Cd3 apc r700 ucht1

Immunophenotyping was performed on peripheral blood samples collected longitudinally and at the study endpoint on whole blood and on mononuclear cells isolated from the tissues of humanized mice. Prior to antibody incubation, Ig-binding sites were blocked. The fluorochrome-conjugated antibodies included CD45-V500 (HI30, BD Biosciences, San Jose, USA), CD3-APC-R700 (UCHT1, BD Biosciences, San Jose, USA), CD4-APC-H7 (RPA-T4, BD Biosciences, San Jose, USA), CD8-FITC (SK1, BD Biosciences, San Jose, USA), CD19-PE-Cy7 (SJ25C1, BD Biosciences, San Jose, USA), CD27-PE (M-T271, BD Biosciences, San Jose, USA), CD38-APC (HB7, BD Biosciences, San Jose, USA), CD45RA-Pacific Blue (F8-11-13, Bio-Rad, Hercules, USA), HLA-DR-PerCP (L243, BD Biosciences, San Jose, USA), mouse IgG1k-APC (MOPC-21, BD Biosciences, San Jose, USA), mouse IgG1k- Pacific Blue (MOPC-21, BD Biosciences, San Jose, USA), mouse IgG1k-PE (MOPC-21, BD Biosciences, San Jose, USA) and mouse IgG2ak-PerCP (X39, BD Biosciences, San Jose, USA). After antibody incubation, blood samples were lysed with t 1× BD FACS lysing solution (BD Biosciences, San Jose, USA). Samples were then analyzed on a BD LSRFortessa instrument (BD Biosciences, San Jose, USA).

The following monoclonal antibodies were used for FACS: CCR7 APC-Cy7 (G043H7, Biolegend, 353211), CD69 PE (FN50, BD, 555531), CD3 APC R700 (UCHT1, BD, 565119), PD-1 BV421 (EH12.2H7, Biolegend, 329920), HLA-DR BV605 (G46-6, BD, 562845), CXCR5 BB515 (RF8B2, BD, 564624), CD14 PE-Cy5 (61D3, abcam, ab25395), CD16 PE-Cy5 (3G8, Biolegend, 3012010), CD19 PE-Cy5 (HIB19, Biolegend, 302210), CD45RA PE CF594 (HI100, BD, 562298), CD8 PE-Cy5.5 (RPA-T8, eBioscience, 35-0088-42), and CD4 PE Cy7 (RPA-T4, Biolegend, 300512).