Journal: Frontiers in pharmacology

Article Title: Transgenic Overexpression of Galectin-3 in Pancreatic β Cells Attenuates Hyperglycemia in Mice: Synergistic Antidiabetic Effect With Exogenous IL-33.

doi: 10.3389/fphar.2021.714683

Figure Lengend Snippet: FIGURE 4 | Transgenically enhanced Gal-3 expression on β-cells decreases the percentage of pro-inflammatory cells in pancreatic islets. Phenotypic characteristics of cells in pancreatic islets. Percentage of pro-inflammatory and regulatory cells. The percentages and representative dot and contour plots of (A) CD4+ cells, (B) CD4+CXCR3+ cells, (C) CD4+CCR6+ cells, (D) CD8+ cells, (E) CD8+CXCR3+ cells (F) CD4+FoxP3+ cells, and (G) CD4+FoxP3+ST2+. Data from two individual experiments with at least 5–7 mice per group are shown as mean ± SD and compared by the paired t-test or Mann–Whitney test.

Article Snippet: Cells were labeled with fluorochrome-conjugated monoclonal antibodies: anti-mouse CD3 (FAB4841F, R&D Systems, Minneapolis, MN), CD4 (130-102-597, MACS Biotech and FAB554F, R&D Systems, Minneapolis, MN), CD8 (130-102-468, MACS Biotech, Bergisch Gladbach,, Germany and FAB116A, R&D Systems, Minneapolis, MN), CCR6 (FAB5906, R&D Systems, Minneapolis, MN), CXCR3 (562152, BD Biosciences, Franklin Lakes, NJ), FoxP3 (130- 093-013, MACS Biotech, Bergisch Gladbach,, Germany), IL-10 (NBP1-00673PCP, NOVUS, Minneapolis, MN), F4/80 (FAB5580A, R&D Systems, Minneapolis, MN), MHCII (12- 53322-81, EBioscience, San Diego, CA), IL-17A (130-102-344, MACS Biotech, Bergisch Gladbach,, Germany), INFγ (RM9001, Invitrogen, Waltham, Mass), CD206 (FAB2535, R&D Systems, Minneapolis, MN), TNFα (IC410F, R&D Systems, Minneapolis, MN), IL-1β (IC4013P, R&D Systems, Minneapolis, MN), CD11b (553312, BD Biosciences, Franklin Lakes, NJ), Galectin 3 (130-101-312, MACS Biotech, Bergisch Gladbach,, Germany), CD45 (561088, BD Biosciences, Frontiers in Pharmacology | www.frontiersin.org November 2021 | Volume 12 | Article 7146834 FIGURE 2 | Transgenically enhanced Gal-3 expression on β-cells improves the parameters of glucoregulation in the MLD–STZ model of type 1 diabetes.

Techniques: Expressing, MANN-WHITNEY

Journal: Frontiers in pharmacology

Article Title: Transgenic Overexpression of Galectin-3 in Pancreatic β Cells Attenuates Hyperglycemia in Mice: Synergistic Antidiabetic Effect With Exogenous IL-33.

doi: 10.3389/fphar.2021.714683

Figure Lengend Snippet: FIGURE 5 | Application of IL-33 (12–18 days) significantly increased the number of Foxp3+ cells in draining pancreatic lymph nodes of TG mice. Differences in number of regulatory and pro-inflammatory cells were observed after administration of multiple low doses of streptozotocin. The percentage, total number, and representative dot plots of (A) CD4+ cells, (B) CD4+IFN-γ+ cells, (C) CD4+Foxp3+ cells, (D) CD4+Foxp3+ST2+ cells, and (E) CD8+ cells. Data from two individual experiments with at least 5–7 mice per group are shown as mean ± SD and compared by the paired t-test or Mann–Whitney test.

