摘要
妊娠期高血压疾病(Hypertensive disorder complicating pregnancy,HDCP)严重威胁母胎健康,是世界范围内导致围产期母胎高发病率和死亡率的主要原因之一。正常妊娠的维持,需要体液免疫与细胞免疫的共同参与。诸多研究表明,在HDCP患者中,存在着不同程度的免疫失衡。有关细胞免疫因素参与HDCP发病机制的研究报道较多,而有关体液免疫因素对其影响的研究较缺乏,有限的研究多聚焦于血清中细胞因子的变化。本研究从胎儿和母体方面了解HDCP患者真实的体液免疫状态,并从抗胎儿免疫角度探讨其在HDCP发病机制中的作用。
目的检测HDCP患者及正常足月剖宫产孕妇外周血中抗HLA抗体,比较母体血循环中抗HLA抗体特异性是否与胎儿HLA基因型一致,评价母体抗HLA抗体是否是被胎儿源性抗原致敏,探讨母体抗胎儿源性体液免疫在HDCP中的作用。
方法选择46例正常足月剖宫产妇女为对照组,60例HDCP患者为病例组;病例组按病程进展分为妊娠期高血压组、轻度子痫前期组和重度子痫前期组,每组各20例。用Flow PRA法对母体血清中抗HLA抗体进行初筛,对于抗HLA抗体阳性者,用Luminex法检测其抗体特异性;同时用SSP-PCR法检测相应孕妇及胎儿的HLA基因型,比较母体外周血中抗HLA抗体类型是否与其胎儿HLA基因型一致。
结果对照组母体外周血抗HLA-Ⅱ类抗体阳性率为6.5%;妊娠期高血压组为20%,轻度子痫前期组为20%,重度子痫前期组为25%。病例组HLA-Ⅱ类抗体阳性率较对照组高,其差异具有统计学意义(P < 0.05)。病例组各组间母体外周血抗HLA-Ⅱ类抗体阳性率随疾病进展加重而升高,但组间差异无统计学意义(P > 0.05)。对照组母体外周血抗HLA-I类抗体阳性率为19.6%,妊娠期高血压组、轻度子痫前期组和重度子痫前期组均为20%,其差异无统计学意义(P > 0.05)。比较母胎HLA基因类型,母体外周血中抗HLA-Ⅱ类抗体类型与其胎儿HLA基因型一致。
结论HDCP患者外周血中抗胎儿HLA-Ⅱ类抗体明显增加,且被胎儿源性HLA致敏。
目的比较HDCP患者及正常足月剖宫产孕妇胎儿脐血中IL-6阳性单核细胞百分比,了解HDCP患者胎儿单核细胞的激活状态。
方法选择46例正常足月剖宫产妇女作为对照组,60例HDCP患者为病例组;病例组按病程进展分为妊娠期高血压组、轻度子痫前期组和重度子痫前期组,每组各20例。采集脐血(EDTA抗凝)经密度梯度离心分离单个核细胞,经双色荧光标记(FITC-CD14和PE-IL-6单克隆抗体)染色、流式细胞技术测定胎儿脐血中IL-6阳性单核细胞百分比。
结果对照组胎儿脐血中IL-6阳性单核细胞百分比为0.78%±0.54%;妊娠期高血压组为4.38%±0.91%,轻度子痫前期为5.73%±2.18%,重度子痫前期为6.29%±2.70%。病例组胎儿脐血中IL-6阳性单核细胞百分比较对照组高,其差异有统计学意义(P < 0.01)。病例组各组间胎儿脐血中IL-6阳性单核细胞百分比随疾病进展加重而升高,各组间差异有统计学意义(P < 0.05)。
结论HDCP患者脐血中胎儿单核细胞是被激活的,且随其严重程度增强。
目的以正常足月剖宫产为对照,比较HDCP妇女外周血中抗胎儿HLA抗体分泌记忆B细胞的频数差异,探讨抗胎儿HLA抗体分泌记忆B细胞在HDCP发病机制中的作用。
方法选择46例正常足月剖宫产妇女作为对照组,60例HDCP患者为病例组;病例组按病程进展分为妊娠期高血压组、轻度子痫前期组、重度子痫前期组,每组各20例。取相应胎儿脐血(EDTA抗凝),采用密度梯度离心法分离脐血中单个核细胞,与IFN-γ共培养72h后制备成B细胞溶胞体,作为胎儿源性抗原来源,包被96微孔硝酸纤维素膜板;加入体外培养并经美洲商陆(Pokeweed mitogen, PWM)预刺激的孕妇外周血单个核细胞(Peripheral blood mononuclear cells,PBMC),采用酶联免疫斑点法(Enzyme-linked Immunospot Assay,ELISPOT)检测HLA抗体分泌记忆B细胞。比较各组记忆B细胞频数(特异性ASC/总ASC百分比)的差异。
结果正常妊娠足月剖宫产组,抗胎儿HLA抗体分泌记忆B细胞频数为0.37%±0.13%;妊娠期高血压组为0.67%±0.11%,轻度子痫前期组为0.79%±0.21%,重度子痫前期组为0.94%±0.27%。HDCP组抗胎儿HLA抗体分泌记忆B细胞频数高于正常妊娠组,其差异有统计学意义(P < 0.01)。HDCP各组间抗胎儿HLA抗体分泌记忆B细胞频数随疾病进展加重而增高,重度子痫前期与妊娠期高血压组比,其差异有统计学意义(P < 0.05);但轻度子痫前期组与妊娠期高血压组相比,其差异无统计学意义(P > 0.05)。
结论HDCP组母体血循环中抗胎儿HLA抗体分泌记忆B细胞频数较正常妊娠组增加,其可能是母体抗胎儿异体免疫增强的原因。
Hypertensive disorder complicating pregnancy (HDCP) is still a serious threaten to maternal-fetal health, which is contributed to the high maternal-fetal perinatal morbidity worldwide. A lot of studies have shown that immune imbalance occurs in HDCP. To maintain normal pregnancy, the coordination of cell-mediated immunity and humoral immunity is needed. Many reports indicated that cell-mediated immunity involved in the pathogenesis of HDCP. And most of them focused on the effects of serum cytokine profiles. However, there is lack of data about the effects of humoral immunity on the pathogenesis of HDCP. In the present study, the actual status of humoral immunity in HDCP was explored in view of both mother and fetus. It aimed to explore the effects of anti-fetal immunity on the pathogenesis of HDCP.
