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OMIP-026

Phenotypic analysis of B and plasma cells in rhesus macaques

Berit Neumann, Sieghart Sopper, Christiane Stahl-Hennnig Cytometry PART A, Volume 87, Issue 9, 800-802 (2015)

PURPOSE: Our purpose was a broad phenotypic analysis of B and plasma cells regarding differentiation status, activation, proliferation, and chemokine receptor expression in rhesus macaques. We developed two staining panels, which were tested on fresh samples of whole blood or peripheral blood mononuclear cells (PBMCs), bone marrow collected from the iliac crest and femur, lymph nodes, spleen, and tonsils (Figure 1, Table 1). A 10-color-panel was developed to mainly focus on B cells, whereas a 11-color panel concentrated on plasmablasts/plasma cells. Both panels are also applicable for whole blood and bone marrow samples from African green monkeys.

CELL TYPE: Fresh whole blood, PBMCs, mononuclear cells of bone marrow, lymph node, spleen, tonsil

MACHINE: BD LSR2

Marker Target Clone Fluorochrome Purpose
CD45 Rhesus macaque V500 Leukocyte (backbone)
CD10 Rhesus macaque HI10a APC-Cy7 Maturation marker (backbone)
CD3 Rhesus macaque SP34-2 Alexa Fluor 700 Lineage (backbone)
CD20 Rhesus macaque L27 PE-Cy7 Lineage (backbone)
CD21 Rhesus macaque B-ly4 FITC Differentiation (B cell)
CD27 Rhesus macaque M-T271 APC Differentiation (B cell)
CD69 Rhesus macaque TP1.55.3 ECD Activation (B cell)
CD80 Rhesus macaque L307.4 PE Activation (B cell)
ccr7 Rhesus macaque G043H7 Brilliant Violet 421 Homing (B cell)
IgD Rhesus macaque polyclonal Biotin Ig (B cell)
CD19 Rhesus macaque J3-119 PE Lineage (plasma cell)
CD27 Rhesus macaque O323 Brilliant Violet 650 Differentiation (plasma cell)
CD38 Rhesus macaque OKT10 APC Differentiation (plasma cell)
CD138 Rhesus macaque DL-101 FITC Plasma (plasma cell)
CD95 Rhesus macaque Dx2 Biotin Activation (plasma cell)
CD184 Rhesus macaque 12G5 PE-CF594 Homing (plasma cell)
Ki-67 Rhesus macaque B56 PerCP-Cy5.5 Proliferation (plasma cell)
Biotin Rhesus macaque Brilliant Violet 570 secondary reagent for CD95 and IgD (plasma cell)