Article Snippet: Cells were labeled with fluorochrome-conjugated monoclonal antibodies: anti-mouse CD3 (FAB4841F, R&D Systems, Minneapolis, MN), CD4 (130-102-597, MACS Biotech and FAB554F, R&D Systems, Minneapolis, MN), CD8 (130-102-468, MACS Biotech, Bergisch Gladbach,, Germany and FAB116A, R&D Systems, Minneapolis, MN), CCR6 (FAB5906, R&D Systems, Minneapolis, MN), CXCR3 (562152, BD Biosciences, Franklin Lakes, NJ), FoxP3 (130- 093-013, MACS Biotech, Bergisch Gladbach,, Germany), IL-10 (NBP1-00673PCP, NOVUS, Minneapolis, MN), F4/80 (FAB5580A, R&D Systems, Minneapolis, MN), MHCII (12- 53322-81, EBioscience, San Diego, CA), IL-17A (130-102-344, MACS Biotech, Bergisch Gladbach,, Germany), INFγ (RM9001, Invitrogen, Waltham, Mass), CD206 (FAB2535, R&D Systems, Minneapolis, MN), TNFα (IC410F, R&D Systems, Minneapolis, MN), IL-1β (IC4013P, R&D Systems, Minneapolis, MN), CD11b (553312, BD Biosciences, Franklin Lakes, NJ), Galectin 3 (130-101-312, MACS Biotech, Bergisch Gladbach,, Germany), CD45 (561088, BD Biosciences, Frontiers in Pharmacology | www.frontiersin.org November 2021 | Volume 12 | Article 7146834 FIGURE 2 | Transgenically enhanced Gal-3 expression on β-cells improves the parameters of glucoregulation in the MLD–STZ model of type 1 diabetes.

Techniques: MANN-WHITNEY

Journal: Frontiers in pharmacology

Article Title: Transgenic Overexpression of Galectin-3 in Pancreatic β Cells Attenuates Hyperglycemia in Mice: Synergistic Antidiabetic Effect With Exogenous IL-33.

doi: 10.3389/fphar.2021.714683

Figure Lengend Snippet: FIGURE 7 | Combined effect of intracellular galectin-3 and IL-33 in STZ-induced diabetes. (A) Overexpression of intracellular galectin-3 on beta cells increases the resistance to apoptosis and STZ-induced release of autoantigen. (B) Exogenous IL-33 increases the number of regulatory T cells and reduces the number of effector CD4+ and CD8+ cells.

Article Snippet: Cells were labeled with fluorochrome-conjugated monoclonal antibodies: anti-mouse CD3 (FAB4841F, R&D Systems, Minneapolis, MN), CD4 (130-102-597, MACS Biotech and FAB554F, R&D Systems, Minneapolis, MN), CD8 (130-102-468, MACS Biotech, Bergisch Gladbach,, Germany and FAB116A, R&D Systems, Minneapolis, MN), CCR6 (FAB5906, R&D Systems, Minneapolis, MN), CXCR3 (562152, BD Biosciences, Franklin Lakes, NJ), FoxP3 (130- 093-013, MACS Biotech, Bergisch Gladbach,, Germany), IL-10 (NBP1-00673PCP, NOVUS, Minneapolis, MN), F4/80 (FAB5580A, R&D Systems, Minneapolis, MN), MHCII (12- 53322-81, EBioscience, San Diego, CA), IL-17A (130-102-344, MACS Biotech, Bergisch Gladbach,, Germany), INFγ (RM9001, Invitrogen, Waltham, Mass), CD206 (FAB2535, R&D Systems, Minneapolis, MN), TNFα (IC410F, R&D Systems, Minneapolis, MN), IL-1β (IC4013P, R&D Systems, Minneapolis, MN), CD11b (553312, BD Biosciences, Franklin Lakes, NJ), Galectin 3 (130-101-312, MACS Biotech, Bergisch Gladbach,, Germany), CD45 (561088, BD Biosciences, Frontiers in Pharmacology | www.frontiersin.org November 2021 | Volume 12 | Article 7146834 FIGURE 2 | Transgenically enhanced Gal-3 expression on β-cells improves the parameters of glucoregulation in the MLD–STZ model of type 1 diabetes.

Techniques: Over Expression

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

Journal: Frontiers in Immunology

Article Title: Herpes Simplex Virus Type 2 Infection-Induced Expression of CXCR3 Ligands Promotes CD4 + T Cell Migration and Is Regulated by the Viral Immediate-Early Protein ICP4

