fluorochrome conjugated mouse anti-sheep cd4-fitc  (Bio-Rad)

Journal: Frontiers in Immunology

Article Title: Recombinant antigen P29 of Echinococcus granulosus induces Th1, Tc1, and Th17 cell immune responses in sheep

doi: 10.3389/fimmu.2023.1243204

Figure Lengend Snippet: CD4 + and CD8 + T cells in PBMCs. Peripheral blood samples were collected at different immunization times, and CD4 + and CD8 + T cells in PBMCs were analyzed using flow cytometry. (A) Proportion of CD4 + and CD8 + T cells in PBMCs of each group at week 8. (B, C) show the trend of the proportion of CD4 + and CD8 + T cells at different times, respectively. (D) The ratio of CD4 + to CD8 + T cells in each group of PBMCs at week 8. (E) The trend of the ratio of CD4 + to CD8 + T cells at different times. Data were obtained from 9 sheep, and results are presented as mean ± SD ( ns , P > 0.05).

Article Snippet: To determine the cellular phenotypes of PBMCs, cells were directly stained with mouse anti-sheep CD4 (Clone: 44.38) and mouse anti-sheep CD8 (Clone: 38.65) antibodies (Bio-Rad AbD Serotec, Munich, USA), and incubated at 4°C for 30 min in the dark, followed by two washes with washing buffer.

Techniques: Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Recombinant antigen P29 of Echinococcus granulosus induces Th1, Tc1, and Th17 cell immune responses in sheep

doi: 10.3389/fimmu.2023.1243204

Figure Lengend Snippet: Cytokines produced in PBMCs by flow cytometry. PBMCs were stimulated with rEg.P29, and the cells were collected and labeled with antibodies to detect IFN-γ and IL-17A production by CD4 + and CD8 + T cells using flow cytometry. (A) Representative flow scatter plots for detecting IFN-γ production by CD4 + and CD8 + T cells. (B, D) show the IFN-γ production by CD4 + and CD8 + T cells in each group at week 8, respectively. (C, E) show the tendency of IFN-γ production by CD4 + and CD8 + T cells at different times, respectively. (F) Representative flow scatter plots for detecting IL-17A production by CD4 + and CD8 + T cells. (G, I) show the IL-17A production by CD4 + and CD8 + T cells in each group of samples at week 8, respectively. (H, J) show the tendency of IL-17A production by CD4 + and CD8 + T cells at different times, respectively. Data were obtained from 9 sheep, and results are presented as mean ± SD (* P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001).

Article Snippet: To determine the cellular phenotypes of PBMCs, cells were directly stained with mouse anti-sheep CD4 (Clone: 44.38) and mouse anti-sheep CD8 (Clone: 38.65) antibodies (Bio-Rad AbD Serotec, Munich, USA), and incubated at 4°C for 30 min in the dark, followed by two washes with washing buffer.

Techniques: Produced, Flow Cytometry, Labeling

Journal: Frontiers in Immunology

Article Title: Recombinant antigen P29 of Echinococcus granulosus induces Th1, Tc1, and Th17 cell immune responses in sheep

doi: 10.3389/fimmu.2023.1243204

Figure Lengend Snippet: Cytokines produced in spleen lymphocytes by flow cytometry. Spleen lymphocytes were obtained after euthanasia of sheep, cells were labeled with antibodies after rEg.P29 stimulation, and IFN-γ and IL-17A production by CD4 + and CD8 + T cells was detected using flow cytometry. (A) Representative flow scatter plots for detecting IFN-γ production by CD4 + and CD8 + T cells. (B, C) show the IFN-γ production by CD4 + and CD8 + T cells in each group, respectively. (D) Representative flow scatter plots for detecting IL-17A production by CD4 + and CD8 + T cells. (E, F) show the IL-17A production by CD4 + and CD8 + T cells in each group, respectively. Data were obtained from 7 sheep, and results are presented as mean ± SD (**** P < 0.0001).

Article Snippet: To determine the cellular phenotypes of PBMCs, cells were directly stained with mouse anti-sheep CD4 (Clone: 44.38) and mouse anti-sheep CD8 (Clone: 38.65) antibodies (Bio-Rad AbD Serotec, Munich, USA), and incubated at 4°C for 30 min in the dark, followed by two washes with washing buffer.

Techniques: Produced, Flow Cytometry, Labeling

Journal: Frontiers in Immunology

Article Title: Recombinant antigen P29 of Echinococcus granulosus induces Th1, Tc1, and Th17 cell immune responses in sheep

doi: 10.3389/fimmu.2023.1243204

Figure Lengend Snippet: Cytokines produced in lymphocytes of mesenteric lymph nodes by flow cytometry. Lymphocytes of mesenteric lymph nodes were obtained after euthanasia of sheep, cells were labeled with antibodies after rEg.P29 stimulation, and IFN-γ, IL-4, and IL-17A production by CD4 + and CD8 + T cells was detected using flow cytometry. (A) Representative flow scatter plots for detecting IFN-γ production by CD4 + and CD8 + T cells. (B, C) show the IFN-γ production by CD4 + and CD8 + T cells in each group, respectively. (D) Representative flow scatter plots for detecting IL-17A production by CD4 + and CD8 + T cells. (E, F) show the IL-17A production by CD4 + and CD8 + T cells in each group, respectively. Data were obtained from 7 sheep, and results are presented as mean ± SD (**** P < 0.0001).

Article Snippet: To determine the cellular phenotypes of PBMCs, cells were directly stained with mouse anti-sheep CD4 (Clone: 44.38) and mouse anti-sheep CD8 (Clone: 38.65) antibodies (Bio-Rad AbD Serotec, Munich, USA), and incubated at 4°C for 30 min in the dark, followed by two washes with washing buffer.

