Production of transforming growth factor β and interleukin-10 in the mouse spleen at early stage of gestation in models of spontaneous and muramylpeptide-dependent abortions

Abstract

Introduction. The maternal immune response is a key determinant of successful pregnancy development. Among the multiple local and systemic mechanisms of immune tolerance formation to the fetoplacental «graft», the regulatory cytokines interleukin-10 (IL-10) and transforming growth factor β (TGFβ) play the leading role. However, the relationship between the production levels of IL-10 and TGFβ in peripheral organs of the immune system in vivo and the frequency of fetal losses in experimental pregnancy failure has not been sufficiently studied.

The aim of the study was to evaluate the production levels of IL-10 and TGFβ in the spleen of pregnant mice in models of spontaneous and muramylpeptide-dependent abortions at early gestational age.

Material and methods. Allogeneic physiological pregnancy was reproduced by crossing CBA/lac females (H-2k) with Balb/c males (H-2d) and the model of spontaneous abortion by mating females with DBA/2J males (H-2d). To model induced and potentiated abortions, CBA/lac females fertilized by Balb/c or DBA/2J males, respectively, were intraperitoneally injected with muramyl dipeptide β-heptylglycoside (C7MDP) at the dose of 20 μg per 1 animal on days 5 and 7 of gestation (DG). Animals were removed from the experiment at the 8th DG. Cytokine-producing cells in spleen sections were detected by immunohistochemical analysis using rabbit polyclonal antibodies to TGFβ and goat polyclonal antibodies to IL-10. The density of TGFβ- and IL-10+ cells was determined by morphometry using Image Scope M software. The digital data were analyzed using Sigma Stat 3.5 software, the differences were considered significant at p < 0.05.

Results. The density of IL-10+- and TGFβ+-cells in the spleen in spontaneous abortions was half that in physiological pregnancy. Under the conditions of baseline high-fertility crosses, injection of C7MDP into pregnant mice (induction of abortion) resulted in an increase in the density of IL-10+-cells, while the density of TGFβ+-cells did not change. In pregnant mice with initially high levels of spontaneous reproductive losses, exposure to C7MDP (abortion potentiation) caused an increase in the density of IL-10+- and TGFβ+-cells. Localization of TGF-β in spleen functional structures did not differ between physiological pregnancy and spontaneous abortions; muramylpeptide-dependent abortions were characterized by an increase in TGFβ-positive structural and functional areas.

Conclusion. The increased incidence of fetal losses compared with physiological pregnancy occurred against the background of decreased IL-10- and TGFβ-producing cell density in the spleen in spontaneous abortions and increased IL-10+-cell density in induced abortions. TGFβ-production was increased in potentiated abortions compared to spontaneous abortions. The increased density of IL-10+-cell distribution in induced abortions and the increased localization of TGF-β+-zones in the spleen in induced and potentiated muramylpeptide-dependent abortions did not abolish the abortogenic effect of C7MDP.

Keywords:immune tolerance; cytokines; muramylpeptide; muramyl dipeptide β-heptylglycoside; spleen; mouse models of pregnancy failure

For citation: Artemieva K.A., Bogdanova I.M., Stepanova I.I., Stepanov A.A., Tikhonova N.B., Boltovskaya M.N., Kalyuzhin O.V., Zemlyakov A.E. Production of transforming growth factor β and interleukin-10 in the mouse spleen at early stage of gestation in models of spontaneous and muramylpeptide-dependent abortions. Immunologiya. 2021; 42 (2): 131–9. DOI: https://doi.org/10.33029/0206-4952-2021-42-2-131-139 (in Russian)

Funding. The work was performed in the framework of the state task of Research Institute of Human Morphology of MSHE of Russia (No. AAAA-A17-117013050049-3).

Conflict of interests. The authors declare no conflict of interests.

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