doi: 10.3389/fimmu.2018.02932

Figure Lengend Snippet: Contribution of HSV-2 infection-induced CXCR3 ligands to CD4 + T cell infiltration into mouse vagina. Seven days prior to HSV-2 challenge, BALB/c mice were injected with progesterone in multiple sites. One day prior to HSV-2 challenge, CXCL9, CXCL10, and CXCL11 neutralizing antibodies were delivered to the vagina of mice, alone or in combination, while isotype matched control IgG was used as the control. Mice were then anesthetized with pentobarbital sodium and challenged intravaginally with 10 μL/ mouse HSV-2 at a concentration of 6 × 10 7 PFU/ml or mock- challenged. Vaginal lavage fluids and cervical-vaginal tissues were collected at day 7 after challenge. (A) HSV-2 infection induces the production of mouse CXCR3 ligands. The protein levels of CXCL9 and CXCL10 ligands in vaginal lavage fluids were measured by CBA, and the protein level of CXCL11 was detected by ELISA. (B) CXCL9 mediates the migration of CD4 + T cells to the vaginal foci of infected mice. CD4 + T cells in infection foci were detected using anti-CD4 Ab by IHC. The scale bar indicates 100 μm. Data shown are mean ± S.D. ( n = 5 mice/group) of three independent experiments (A) . *** p < 0.001. One representative out of three independent experiments is shown (B) .

Article Snippet: One day prior to challenge, the neutralizing Abs against CXCL9 (2 μg, R&D Systems, MAB554, USA), CXCL10 (2 μg, R&D Systems, MAB554, USA), and CXCL11 (2 μg, R&D Systems, MAB554, USA) were delivered to the vagina of mice, respectively, or in combination.

Techniques: Infection, Injection, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay, Migration

Journal: Frontiers in Immunology

Article Title: Herpes Simplex Virus Type 2 Infection-Induced Expression of CXCR3 Ligands Promotes CD4 + T Cell Migration and Is Regulated by the Viral Immediate-Early Protein ICP4

doi: 10.3389/fimmu.2018.02932

Figure Lengend Snippet: HSV-2 infection induces the production of CXCR3 ligands in human cervical epithelial cells. (A) HSV-2 infection activates the promoters of human CXCR3 ligands. ME180 cells in 24-well plates were co-transfected with 150 ng CXCL9-Luc, CXCL10-Luc or CXCL11-Luc, and 15 ng internal control plasmid phRL-TK. At 4 h post-transfection, cells were infected with HSV-2 or ultraviolet-inactivated HSV-2 (UV-HSV-2) at an MOI of 1 for 24 h. DLR assay was performed. Values for the samples were normalized using Renilla luciferase values and expressed as fold increase of the value induced in mock-infected samples. (B) HSV-2 infection induces the mRNA production of CXCR3 ligands. ME180 cells in 6-well plates were infected with HSV-2 at an MOI of 1 for 24 h. Cells were harvested and total RNA was extracted. The expression of CXCR3 ligands and GAPDH was evaluated by relative real-time quantitative PCR. The Ct values of GAPDH among all groups were equable and not overloaded. mRNA copies of CXCR3 ligands were normalized using GAPDH and expressed as fold increase of the value for the mock-infected control. (C) HSV-2 infection induces the production of CXCR3 ligands. As depicted in (B) , cell supernatants were collected, and the protein level of CXCR3 ligands was measured by CBA. Data shown are mean ± S.D. of three independent experiments (A, B, and C). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: One day prior to challenge, the neutralizing Abs against CXCL9 (2 μg, R&D Systems, MAB554, USA), CXCL10 (2 μg, R&D Systems, MAB554, USA), and CXCL11 (2 μg, R&D Systems, MAB554, USA) were delivered to the vagina of mice, respectively, or in combination.

Techniques: Infection, Transfection, Control, Plasmid Preparation, Luciferase, Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: Herpes Simplex Virus Type 2 Infection-Induced Expression of CXCR3 Ligands Promotes CD4 + T Cell Migration and Is Regulated by the Viral Immediate-Early Protein ICP4

