * P <0.05, ** P <0.001. TILs – tumor infiltrating leucocytes.

Article Snippet: PD-L1 (28-8) (Roche Diagnostics, Basel, Switzerland), CD8 (OTI7C10; OriGene Technologies, Inc., Beijing, China) and CD4 (OTI1D6; OriGene Technologies, Inc.) monoclonal antibodies were used to stain the TMAs.

Techniques: Expressing

Journal: Materials Today Bio

Article Title: A smart all-in-one strategy based on hybrid bacteria with targeting drug delivery and spatiotemporal drug release to boost the synergistic therapeutic efficacy against TNBC

doi: 10.1016/j.mtbio.2025.101849

Figure Lengend Snippet: Evaluation of antitumor immune responses and biosafety evaluation. (A) FCM analysis and quantitative analysis of (B) CD4 + T cells and (C) CD8 + T cells in tumor tissues. The levels of (D) IL-6, (E) IFN-γ and (F) TNF-α in the serum of mice quantified by ELISA at the end of treatment. (G) Body weight of mice after various treatments (n = 4). (H) H&E staining of the major organs (heart, liver, spleen, lung and kidney) of mice after various treatments (Scale bars: 100 μm). Data are presented as mean ± SD (ns, not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: Cell counting kit-8 (CCK-8), PE Anti-mouse CD3, FITC Anti-mouse CD8a and APC Anti-mouse CD4 were obtained from Elabscience Biotechnology Co., Ltd (Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Neuroinflammation

Article Title: Promoted CD4 + T cell-derived IFN-γ/IL-10 by photobiomodulation therapy modulates neurogenesis to ameliorate cognitive deficits in APP/PS1 and 3xTg-AD mice

doi: 10.1186/s12974-022-02617-5

Figure Lengend Snippet: PBMT treatment of lymph nodes affected the activation of CD4 + T cells, the expression of IFN-γ/IL-10 in CD4 + T cells, and the recruitment of CD4 + T cells to the brain of APP/PS1 and 3xTg-AD mice. A , B Flow cytometry was used to detect the number of CD4 + IFN-γ + T cells and CD4 + IL-10 + T cells in APP/PS1 and 3xTg-AD mouse brain tissues after PBMT treatment ( n = 3 per group) ( A ) and its quantitative analyses ( B ). C , D Representative images ( C ) and quantitative analyses ( D ) of CD4 + T cells in cortex regions from PBMT-treated or un-treated APP/PS1 and 3xTg-AD groups. Nuclei was stained by DAPI. Scale bar: 50 μm, ( n = 4 per group). E Western blotting analysis and quantification of CD4 protein expression in APP/PS1 and 3xTg-AD mouse brains with or without PBMT, ( n = 3 per group). F The number of CD4 + CD69 + T cells in the brain tissue of six groups was detected by flow cytometry. The statistical analysis of F was provided in Additional file : Fig. S3A. All quantifications are presented as mean ± SEM and were analyzed by One-way ANOVA test; * p < 0.05 versus WT group; ### p < 0.001, ## p < 0.01, # p < 0.05 versus indicated group

Article Snippet: The primary, secondary antibodies, and their dilutions were as follows: anti-Nestin (Proteintech, 19483-1-AP, 1:100), anti-Tuj1 (Proteintech, 66375-1-lg, 1:300); anti-GFAP (CST, 3670, 1:300); anti-Iba-1 (CST, 17198, 1:300); anti-Aβ (Biolegend, 109902, 1:300); anti-CD4 (RD, MAB554-SP, 1:50); anti-PSD95 (Proteintech, 20665-1-AP, 1:300); anti-Lamp1 (Abcam, ab208943, 1:100); Goat anti-Mouse IgG H&L (Alexa Fluor ® 488) (Abcam, ab150113, 1:400); Goat anti-Rabbit IgG H&L (Alexa Fluor ® 555) (Abcam, ab150078, 1:400); Goat anti-Rat IgG H&L (Alexa Fluor ® 647) (Abcam, ab150159, 1:400), and DAPI for cells nuclear staining (Sigma, D9542-1MG).

