OMIP-026
Phenotypic analysis of B and plasma cells in rhesus macaques
Cytometry PART A, Volume 87, Issue 9, 800-802 (2015)
Berit Neumann, Sieghart Sopper, Christiane Stahl-Hennnig
Description: 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. 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.
Species: African green monkey, Rhesus macaque
Tissue: Bone marrow, Lymph node, Peripheral blood, Spleen, Tonsil
Cell Subset: Plasma cell/ asc, B cell, B memory
Keyword: Activation, Lysed whole blood (lwb), Proliferation
Machine: BD LSR2
Panel details
| Marker | Species | Clone | Fluorophore | Purpose | Co-expressions groups | Antigen Density |
|---|---|---|---|---|---|---|
| CD3 | Rhesus macaque | SP34-2 | Alexa Fluor 700 | Lineage (backbone) | High | |
| CD20 | Rhesus macaque | L27 | PE-Cy7 | Lineage (backbone) | B cell | Med |
| CD21 | Rhesus macaque | B-ly4 | FITC | Differentiation (B cell) | B cell | Low |
| CD27 | Rhesus macaque | M-T271 | APC | Differentiation (B cell) | B cell | Med |
| CD69 | Rhesus macaque | TP1.55.3 | ECD | Activation (B cell) | B cell | Med |
| CD80 | Rhesus macaque | L307.4 | PE | Activation (B cell) | B cell | Low |
| CD19 | Rhesus macaque | J3-119 | PE | Lineage (plasma cell) | plasma cell, B cell | Low |
| CD27 | Rhesus macaque | O323 | Brilliant Violet 650 | Differentiation (plasma cell) | plasma cell | Med |
| CD138 | Rhesus macaque | DL-101 | FITC | Plasma (plasma cell) | plasma cell | Med |
| Ki-67 | Rhesus macaque | B56 | PerCP-Cy5.5 | Proliferation (plasma cell) | B cell, apoptosis | Low |
| Biotin | Rhesus macaque | Brilliant Violet 570 | secondary reagent for CD95 and IgD (plasma cell) | B cell | ||
| CCR7 | Rhesus macaque | G043H7 | Brilliant Violet 421 | Homing (B cell) | B cell | Low |
| IgD | Rhesus macaque | polyclonal | Ig (B cell) | B cell | High | |
| CD95 | Rhesus macaque | Dx2 | Activation (plasma cell) | B cell, apoptosis | Low | |
| CD184 | Rhesus macaque | 12G5 | PE-CF594 | Homing (plasma cell) | plasma cell | Med |
| CD45 | Rhesus macaque | D058-128 | V500 | Leukocyte (backbone) | B cell, plasma cell | High |
| CD38 | Rhesus macaque | OKT10 | APC | Differentiation (plasma cell) | plasma cell, B cell | High |
| CD10 | Rhesus macaque | HI10a | APC-Cy7 | Maturation marker (backbone) | High |
