Journal: Poultry science

Article Title: The effects of arginine and branched-chain amino acid supplementation to reduced-protein diet on intestinal health, cecal short-chain fatty acid profiles, and immune response in broiler chickens challenged with Eimeria spp.

doi: 10.1016/j.psj.2023.102773

Figure Lengend Snippet: Figure 3. Effects of reduced-protein diets supplemented with argi- nine or branched chain amino acids on cecal tonsil CD25+CD4+ cell (Tregs) percentage in broiler chickens challenged with Eimeria spp. Tregs was presented as percentage of CD4+ cells on d 21 and 28. PC, positive control with 20.0% crude protein; NC, negative control with 17.5% crude protein; ARG and BCAA, negative control supplemented with arginine or BCAA. The error bars represent the SEM values. (A), Bars without a common letter differ significantly. P-values: P-challenge < 0.001, P-diet = 0.646, P-interaction = 0.010. (B), The asterisk (*) denoted significant difference. P-values: P-challenge < 0.001, P- diet = 0.507, P-interaction = 0.089.

Article Snippet: For CD4+CD25+ cell analysis, the cells were incubated with 1 mg/mL mouse antichicken CD25 antibody (Shanmugasundaram and Selvaraj, 2011) and unlabeled mouse IgG at 1:100 dilution in a 96-well plate for 50 min at 4°C, followed by incubation with a 1:250 dilution of FITC-conjugated mouse antichicken CD4 (Southern Biotech) for 20 min at 4°C.

Techniques: Positive Control, Negative Control

Journal: Scientific Reports

Article Title: Regulation of CD4 + CD8 − CD25 + and CD4 + CD8 + CD25 + T cells by gut microbiota in chicken

doi: 10.1038/s41598-018-26763-0

Figure Lengend Snippet: Numbers of CD4 + CD8 − CD25 + and CD4 + CD8 + CD25 + T cells were reduced in cecal tonsils of ABX-treated chickens. Chickens were given water containing antibiotics at hatching for 3 weeks, and cecal tonsils were taken. Single cells from cecal tonsils were stained with anti-chicken TCRγδ, CD3, CD4, CD8α, and CD25 antibodies. The cells were pre-gated for CD3 + γδTCR − cells. Changes in ( A ) the total number of cells, the percentages of ( B ) CD4 + CD8 − CD25 + and ( C ) CD4 + CD8 + CD25 + T cells, and the absolute numbers of ( D ) CD4 + CD8 − CD25 + and ( E ) CD4 + CD8 + CD25 + T cells are shown. ( A – E ) Data were obtained from six chickens in each group and presented as mean ± SD. Asterisks indicate significant differences between Con and ABX. Data are representative of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: To examine CD4 + subtypes of T cells, anti-chicken CD3-Percific Blue (clone CT-4), CD4-FITC, CD8α-SPRD (clone CT-8), TCRgd-PE (clone TCR1), CD5-biotin (clone 2-191) (all from Southern Biotec), and CD25-Alexa Fluor ® 647 (AbD Serotec) antibodies and Brilliant Violet 605 streptavidin (BioLegend, San Diego, CA) were used.

Techniques: Staining

Journal: Scientific Reports

Article Title: Regulation of CD4 + CD8 − CD25 + and CD4 + CD8 + CD25 + T cells by gut microbiota in chicken

doi: 10.1038/s41598-018-26763-0

Figure Lengend Snippet: Expression of IL-10 and IFN-γ mRNA among CD4 + T cell subsets in cecal tonsils of ABX-treated chickens. Chickens were given water containing antibiotics at hatching for 7 days, and cecal tonsils were taken. Single cells from cecal tonsils were stained with anti-chicken CD4, CD8α, and CD25 antibodies. Each subset of CD4 + T cells was sorted using an ARIA II FACS sorter. The mRNA was extracted from each subset, and the expression levels of ( A ) IL-10 and ( B ) IFN- γ were determined by RT-qPCR. Data were obtained from three chickens in each group and presented as the mean ± SD. Asterisks indicate significant differences between Con and ABX. * P < 0.05 and ** P < 0.01.

Article Snippet: To examine CD4 + subtypes of T cells, anti-chicken CD3-Percific Blue (clone CT-4), CD4-FITC, CD8α-SPRD (clone CT-8), TCRgd-PE (clone TCR1), CD5-biotin (clone 2-191) (all from Southern Biotec), and CD25-Alexa Fluor ® 647 (AbD Serotec) antibodies and Brilliant Violet 605 streptavidin (BioLegend, San Diego, CA) were used.

Techniques: Expressing, Staining, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Regulation of CD4 + CD8 − CD25 + and CD4 + CD8 + CD25 + T cells by gut microbiota in chicken

doi: 10.1038/s41598-018-26763-0

Figure Lengend Snippet: Changes in microbial number (colony forming units; CFU), CD4 + CD8 − CD25 + , and CD4 + CD8 + CD25 + T cells in cecal tonsils of ABX-treated chickens after co-housing with control chickens. Chickens were treated with ABX at hatching for 7 days and then co-housed with control chickens for 7 days. ( A ) CFU was measured from cecal contents (1 mg/ml) at 1/4 (6 hours), 1, 3, and 5 days after co-housing. ( B ) Proportions and numbers of CD4 + CD8 − CD25 + and CD4 + CD8 + CD25 + T cells in cecal tonsils were analyzed by flow cytometry after co-housing. Data were obtained from more than six chickens in each group and presented as the mean ± SD. Significant differences between Con and ABX are shown by asterisks, and data are representative of three independent experiments. * P < 0.05 and *** P < 0.001.