Techniques: Activation Assay, Expressing, Flow Cytometry, Staining, Western Blot

Journal: Journal of Neuroinflammation

Article Title: Promoted CD4 + T cell-derived IFN-γ/IL-10 by photobiomodulation therapy modulates neurogenesis to ameliorate cognitive deficits in APP/PS1 and 3xTg-AD mice

doi: 10.1186/s12974-022-02617-5

Figure Lengend Snippet: PBMT-induced ROS generation in CD4 + T cells activated JAK2/STAT4/STAT5 signal pathway to upregulate the expression of IFN-γ/IL-10. A CD4 antibody was used to staining the CD4 + T cells, and then the flow cytometry was used to detect and quantitative analyze the generation of ROS (revealed by DCF) in WT, APP/PS1, and 3xTg-AD CD4 + T cells with or without PBMT ( n = 4 per group). Cells were pre-incubated with N-acetyl cysteine (NAC, 1 mM) before PBMT. B , C Western blotting analysis ( B ) and quantification ( C ) of the expression of p-JAK2, p-STAT4, p-STAT5, T-bet, Foxp3 in T cells from WT, APP/PS1, and 3xTg-AD with or without PBMT ( n = 3 per group). Some cells were pre-incubated with NAC or TG-101348 (JAK2 inhibitor, MCE, HY-10409, 1 μM) before PBMT. D , E The concentration of IFN-γ ( D ) and IL-10 ( E ) in T cells from WT, APP/PS1, and 3xTg-AD conditioned medium (CM) were detected by ELISA after PBMT ( n = 4 per group). All quantifications are presented as mean ± SEM and were analyzed by One-way ANOVA test; *** p < 0.001, ** p < 0.01, * p < 0.05 versus control group; ### p < 0.001, ## p < 0.01, # p < 0.05 versus indicated group

Article Snippet: The primary, secondary antibodies, and their dilutions were as follows: anti-Nestin (Proteintech, 19483-1-AP, 1:100), anti-Tuj1 (Proteintech, 66375-1-lg, 1:300); anti-GFAP (CST, 3670, 1:300); anti-Iba-1 (CST, 17198, 1:300); anti-Aβ (Biolegend, 109902, 1:300); anti-CD4 (RD, MAB554-SP, 1:50); anti-PSD95 (Proteintech, 20665-1-AP, 1:300); anti-Lamp1 (Abcam, ab208943, 1:100); Goat anti-Mouse IgG H&L (Alexa Fluor ® 488) (Abcam, ab150113, 1:400); Goat anti-Rabbit IgG H&L (Alexa Fluor ® 555) (Abcam, ab150078, 1:400); Goat anti-Rat IgG H&L (Alexa Fluor ® 647) (Abcam, ab150159, 1:400), and DAPI for cells nuclear staining (Sigma, D9542-1MG).

Techniques: Expressing, Staining, Flow Cytometry, Incubation, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Neuroinflammation

Article Title: Promoted CD4 + T cell-derived IFN-γ/IL-10 by photobiomodulation therapy modulates neurogenesis to ameliorate cognitive deficits in APP/PS1 and 3xTg-AD mice

doi: 10.1186/s12974-022-02617-5

Figure Lengend Snippet: Schematic representation for promoted CD4 + T cell-derived IFN-γ/IL-10 by photobiomodulation therapy modulates neurogenesis to ameliorate cognitive deficits in APP/PS1 and 3xTg-AD mice

Article Snippet: The primary, secondary antibodies, and their dilutions were as follows: anti-Nestin (Proteintech, 19483-1-AP, 1:100), anti-Tuj1 (Proteintech, 66375-1-lg, 1:300); anti-GFAP (CST, 3670, 1:300); anti-Iba-1 (CST, 17198, 1:300); anti-Aβ (Biolegend, 109902, 1:300); anti-CD4 (RD, MAB554-SP, 1:50); anti-PSD95 (Proteintech, 20665-1-AP, 1:300); anti-Lamp1 (Abcam, ab208943, 1:100); Goat anti-Mouse IgG H&L (Alexa Fluor ® 488) (Abcam, ab150113, 1:400); Goat anti-Rabbit IgG H&L (Alexa Fluor ® 555) (Abcam, ab150078, 1:400); Goat anti-Rat IgG H&L (Alexa Fluor ® 647) (Abcam, ab150159, 1:400), and DAPI for cells nuclear staining (Sigma, D9542-1MG).

Techniques: Derivative Assay