Fig. 9

Validation of the reduction in nonclassical monocytes via flow cytometry. A Gating scheme for identification and analysis of monocyte subtypes. PBMCs were gated as follows: debris was excluded, non-viable cells were excluded, then doublets were excluded. Next, monocytes were gated based on their high side scatter and CD14 expression. B Monocyte subtypes were gated based on their characteristic CD14 and CD16 expression, with classical monocytes having high CD14 expression and low CD16 expression, intermediate monocytes having high CD14 expression and moderate-to-high CD16 expression, and NC monocytes having low CD14 expression and high CD16 expression. C Quantification of the frequency of NC (left), intermediate (center), and classical monocytes (right), either as a percentage of PBMCs (top row) or all monocytes (bottom row). NC monocytes were reduced in MAPT pathogenic variant carriers as a fraction of PBMCs (upper left, p = 0.02) and as a fraction of monocytes (lower left, p = 0.05). Intermediate monocytes (center) showed a trend toward reduction relative to both PBMCs and monocytes. Classical monocytes (right) showed no change as a fraction of PBMCs but were significantly increased in MAPT pathogenic variant carriers as a fraction of all monocytes