Objective To detect the anti-HLA antibodies in peripheral blood of women with HDCP and normal term pregnancy, and compare if the antibody specificity was consistent with the HLA genotype of fetus. To evaluate if the HLA antibodies were activated by fetal-derived antigens, illustrating the possible role of maternal anti-fetal immunity in the pathogenesis of HDCP.
Methods 60 patients with HDCP were divided into 3 groups, including gestational hypertension group, light pre-eclampsia group and severe pre-eclampsia group. 46 normal term pregnant women were selected as control group. Flow PRA was applied to screen the HLA antibodies in serum. For those women with positive results, the antibody specificity was further determinated by Luminex assay. Meanwhile, the HLA phenotypes of the mother and fetus were detected by SSP-PCR. It was compared if the HLA antibody specificity was in consistent with the HLA genotypes.
Results The positive rate of anti-HLA classⅡantibody in maternal peripheral blood of normal term pregnancy was 6.5%, while it was 20% in gestational hypertension group, 20% in pre-eclampsia group and 25% in severe pre-eclampsia group. The difference was statistically significant (P < 0.05). The positive rate of anti-HLA class I in maternal peripheral blood of normal term pregnancy was 19.6%, while it was 20% in gestational hypertension group, pre-eclampsia group and severe pre-eclampsia group, respectively. There was no significant difference (P > 0.05). The fetal HLA genotypes were in good consistence with the specificity of anti-HLA class II antibody in maternal peripheral blood.
Conclusion The positive rate of the anti-HLA classⅡantibodies in HDCP groups was increased, which was activated by the fetal-derived antigens.
Objective To compare the percentage of IL-6 positive monocytes in fetal cord blood of normal term pregnant women and patients with HDCP, finding out the activation status of fetal monocytes.
Methods 60 patients with HDCP were divided into 3 groups, including gestational hypertension group, light pre-eclampsia group and severe pre-eclampsia group. 46 normal term pregnant women were selected as control group. The cord mononuclear cells (CBMC) were isolated by density gradient centrifugation and stained with FITC-CD14 and PE-IL-6 monoclonal antibodies. The percentage of IL-6 positive monocytes in cord blood was determined by flow cytometry.
