Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: High-Dimensional Flow Cytometry Analysis of Regulatory Receptors on Human T Cells, NK Cells, and NKT Cells

doi: 10.1007/978-1-0716-0849-4_14

Figure Lengend Snippet: Antibodies and reagents

Article Snippet: 2.5 μl 15 CD4 L3T4 BUV805 BD Biosciences Lymph.

Techniques:

Journal: British Journal of Cancer

Article Title: Treatment with interleukin-2 in malignant pleural mesothelioma: immunological and angiogenetic assessment and prognostic impact

doi: 10.1038/sj.bjc.6605438

Figure Lengend Snippet: Correlations between immunological and angiogenetic parameters in the IL-2-treated patients’ group and in the untreated patients’ group

Article Snippet: For TIL-immunohistochemical staining, sections were incubated with the following antibodies: mouse anti-human Foxp3 monoclonal antibody (clone 236A/E7 diluted 1 : 300; Abcam, Cambridge, UK) , mouse anti-human CD8 monoclonal antibody (clone C8/144B, ready to use for the Ventana automated slide stainer; Ventana) , and mouse anti-human CD4 monoclonal antibody (clone 1F6 diluted 1 : 20; Diagnostic BioSystem, Pleasanton, CA, USA).

Techniques:

Journal: British Journal of Cancer

Article Title: Treatment with interleukin-2 in malignant pleural mesothelioma: immunological and angiogenetic assessment and prognostic impact

doi: 10.1038/sj.bjc.6605438

Figure Lengend Snippet: Association of immunological and angiogenetic variables with clinicopathologic parameters

Article Snippet: For TIL-immunohistochemical staining, sections were incubated with the following antibodies: mouse anti-human Foxp3 monoclonal antibody (clone 236A/E7 diluted 1 : 300; Abcam, Cambridge, UK) , mouse anti-human CD8 monoclonal antibody (clone C8/144B, ready to use for the Ventana automated slide stainer; Ventana) , and mouse anti-human CD4 monoclonal antibody (clone 1F6 diluted 1 : 20; Diagnostic BioSystem, Pleasanton, CA, USA).

Techniques:

Journal: British Journal of Cancer

Article Title: Treatment with interleukin-2 in malignant pleural mesothelioma: immunological and angiogenetic assessment and prognostic impact

doi: 10.1038/sj.bjc.6605438

Figure Lengend Snippet: Univariate analysis of overall survival and time to progression in MPM patients treated with IL-2

Article Snippet: For TIL-immunohistochemical staining, sections were incubated with the following antibodies: mouse anti-human Foxp3 monoclonal antibody (clone 236A/E7 diluted 1 : 300; Abcam, Cambridge, UK) , mouse anti-human CD8 monoclonal antibody (clone C8/144B, ready to use for the Ventana automated slide stainer; Ventana) , and mouse anti-human CD4 monoclonal antibody (clone 1F6 diluted 1 : 20; Diagnostic BioSystem, Pleasanton, CA, USA).

Techniques:

Journal: Frontiers in Immunology

Article Title: Identification of a DRB3*011:01-restricted CD4 + T cell response against bovine respiratory syncytial virus fusion protein

doi: 10.3389/fimmu.2023.1040075

Figure Lengend Snippet: Stimulation of CD4 + T cells with synthetic, overlapping peptides from the BRSV-375 F protein. CD4 + T cells from dairy cows were positively selected and cultured with autologous antigen presenting cells loaded with synthetic peptide. T cell activation was assessed via IFN-γ secretion using ELISpot assays. Only cells from DRB3*011:01 positive animals responded to stimulation with five out of 88 peptides tested. T cell responses from animals 1141 (A) , 1310 (B) , 2938 (C) , and 2942 (D) displayed a measurable T cell response. Peptides sharing N-terminal amino acid with BRSV-375 F protein residue 253 generated a slight response while those with N-terminal amino acids matching residues 257-260 displayed a larger response. T cells were stimulated with PHA at a concentration of 5μg/mL as a positive control, and cRPMI media as a negative control. Data presented as mean ± SEM, p < 0.05 was considered significant and indicated by asterisk.

Article Snippet: Cell suspensions were labeled with the same mouse anti-bovine CD4 monoclonal antibody as above followed by goat anti-mouse IgG1-AlexaFluor 488 then analyzed on a FACS Symphony custom flow cytometer (BD Biosciences, Franklin Lakes, New Jersey, USA).

