cd4 percp cy 5 5  (Bio-Rad)

Journal: Vaccines

Article Title: Modulation of T-Cell-Dependent Humoral Immune Response to Influenza Vaccine by Multiple Antioxidant/Immunomodulatory Micronutrient Supplementation

doi: 10.3390/vaccines12070743

Figure Lengend Snippet: The supplementation with antioxidants/immunomodulators increased the frequency of GC CD4+ T cells in dLNs and spleens from mice injected with one dose of QIV. ( a ) The graph bars display the percentages of GC CD4+CXCR5+Bcl6+ T cells among (upper row) draining lymph node (dLN) lymphocytes and (lower row) splenocytes from mice injected with one dose (1D) and two doses (2D) of QIV (Vx), and administered with saline (Sal) or supplement (Suppl) determined by flow cytometry. The analyses were performed 14 days after injection of the first and second doses of QIV. The flow cytometry dot plots indicate the gating strategy for GC CD4+CXCR5+Bcl6+ T cells among (upper row) dLN lymphocytes and (lower row) splenocytes. (Right) CXCR5 vs. Bcl6 staining of CD4+ cells gated as indicated in (left) flow cytometry dot plot. CD4+ cells were gated within lymphocyte gates shown in a. ( b ) The graph bars display the proliferative index of GC CD4+CXCR5+ T cells, i.e., the increase in frequency of proliferating Ki67+ cells among GC CD4+CXCR5+ T cells in the presence of QIV antigens over that in their absence in cultures of (upper row) dLN lymphocytes and (lower row) splenocytes from mice injected with 1D and 2D of QIV (Vx) and administered with Supp or Sal in response to restimulation with QIV antigens. Flow cytometry dot plots indicate Ki67+ staining of CD4+ CXCR5+ (upper row) dLN lymphocytes and (lower row) splenocytes from lymphocyte gates shown in Fig 3b. Data are presented as median and interquartile range (IQR) (individual data points are incorporated into bars). n = 6 mice/group. * p ≤ 0.05, ** p ≤ 0.01.

Article Snippet: The following antibodies: fluorescein isothiocyanate (FITC)-conjugated anti-B220 (CD45R, clone RA3-6B2), FITC-conjugated anti-FoxP3 (clone FJK-16s), and phycoerythrin (PE)-conjugated anti-CD95 (clone 15A7) were obtained from eBiolegend (Carlsbad, CA, USA); PE-conjugated anti-CD4 antibody (clone RM4-5) was purchased from Biolegend (San Diego, CA, USA); peridinin chlorophyll protein cyanine (PerCP-Cy™5.5)-conjugated anti-CD4 antibody (clone RM4-5), perCP-Cy™5.5- or PE-conjugated anti-mouse CXCR5 (Clone 2G8), and Alexa Fluor (AF) 647-conjugated anti-human/mouse Bcl6 (K112-91) were acquired from BD Biosciences Pharmingen (Mountain View, CA, USA) and were used for immunostaining and flow cytometric analysis (FCA).

Techniques: Injection, Saline, Flow Cytometry, Staining

Journal: Vaccines

Article Title: Modulation of T-Cell-Dependent Humoral Immune Response to Influenza Vaccine by Multiple Antioxidant/Immunomodulatory Micronutrient Supplementation

doi: 10.3390/vaccines12070743

Figure Lengend Snippet: Supplementation with antioxidants/immunomodulators increased the frequency of Tfh cells and affected Tfr/Tfh and Tfr/GC B cell ratios in dLNs and spleens from mice injected with one dose of QIV. ( a ) Bar graphs indicate the frequency of CD4+CXCR5+Bcl6+FoxP3- (Tfh) cells and CD4+CXCR5+Bcl6+FoxP3+ (Tfr) cells among (upper row) draining lymph node (dLN) lymphocytes and (lower row) splenocytes from mice injected with one dose (1D) and two doses (2D) of QIV (Vx), and treated with saline (Sal) or supplement (Suppl). The analyses were performed 14 days after the first and second QIV injections. Flow cytometry dot plots indicate FoxP3 staining (R1 = FoxP3- cells; R2 = FoxP3+ cells) of CD4+CXCR5+ Bcl6+ (upper row) dLN lymphocytes and (lower row) splenocytes gated as shown in a. Bar graphs indicate ( b ) Tfr/Tfh cell ratio, ( c ) Tfr/GC B cell ratio in dLN lymphocytes and splenocytes from mice injected with 1D and 2D of QIV (Vx) and administered with saline (Sal) or supplement (Suppl). Data are presented as median and interquartile range (IQR) (individual data points are incorporated into bars). n = 6 mice/group. * p ≤ 0.05, ** p ≤ 0.01.

