Soluble HLA-G blood levels are not increased during ongoing pregnancy in women with a history of recurrent pregnancy loss.

Recurrent pregnancy loss (RPL) affects 1-2 % of couples who are trying to conceive. At some point, some couples do maintain a healthy pregnancy to term, but the underlying mechanism of RPL remains elusive. Human leukocyte antigen (HLA)-G is an immune modulatory molecule. Our group previously showed increased HLA-G levels in the decidua of term pregnancies after RPL, while other studies showed reduced soluble HLA-G (sHLA-G) blood levels in women with RPL. This led us to investigate sHLA-G levels in blood of women with RPL who had either a subsequent pregnancy loss (RPL-pregnancy loss) or a healthy term pregnancy (RPL-live birth), and compare these to healthy control pregnancies and non-pregnant controls. Soluble HLA-G concentrations were quantified by ELISA. Women with healthy term pregnancy had increased sHLA-G levels compared to non-pregnant controls. In contrast, RPL-live birth women at term did not have increased blood sHLA-G levels. Soluble HLA-G levels remained stable between first and third trimester. Interestingly, when comparing first trimester samples of RPL-live birth to RPL-pregnancy loss, sHLA-G levels also did not significantly differ. High sHLA-G levels in blood seem not to be crucial for an ongoing healthy pregnancy after RPL. However, since it was previously shown that women with RPL-live birth have increased HLA-G levels in term decidua compared to control pregnancies, the current data suggest that local and systemic immune regulation are not necessarily in concert. Further study of the contribution of fetus-derived HLA-G and HLA-G of maternal origin may provide more insight in the pathophysiology of RPL.

Effect of seminal plasma on dendritic cell differentiation in vitro depends on the serum source in the culture medium.

Dendritic cells (DCs) are key in shaping immune responses and are recruited to the human cervix after coitus by seminal plasma (SP). SP has been shown to skew the differentiation of monocyte-derived DCs towards an anti-inflammatory profile when cultured in medium containing fetal calf serum (FCS). Here, we confirmed that SP skewed DCs cultured in fetal bovine serum (FBS) towards a tolerogenic profile. To create a setting more similar to the in vivo situations in humans, we tested the immune regulatory effect of SP on DCs in cell cultures containing human serum (HS). SP-DCs cultured in HS did show increased CD14 and decreased CD1a, indicating an inhibited maturation phenotype. Gene expression of TGF-ß and IL-10 and IL-10 protein expression were elevated in LPS-activated SP-DCs, whereas IL-12p70 protein levels were decreased compared to LPS-activated control DCs. In contrast to FBS culture conditions, in the presence of HS co-cultures of SP-DCs with allogeneic peripheral blood mononuclear cells (PBMCs) did not result in decreased T cell proliferation and inflammatory cytokine production. Thus, under HS culture conditions SP can skew the differentiation of monocyte-derived DCs phenotypically towards alternatively activated DCs, but this immune regulatory phenotype is functionally less pronounced compared to SP-treated DCs cultured in FBS containing medium. These findings highlight the importance of the source of the serum that is used in SP treated cell cultures in vitro.