Senior Scientist Standard BioTools Markham ON, Ontario, Canada
Disclosure(s):
Michael Cohen, PhD: No relevant disclosure to display
Introduction/Rationale: Immunosenescence disrupts surveillance and contributes to age-related diseases, yet its mechanisms remain incompletely understood. Deep immune profiling in mouse models is essential to elucidate age-related modulators of disease. High-parameter cytometry with CyTOF™ technology enables identification of 50-plus markers at the single-cell level, providing exceptional resolution without spectral unmixing or compensation. This study leverages CyTOF technology to characterize aging-associated immune alterations in spleen and lung tissues of aged and young adult mice.
Methods: Aged (75–76 weeks) and young adult (6–8 weeks) C57BL/6J mice were evaluated. Spleen and lung tissues were processed and stained using the 33-antibody Maxpar™ OnDemand Mouse Immune Profiling Panel Kit for spleen and an expanded customized 50-parameter panel for lung. Live-cell CD45 barcoding enabled pooled sample analysis. Data was acquired on a Helios™ CyTOF instrument and analyzed using Maxpar Pathsetter software for automated identification of 38 splenic and 55 pulmonary immune subsets.
Results: In spleen, aged mice showed reduced frequencies of CD4 and CD8 T cells, NK cells and γδ T cells, with increased PD-1+ T cells and B cells. NK cell maturation was impaired, with altered expression of PD-L1, CD11c and CD49b. In lungs, aged mice exhibited higher frequencies of exhausted CD8 T cells (PD-1+TIM-3+), cytotoxic T cells (granzyme B+) and monocyte-derived dendritic cells, alongside a pro-inflammatory alveolar macrophage phenotype. Overall, 26 splenic and 28 pulmonary subsets showed significant age-related differences.
Conclusion: This study demonstrates the power of CyTOF immune profiling with automated analysis to uncover aging-associated immune alterations. The Mouse Immune Profiling Panel and Maxpar Pathsetter software provide a scalable and reproducible solution for deep phenotyping in mouse models, facilitating mechanistic insights into immunosenescence and supporting translational research in aging and immune modulation.