Results The percentage of IL-6 positive monocytes in cord blood of normal group was 0.78%±0.54%; while it was 4.38%±0.91% in the gestational hypertension group, 5.73%±2.18% in light pre-eclampsia and 6.29%±2.70% in severe pre-eclampsia. The difference was statistically significant (P < 0.01). Moreover, the percentage of IL-6 positive monocytes was increased along with the progression of the disease (P < 0.05).
Conclusion The fetal monocytes were activated in patients with HDCP and increased with the progression of the disease.
Objective To explore the role of anti-fetal HLA antibody-secreting memory B cells in HDCP.
Methods 60 patients with HDCP were divided into 3 groups, including gestational hypertension group, light pre-eclampsia group and severe pre-eclampsia group. 46 normal term pregnant women were selected as control group. The CBMC were isolated by density gradient centrifugation. The B cell lysate was obtained by stimulating the CBMC with IFN-γfor 72 h, which was applied as the fetal antigens. The PBMC were stimulated with PWM and cultured in vitro, which owned the ability to produce HLA antibodies. The frequency of anti-HLA antibody secreting memory B cells were detected by ELISPOT assay.
Results In control group, the frequency of anti-fetal-HLA secreting memory B cell was 0.37%±0.13%; while it was 0.67%±0.11% in gestational hypertension group, 0.79%±0.21% in light pre-eclampsia group and 0.94%±0.27% in severe pre-eclampsia group. The difference was statistically significant (P < 0.01). The difference between gestational hypertension group and severe pre-eclampsia group was statistically significant (P < 0.05). However, there was no statistically significant between gestational hypertension group and light pre-eclampsia group (P > 0.05).
Conclusion The frequency of anti-fetal HLA antibody secreting B cells was increased in HDCP, which might be the cause of enhanced humoral immunity.
引文
[1] Redman CW, Sargent IL. Latest advances in understanding preeclampsia. Science, 2005, 308: 1592 ~ 1594
[2] Sibai B, Dekker G, Kupferminc M. Pre-eclampsia. Lancet, 2005, 365: 785 ~ 799
[3] Khan KS, Wojdyla D, Say L, et al. WHO analysis of causes of maternal death: a systematic review. Lancet, 2006, 367: 1066 ~ 1074
[4] Ling Z, Yan W, Aihua L. Quantitative abnormalities of fetal trophoblast cells in maternal circulation in preeclampsia. Prenat Diagn, 2008, 28: 1160 ~ 1166
[5] Billingham RE, Brent L, Medawar PB. Active acquired tolerance of foreign cells. Natrue, 1953, 172: 603 ~ 606
[6] Choudhury SR, Knapp LA. Human reproductive failureⅠ: immunological factors. Human Reproduction Update, 2000, 7: 113 ~ 134
[7]魏军,刘岿然,隋丽红.子痫前期患者外周血单个核细胞分泌Th1,Th2型细胞因子功能变化研究.中国现代医学杂志,2005,15:2414 ~ 2415
[8] Mavrou A, Kouvidi E, Antsaklis A, et al. Identification of nucleated red blood cells in maternal circulation: A second step in screening for fetal aneuploidies and pregnancy complications. Prenat Diagn, 2007, 27: 150 ~ 153
[9] Hoskin DW, et al. Specific maternal anti-fetal lymphoproliferative responses and their regulation by natural immunosuppressive factors. Clin Exp Immunol, 1989, 76: 262 ~ 267
[10] Thellin O, Coumans B, Zorzi W, et al. Tolerance to the foeto-placental‘graft’: ten ways to support a child for nine months. Curr Opin Immunol, 2000, 12: 731 ~ 737
[11] Rogenhofer N, Toth B, Kiessig S,et al. Enzyme linked immunosorbent assay (ELISA) as screening method for anti-paternal allo-antibodies in patients with recurrent pregnany loss (RPL). Eur J Obstet Gynecol Biol, 2008, 136: 155 ~ 159
