Comparable vitamin D3 metabolism in the endometrium of patients with recurrent spontaneous abortion and fertile controls.

Vitamin D exerts important roles during pregnancy, and its deficiency may be associated with several pregnancy complications, including pregnancy loss, yet no data are available for molecules involved in vitamin D metabolism in patients with unexplained recurrent spontaneous abortion. In this study, we investigated possible difference in endometrial expression of vitamin D3 receptor (VDR), 1α-hydroxylase (CYP27B1), and 24-hydroxylase (CYP24A1) in women with recurrent spontaneous abortion (n = 8) and healthy controls (n = 8). Gene expression of VDR, CYP27B1, and CYP24A1 was determined by real-time PCR, while VDR and CYP27B1 proteins were localized by immunohistochemistry and their abundance was validated by Western blot. We found that both patient and control groups expressed comparable levels of endometrial VDR, CYP27B1, and CYP24A1 transcripts. In line with the gene-expression results, CYP27B1 and different isoforms of VDR protein were present at the same abundance in the endometria of both groups. No significant alteration in VDR and CYP27B1 immunoreactivity pattern was found in the endometrium of patients compared to fertile controls, however. The results of the present study, therefore, do not support the hypothesis of differential expression of key molecules involved in vitamin D3 metabolism in the endometrium of recurrent spontaneous abortion patients and fertile controls.

Effects of 1,25(OH)2 vitamin D3 on cytokine production by endometrial cells of women with recurrent spontaneous abortion.

OBJECTIVE: To investigate immunomodulatory effect of 1,25(OH)2 vitamin D3 (1,25(OH)2D3) on cytokine production by endometrial cells of women with unexplained recurrent spontaneous abortion (URSA). DESIGN: In vitro study. SETTING: Academic research center. PATIENT(S): Patients with URSA and healthy controls. INTERVENTION(S): Treatment with 1,25(OH)2D3. MAIN OUTCOME MEASURE(S): Production of interferon γ (IFN-γ), interleukin-10 (IL-10), transforming growth factor ß (TGF-ß), IL-17, IL-6, and IL-8 by whole endometrial cells (WECs) and endometrial stromal cells in the presence and absence of 1,25(OH)2D3 and 1α-hydroxylase activity of these cell populations were measured in patients with URSA and healthy controls. RESULT(S): 1,25(OH)2D3 interfered with production of cytokines by WECs of the control and URSA groups, except IL-8 which was increased in URSA group. In endometrial stromal cells, 1,25(OH)2D3 down-regulated cytokine production as well with stimulatory effect on the production of TGF-ß in patients with URSA. Cytokine profile of WECs from patients with URSA skewed toward TH2 phenotype after treatment with 1,25(OH)2D3. Endometrial cells of both groups had comparable capacity to produce 1,25(OH)2D3. CONCLUSION(S): Considering the complex network of immunoregulation at the fetomaternal interface, potential beneficial effects of vitamin D3 in patients with URSA need to be investigated in clinical practice. Comparable levels of 1,25(OH)2D3 production and similar trend of cytokine expression by WECs of URSA and control groups after vitamin D3 treatment reflect the same local metabolic machinery of this hormone.

Vitamin D3 receptor is expressed in the endometrium of cycling mice throughout the estrous cycle.

OBJECTIVE: To investigate the expression and localization of vitamin D(3) receptor (VDR) in reproductive organs of cycling mice. DESIGN: Experimental animal study. SETTING: Academic research center. ANIMAL(S): Mature (8 to 12 weeks old) cycling female Balb/c mice. INTERVENTION(S): Reproductive tissue, including endometrium, ovary, and fallopian tubes, were collected at each phase of estrous cycle to examine VDR expression. MAIN OUTCOME MEASURE(S): Expression of VDR messenger (mRNA) was determined by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR). The presence and localization of VDR was assessed by immunohistochemistry, and the intensity of VDR expression was quantified with U.S. National Institutes of Health image-analysis software. RESULT(S): The VDR mRNA was expressed in the endometrium throughout the estrous cycle. The relative expression of VDR mRNA at the estrus phase was more prominent compared with the other phases. Immunohistochemical analysis revealed that dendritic cells, macrophages, and luminal and glandular epithelial cells of the endometrium, granulosa, and cumulus oophorus cells of the ovary and fallopian epithelial cells strongly express VDR, particularly during the estrus phase. CONCLUSION(S): Our findings have demonstrated, for the first time, that VDR is present and differentially expressed in murine reproductive organs throughout the estrous cycle. Further studies are required to evaluate the functional immunologic role of VDR.

The Predictive Value of HLA-DR Matching and Cytokine Gene Polymorphisms in Renal Allograft Acute Rejection: a Living-unrelated Donor (LURD) Study.

BACKGROUND: In addition to Human Leukocyte Antigens (HLA) compatibility, gene polymorphism in cytokines might be important in the quality of allogeneic immune response. OBJECTIVES: In this study HLA-DR matching and a number of cytokine gene polymorphisms have been evaluated in occurrence of acute rejection after living-unrelated donor (LURD) kidney transplantation. METHODS: A total of 42 renal transplants that were performed at Hashemi Nejad Kidney Hospital (Tehran/Iran) and then followed up for 3 months post-transplantation were included in our study. Using PCR-SSP, HLA-DR alleles (DR1-18) of recipient and donor pairs and gene polymorphisms in TNF-, TGF-1, IL-10, IL-6, and IFN- of recipients were determined. RESULTS: Acute rejection was observed in 11(26.2%) of 42 renal recipients. The frequency of one and two HLA-DR mismatches in rejector and nonrejector groups were 2(18.2%), 9(81.8%) and 13(41.9%), 17(54.8%) respectively. HLA-DR incompatibility was not significantly higher in rejector (1.820.40) than nonrejector (1.520.57) recipients (P=0.069) and more than half of nonrejectors had completely mismatched HLA-DR antigens with their donors. Polymorphisms associated with all the cytokines mentioned above were found to have no correlation with acute rejection. CONCLUSION: Regarding to high frequency of HLA-DR mismatching in nonrejector group, we can postulate that the immunogenecity of LURD renal transplant is not similar to the cadaveric one, and predictive value of HLA-DR mismatching for acute rejection is not as reliable as in cadaveric transplant. Although, there is no doubt about influence of HLA compatibility on allograft survival. In addition, we failed to demonstrate further association between combined recipient cytokine genotypes and HLA-DR matching with acute rejection in this study. Thus, we conclude that a more detailed immunogenetic profile is necessary to predict transplant outcome or immunosuppression tailoring.