The effect of human amniotic epithelial cell on dendritic cell differentiation of peripheral blood monocytes: An experimental study.

BACKGROUND: The amniotic membrane plays an important role in maintaining a healthy pregnancy. The main population cells from amniotic membrane include human amnion epithelial cells (hAECs) which have been shown to possess immunomodulatory properties. OBJECTIVE: The proximity of hAECs with monocyte leads to the generation of tollerogenic dendritic cells. MATERIALS AND METHODS: hAECs were obtained from normal pregnancy. Peripheral blood monocytes were isolated by anti-CD14 MACS method. Co-cultures of monocytes and hAECs were established in Transwell chambers supplemented with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) in the absence and presence of lipopolysaccharide (LPS) to produce immature and mature DCs, respectively. Immunophenotyping of the obtained DCs was done through flow cytometry and the production of cytokines was measured by ELISA. Mixed leukocyte Reaction (MLR) was also performed for the functional assessment of DCs. RESULTS: Immunophenotyping of [hAECs - Immature DC (iDC)] and [hAECs - iDC] + LPS cells revealed that the expression of CD1a, CD80, CD86, CD40, HLA-DR, and CD83 markers showed no significant difference as compared with the control group (iDCs and mDCs alone). In the [hAECs-iDCs] + LPS cells, the percentage of CD14 cells at the ratio of 1:2.5 showed significant differences compared to the control group. The production of IL-10 and IL-12 showed no significant difference in any of the cultures as compared to the control groups. Also, co-cultured DCs did not inhibit proliferation of lymphocyte. CONCLUSION: Our findings show that factors secreted from cultured hAECs are unable to generate of tollerogenic dendritic cells. To achieve a better understanding of other mechanisms more investigations are needed.

Assessing the potential of HSPA2 and ADAM2 as two biomarkers for human sperm selection.

Selection of the best sperm, with the least defects, is a critical factor in the success of ART especially in male factor infertility. This study assessed the potential Heat shock protein (HSPA2) and metallopeptidase domain2 (ADAM2) biomarkers for sperm selection. Sperm were obtained from 72 asthenoteratozoospermic and 42 normospermic ejaculates. The semen characteristic, DNA fragmentation (DFI), chromatin maturation index (CMI), ADAM2 and HSPA2 levels on sperm, and their correlation with embryo quality were assessed in both groups. Results showed the significant reduction in HSPA2 and ADAM2 in asthenoteratozoospermic compared to normazoospermic ejaculates regarding the cut-off value of 14 and 13% for these two biomarkers. The specificity of HSPA2 and ADAM2 separately, and the combination of these two biomarkers, were 95.2, 90.5 and 93.5%, respectively, for sperm from normozoospermic ejaculates. However, they were 48.6, 50.0 and 54.5% for asthenoteratozoospermic ones. A significant correlation was observed with HSPA2, ADAM2 and a combination of these two biomarkers with CMI, DFI and embryo quality. Although a combination of these two biomarkers have the potential to be a good choice for selecting sperm with the lowest level of chromatin damage, it seems that selection according to HSPA2 has priority over ADAM2 or a combination of the two.

Mesenchymal Stem Cells Upregulate the Expression of PD-L1 But Not VDR in Dendritic Cells.

OBJECTIVE: Mesenchymal stem cells (MSCs) show immunomodulatory functions. But the exact mechanism underlying these activities of MSCs is still not completely understood. There have been a few studies which have assessed the effects of these cells on dendritic cells (DCs) function. Given the importance of programmed cell death receptor-1 (PD-L1) and vitamin D receptor (VDR) expression in induction of tolerance in DCs, we were encouraged to investigate if one of the immunomodulatory functions of MSCs could be inducing upregulation of PD-L1 and VDR on DCs or not. METHODS: DCs were co-cultured with MSCs or treated with them in transwell plates in the presence or absence of Lipopolysaccharide (LPS). Expression of PD-L1 and VDR mRNA and proteins in treated DCs were assessed by Real-time PCR and Western blot techniques. Furthermore, treated DCs were co-cultured with allogeneic T-cells, and T-cell proliferation and cytokine secretions in co-culture supernatants were assessed. RESULTS: The results showed that PD-L1 but not VDR expression is significantly upregulated in the DCs co-cultured with MSCs. Furthermore, cell-to-cell contact and also presence of maturation inducers like LPS is necessary for this function. Moreover, our results indicated that MSCs could induce tolerogenic DCs (TolDCs) which could decrease the secretion of IL-2 by T-cells and inhibit T-cell proliferation as well as increase secretion of IL-10. CONCLUSIONS: Overall, our results show that MSCs may have several suppressive effects on immune responses by induction of TolDCs expressing more PD-L1 immunomodulatory molecule and change the cytokines profile of DCs and T-cells.

Investigating Association of Three Polymorphisms of Coagulation Factor XIII and Recurrent Pregnancy Loss.

PROBLEM: among important suspected causes of thrombophilia in women with recurrent pregnancy loss (RPL) are the polymorphisms of coagulation factor XIII (FXIII) gene. We performed a case-control study on the association between three polymorphisms of factor XIII (FXIII G103T, FXIII A614T and FXIII C1694T) and RPL in Iranian women. METHOD OF STUDY: DNA samples from peripheral blood of 100 female patients with at least two recurrent abortions, as case group, and 100 healthy women with history of at least two successful deliveries were subjected to PCR-RFLP, and the frequencies of the polymorphisms were calculated and compared between the two groups. RESULTS: the prevalence of FXIII G103T polymorphism was 29% in the case group and 17% in the control group (P = 0.158). The frequencies of FXIII A614T and FXIII C1694T were 84% and 66% in the case group and 48% and 31% in the control group (P <0.001 and P < 0.001), respectively. The two latter polymorphisms are associated with RPL in Iranian women and increase the risk of RPL. A correlation was also found between FXIII A614T and FXIII C1694T polymorphisms (P < 0.001). CONCLUSION: we suggest the evaluation of FXIII A614T and FXIII C1694T polymorphisms in women with RPL.