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Multiplex determination of murine seminal fluid cytokine profiles

Perinatal Research Group, Pathology and Tumour Biology, Leeds Institute of Molecular Medicine, Level 4, JIF Building, St James’s University Hospital, Beckett Street, Leeds LS9 7TF, UK and ¹ Molecular Medicine Unit, Clinical Sciences Building, St James’s University Hospital, Beckett Street, Leeds LS9 7TF, UK

Correspondence should be addressed to N M Orsi; Email: n.m.orsi{at}leeds.ac.uk

Seminal fluid is known to be responsible for orchestrating mating-induced immunomodulation. Central to this process are numerous cytokines that modulate uterine leukocyte recruitment and trafficking. Despite this, a comprehensive analysis of the cytokine profile of murine seminal fluid is lacking. This study addressed this issue by using multiplex immunoassays to characterise the profile of interleukin (IL)-1 , IL-1ß , IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-17, eotaxin, granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), interferon (IFN)-, keratinocyte-derived chemokine (KC), monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1 , MIP-1ß , regulated upon activation normal T-cell expressed and secreted (RANTES), and tumour necrosis factor (TNF)- in fluid drawn from the seminal vesicles of single mice (n = 18). Their levels and ratios were compared with those found in serum. IL-1 , IL-1ß , IL-2, IL-5, IL-9, IL-12 (p40), IL-12 (p70), IL-13, IL-17, GM-CSF, IFN-, MCP-1 and TNF- levels were significantly higher in serum; IL-4, G-CSF, eotaxin, KC and RANTES exhibited the opposite trend. Based on these findings, we propose a model of mating-induced immunomodulation that implicates seminal eotaxin, RANTES and MIP-1 in the relocation and concentration of extravasated migrating endometrial eosinophils to the luminal epithelium. Furthermore, KC may participate in uterine neutrophil chemotaxis and activation. Eotaxin and MIP- , together with IL-1ß and IL-9, may also enhance further cytokine synthesis for endometrial antigen-presenting cell recruitment for processing paternal ejaculate antigens. IL-4 and G-CSF could also minimise deleterious cell-mediated immunity and modulate IFN- production, thereby supporting the establishment of pregnancy.