Techniques: Cell Culture, Activation Assay, Enzyme-linked Immunospot, Generated, Concentration Assay, Positive Control, Negative Control

Journal: Frontiers in Immunology

Article Title: Identification of a DRB3*011:01-restricted CD4 + T cell response against bovine respiratory syncytial virus fusion protein

doi: 10.3389/fimmu.2023.1040075

Figure Lengend Snippet: Identification of the minimum peptide of a CD4 + T cell epitope in BRSV-375 F protein. C-terminal truncated peptides were used to stimulate CD4 + T cells in culture with autologous DRB3*011:01 antigen presenting cells. T cell stimulation was observed for peptides greater than 8 amino acids in length. The 9-mer minimum peptide sequence of INDMPITND was identified. T cells were stimulated with PHA at a concentration of 5μg/mL as a positive control, and cRPMI media as a negative control. Cows 1141 (A) and 2942 (D) had a significantly higher response to peptides containing the core INDMPITND sequence while T cells from 1310 (B) , 2938 (C) , 1503 (E) responded to these peptides, the results were not significant. T cells collected from cow 1613 (F) did not produce an observable response to peptide stimulation. Data presented as mean ± SEM, p < 0.05 was considered significant and indicated by asterisk.

Article Snippet: Cell suspensions were labeled with the same mouse anti-bovine CD4 monoclonal antibody as above followed by goat anti-mouse IgG1-AlexaFluor 488 then analyzed on a FACS Symphony custom flow cytometer (BD Biosciences, Franklin Lakes, New Jersey, USA).

Techniques: Cell Stimulation, Sequencing, Concentration Assay, Positive Control, Negative Control

Journal: Frontiers in Immunology

Article Title: Identification of a DRB3*011:01-restricted CD4 + T cell response against bovine respiratory syncytial virus fusion protein

doi: 10.3389/fimmu.2023.1040075

Figure Lengend Snippet: Inhibition of BoLA DR, but not BoLA DQ, significantly reduces CD4 + T cell activation. Antigen presenting cells were incubated with antibodies against BoLA-DR or BoLA-DQ molecules prior to stimulation with synthetic overlapping peptides (A) or C-terminal truncated peptides (B) . Antibodies raised against MHC class II molecules were provided as mouse ascites fluid. A reduction of in T cell stimulation was observed only with following pre-treatment with anti-BoLA DR antibodies (A) . Antibodies were purified to ensure the reduction in T cell stimulation was due to antibody binding and not background from ascites fluid (B) . Anti-bovine IL-4 antibody was used as an isotype control. Cells were cultured in cRPMI as a media only control. Data is presented as mean ± SEM, p < 0.05 was considered significant and indicated by asterisk.

Article Snippet: Cell suspensions were labeled with the same mouse anti-bovine CD4 monoclonal antibody as above followed by goat anti-mouse IgG1-AlexaFluor 488 then analyzed on a FACS Symphony custom flow cytometer (BD Biosciences, Franklin Lakes, New Jersey, USA).

Techniques: Inhibition, Activation Assay, Incubation, Cell Stimulation, Purification, Binding Assay, Cell Culture

Journal: Frontiers in Immunology

Article Title: Identification of a DRB3*011:01-restricted CD4 + T cell response against bovine respiratory syncytial virus fusion protein

doi: 10.3389/fimmu.2023.1040075

Figure Lengend Snippet: CD4 + T cell response to computationally predicted peptides presented by artificial antigen presenting cells (aAPC). Synthetic peptides computationally predicted to bind strongly to select BoLA-DRB3 alleles used to stimulate CD4 + T cells after loading to aAPCs expressing BoLA-DR and CD80 molecules. DRB3*011:01 predicted peptides, with a core sequence of INDMPITND, were observed to induce T cell activation (B) while those predicted to bind to *001:01 (A) , *012:01 (C) , and *014:01:01 (D) did not. Cells were stimulated with 5μg/mL PHA as a positive control. Data presented as mean ± SEM, p < 0.05 was considered significant and indicated by asterisk.

Article Snippet: Cell suspensions were labeled with the same mouse anti-bovine CD4 monoclonal antibody as above followed by goat anti-mouse IgG1-AlexaFluor 488 then analyzed on a FACS Symphony custom flow cytometer (BD Biosciences, Franklin Lakes, New Jersey, USA).

Techniques: Expressing, Sequencing, Activation Assay, Positive Control