Comprehensive Immunophenotyping of Nonhuman Primate Cells

Flow cytometric analysis was performed on PBMCs and RB-derived cells according to standard procedures using a panel of mAbs that others and we have shown to be cross-reactive with RM immune cells (10 (link), 19 (link)) (Supplementary Table 2). The following Abs were used: anti–CD4-APCCy7 (clone OKT4), anti–HLA-DR-BV711 (clone L243), and anti-CD20 PerCpCy5.5 (clone 2H7) all from Biolegend, San Diego, CA, USA; anti–CD95-CF594 (clone DX2), anti–beta7-PECy5 (clone FIB504), anti–CCR7-PECy7 (clone 3D12), anti–Ki67-Alexa700 (clone B56), anti–CD3-BUV395 (clone SP34-2), anti–CD8-BUV496 (clone RPA-T8), anti–CD56-BV605 (clone B159), and anti–CD16-BV650 (clone 3G8) all from Becton–Dickinson, BD Biosciences, San Jose, CA, USA; anti–NKG2A-APC (clone Z199), from Beckman Coulter, Brea, CA, USA; Aqua Live/Dead amine dye-AmCyan from ThermoFisher Scientific, Invitrogen, Waltham, MA, USA; anti–CD38-FITC (clone AT-1) from STEMCELL Technologies, Vancouver, British Columbia, Canada; and anti–α4β7-PE (clone Act-1) obtained from the NIH Non-human Primate Reagent Resource, University of Massachusetts Medical School. Flow cytometric acquisition was performed on at least 100,000 CD3⁺ T cells on a BD LSRII Flow Cytometer driven by BD FACSDiva software. Analyses of the acquired data were performed by FlowJo software, Tree Star, Inc., Ashland, OR, USA.

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Pino M., Uppada S.B., Pandey K., King C., Nguyen K., Shim I., Rogers K., Villinger F., Paiardini M, & Byrareddy S.N. (2020). Safety and Immunological Evaluation of Interleukin-21 Plus Anti-α4β7 mAb Combination Therapy in Rhesus Macaques. Frontiers in Immunology, 11, 1275.

Publication 2020

anti–NKG2A-APC (clone Z199), BD LSRII Flow Cytometer

Amine Anti beta7 Anti cd3 Cells Clone Cross reactive Fitc Flow cytometric Hla dr Human resource Mabs Primate Stemcell T cells Tree

Corresponding Organization : University of Nebraska Medical Center

Other organizations : University of Louisiana at Lafayette

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Variable analysis

independent variables

Flow cytometric analysis was performed on PBMCs and RB-derived cells

dependent variables

Expression of CD4, HLA-DR, CD20, CD95, beta7, CCR7, Ki67, CD3, CD8, CD56, CD16, NKG2A, CD38, and alpha4beta7

control variables

Standard procedures for flow cytometric analysis were used
A panel of monoclonal antibodies (mAbs) that are known to be cross-reactive with RM immune cells was used

controls

Positive control: None specified
Negative control: None specified

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