Effect of pregnant mouse serum on induction of indolamine 2, 3- dioxygenase in dendritic cells

Nikoo, Shoreh; Moazzeni, Seyyed Mohammad; Bozorgmehr, Mahmood; Zarnani, Amir Hassan

Effect of pregnant mouse serum on induction of indolamine 2, 3- dioxygenase in dendritic cells

Authors

Shoreh Nikoo ¹

Seyyed Mohammad Moazzeni ¹

Mahmood Bozorgmehr ¹

Amir Hassan Zarnani ²

¹ Immunology Department, Faculty of Medical Science, Tarbiat Modarres University, Tehran, Iran

² Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran; Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran

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

Keywords

Allogenic MLR

Dendritic cell

HPLC

IDO

Indolamine 2,3- dioxygenase

Pregnancy

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