Alteration of Th17 and Treg cells in patients with unexplained recurrent spontaneous abortion before and after lymphocyte immunization therapy

Reproductive Biology and Endocrinology

Yu-Sheng Liu¹  Hong-Bing Luan¹  Gui-Xiang Zhou¹  Bo Xu¹  Qing Li¹  Ying-Xin Zhang¹  Li-Hua Luo¹  Li Wu¹ 

[1] Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, Anhui, China

关键词: Lymphocyte immunization therapy;    Unexplained recurrent spontaneous abortion;    Treg cells;    Th17 cells;   

Others  :  1139781
DOI  :  10.1186/1477-7827-12-74

 received in 2014-04-07, accepted in 2014-07-23,  发布年份 2014

【 摘 要 】

Background

Several types of T cells have been associated with the pathogenesis of unexplained recurrent spontaneous abortion (URSA), including Th1/Th2/Th17/Tregs cell. It has been appreciated that immunotherapy with paternal or third party lymphocytes is an effective method of treatment for URSA patients. The balance of Th1/Th2 cells could be maintained and an increase of Treg cells would be beneficial after immunotherapy; however, the mechanism by which the Th17/Treg balance affects URSA has not yet been fully elucidated.

Methods

Here, we used flow cytometry, liquid chip technology and quantitative real-time PCR (qPCR) methods to characterize Th17/Treg cell populations after immunotherapy. We found that after immunotherapy in URSA patients, the percentage of Th17 cells decreased and the percentage of Treg cells in peripheral blood mononuclear cells (PBMC) increased, as detected by flow cytometry.

Results

Immunotherapy may induce a decrease in the Th17/Treg ratio and the Treg bias, which may be beneficial for the maintenance of pregnancy. The expression level of ROR gamma t, a transcription factor found in Th17 cells, decreased and the expression of the Treg-specific transcription factor Foxp3 increased in peripheral blood as detected by qPCR. Immunotherapy may induce a decrease in the ratio of ROR gamma t to Foxp3 and a Treg cell bias, which would be beneficial for pregnancy maintenance. The secretion of the Treg-associated cytokine TGF-beta, as well as Th2 cytokines, was increased in serum, while the secretion of Th17-associated cytokine IL-17A and Th1 cytokine production was decreased. The Th1/Th2 cytokine ratio significantly decreased. Similarly, the Th17/Treg ratio significantly decreased in the total patient after immunotherapy.

Conclusions

These results indicate that in patients with URSA, immunotherapy with mononuclear cells derived from the baby’s father could affect both Th1/Th2 and Th17/Treg balance, and we found that the Th2 and Treg bias would be beneficial for pregnancy, which may lead to a balancing of the Th17/Treg ratio in URSA patients after immunotherapy.

