Age-associated inflammatory monocytes are increased in menopausal females and reversed by Hormone Replacement Therapy

Biological sex is a crucial, but poorly understood variable in age-related susceptibility to infections and chronic inflammation - inflammageing. Monocytes are important immune cells responsible for initiating and resolving inflammatory responses to infection. While changes in monocyte populations result in increased susceptibility to infection, there is limited research on the impact of age and sex on human monocyte phenotype and function. The aim of this work was to dissect the impact of increasing age and biological sex on human monocyte phenotype and function. Here we show that older females have increased inflammatory intermediate and non-classical monocytes compared to young. These monocyte subsets were the most inflammatory ex vivo and their frequency correlated with markers of systemic inflammation. Proteomic analysis of sorted monocyte populations demonstrated that the three human monocyte subsets have largely distinct phenotypes. Additionally, proteomic analysis identified key age-associated protein pathways, including complement cascade and phagocytosis, downregulated in monocytes from older compared to younger individuals. We confirmed the proteomics findings showing that circulating C3 concentrations were reduced with age in females but not males. This decrease in complement in older females resulted in reduced monocyte phagocytosis. Crucially, we demonstrate that in peri/menopausal females, Hormone Replacement Therapy (HRT) reversed this expansion in intermediate monocytes and decreased circulating CRP as compared to age matched controls. Importantly peri/menopausal females on HRT had increased C3 serum concentrations and significantly improved monocyte phagocytosis. The data presented here indicate the importance of menopause in ageing monocyte phenotype and function. This data highlights the potential use of HRT in restoring monocyte function in females during ageing.