JRI 

Shoreh Nikoo
- Immunology Department, Faculty of Medical Science, Tarbiat Modarres University, Tehran, Iran
Seyyed Mohammad Moazzeni Corresponding Author
- Immunology Department, Faculty of Medical Science, Tarbiat Modarres University, Tehran, Iran
Mahmood Bozorgmehr
- Immunology Department, Faculty of Medical Science, Tarbiat Modarres University, Tehran, Iran
Amir Hassan Zarnani
1- Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
2- Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran

Received: 10/1/2006 Accepted: 10/1/2006 - Publisher : Avicenna Research Institute

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Abstract

Introduction: Several studies have shown that many factors are involved in the maternal tolerance to the fetus. Indolamine 2, 3- dioxygenase (IDO) enzyme which catabolizes tryptophan is one of the factors that have been reported to play an important role leading to a successful pregnancy. The objective of this study was to evaluate the effects of pregnant mouse serum on the induction of indolamine 2, 3 dioxygenase in dendritic cells (DCs) which may be used as a basis for practical studies on the immunological bases of recurrent abortions. Materials & Methods: Allogenic pregnant mice sera were collected in mid-pregnancy. DCs were isolated from Balb/c mouse spleen through a three-step method, including: Collagenase digestion of spleen tissue, low density cells separation via the Nycodenz density gradient centri-fugation and plastic adherence. T cells were isolated from C57BL/6 mouse lymph nodes through nylon wool method. As stimulator cells, pregnant and non-pregnant mice sera treated DCs were irradiated and co-cultured with purified T cells (allogenic MLR). 1-methyl-tryptophan (1-MT), as the specific inhibitor of IDO, was added to some wells of MLR assay in different concentrations and T cells proliferation response was measured by 3H-Thymidine incorporation. The MLR supernatant was also analyzed by HPLC for its tryptophan and kynurenin (Trp metabolite) content. All tests were repeated for 5 times. Man-Whitneys non- parametric test was used to evaluate the differences among groups. Confidence interval was 95% and p-values<0.05 were regarded as significantresults: the results showed the ability of pregnant mice sera to reduce the dendritic cells ability in t cell proliferation induction compared to non-pregnant mice sera but addition of 1-mt did not have any significant effect on this inhibition. additionally, ido metabolites concentration assessment in the presence or absence of 1-mt, through hplc method, did not show any significant difference.conclusion: there are many factors in pregnant mice sera such as progesterone, il-10, vit d3, etc. which might cause inhibition of t lymphocyte proliferation response in allogenic mlr through affecting dcs efficiency. although it seems that ido expression by dcs is not respon-sible for decrease in t cell proliferation after treatment of dcs by pregnant mice sera, thus some other mechanisms might be responsible for this phenomenon which their identification needs more investigation.< pan>


Keywords: Pregnancy, Dendritic cell, IDO, Allogenic MLR, HPLC, Indolamine 2,3- dioxygenase


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