Cytokine changes and embryo attachment in mouse endometrial cells following treated with peripheral blood mononuclear cells (PBMCs) expressing ectopic hCG, and hCG-activated PBMCs

Abstract

Objective: To compare the effect of human chorionic gonadotropin (hCG)-producing peripheral blood mononuclear cells (PBMCs) and PBMCs activated by hCG in vitro and expressions of related immune genes in mouse implantation.

Methods: hCG-producing PBMCs (transfected PBMC) and PBMCs activated by hCG in vitro were introduced into isolated mouse endometrial cells, while cell cultures were divided into four groups: the control, PBMC, transfected, and activated PBMC groups. The expression of studied genes (IL-lβ, IL-6, Lif, and Vegf) was evaluated and blastocyst attachment on the cocultured cells (isolated endometrial cells and PBMC cells) was monitored in all four groups.

Results: Data showed that expression decreased in the PBMC group compared to the treated PBMC (transfected and activated PBMCs) and increased in transfected PBMC compared to the activated PBMC. Attachment and migration of blastocysts were dramatically enhanced in the transfected PBMC group compared to the activated PBMC group (P<0.05).

Conclusions: Use of hCG-producing PBMCs (transfected PBMC) has more influence on endometrial receptivity.

Keywords

Peripheral blood mononuclear cells / hCG / Embryo attachment / Immune response / Immune cells / In vitro / Expression gene

Conflict of interest statement

The authors declare no conflicts of interest.

Funding

This study received no extramural funding.

Authors’ contributions

Mohammad Salehi, Mojgan Bandehpour and Delsuz Rezaee contributed to project administration, study conceptualization and methodology. Bahram Kazemi, Saiyad Bastaminejad and Sajad Najafi performed the review, and editing of manuscript and analysis. Delsuz Rezaee performed the experiment, original manuscript writing, and submitted the data for analysis.

References

[1] Yu N, Zhang B, Xu M, Wang S, Liu R, Wu J, et al. Intrauterine administration of autologous peripheral blood mononuclear cells (PBMCs) activated by hCG improves the implantation and pregnancy rates in patients with repeated implantation failure: A prospective randomized study. Am J Reprod Immunol 2016; 76(3):212-216.

[2] Zeyneloglu HB, Onalan G. (eds.) Remedies for recurrent implantation failure. Semin Reprod Med 2014; 32(04):297-305.

[3] Yu N, Yang J, Guo Y, Fang J, Yin T, Luo J, et al. Intrauterine administration of peripheral blood mononuclear cells (PBMCs) improves endometrial receptivity in mice with embryonic implantation dysfunction. Am J Reprod Immunol 2014; 71(1):24-33.

[4] Fujiwara H. Do circulating blood cells contribute to maternal tissue remodeling and embryo-maternal cross-talk around the implantation period? Mol Hum Reprod 2009; 15(6):335-343.

[5] Leo AJ, Grande DA. Mesenchymal stem cells in tissue engineering. Cells Tissues Organs 2006; 183(3):112-122.

[6] Yoshioka S, Fujiwara H, Nakayama T, Kosaka K, Mori T, Fujii S. Intrauterine administration of autologous peripheral blood mononuclear cells promotes implantation rates in patients with repeated failure of IVF-embryo transfer. Hum Reprod 2006; 21(12):3290-3294.

[7] Yu N, Yan W, Yin T, Wang Y, Guo Y, Zhou D, et al. hCG-activated human peripheral blood mononuclear cells (PBMC) promote trophoblast cell invasion. PloS One 2015; 10(6):e0125589.

[8] Madkour A, Bouamoud N, Louanjli N, Kaarouch I, Copin H, Benkhalifa M, et al. Intrauterine insemination of cultured peripheral blood mononuclear cells prior to embryo transfer improves clinical outcome for patients with repeated implantation failures. Zygote 2016; 24(1):58-69.

[9] Dehghan Z, Mohammadi-Yeganeh S, Rezaee D, Salehi M. MicroRNA-21 is involved in oocyte maturation, blastocyst formation, and preimplantation embryo development. Dev Biol 2021; 480:69-77.

[10] Rezaee D, Bandehpour M, Kazemi B, Salehi M. Role of intrauterine administration of transfected peripheral blood mononuclear cells by GM-CSF on embryo implantation and pregnancy rate in mice. Mol Hum Reprod 2020; 26(2):101-110.

[11] Rezaee D, Bandehpour M, Kazemi B, Hosseini S, Dehghan Z, Bastaminejad S, et al. Effects of human chorionic gonadotropin-producing peripheral blood mononuclear cells on the endometrial receptivity and implantation sites of the mouse uterus. Clin Exp Reprod Med 2022; 49(4):248-258.

[12] Carson DD, Bagchi I, Dey SK, Enders AC, Fazleabas AT, Lessey BA, et al. Embryo implantation. Dev Biol 2000; 223(2):217-237.

[13] Mor G, Cardenas I. The immune system in pregnancy: A unique complexity. Am J Reprod Immunol 2010; 63(6):425-433.

[14] Okitsu O, Kiyokawa M, Oda T, Miyake K, Sato Y, Fujiwara H. Intrauterine administration of autologous peripheral blood mononuclear cells increases clinical pregnancy rates in frozen/thawed embryo transfer cycles of patients with repeated implantation failure. J Reprod Immunol 2011; 92(1-2):82-87.

[15] Cole LA. Biological functions of hCG and hCG-related molecules. Reprod Biol Endocrinol 2010; 8(1):1-14.

[16] Makrigiannakis A, Vrekoussis T, Zoumakis E, Kalantaridou SN, Jeschke U. The role of hCG in implantation: A mini-review of molecular and clinical evidence. Int J Mol Sci 2017; 18(6):1305-1313.

[17] Kosaka K, Fujiwara H, Tatsumi K, Yoshioka S, Sato Y, Egawa H, et al. Human chorionic gonadotropin (hCG) activates monocytes to produce interleukin-8 via a different pathway from luteinizing hormone/hCG receptor system. J Clin Endocrinol Metab 2002; 87(11):5199-5208.

[18] Schumacher A, Brachwitz N, Sohr S, Engeland K, Langwisch S, Dolaptchieva M, et al. Human chorionic gonadotropin attracts regulatory T cells into the fetal-maternal interface during early human pregnancy. J Immunol 2009; 182(9):5488-5497.

[19] Kosaka K, Fujiwara H, Tatsumi K, Yoshioka S, Higuchi T, Sato Y, et al. Human peripheral blood mononuclear cells enhance cell-cell interaction between human endometrial epithelial cells and BeWo-cell spheroids. Hum Reprod 2003; 18(1):19-25.

[20] Mor G, Cardenas I, Abrahams V, Guller S. Inflammation and pregnancy: The role of the immune system at the implantation site. Ann N Y Acad Sci 2011; 1221(1):80-87.

[21] Ideta A, Hayama K, Nakamura Y, Sakurai T, Tsuchiya K, Tanaka S, et al. Intrauterine administration of peripheral blood mononuclear cells enhances early development of the pre-implantation bovine embryo. Mol Reprod Dev 2010; 77(11):954-962.

[22] Hashii K, Fujiwara H, Yoshioka S, Kataoka N, Yamada S, Hirano T, et al. Peripheral blood mononuclear cells stimulate progesterone production by luteal cells derived from pregnant and non-pregnant women: Possible involvement of interleukin-4 and 10 in corpus luteum function and differentiation. Hum Reprod 1998; 13(10):2738-2744.