Article Snippet: The following antibodies: fluorescein isothiocyanate (FITC)-conjugated anti-B220 (CD45R, clone RA3-6B2), FITC-conjugated anti-FoxP3 (clone FJK-16s), and phycoerythrin (PE)-conjugated anti-CD95 (clone 15A7) were obtained from eBiolegend (Carlsbad, CA, USA); PE-conjugated anti-CD4 antibody (clone RM4-5) was purchased from Biolegend (San Diego, CA, USA); peridinin chlorophyll protein cyanine (PerCP-Cy™5.5)-conjugated anti-CD4 antibody (clone RM4-5), perCP-Cy™5.5- or PE-conjugated anti-mouse CXCR5 (Clone 2G8), and Alexa Fluor (AF) 647-conjugated anti-human/mouse Bcl6 (K112-91) were acquired from BD Biosciences Pharmingen (Mountain View, CA, USA) and were used for immunostaining and flow cytometric analysis (FCA).

Techniques: Injection, Saline, Flow Cytometry, Staining

Journal: Vaccines

Article Title: Modulation of T-Cell-Dependent Humoral Immune Response to Influenza Vaccine by Multiple Antioxidant/Immunomodulatory Micronutrient Supplementation

doi: 10.3390/vaccines12070743

Figure Lengend Snippet: The supplementation with antioxidants/immunomodulators increased the frequency of IL-21+ cells among CD4+ cells from dLNs and spleens of mice injected with one dose of QIV. ( a ) The bar graphs display the percentage of IL-21+ cells among freshly isolated CD4+ (upper row) draining lymph node (dLN) lymphocytes and (lower row) splenocytes from mice injected with one dose (1D) and two doses (2D) of QIV (Vx), and administered with saline (Sal) or supplement (Suppl) 14 days after the first and the second doses of QIV. Flow cytometry dot plots indicate the gating strategy for flow cytometry analysis of IL-21+ cells among CD4+ cells in freshly isolated (upper row) dLN lymphocytes and (lower row) splenocytes from lymphocytes gated as shown in a. ( b ) The bar graphs indicate the percentage of increase in the frequency of IL-21+ cells among CD4+ lymphocytes in QIV-supplemented over QIV-free (upper row) dLN lymphocyte and (lower row) splenocyte cultures from mice injected with 1D and 2D of QIV (Vx), and administered with Sal or Suppl. Flow cytometry dot plots indicate IL-21 staining of CD4+ cells from lymphocytes gated as presented in Fig 3b. Data are presented as median and interquartile range (IQR) (individual data points are incorporated into bars). n = 6 mice/group. ** p ≤ 0.01.

Article Snippet: The following antibodies: fluorescein isothiocyanate (FITC)-conjugated anti-B220 (CD45R, clone RA3-6B2), FITC-conjugated anti-FoxP3 (clone FJK-16s), and phycoerythrin (PE)-conjugated anti-CD95 (clone 15A7) were obtained from eBiolegend (Carlsbad, CA, USA); PE-conjugated anti-CD4 antibody (clone RM4-5) was purchased from Biolegend (San Diego, CA, USA); peridinin chlorophyll protein cyanine (PerCP-Cy™5.5)-conjugated anti-CD4 antibody (clone RM4-5), perCP-Cy™5.5- or PE-conjugated anti-mouse CXCR5 (Clone 2G8), and Alexa Fluor (AF) 647-conjugated anti-human/mouse Bcl6 (K112-91) were acquired from BD Biosciences Pharmingen (Mountain View, CA, USA) and were used for immunostaining and flow cytometric analysis (FCA).

Techniques: Injection, Isolation, Saline, Flow Cytometry, Staining

Journal: Nutrients

Article Title: Glycation of Whey Proteins Increases the Ex Vivo Immune Response of Lymphocytes Sensitized to β-Lactoglobulin

doi: 10.3390/nu15143110

Figure Lengend Snippet: T cell profiles in tissues of mice immunized with β-lactoglobulin (β-lg) via intragastric (i-g) and intraperitoneal (i-p) routes: ( A ) CD4 + , ( B ) CD8 + , ( C ) CD4 + CD25 + , and ( D ) CD4 + CD25 + FoxP3 + lymphocytes in mesenteric lymph nodes (MLN), Peyer patches (PP), spleen (SPL), head and neck lymph nodes (HNLN), and peripheral blood mononuclear cells (PMBC). Results are presented as the means of the groups ( n = 8) ± SD. Statistical differences between groups are denoted in the following manner: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: After 120 h of incubation at 37 °C and 5% CO 2 , lymphocytes were additionally washed and stained with rat anti-mouse CD4 antigen (PerCP-Cy 5.5, Clone RM4-5, BD Pharmingen, San Diego, CA, USA) and propidium iodide (BD Pharmingen).

