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Updated: Jun 11, 2026
Multicolor Flow Cytometry Analyses of Cellular Immune Response in Rhesus Macaques
Published on: April 22, 2010
Immunophenotyping of lymphocyte, monocyte and dendritic cell subsets in normal rhesus macaques by 12-color flow
Patrick Autissier1, Caroline Soulas, Tricia H Burdo
1Department of Biology, Boston College, Higgins Hall 468, 140 Commonwealth Avenue, Chestnut Hill, MA 02467, USA.
Insights
This study developed a 12-color flow cytometry panel to simultaneously analyze rhesus macaque immune cells. This comprehensive panel aids in understanding immune responses during infectious diseases like SIV.
Area of Science:
- Immunology
- Primate Research
- Flow Cytometry
Background:
- Monitoring immune cell dynamics in rhesus macaques is vital for infectious disease research.
- Existing methods often require multiple panels, limiting simultaneous analysis.
Purpose of the Study:
- To develop and validate a single 12-color flow cytometry panel for comprehensive immune cell phenotyping in rhesus macaques.
- To enable simultaneous analysis of lymphocyte, monocyte, and dendritic cell subsets.
Main Methods:
- A 12-color flow cytometry panel was designed using a cocktail of 12 antibodies.
- Blood samples from healthy rhesus macaques were analyzed.
- Data were compared against smaller, lineage-specific panels.
Main Results:
- The panel successfully identified major immune cell subsets, including T lymphocytes (CD4+, CD8+), B cells, NK cells, NKT cells, and monocyte subsets.
- Four distinct Lin-HLA-DR+ subsets were identified: CD34+ hematopoietic stem cells, plasmacytoid dendritic cells (DCs), and two types of myeloid DCs (CD11c+ CD16+ and CD11c(-/dim) CD1c+).
- The 12-color panel provided comparable and comprehensive data to smaller panels.
Conclusions:
- A single, multiparameter flow cytometry panel allows for the simultaneous enumeration of key immune cell populations in rhesus macaques.
- This approach offers a broader perspective on immune responses, particularly during Simian Immunodeficiency Virus (SIV) infection.
- It facilitates a better understanding of individual cell types' roles in AIDS pathogenesis.
Abstract:
Monitoring changes in rhesus macaque immune cell populations during infectious disease is crucial. The aim of this work was to simultaneously analyze the phenotype of rhesus macaque lymphocyte, monocyte and dendritic cell (DC) subsets using a single 12-color flow cytometry panel. Blood from healthy non-infected rhesus macaques was labeled with a cocktail of 12 antibodies. Data were compared to three smaller lineage specific panels and absolute and relative percentages of cells were compared. Our 12-color panel allows for the identification of the following major subsets: CD4+ and CD8+ T lymphocytes, B lymphocytes, natural killer (NK) cells, natural killer T (NKT) cells, monocyte subsets and four non-overlapping Lin-HLA-DR+ cell subsets: CD34+ hematopoietic stem cells, CD11c- CD123+ plasmacytoid DC, CD11c+ CD16+ and CD11c(-)(/dim) CD1c+ myeloid DC. The development of a multiparameter flow cytometry panel will allow for simultaneous enumeration of mature lymphocyte, NK cells, monocyte and DC subsets. Studying these major players of the immune system in one panel may give us a broader view of the immune response during SIV infection and the ability to better define the role of each of these individual cell types in the pathogenesis of AIDS.