[12] Steinborn A, Seidl C, Sayehli C, et al. Anti-fetal immune response mechanism may beinvolved in the pathogenesis of placental abruption. Clin Immunol, 2004, 110: 45 ~ 54
[13] Schiessl B. Inflammatory response in preeclampsia. Mol Aspects Med, 2007, 28: 210 ~ 219
[14] Pandey KM, Thakur S, Agrawal S. Lymphocyte immunotherapy and its probable mechanism in the maintance of pregnancy in women with recurrent spontaneous abortion. Arch Gynecol Obstet, 2004, 269: 161 ~ 172
[15] Steinborn A, Saran G, Schneider A, et al. The presence of gestational diabetes is associated with increased detection of anti-HLA-classⅡantibodies in the maternal circulation. Am J Reprod Immunol, 2006, 56: 124 ~ 134
[16] Steinborn A, Sohn C, Sayehli C, et al. Preeclampsia, a pregnancy-specific disease, is associated with fetal monocyte activation. Clin Immunol, 2001, 100: 305 ~ 313
[1] Steinborn A, Sohn C, Sayehli C, et al. Preeclampsia, a pregnancy-specific disease is associated with fetal monocyte activation. Clin Immunol, 2001, 100: 305 ~ 313
[2] Steinborn A, Seidl C, Sayehli C, et al. Anti-fetal immune response mechanisms may be involved in the pathogenesis of placental abruption. Clin Immunol, 2004, 110: 45 ~ 54
[3] Ling ZD, Ziltener HJ, Webb BT, et al. Aggregated immunoglobulin and Fc fragment of IgG induce IL-6 release from human monocytes. Cell Immunol, 1990, 129: 95 ~ 103
[4] Krutmann J, Kirnbauer R, Kock A, et al.Cross-linking Fc receptors on monocytes triggers IL-6 production: role in anti-CD3-induced T cell activation. J Immunol, 1990, 145: 1337 ~ 1342
[5] Saito S, Sasaki Y, Sakai M. CD4+CD25high regulatory T cells in human p pregnancy. J Reprod Immunol, 2005, 65: 111 ~ 120
[6] Heikkinen J, Mottonen M, Alanen A, et al. Phenotypic characterization of regulatory T cells in the human deciduas. Clin Exp Immunol, 2004, 136: 373 ~ 378
[7] Sasaki Y, Sakai M, Miyazaki S, et al. Decidual and peripheral blood CD4+CD25+ regulatory T cells in early pregnancy subjects and spontaneous abortion cases. Mol Hum Rep rod, 2004, 10: 347 ~ 353
[8] Somerset DA, Zheng Y, Kilby MD, et al. Normal human pregnancy is associated with an elevation in the immune suppressive CD25+CD4+ regulatory T-cell subset. Immunology, 2004, 112: 38 ~ 43
[9] Hunt JS, Petroff MG, McIntire RH, et al. HLA-G and immune tolerance in pregnancy. FASEB J, 2005, 19: 681 ~ 693
[10] Mori T, Guo MW, Sato E, et al. Molecular and immunological approaches to mammalian fertilization. J Reprod Immunol, 2000, 47: 139 ~ 158
[11] Martin-Villa JM, Luque I, Martinez-Quiles N,et al. Diploid expression of human leukocyte antigen class I and class II molecules on spermatozoa and their cyclic inversecorrelation with inhibin concentration. Biol Reprod, 1996, 55: 620 ~ 629
[12] Sakaguchi S. Naturally arising CD4+ regulatory T cells for immunologic self-tolerance and negative control of immune responses. Annu. Rev Immunol, 2004, 22: 531 ~ 562
[13] Dekker GA, Robillard PY. Preeclampsia: A Couple’s Disease with Maternal and Fetal Manifestations. Current Pharmaceutical Design, 2005, 11: 699 ~ 710
[14] Ranella A, Vassiliadis S, Mastora C, et al. Constitutive intracellular expression of human leukocyte antigen (HLA)-DO and HLA-DR but not HLA-DM in trophoblast cells. Hum Immunol, 2005, 66: 43 ~ 55
[15] Adams KM, Yan Z, Stevens AM, et al. The changing maternal“self”hypothesis: a mechanism for maternal tolerance of the fetus. Placenta, 2007, 28: 378 ~ 382
