At least 100 cells were differentiated by light microscopy based on conventional morphologic criteria. Neutrophils displayed a multilobed nucleus and a fine pink staining. Eosinophils are characterized by the bilobed nucleus and deep pink staining of the cytoplasm. Lymphocytes have got a large, round, deeply blue nucleus. Monocytes were identified by the kidney shaped or bilobed nucleus. Cell-free BAL supernatant was collected for cytokine and MMP-9 Navitoclax nmr detection. Mice were injected i.p. with 1 mL of 3% thioglycollate media (BD Biosciences) or PBS as control. At indicated time points peritoneal lavage was collected. Cells in the lavage fluid were counted and
cell differentials were determined on slide preparations stained with Diff Quik (Dade Behring, Marburg, Germany). Cells were differentiated as described above. Cell-free peritoneal fluid was collected. Peritoneal tissue was dissected for histological studies. We greatly appreciate the PD-0332991 price technical assistance of Mr. Danny Gutknecht. We thank Manuela Ackermann for performing i.v. injection and Jutta Jahns for irradiation of mice. This work was
supported by the Deutsche Forschungsgemeinschaft to Anja Saalbach and Ulf Anderegg (SA 683/2-1). Conflict on interest: The authors declare no financial or commercial conflict of interest. Detailed facts of importance to specialist readers are published as ”Supporting Information”. Such documents are peer-reviewed, but not copy-edited or typeset. They are made available as submitted by the authors.
“
“Successful embryo implantation occurs followed by a local inflammatory/T helper type 1 (Th1) response, subsequently redirected towards a tolerogenic predominant profile. The lack of control of this initial local inflammatory response may be an underlying cause of early pregnancy complications as recurrent spontaneous abortions (RSA). Considering that Cyclin-dependent kinase 3 vasoactive intestinal peptide (VIP) mediates anti-inflammatory and tolerogenic effects in several conditions we hypothesized that VIP might contribute to tolerance towards trophoblast antigens during the early interaction of maternal leucocytes and trophoblast cells. In this study we investigated VIP/VPAC system activity and expression on maternal peripheral blood mononuclear cells (PBMCs) after interaction with immortalized trophoblast cells (Swan-71 cell line) as an in-vitro model of feto–maternal interaction, and we analysed whether it modulates maternal regulatory T cell (Treg)/Th1 responses. We also investigated the contribution of the endogenous VIP/VPAC system to RSA pathogenesis. VIP decreased T-bet expression significantly, reduced monocyte chemotactic protein-1 (MCP-1) and nitrite production in co-cultures of PBMCs from fertile women with trophoblast cells; while it increased the frequency of CD4+CD25+ forkhead box protein 3 (Foxp3)+ cells, transforming growth factor (TGF)-β expression and interleukin (IL)-10 secretion.