Techniques:

Journal: Nutrients

Article Title: Glycation of Whey Proteins Increases the Ex Vivo Immune Response of Lymphocytes Sensitized to β-Lactoglobulin

doi: 10.3390/nu15143110

Figure Lengend Snippet: Mesenteric lymph nodes’ T lymphocyte profiles. ( A ) CD4 + , ( B ) CD4 + CD25 + , and CD4 + FoxP3. ( C ) CD8+, and ( D ) CD4 + CD8 + populations. Cells were stimulated with α-lactalbumin (α-la) and β-lactoglobulin (β-lg), and after glycation, antigens with glucose (α-la/G; β-lg/G) or lactose (α-la/L; β-lg/L), cells grown in medium only (C) and stimulated with Concanavalin A served as controls (10 μg/mL) (ConA). Lymphocytes were isolated from mice ( n = 8) immunized with β-lg intragastrically (black bars) or intraperitoneally (grey bars). Results are presented as the means of the groups ± SD. Different superscript letters denote statistically significant differences ( p < 0.05).

Article Snippet: After 120 h of incubation at 37 °C and 5% CO 2 , lymphocytes were additionally washed and stained with rat anti-mouse CD4 antigen (PerCP-Cy 5.5, Clone RM4-5, BD Pharmingen, San Diego, CA, USA) and propidium iodide (BD Pharmingen).

Techniques: Isolation

Journal: Nutrients

Article Title: Glycation of Whey Proteins Increases the Ex Vivo Immune Response of Lymphocytes Sensitized to β-Lactoglobulin

doi: 10.3390/nu15143110

Figure Lengend Snippet: Splenocyte T lymphocyte profiles: ( A ) CD4 + , ( B ) CD25 + , and ( C ) CD8 + after stimulation with α-lactalbumin (α-la) and β-lactoglobulin (β-lg) and after glycation of antigens with glucose (α-la/G; β-lgG) or lactose (α-la/L), β-lg/L). Cells grown in medium only (C) and stimulated with Concanavalin A (10 μg/mL) (ConA) served as controls. Splenocytes were isolated from mice ( n = 8) immunized intragastrically (black bars) and intraperitoneally (grey bars) immunized with β-lg. Results are presented as the means of the groups (n=8) ± SD. Different superscript letters denote statistically significant differences ( p < 0.05).

Article Snippet: After 120 h of incubation at 37 °C and 5% CO 2 , lymphocytes were additionally washed and stained with rat anti-mouse CD4 antigen (PerCP-Cy 5.5, Clone RM4-5, BD Pharmingen, San Diego, CA, USA) and propidium iodide (BD Pharmingen).

Techniques: Isolation

Journal: Cell Reports Medicine

Article Title: An immunometabolism subtyping system identifies S100A9 + macrophage as an immune therapeutic target in colorectal cancer based on multiomics analysis

doi: 10.1016/j.xcrm.2023.100987

Figure Lengend Snippet:

Article Snippet: The LIVE/DEAD Fixable Violet dead cell staining kit (1:1000), CD4-PerCP-Cy™5.5 (Clone: RM4-5; 561115; 1:600), CD8-BV510 (Clone: 53–6.7; 563068; 1:600), CD11b-Buv395 (Clone: OX-42; 743983; 1:600), CD45-BV785(Clone: HI30; 563716; 1:600), CD11b-Buv395(Clone: M1/70; 563553; 1:600), CD4-PerCP-Cy5.5(Clone: GK1.5; 552051; 1:600), and TCF-7-PE (Clone: S33-966; 564217; 1:600) were purchased from BD Bioscience (USA, CA).

Techniques: Staining, Sequencing, Chromatography, RNA Sequencing Assay, Software

Journal: Nutrients

Article Title: Probiotics Prevent Hypertension in a Murine Model of Systemic Lupus Erythematosus Induced by Toll-Like Receptor 7 Activation

doi: 10.3390/nu13082669

Figure Lengend Snippet: Effects of probiotic treatments on T-cell polarization in mesenteric lymph nodes from imiquimod-treated mice. ( A ) Regulatory T cells (Treg; CD4+ FoxP3+), ( B ) Th17 (CD4+ IL17a+), and ( C ) Th1 (CD4+ interferon-γ+ [IFN-γ+]) cells measured in mesenteric lymphoid nodes from all experimental groups. Groups: control (Ctrl), Imiquimod (IMQ), IMQ treated with Lactobacillus fermentum CECT5716 (LC40), and IMQ-treated with Bifidobacterium breve CECT7263 (BFM). Results are expressed as mean ± SEM. * p < 0.05 compared with the Ctrl group. # p < 0.05 compared with the IMQ group.

Article Snippet: Next, cells were put into polystyrene test tubes for surface staining using anti-CD4 (PerCP-Cy, clone RM4-5; BD Biosciences), anti-CD45 (APE-eFluor 780, clone 30-F11; BD Biosciences), anti-B220 (allophycocyanin, cloneRA3-6B2; BD Biosciences, CA, USA), anti-CD25 (PE-Vio 770, clone 7D4; Miltenyi Biotec) for 20 min at 4 °C in the dark.

Techniques: