Sharkey, D.J.Lyons, H.E.Chan, H.Y.Robertson, S.A.Álvarez-Rodríguez, M.2025-12-012025-12-012025Spermatology: Methods and Protocols, 2025 / Álvarez-Rodríguez, M. (ed./s), vol.2897, Ch.45, pp.663-673978-1-0716-4406-5https://hdl.handle.net/2440/148364Series Editor John M. Walker, Print ISSN 1064-3745 ; Electronic ISSN 1940-6029.Seminal fluid introduced into the female reproductive tract following unprotected coitus initiates an inflammation-like response in the cervical tissues that stimulates immune tolerance to male seminal fluid alloantigens and promotes immune adaptation for pregnancy. This response commences when seminal fluid factors induce genes encoding pro-inflammatory cytokines and chemokines including CSF2, IL6, and CXCL8, that in turn cause recruitment of macrophages, dendritic cells, and T cells into the epithelial and stromal layers. Various signaling agents present in seminal plasma provoke the post-coital pro-inflammatory activation and regulate the quality of immune response. However, the factors identified to date do not account for all the biological activity within seminal fluid, implying sperm-associated factors may also contribute. In this chapter, we report methods for evaluating the effects of human seminal fluid components (seminal plasma and sperm) on the female reproductive tract immune response in primary and immortalized ectocervical epithelial cells in vitro.en© The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2025seminal fluid; seminal plasma; sperm; cervix; inflammation; cytokines; humanGenitalia, FemaleCervix UteriSpermatozoaEpithelial CellsSemenHumansCytokinesFemaleMaleBook chapter10.1007/978-1-0716-4406-5_45736950Sharkey, D.J. [0000-0002-4831-7950]Lyons, H.E. [0000-0001-5305-2654]Robertson, S.A. [0000-0002-9967-0084]