doi: 10.3389/fimmu.2018.02932

Figure Lengend Snippet: HSV-2 infection-induced CXCL9 plays a predominant role in mediating CD4 + T cell migration. (A) The concentrations of CXCR3 ligands in the supernatants of ME180 cells infected with HSV-2 or mock-infected with DMEM were detected by CBA. (B,C) CXCL9 induced by HSV-2 recruits the migration of PBMCs (B) and CD4 + T cells (C) . ME180 cells in 6-well plates were infected with HSV-2 at an MOI of 1 for 24 h. Cell supernatants were collected and added to the lower chamber of transwell plates in the absence or presence of anti-CXCL9, –CXCL10, or/and –CXCL11 neutralizing Ab or control Ab for 1 h. (D) Neutralization of CXCR3 reduces the migration of CD4 + T cells induced by HSV-2 infection. ME180 cells in 6-well plates were infected with HSV-2 at an MOI of 1 for 24 h. Cell supernatants were collected and added to the lower chamber of transwell plates. The activated CD4 + T cells were incubated with RPMI 1,640 medium containing anti-CXCR3 neutralizing Ab for 1 h and placed in the upper chamber. (E) Recombinant CXCL9 significantly induces the migration of CD4 + T cells. DMEM containing recombinant CXCL9, CXCL10, or CXCL11 (48 pg/mL, 55 pg/mL and 175 pg/mL, respectively; the lowest concentration induced by HSV-2 infection) was added to the lower chamber of transwell plates. (F,G) Recombinant CXCL10 or CXCL11 mediates the migration of CD4 + T cells in a dose-dependent manner. DMEM containing recombinant CXCL10 or CXCL11 was added to the lower chamber of transwell plates. CXCL10 or CXCL11 was started from 55 pg/mL and 175 pg/mL, respectively, at a concentration gradient of two times. The activated CD4 + T cells were placed in the upper chamber. After 2 h incubation, cells migrated to lower chambers were collected and counted using an automatic cell counter. Cells migration was expressed as percentage of input. Input cells in the upper chamber were 5 × 10 5 . Data shown are mean ± S.D. of three independent experiments (A–G) . ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: One day prior to challenge, the neutralizing Abs against CXCL9 (2 μg, R&D Systems, MAB554, USA), CXCL10 (2 μg, R&D Systems, MAB554, USA), and CXCL11 (2 μg, R&D Systems, MAB554, USA) were delivered to the vagina of mice, respectively, or in combination.

Techniques: Infection, Migration, Control, Neutralization, Incubation, Recombinant, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Herpes Simplex Virus Type 2 Infection-Induced Expression of CXCR3 Ligands Promotes CD4 + T Cell Migration and Is Regulated by the Viral Immediate-Early Protein ICP4

doi: 10.3389/fimmu.2018.02932

Figure Lengend Snippet: HSV-2 ICP4 promotes the production of human CXCR3 ligands. (A) ICP4 induces the activation of CXCR3 ligand promoters. ME180 cells in 24-well plates were transfected with 300 ng expression plasmid of HSV-2 gene or empty vector together with 150 ng CXCR3 ligand reporter and 15 ng phRL-TK. At 24 h post-transfection, DLR assay was performed. Values for the samples were normalized using Renilla luciferase values and expressed as fold increase of the value induced in cells transfected with empty vector. (B) The expression of HSV-2 genes was detected using anti-Flag Ab by Western Blot. ME180 cells were transfected with 3 μg HSV-2 gene expression plasmid for 24 h. The proteins were collected and detected using mouse anti-Flag Ab. (C) ICP4 induces the mRNA production of CXCR3 ligands. ME180 cells in 6-well plates were transfected with 3 μg ICP4 expression plasmid for 24 h. Cells were harvested and total RNA was extracted. The expression of CXCR3 ligands and GAPDH gene was evaluated by relative real-time quantitative PCR. The Ct values of GAPDH among all groups were equable and not overloaded. mRNA copies of CXCR3 ligands were normalized using GAPDH and expressed as fold increase of the value for the empty vector-transfected control. (D) ICP4 induces the production of CXCR3 ligands. As depicted in (C) , cell supernatants were collected, and the protein levels of CXCR3 ligands were measured by CBA. (E,F) CXCL9 induced by ICP4 recruits the migration of PBMCs (E) and CD4 + T cells (F) . ME180 cells in 6-well plates were transfected with 3 μg ICP4 expression plasmid for 24 h. Cell supernatants were collected and added to the lower chamber of transwell plates in the absence or presence of anti-CXCL9, –CXCL10, or/and –CXCL11 neutralizing Ab or control Ab for 1h. (G) Neutralization of CXCR3 reduces the migration of CD4 + T cells induced by ICP4. ME180 cells in 6-well plates were infected with HSV-2 at an MOI of 1 for 24 h. Cell supernatants were collected and added to the lower chamber of transwell plates. The activated CD4 + T cells were incubated with RPMI 1,640 medium containing anti-CXCR3 neutralizing Ab for 1 h and placed in the upper chamber. As depicted in Figure , cells migrated to lower chambers were counted. Cells migration was expressed as percentage of input. One representative out of three independent experiments is shown (B) . Data shown are mean ± S.D. of three independent experiments (A,C–G) . ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: One day prior to challenge, the neutralizing Abs against CXCL9 (2 μg, R&D Systems, MAB554, USA), CXCL10 (2 μg, R&D Systems, MAB554, USA), and CXCL11 (2 μg, R&D Systems, MAB554, USA) were delivered to the vagina of mice, respectively, or in combination.