Article Snippet: To examine CD4 + subtypes of T cells, anti-chicken CD3-Percific Blue (clone CT-4), CD4-FITC, CD8α-SPRD (clone CT-8), TCRgd-PE (clone TCR1), CD5-biotin (clone 2-191) (all from Southern Biotec), and CD25-Alexa Fluor ® 647 (AbD Serotec) antibodies and Brilliant Violet 605 streptavidin (BioLegend, San Diego, CA) were used.

Techniques: Control, Flow Cytometry

Journal: Scientific Reports

Article Title: Regulation of CD4 + CD8 − CD25 + and CD4 + CD8 + CD25 + T cells by gut microbiota in chicken

doi: 10.1038/s41598-018-26763-0

Figure Lengend Snippet: Changes in CD4 + CD8 − CD25 + and CD4 + CD8 + CD25 + T cells in chickens treated with acetate. SCFAs (acetate 50 mM, butyrate 30 mM, propionate 10 mM) and/or ABX in drinking water were administered to chickens at hatching for 7 days. The numbers of ( A , C , E ) CD4 + CD8 − CD25 + and ( B , D , F ) CD4 + CD8 + CD25 + T cells in cecal tonsils were calculated. ( G ) Subsets of CD4 + T cells, B cells (Bu-1 + ), and APCs (KUL01 + , MHC class II (MHC2) + KUL01 − Bu-1 − ) were sorted using an ARIA II FACS sorter. mRNA was extracted from each subset, and the expression level of GPR43 was determined by RT-qPCR. ( A – G ) Data were obtained from more than three chickens in each group and presented as the mean ± SD. Different characters indicate significant differences at P < 0.05. The figures are representative of three independent experiments.

Article Snippet: To examine CD4 + subtypes of T cells, anti-chicken CD3-Percific Blue (clone CT-4), CD4-FITC, CD8α-SPRD (clone CT-8), TCRgd-PE (clone TCR1), CD5-biotin (clone 2-191) (all from Southern Biotec), and CD25-Alexa Fluor ® 647 (AbD Serotec) antibodies and Brilliant Violet 605 streptavidin (BioLegend, San Diego, CA) were used.

Techniques: Expressing, Quantitative RT-PCR

Journal: The Journal of biological chemistry

Article Title: The epithelial sodium-hydrogen antiporter Na+/H+ exchanger 3 accumulates and is functional in recycling endosomes.

doi: 10.1074/jbc.273.4.2035

Figure Lengend Snippet: FIG. 4. Comparison of the distribu- tion of NHE-3HA with markers of the Golgi cisternae, TGN, and lysosomes. A–C, NHE3-HA-transfected cells were fixed, permeabilized, and stained using monoclonal anti-HA antibodies, as in Fig. 2. D, the cells in A were also immuno- stained with polyclonal antibodies to a-mannosidase II, a marker of medial and trans-Golgi cisternae (25). E, the cells in B were stably transfected with a TGN38- CD25 chimera, as described under “Mate- rials and Methods.” This chimeric marker of the TGN (26) was labeled using anti- CD25 antibodies. F, prior to fixation, the cells in C were allowed to internalize fix- able fluoresceinated dextran for 12 h. Af- ter a 4-h chase, to clear the dextran from the endosomal compartment, the cells were fixed and immunostained. Insets near the top right corners of A–E are mag- nifications of the areas demarcated by dotted lines. Confocal images are repre- sentative of at least three experiments.

Article Snippet: FITC-labeled anti-T-cell antigen (CD25) Ab was from Serotec.

Techniques: Comparison, Transfection, Staining, Marker, Stable Transfection, Labeling

Journal: Toxins

Article Title: Exposure to Subclinical Doses of Fumonisins, Deoxynivalenol, and Zearalenone Affects Immune Response, Amino Acid Digestibility, and Intestinal Morphology in Broiler Chickens

doi: 10.3390/toxins17010016

Figure Lengend Snippet: Effect of combined doses of mycotoxins on cecal tonsils CD4+, CD8+ ratio, and Tregs percentage. Flow cytometry was used to analyze the percentages of CD4+, CD8+ T, and CD4+CD25+ T regulatory cells in the cecal tonsils. Means within a row with no common superscript differ significantly ( p < 0.05) ( n = 6).

Article Snippet: For the T-regulatory cell (Tregs) percentages, cells (1 × 10 6 ) were incubated with 10 μg/mL of phycoerythrin-conjugated mouse anti-chicken CD25 antibody [ ], 1:250 dilution of FITC-d- conjugated mouse anti-chicken CD4+ (Southern Biotechnology Associates, Birmingham, AL, USA), and a 1:200 dilution of unlabeled mouse IgG for 45 min.

Techniques: Flow Cytometry, Control

Journal: Toxins

Article Title: Exposure to Subclinical Doses of Fumonisins, Deoxynivalenol, and Zearalenone Affects Immune Response, Amino Acid Digestibility, and Intestinal Morphology in Broiler Chickens

doi: 10.3390/toxins17010016

Figure Lengend Snippet: Effect of combined doses of mycotoxins on spleen CD4+, CD8+, and Tregs percentage. CD4+ and CD8+ T cells and CD4 + CD25 + T regulatory cell percentages in splenocytes were analyzed by flow cytometry. Means within a row with no common superscript differ significantly ( p < 0.05) ( n = 6).

Article Snippet: For the T-regulatory cell (Tregs) percentages, cells (1 × 10 6 ) were incubated with 10 μg/mL of phycoerythrin-conjugated mouse anti-chicken CD25 antibody [ ], 1:250 dilution of FITC-d- conjugated mouse anti-chicken CD4+ (Southern Biotechnology Associates, Birmingham, AL, USA), and a 1:200 dilution of unlabeled mouse IgG for 45 min.

Techniques: Flow Cytometry, Control