【 授权许可】

   
2014 Wu et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Sierra S, Stephenson M: Genetics of recurrent pregnancy loss. Semin Reprod Med 2006, 24:17-24.
• [2]Erlebacher A: Why isn’t the fetus rejected? Curr Opin Immunol 2001, 3:590-593.
• [3]Raghupathy R, Makhseed M, Azizieh F, Hassan N, Al-Azemi M, Al-Shamali E: Maternal Th1 and Th2 type reactivity to placental antigens in normal human pregnancy and unexplained recurrent spontaneous abortions. Cell Immunol 1999, 196:122-130.
• [4]Bettelli E, Carrier Y, Gao W, Korn T, Strom TB, Oukka M, Weiner HL, Kuchroo VK: Reciprocal developmental pathways for the generation of pathogenic effector or TH17 and regulatory T cells. Nature 2006, 441:235-238.
• [5]Mucida D, Park Y, Kim G, Turovskaya O, Scott I, Kronenberg M, Cheroutre H: Reciprocal Th17 and regulatory T cell differentiation mediated by retinoic acid. Science 2007, 317:256-260.
• [6]Afzali B, Lombardi G, Lechler RI, Lord GM: The role of T helper 17 (Th17) and regulatory T cells (Treg) in human organ transplantation and autoimmune disease. Clin Exp Immunol 2007, 1481:32-46.
• [7]Jin D, Zhang L, Zheng J, Zhao Y: The inflammatory Th17 subset in immunity against self and non-self antigens. Autoimmunity 2008, 41:154-162.
• [8]Lee SK, Kim JY, Hur SE, Kim CJ, Na BJ, Lee M, Gilman-Sachs A, Kwak-Kim J: An imbalance in interleukin-17-producing T and Foxp3+ regulatory T cells in women with idiopathic recurrent pregnancy loss. Hum Reprod 2011, 11:2964-2971.
• [9]Somerset DA, Zheng Y, Kilby MD, Sansom DM, Drayson MT: Normal human pregnancy is associated with an elevation in the immune suppressive CD25 + CD4 + regulatory T-cell subset. Immunology 2004, 112:38-43.
• [10]Sasaki Y, Sakai M, Miyazaki S, Higuma S, Shiozaki A, Saito S: Decidual and peripheral blood CD4 + CD25+ regulatory T cells in early pregnancy subjects and spontaneous abortion cases. Mol Hum Reprod 2004, 10:347-353.
• [11]Aluvihare VR, Kallikourdis M, Betz AG: Regulatory T cells mediate maternal tolerance to the fetus. Nat Immunol 2004, 5:266-271.
• [12]Wang WJ, Hao CF, Yi-Lin , Yin GJ, Bao SH, Qiu LH, Lin QD: Increased prevalence of T helper 17 (Th17) cells in peripheral blood and decidua in unexplained recurrent spontaneous abortion patients. J Reprod Immunol 2010, 84:164-170.
• [13]Liu YS, Wu L, Tong XH, Wu LM, He GP, Zhou GX, Luo LH, Luan HB: Study on the relationship between Th17 cells and unexplained recurrent spontaneous abortion. Am J Reprod Immunol 2011, 65:503-511.
• [14]Mowbray JF, Gibbings C, Liddell H, Reginald PW, Underwood JL, Beard RW: Controlled trial of treatment of recurrent spontaneous abortion by immunization with paternal cells. Lancet 1985, 8435:941-943.
• [15]Unander AM, Lindholm A: Transfusion of leucocyte-rich erythrocyte concentrates: a successful treatment in selected cases of habitual abortion. Am J Obstet Gynecol 1986, 154:516-520.
• [16]Gatenby PA, Cameron K, Simes RJ, Adelstein S, Bennett MJ, Jansen RP, Shearman RP, Stewart GJ, Whittle M, Doran TJ: Treatment of recurrent spontaneous abortion by immunization with paternal lymphocytes: results of a controlled trial. Am J Reprod Immunol 1993, 29:88-94.
• [17]Lin Q, Zhao A, Zhou H: Immunotherapy of primary habitual abortion. Zhonghua Fu Chan Ke Za Zhi 1996, 31:351-352.
• [18]Jin LP, Li DJ, Zhang JP, Wang MY, Zhu XY, Zhu Y, Meng Y, Yuan MM: Adoptive transfer of the paternal antigen-hyporesponsive T cells induces maternal tolerance to the allogeneic fetus in abortion prone matings. J Immunol 2004, 173:3612-3619.
• [19]Szpakowski A, Malinowski A, Głowacka E, Wilczyński JR, Kolasa D, Dyński M, Tchórzewski H, Zeman K, Szpakowski M: The influence of paternal lymphocyte immunization on the balance of Th1⁄Th2 type reactivity in women with unexplained recurrent spontaneous abortion. Ginekol Po 2000, 71:586-592.
• [20]Yokoo T, Takakuwa K, Ooki I, Kikuchi A, Tamura M, Tanaka K: Alteration of TH1 and TH2 cells by intracellular cytokine detection in patients with unexplained recurrent abortion before and after immunotherapy with the husband’s mononuclear cells. Fertil Steril 2006, 85:1452-1458.
• [21]Qiu L: Study on changes of serumT helper cell type 1 and type 2 cytokines after active immunotherapy in women with unexplained habitual abortion. Zhonghua Fu Chan Ke Za Zhi 2001, 36:408-410.
• [22]Yang H, Qiu L, Di W, Zhao A, Chen G, Hu K, Lin Q: Proportional change of CD4 + CD25+ regulatory T cells after lymphocyte therapy in unexplained recurrent spontaneous abortion patients. Fertil Steril 2009, 92:301-305.