Techniques: Produced, Flow Cytometry, Labeling

Journal: Frontiers in Immunology

Article Title: Recombinant antigen P29 of Echinococcus granulosus induces Th1, Tc1, and Th17 cell immune responses in sheep

doi: 10.3389/fimmu.2023.1243204

Figure Lengend Snippet: Proliferation of CD4 + and CD8 + T cells. PBMCs were labeled with CFSE, stimulated in vitro with rEg.P29, and the decrease in CFSE fluorescence intensity of labeled cells was detected using flow cytometry to assess the proliferation of CD4 + and CD8 + T cells. (A) Histogram plots of CFSE fluorescence of lymphocytes, CD4 + , and CD8 + T cells. (B–D) represent the proliferation frequencies of lymphocytes, CD4 + , and CD8 + T cells, respectively. Data were obtained from 5 sheep, and results are presented as mean ± SD (**** P < 0.0001).

Article Snippet: To determine the cellular phenotypes of PBMCs, cells were directly stained with mouse anti-sheep CD4 (Clone: 44.38) and mouse anti-sheep CD8 (Clone: 38.65) antibodies (Bio-Rad AbD Serotec, Munich, USA), and incubated at 4°C for 30 min in the dark, followed by two washes with washing buffer.

Techniques: Labeling, In Vitro, Fluorescence, Flow Cytometry

Journal: PLoS ONE

Article Title: CXCR4 Is Dispensable for T Cell Egress from Chronically Inflamed Skin via the Afferent Lymph

doi: 10.1371/journal.pone.0095626

Figure Lengend Snippet: Lymphocytes traveling in afferent lymph draining uninflamed or chronically inflamed skin (>d15 post induction of inflammation with CFA) were collected from sheep, and chemotaxis was tested toward mouse CCL21 and human CXCL12 in a Transwell chemotaxis assay. Migration of CD4, CD8 and γδ T cells is expressed as the percentage of each T cell subset that migrated to the lower chamber. Data points represent the mean migration of T cells from individual animals based on triplicate wells for each concentration, and the mean ± SEM for all animals analyzed is shown. N = 5–7 animals per condition and T cell subset.

Article Snippet: The following mouse anti-sheep mAbs were used: CD4 (clone 44.38; Serotec), CD8 (clone 38.65; Serotec), and γδ TCR (clone 86D; VMRD).

Techniques: Chemotaxis Assay, Migration, Concentration Assay

Journal: PLoS ONE

Article Title: CXCR4 Is Dispensable for T Cell Egress from Chronically Inflamed Skin via the Afferent Lymph

doi: 10.1371/journal.pone.0095626

Figure Lengend Snippet: ( A–C ) Adoptive transfer exit experiments with fetal liver chimeric mice as a source of donor splenocytes. As depicted in the experimental scheme ( A ), fetal liver chimeras were generated by reconstituting lethally irradiated CD45.1 + wildtype recipient mice with fetal liver cells from (CD45.2 + ) Cxcr4 −/− Ccr 7 −/− or (CD45.2 + ) Ccr7 −/− or (CD45.1 + ) wildtype donors. Splenocytes from the fetal liver chimeras were labeled with eFlour670 and CFSE, so that the combination of congenic maker and fluorescent label uniquely marked cells of each genotype. Labeled splenocytes of all three genotypes were mixed and co-injected into chronically inflamed footpads. ( B ) Analysis of migrated lymphocyte subsets recovered in the draining popliteal lymph nodes 12 h post cell transfer. Data is presented as the ratio of migrated to input cells for Ccr7 −/− or Cxcr4 −/− Ccr 7 −/− relative to wildtype cells of each subset. Connected lines represent individually analyzed recipient mice. Horizontal, non-connecting lines indicate the mean of each group. ( C ) Flow cytometric analysis of memory (CD45RB l °) versus naïve (CD45RB hi ) phenotype CD4 and CD8 T cells in injected and migrated populations for each genotype. Data combined from 2 experiments analyzing 5–10 recipient mice per experiment ( B ) or the cell phenotype of one out two experiment with similar results ( C ) is shown. WT, wildtype.

Article Snippet: The following mouse anti-sheep mAbs were used: CD4 (clone 44.38; Serotec), CD8 (clone 38.65; Serotec), and γδ TCR (clone 86D; VMRD).

Techniques: Adoptive Transfer Assay, Generated, Irradiation, Labeling, Injection

Journal: STAR Protocols

Article Title: Multiparameter flow cytometry assay to analyze the pulmonary T cell profiles in the ovine model of respiratory syncytial virus infection

doi: 10.1016/j.xpro.2022.101688

Figure Lengend Snippet:

Article Snippet: Anti-CD4 (Clone 44.38; mouse IgG2a; 1:400 dilution) , Bio-Rad , Cat# MCA2213GA.

Techniques: Virus, Recombinant, Sterility, Staining, Cell Culture, Hood, Flow Cytometry, Software

Journal: STAR Protocols

Article Title: Multiparameter flow cytometry assay to analyze the pulmonary T cell profiles in the ovine model of respiratory syncytial virus infection

doi: 10.1016/j.xpro.2022.101688

Figure Lengend Snippet:

Article Snippet: Anti-CD4 (Clone 44.38; mouse IgG2a; 1:400 dilution) , Bio-Rad , Cat# MCA2213GA.

Techniques: Virus, Recombinant, Sterility, Staining, Cell Culture, Hood, Flow Cytometry, Software