anti-ccr7–pe-cy7 3d12  (Becton Dickinson)

Journal: Nature

Article Title: A common allele of HLA is associated with asymptomatic SARS-CoV-2 infection

doi: 10.1038/s41586-023-06331-x

Figure Lengend Snippet: a – c , Ex vivo combinatorial tetramer analysis for the four indicated peptides was performed in nine pre-pandemic donor samples. a , b , The number of donors with detectable tetramer + CD8 + T cells ( a ) and their frequencies ( b ) are shown. c , The proportion of naive (CD45RA + CCR7 + ) and memory (combination of CD45RA − CCR7 − , CD45RA − CCR7 + , CD45RA + CCR7 − ) cells among tetramer + CD8 + T cells (based on n = 9 samples). d , Phenotypic analysis after TAME of ex vivo tetramer + NQK-Q8-specific (tet-Q8) and NQK-A8-specific (tet-A8) T cells in 7 and 6 donors, respectively. Cell types were defined as follows: T naive (CD45RA + CCR7 + CD95 − ); T SCM (stem cell memory, CD45RA + CCR7 + CD95 + ); T CM (central memory, CD45RA − CCR7 + ); T EM (effector memory, CD45RA − CCR7 − ); T EMRA (terminally differentiated, CD45RA + CCR7 − ). Data are mean ± s.e.m.

Article Snippet: After enrichment, cells were stained with an antibody panel including anti-CD3-BV480 (dilution 1:100), anti-CD8-PerCP-Cy5.5 (1:50), anti-CD4-BV650 (1:100), anti-CD14-APCH7 (1:200), anti-CD19-APCH7 (1:100), anti-CD45RA-FITC (1:100), anti-CD27-APC (1:100), anti-CCR7-PE-Cy7 (1:50), anti-CD95-BV421 (1:50), anti-PD1-BV605 (1:100) (all BD Biosciences) and Live/Dead Fixable Near-IR Dead Cell Stain (1:1,000) (Life Technologies) (Supplementary Fig. ).

Techniques: Ex Vivo

Journal: Nature

Article Title: A common allele of HLA is associated with asymptomatic SARS-CoV-2 infection

doi: 10.1038/s41586-023-06331-x

Figure Lengend Snippet: ( a,b ) FACS plots of NQK-A8-tetramer + CD8 + T cells following tetramer magnetic enrichment ( a ) and NQK-Q8-tetramer + CD8 + T cells ( b ). The NQK-A8- and NQK-Q8- tetramer + CD8 + T cells shown as blue dots were plotted versus CD3 + T cells shown as grey dots and gated on CCR7 and CD45RA to determine memory status, following tetramer magnetic enrichment for each donor. ( c ) The percentage of tetramer + of CD8 + T cells after tetramer magnetic enrichment is reported for each donor with the purple bar for the Tet-Q8 tetramer and the orange bar for the Tet-A8 tetramer (n = 7 and n = 6 biologically independent samples, respectively). Data are presented as median values with IQR (interquartile range). The individual percentage for each donor is indicated on panels a and b . ( d ) The proportion of T naïve (CD45RA + CCR7 + CD95 - ); T SCM (stem cell memory, CD45RA + CCR7 + CD95 + ); T CM (central memory, CD45RA - CCR7 + ); T EM (effector memory, CD45RA - CCR7 - ); T EMRA (terminally differentiated, CD45RA + CCR7 - ) cells in different donors.

Article Snippet: After enrichment, cells were stained with an antibody panel including anti-CD3-BV480 (dilution 1:100), anti-CD8-PerCP-Cy5.5 (1:50), anti-CD4-BV650 (1:100), anti-CD14-APCH7 (1:200), anti-CD19-APCH7 (1:100), anti-CD45RA-FITC (1:100), anti-CD27-APC (1:100), anti-CCR7-PE-Cy7 (1:50), anti-CD95-BV421 (1:50), anti-PD1-BV605 (1:100) (all BD Biosciences) and Live/Dead Fixable Near-IR Dead Cell Stain (1:1,000) (Life Technologies) (Supplementary Fig. ).

Techniques:

Journal: Frontiers in Immunology

Article Title: TMEM123 a key player in immune surveillance of colorectal cancer

doi: 10.3389/fimmu.2023.1194087

Figure Lengend Snippet: Table 1 highlights the reagents, recombinant DNA and oligonucleotides, cell lines, software, and source data essential to reproduce results presented in the manuscript.

Article Snippet: anti-human-CCR7 PE-Cy7 , BD Biosciences , Cat #560922; 3D12.

Techniques: Recombinant, Software, Antibody Labeling, Activation Assay, Labeling, Protease Inhibitor, Staining, In Vivo, Transfection, Blocking Assay, SYBR Green Assay, Fractionation, Bicinchoninic Acid Protein Assay, Western Blot, Membrane, Expressing, TA Cloning, Plasmid Preparation, Sequencing, Control

Journal: Haematologica

Article Title: Complement protein C3a enhances adaptive immune responses towards FVIII products

doi: 10.3324/haematol.2022.281762

Figure Lengend Snippet: Recombinant FVIII co-stimulation with lipopolysaccharide plus plasma mainly induces proliferation of central memory T cells. Dendritic cells (DC) from HLA-DRB1*11-positive donors were treated with recombinant (r)FVIII (1 IU/mL), lipopolysaccharide (LPS) (0.1 mg/mL), and plasma (2.5 mL/mL). After 24 hours, carboxyfluorescein succinimidyl ester (CFSE)-labeled autologous CD4 + T cells were added. On day 9 of co-culture, T cells were harvested and stained with HLA-matched FVIII-specific tetramer and an antibody mix containing anti-CD3, anti-CD4, anti-CD45RA, anti-CD45RO, and anti-CCR7 antibodies. The CD4 + T cells were defined as 4 subsets: central memory (CM) T cells (CD45RO + CD45RA - CCR7 + ), effector memory (EM) T cells (CD45RO + CD45RA - CCR7 - ), naïve T cells (CD45RO - CD45RA + CCR7 + ), and effector T cells (TE) (CD45RO - CD45RA + CCR7 - ). (A) Data from 1 representative donor are shown. (B) Percentage distribution of CD4 + T-cell subtypes from proliferated CD4 + T cells (n=17, 5 donors from 7 independent experiments) and (C) from proliferated CD4 + FVIII-specific T cells (n=7, 6 donors from 2 independent experiments) are summarized. Error bars indicate the mean ± standard error of the mean. SSC: side scatter; FSC: forward scatter.

Article Snippet: T-cell surface markers were stained for 20 min at 4°C using the following antibodies: anti-CD3-AmCyan (clone SK7, BD Pharmingen), anti-CD4-PacBlue, anti-CD4-APC (both clone RPA-T4, BD Pharmingen), anti-CD45RA-PerCP (clone HI100, BioLegend), anti-CD45RO-AF700 (clone IV N31, BioLegend), and anti-CCR7-PE-Cy7 (clone 3C12, BD Pharmingen).

Techniques: Recombinant, Labeling, Co-Culture Assay, Staining