Development of a 40+ color full spectrum flow cytometry panel for in-depth immunophenotyping of chief cell subsets in non-human primate peripheral blood mononuclear cells

Abstract Leveraging full-spectrum technology, we designed and optimized a 40+ color flow cytometry panel for the analysis of non-human primate peripheral blood mononuclear cells (PBMCs). Non-human primate models of disease allow for the closest approximation of human outcomes to vaccination, disease, and treatment due to similarities in physiological host responses, which increase predictive accuracy. This first panel of its kind achieves comprehensive profiling of immune subpopulations by the inclusion of surface, intracellular, and cytokine markers representative of differentiation, activation, functionality, and proliferation to deeply interrogate the immune response to infectious disease and vaccination. The optimized panel is additionally appropriate for the discovery of new immune phenotypes important for protection against pathogens. After initial optimization for use with cynomolgus macaque (Macaca fascicularis) and rhesus macaque (Macaca mulatta) PBMCs, the panel was further adapted for use with African green monkey (Chlorocebus aethiops) PBMCs to allow for cross-comparison between three frequently implemented non-human primate models of infectious disease. This study was supported by the US Army Medical Research Acquisition Activity contract number W81XWH1910027 to TWG and Department of Health and Human Services, National Institutes of Health grant number UC7AI094660 for BSL-4 operations support of the Galveston National Laboratory.