• [23]Ivanov II, McKenzie BS, Zhou L, Tadokoro CE, Lepelley A, Lafaille JJ, Cua DJ, Littman DR: The orphan nuclear receptor ROR-gammat directs the differentiation program of pro-inflammatory IL-17+ T helper cells. Cell 2006, 126:1121-1133.
• [24]Kim JM, Rudensky A: The role of the transcription factor Foxp3 protein in the development of regulatory T cells. Microbes Infect 2004, 6:745-751.
• [25]Seol HJ, Oh MJ, Lim JE, Jung NH, Yoon SY, Kim HJ: The role of CD4 + CD25bright regulatory T cells in the maintenance of pregnancy, premature rupture of membranes, and labor. Yonsei Med J 2008, 3:366-371.
• [26]Tilburgs T, Roelen DL, van der Mast BJ, de Groot-Swings GM, Kleijburg C, Scherjon SA, Claas FH: Evidence for a selective migration of fetus specific CD4+CD25bright regulatory T cells from the peripheral blood to the decidua in human pregnancy. J Immunol 2008, 8:5737-5745.
• [27]Ober C, Karrison T, Odem RR, Barnes RB, Branch DW, Stephenson MD, Baron B, Walker MA, Scott JR, Schreiber JR: Mononuclear-cell immunization in prevention of recurrent miscarriages: A randomised trial. Lancet 1999, 354:365-369.
• [28]Pandey MK, Agrawal S: Induction of MLR-Bf and protection of fetal loss: a current double blind randomized trial of paternal lymphocyte immunization for women with recurrent spontaneous abortion. Int Immunopharmacol 2004, 4:289-298.
• [29]Worldwide collaborative observational study and meta-analysis on allogeneic leucocyte immunotherapy for recurrent spontaneous abortions. Recurrent Miscarriage Immunotherapy Trialists Group Am J Reprod Immunol 1994, 32:55-72.
• [30]Makhseed M, Raghupathy R, Azizieh F, Omu A, Al-Shamali E, Ashkanani L: Th1 and Th2 cytokine profiles in recurrent aborters with successful pregnancy and with subsequent abortions. Hum Reprod 2001, 10:2219-2226.
• [31]Makhseed M, Raghupathy R, Azizieh F, Al-Azemi MM, Hassan NA, Bandar A: Mitogen-induced cytokine responses of maternal peripheral blood lymphocytes indicate a differential Th-type bias in normal pregnancy and pregnancy failure. Am J Reprod Immunol 1999, 5:273-281.
• [32]Wilczyński JR, Radwan P, Tchórzewski H, Banasik M: Immunotherapy of patients with recurrent spontaneous miscarriage and idiopathic infertility: does the immunization-dependent Th2 cytokine overbalance really matter? Arch Immunol Ther Exp (Warsz) 2012, 60:151-160.
• [33]Kozma N, Halasz M, Polgar B, Poehlmann TG, Markert UR, Palkovics T, Keszei M, Par G, Kiss K, Szeberenyi J, Grama L, Szekeres-Bartho J: Progesterone-induced blocking factor activates STAT6 via binding to a novel IL-4 receptor. J Immunol 2006, 176:819-826.
• [34]Szekeres-Bartho J, Wegmann TG: A progesterone-dependent immunomodulatory protein alters the Th1/Th2 balance. J Reprod Immunol 1996, 31:81-95.
• [35]Piccinni MP, Giudizi MG, Biagiotti R, Beloni L, Giannarini L, Sampognaro S, Parronchi P, Manetti R, Annunziato F, Livi C: Progesterone favors the development of human T helper cells producing Th2-type cytokines and promotes both IL-4 production and membrane CD30 expression in established Th1 cell clones. J Immunol 1995, 155:128-133.
• [36]Nakashima A, Ito M, Shima T, Bac ND, Hidaka T, Saito S: Accumulation of IL-17-positive cells in decidua of inevitable abortion cases. Am J Reprod Immunol 2010, 64:4-11.
• [37]Arruvito L, Billordo A, Capucchio M, Prada ME, Fainboim L: IL-6 trans-signaling and the frequency of CD4 + FOXP3+ cells in women with reproductive failure. J Reprod Immunol 2009, 82:158-165.
• [38]Chen W, Jin W, Hardegen N, Lei KJ, Li L, Marinos N, McGrady G, Wahl SM: Conversion of peripheral CD4 + CD25 naive T cells to CD4 + CD25+ regulatory T cells by TGF-betainduction of transcription factor Foxp3. J Exp Med 2003, 198:1875-1886.
• [39]Zheng SG, Wang J, Horwitz DA, Cutting edge: Foxp3 + CD4 + CD25+ regulatory T cells induced by IL-2 and TGF-beta are resistant to Th17 conversion by IL-6. J Immunol 2008, 180:7112-7116.
• [40]Xu L, Kitani A, Fuss I, Strober W, Cutting edge: Regulatory T cells induce CD4 + CD25-Foxp3- T cells or are self-induced to become Th17 cells in the absence of exogenous TGF-beta. J Immunol 2007, 178:6725-6729.
• [41]Zheng SG, Gray JD, Ohtsuka K, Yamagiwa S, Horwitz DA: Generation ex vivo of TGF-beta-producing regulatory T cells from CD4 + CD25- precursors. J Immunol 2002, 169:4183-4189.
• [42]Zheng SG, Wang JH, Gray JD, Soucier H, Horwitz DA: Natural and induced CD4 + CD25+ cells educate CD4 + CD25- cells to develop suppressive activity: the role of IL-2, TGF-beta, and IL-10. J Immunol 2004, 172:5213-5221.