hla a2 pe cy7  (R&D Systems)

Journal: bioRxiv

Article Title: Hypoimmunogenic hPSC-derived cardiac organoids for immune evasion and heart repair

doi: 10.1101/2025.04.09.648007

Figure Lengend Snippet: a-d, Bar charts of flow cytometry data showing expression of HLA-A2, HLA-BC, and HLA-E as percent positive expression in ( a ) hPSC-CMs, ( b ) hPSC-cFbs, ( c ) hPSC-ECs, and ( d ) hPSC-PCs by wild-type hPSC-derived cells (red) and UDET hPSC-derived cells (blue). e , Timeline of T cell cytotoxicity assay to measure T cell-mediated lysis of target cells. f-i , Percent lysis as quantified from LDH assay following exposure of activated CD8+ T cells to wild-type (red) and UDET (blue)-derived ( f ) hPSC-CMs, ( g ) hPSC-cFbs, ( h ) hPSC-ECs, and ( i ) hPSC-PCs. j , Timeline of NK cytotoxicity assay to measure NK cell-mediated lysis of target cells. k-n , Percent lysis as quantified from LDH assay following exposure of NK cells to wild-type (red) and UDET (blue)-derived ( k ) hPSC-CMs, ( l ) hPSC-cFbs, ( m ) hPSC-ECs, and ( n ) hPSC-PCs. Data shown from one donor representative of 2 biological replicates with 3-5 technical replicates. nd=no difference, *=p<0.05.

Article Snippet: Pre-conjugated antibodies used: B2M-APC (1:100; Biolegend, 316312, clone 2M2), HLA-A/B/C-PE (1:40; Biolegend, 311406, clone W6/32), HLA-E-APC (1:40; Biolegend, 342606, clone 3D12), HLA-G-APC (1:100; Biolegend, 335909, clone 87G), HLA-E-APC (1:50; Invitrogen, 17995342, clone 3D12HLA-E), HLA-BC-APC (1:50; Invitrogen, 17593542, clone B1.23.2), HLA-A2-PE-Cy7, clone BB7.2), CD25-APC (1:50; R&D Systems, FAB1020A, clone 24212), CD45-BV421 (1:50; Biolegend, 304032, clone HI30), CD3-BV605 (1:50; Biolegend, 344836, clone SK7), CD8-APC (1:50; Biolegend, 344722, clone SK1), CD4-PE (1:50; Biolegend, 980804, clone SK3).

Techniques: Flow Cytometry, Expressing, Derivative Assay, Cytotoxicity Assay, Lysis, Lactate Dehydrogenase Assay

Journal: Frontiers in Immunology

Article Title: POTN: A Human Leukocyte Antigen-A2 Immunogenic Peptides Screening Model and Its Applications in Tumor Antigens Prediction

doi: 10.3389/fimmu.2020.02193

Figure Lengend Snippet: Overview of the immunogenicity and HLA-A2 binding affinity of candidate peptides predicted by the POTN model.

Article Snippet: In brief, T2 cells (500 μL, 1 × 10 6 cells/mL) were incubated with the peptide (25 μg, 50 μg/mL; dissolved in DMSO at a concentration of 10 mg/mL) in serum-free IMDM medium, supplemented with human β2-microglobulin (3 µg/mL, Merck, USA) at 37°C for 18 h. The T2 cells were washed twice and incubated with the anti-human HLA-A2-PE-cy7 antibody (BB7.2, eBioscience, USA) at 4°C for 30 min.

Techniques: Binding Assay

Journal: Frontiers in Immunology

Article Title: POTN: A Human Leukocyte Antigen-A2 Immunogenic Peptides Screening Model and Its Applications in Tumor Antigens Prediction

doi: 10.3389/fimmu.2020.02193

Figure Lengend Snippet: Binding affinity and ICS assay for the peptides identified using the POTN model. (A) Identified peptides categorized based on binding affinity to HLA-A2. ICS assay for each peptide in (B) donor 1, (C) donor 2, (D) donor 3, (E) donor 4, and (F) donor 5, (G) Number of immunogenic peptides in each group (e.g., “1/5” indicates that in the group, one peptide elicited T cell response in one donor [1/5]; “2/5” indicates that in the group, one peptide elicited T cell response in two donors [2/5]). (H) Number of the donors in which an immune response was elicited by the identified peptides. (I) Enrichment curves.

Article Snippet: In brief, T2 cells (500 μL, 1 × 10 6 cells/mL) were incubated with the peptide (25 μg, 50 μg/mL; dissolved in DMSO at a concentration of 10 mg/mL) in serum-free IMDM medium, supplemented with human β2-microglobulin (3 µg/mL, Merck, USA) at 37°C for 18 h. The T2 cells were washed twice and incubated with the anti-human HLA-A2-PE-cy7 antibody (BB7.2, eBioscience, USA) at 4°C for 30 min.

Techniques: Binding Assay