Techniques: Activation Assay, Transfection, Expressing, Plasmid Preparation, Luciferase, Western Blot, Gene Expression, Real-time Polymerase Chain Reaction, Control, Migration, Neutralization, Infection, Incubation

Journal: Frontiers in Immunology

Article Title: Herpes Simplex Virus Type 2 Infection-Induced Expression of CXCR3 Ligands Promotes CD4 + T Cell Migration and Is Regulated by the Viral Immediate-Early Protein ICP4

doi: 10.3389/fimmu.2018.02932

Figure Lengend Snippet: HSV-2 ICP4 regulates the expression of CXCR3 ligands via the p38 MAPK signaling pathway. (A–C) HSV-2 regulates the expression of CXCL9 (A) , CXCL10 (B) , and CXCL11 (C) via p38/MAPK signaling pathway. ME180 cells in 24-well plates were co-transfected with 150 ng CXCR3 ligand reporter and 15 ng phRL-TK. At 4 h post-transfection, cells were infected with HSV-2 at an MOI of 1 and supplemented with inhibitor PD98059, SP600125, or SB203580. DLR assay was performed at 24 h post-transfection. Values for the samples were normalized using Renilla luciferase values and expressed as fold increase of the value induced in mock-infected samples. (D–F) HSV-2 ICP4 regulates the expression of CXCL9 (D) , CXCL10 (E) , and CXCL11 (F) via p38/MAPK signaling pathway. ME180 cells in 24-well plates were co-transfected with 300 ng empty vector or ICP4 expression plasmid together with 150 ng CXCR3 ligand reporter and 15 ng phRL-TK. At 4 h post-transfection, cells were cultured in complete DMEM supplemented with inhibitor PD98059, SP600125, or SB203580. DLR assay was performed at 24 h post-transfection. Values for the samples were normalized using Renilla luciferase values and expressed as fold increase of the value induced in cells transfected with empty vector. (G) ICP4 activates p38 MAPK signaling pathway. ME180 cells were transfected with 3 μg ICP4 expression plasmid. The protein level of p38, phospho-p38 (p-p38) or phospho-C/EBP-β (p-C/EBP-β) was detected by Western Blot. Data shown are mean ± S.D. of three independent experiments (A–F) . ns, not significant, *** p < 0.001. One representative out of three independent experiments is shown (G) .

Article Snippet: One day prior to challenge, the neutralizing Abs against CXCL9 (2 μg, R&D Systems, MAB554, USA), CXCL10 (2 μg, R&D Systems, MAB554, USA), and CXCL11 (2 μg, R&D Systems, MAB554, USA) were delivered to the vagina of mice, respectively, or in combination.

Techniques: Expressing, Transfection, Infection, Luciferase, Plasmid Preparation, Cell Culture, Western Blot

Journal: Composites Part B: Engineering

Article Title: Killing three birds with one stone: Tumor-membrane-decorated Prussian blue nanovaccines for synergistic management of skin tumors, radiation dermatitis and wounds

doi: 10.1016/j.compositesb.2023.110900

Figure Lengend Snippet: Fig. 9. Mechanistic analysis of PB nanozymes and PBVac for cancer therapy. Representative flow-cytograms and quantitative analysis of (A) CD3+CD4+ T cells and (B) CD3+CD8+ T cells in the tumors. Data are the mean ± SD (n = 3). *P < 0.05; **P < 0.01; ***P < 0.001; n. s. (not significant).

Article Snippet: Cell staining buffer and APC anti-mouse CD3 antibody (E-AB-F1013E), PE/Cyanine7 anti-mouse CD4 antibody [GK1.5] (E-AB-F1097H), and PerCP/Cyanine5.5 Anti-mouse CD8a antibody [53–6.7] (E-AB-F1104J) was provided by Elabscience Biotechnology (Wuhan, China).

Techniques: