Paper Information

Journal:   INTERNATIONAL JOURNAL OF FERTILITY AND STERILITY   SUMMER 2013 , Volume 7 , Number SUPPLEMENT 1; Page(s) 108 To 108.
 
Paper: 

COMPARISON BETWEEN THE EFFECT OF 17-BESTRADIOL AT PREOVULATORY AND PREGNANCY LEVELS FOR IN VITRO GENERATION OF REGULATORY T CELLS

 
 
Author(s):  IDALI F., VAHEDIAN Z., MIRZADEGAN E., MAHDAVIPOUR M., ZARNANI A.H., JEDDI TEHRANI M., FATEMI R.*
 
* REPRODUCTIVE IMMUNOLOGY RESEARCH CENTER, AVICENNA RESEARCH INSTITUTE, ACECR, TEHRAN, IRAN
 
Abstract: 

Background: Naturally occurring CD4+CD25+regulatory T cells (Treg) exert an important role in mediating maternal tolerance to the fetus during pregnancy. During pregnancy the elevated number of regulatory T cells (Tregs) correlates with 17-b oestradiol (E2) level. The role of maternal hormones in successful pregnancy is well known. Maternal hormones through re-engineering of the maternal immune response provide protection for the semi-allogeneic fetus, while any changes in maternal tolerance may lead to abortion, and reproductive abnormality.
The aim of this study was to compare the effect of E2 at preovulatory and pregnancy levels for generation of Tregs in vitro.
Materials and Methods: We treated the magnetic bead separated peripheral blood naive T cells (n=4) with E2 and anti-CD28 antibody in anti-CD3 coated plates for 96hours at 37oc and 5%Co2. Two different concentrations of E2 (pregnancy and preovulatory) were used.
Naïve T cells with no treatment were termed as control cells. The phenotype of induced Tregs was investigated through analysis of CD25 and Foxp3 expression. XTT assay was used to monitor PHA-mediated proliferation of conditioned T cells and their effect on proliferation of autologous/allogeneic PBMC in co-culture experiments.
ELISA technique was applied for measurement of cytokines: IL-10, TNF-a and IFN-g in cell supernatants of a secondary culture in the presence or absence of PHA.
Results: After 4 days of incubation, CD25 and Foxp3 were detected by flow cytometry. In comparison to E2preg, E2preov-conditioned T cells showed significantly higher frequencies of Foxp3 expression. E2preov-conditioned T cells showed a significantly reduced proliferation capacity in comparison to other groups of conditioned T cells. Decreased TNF-a level and IFN-g/ IL-10 ratio were noted especially in cultures with preovulatory concentration of E2. In addition, production of TNF-a by E2 preovulatory treated cells was significantly lower than that by E2 pregnancy in the presence or absence of PHA.
Conclusion: Our data suggest estrogen as a good candidate for in vitro generation of Tregs. This data might along with the development of new therapeutic methods for treatment of autoimmune diseases and pregnancy complications, reduce abortion incidence in the future.

 
Keyword(s): REGULATORY T CELLS, 17-BESTRADIOL, PREOVOLATORY, PREGNANCY
 
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