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OMIP-012

Phenotypic and numeric determination of human leukocyte reconstitution in humanized mice

Brian R. Long, Cheryl A. Stoddart Cytometry PART A, Volume 81A, Issue 8, 646-648 (2012)

PURPOSE: This panel was developed to determine both the frequency and absolute number of human leukocytes and leukocyte subsets present in the peripheral blood of humanized mice. The panel also provides information concerning the activation state of peripheral leukocytes by cell surface staining for HLA-DR and CD38, relevant to studies of HIV disease pathogenesis (1–3). This panel has been used with EDTA anticoagulated whole blood in conjunction with bead-based enumeration for quantitative assessment of human cell chimerism. This panel also works well for staining of peripheral blood mononuclear cells (PBMCs) prepared by density gradient centrifugation and for dispersed splenocytes. The multicolor panel described here has been used in studies of humanized mouse reconstitution (4) and longitudinal studies of HIV pathogenesis in NSG-BLT mice (5).

CELL TYPE: Whole blood, Ficoll-separated PBMC, splenocytes

MACHINE: BD LSR2

Marker Target Clone Fluorochrome Purpose
CD19 Human HIB19 APC-Cy7 human B cell
CD45 Human HI30 Alexa Fluor 700 human leukocyte
CD45 Mouse 30-F11 APC excusion of mouse leukocyte
CD56 Human HCD56 PE-Cy7 human NK
CD3 Human UCHT1 ECD human T cell
CD38 Human HIT2 PE activation
HLA-DR Human L243 FITC activation
CD8 Human 3B5 Qdot 605 human CD8
CD4 Human RPA-T4 Pacific Blue human CD4
Isotype Ctrl Mouse MOPC-21 PE set gate for CD38
CD94 Human DX22 FITC NK
CD16 Human 3G8 PerCP NK
CD314 Human 1D11 PE NK
CD159c Human 134591 APC NK