Deep phenomics of African green monkey and Baboon PBMCs using a 30-marker spectral flow cytometry panel

Vaccines are the best way to prevent and combat diseases. Non-human primate (NHP) models have proven to be the gold standard model for studying the efficacy of certain vaccine candidates prior to human trials. Cell-mediated immunity serves as a surrogate marker for predicting vaccine efficacy. Despite the availability of deep-immunophenotyping panels for T cells in the peripheral blood of humans, their application to NHPs, particularly African Green Monkeys (AGMs) and baboons, remains limited. Notably, AGMs are valuable for HIV and COVID-19 research, while baboons are utilized for pertussis research as a natural infection model and vaccine trial. To overcome such limitation, using spectral flow cytometry, we developed a 30-parameter panel to profile major immune cell subsets (B cells, NK cells, DCs, monocytes) either in AGM or baboon PBMCs. We also profiled T cells, memory phenotypes, functional cytokines by tracking established Th1 markers IFN{gamma}, TNF and IL-2; Th2 markers IL-4/5/13 and the Th17 marker, IL-17, activation-induced markers (AIMs) and FoxP3+ Tregs in both AGM and baboon PBMCs in steady state as well as in mitogen stimulated conditions. For AIMs, CD69, CD134 (OX40) and CD137 (4-1BB) are included and validated in both NHP species. This panel holds potential for exploratory analysis of the impact of these functional cytokines, AIMs, or Tregs on host immunity following infection or vaccination. The panels high level of customization empowers researchers to tailor it to their specific research needs.