Seminal fluid contains signalling molecules including transforming growth factor-beta (TGFβ) which contribute to establishing fetomaternal tolerance required for embryo implantation and placental development. T cells are recruited in response to seminal fluid to regulate key immune events during the peri-implantation period. The impact of seminal fluid on T cell receptor (TCR) gene expression in the endometrium during the peri-implantation period has not previously been defined. In this study, high-throughput RNA sequencing was used to assess the RNA profile in the endometrium of C57Bl/6 females mated with either intact (INT), vasectomised (VAS) or seminal-vesicle-deficient and vasectomised (SVX/VAS) BALB/c males on day (d) 3.5 post-coitus (pc) (n=3-4/group). Samples were run on the Illumina HiSeq2000 platform to a depth of 30 million reads per sample and differential gene expression analysis was carried out using R/Bioconductor packages limma and edgeR. TCR signalling pathway genes were prominent amongst the genes most differentially regulated between groups. In INT-mated females, TCRs, Trbc2, Trdc, Tcrg-C1 and Tcrg-C2, their co-receptors, Cd8a, and TCR signalling pathway-related genes, Cd3d, Cd3e and Cd3g, were all induced to a higher level compared to SVX/VAS-mated females (FC>1.5, P<0.05). These endometrial gene expression changes required exposure to the seminal plasma portion of seminal fluid, as seminal plasma exposure alone in VAS mating achieved a similar gene induction level as in INT mating. This data suggests seminal fluid exposure regulates events that result in elevated T cell receptor signalling pathway expression, presumably resulting from T-cell recruitment and/or proliferation. This study highlights the importance of seminal fluid exposure in eliciting an appropriate T cell response during the peri-implantation period. The results are consistent with previous studies identifying seminal fluid contact as instrumental for inducing T-cell tolerance.