[16] Steinman RM, Hawiger D, Nussenzweig MC. Tolerogenic dendritic cells. Annu Rev Immunol, 2003, 21: 685 ~ 711
[17] Bouteiller PL, Pizzato N, Barakonyi A, et al. HLA-G, pre-eclampsia, immunity and vascular events. J Reprod Immunol, 2003, 59: 219 ~ 234
[18] Aluvihare VR, Kallikourdis M, Betz AG. Regulatory T cells mediate maternal tolerance to the fetus. Nat Immunol, 2004, 5: 266 ~ 271
[19] Sasaki Y, Darmochwal-Kolarz D, Suzuki D, et al. Proportion of peripheral blood- and decidual CD4+CD25bright regulatory T cells in pre-eclampsia. Clin Exp Immunol, 2007, 149: 139 ~ 145
[20] Paeschke S, Chen F, Horn N, et al. Pre-eclampsia is not associated with changes in the levels of regulatory T cells in peripheral blood. Am J Reprod Immunol, 2005, 54: 384 ~ 389
[1] Sibai BM. Immunologic aspects of preeclampsia. Clin Obstet Gynecol, 1991, 34: 27 ~ 34
[2] Steinborn A, Seidl C, Sayehli C, et al. Anti-fetal immune response mechanism may be involved in the pathogenesis of placental abruption. Clin Immunol, 2004, 110: 45 ~ 54
[3] Barbara S. Inflammatory response in preeclampsia. Mol Aspects Med, 2007, 28: 210 ~ 219
[4] Darmochwal-Kolarz D, Rolinski J, Leszczynska-Gorzelak B, et al. The expressions of intracellular cytokines in the lymphocytes of preeclamptic patients. Am J Reprod Immunol, 2002, 48: 381 ~ 386
[5] Azizieh F, Raghupathy R, Makhseed M: Maternal cytokine production patterns in women with pre-eclampsia. Am J Reprod Immunol, 2005, 54: 30 ~ 37
[6] Sakai M, Tsuda H, Tanebe K,et al. Interleukin-12 secretion by peripheral blood mononuclear cells is decreased in normal pregnant subject and increased in preeclamptic patients. Am J Reprod Immunol, 2002, 47: 91 ~ 97
[7] Steinborn A, Saran G, Schneider A, et al. The Presence of Gestational Diabetes is Associated with Increased Detection of Anti-HLA-class II Antibodies in the Maternal Circulation. Am J Reprod Immunol, 2006, 56: 124 ~ 134
[8] Salafia CM, Pezzullo JC, Lopez-Zeno JA, et al. Placental pathologic features of preterm preeclampsia, Am J Obstet Gynecol, 1995, 173: 1097 ~ 1105
[9] Steinborn A, Sohn C, Sayehli C, et al. Preeclampsia, a pregnancy-specific disease, is associated with fetal monocyte activation. Clin Immunol, 2001, 100: 305 ~ 313
[10] Wegmann TG, Lin H, Guilbert L, et al. Bidirectional cytokine interactions in thematernal-fetal relationship: Is successful pregnancy a Th2 phenomenon? Immunol Today, 1993, 14: 353 ~ 356
[11] Hunt JS, Hsi BL. Evasive strategies of trophoblast cells: selective expression of membrane antigens. Am J Reprod Immunol, 1990, 23: 57 ~ 63
[12] Riteau B, Rouas-Freiss N, Menier C, et al. HLA-G2, -G3, and -G4 isoforms expressed as nonmature cell surface glycoproteins inhibit NK and antigen-specific CTL cytolysis. J Immunol, 2001, 166: 5018 ~ 5026
[13] Steinborn A, Rebmann V, Scharf A, et al. Placental abruption is associated with decreased maternal plasma levels of soluble HLA-G. J Clin Immunol, 2003, 23: 307 ~ 314
[1] Sibai BM. Immunologic aspects of preeclampsia. Clin Obstet Gynecol, 1991, 34: 27 ~ 34
[2] Steinborn A, Sohn C, Sayehli C, et al. Preeclampsia, a pregnancy-specific disease, is associated with fetal monocyte activation. Clin Immunol, 2001, 100: 305 ~ 313
[3] Steinborn A, Seidl C, Sayehli C, et al. Anti-fetal immune response mechanisms may be involved in the pathogenesis of placental abruption. Clin Immunol, 2004, 110: 45 ~ 54
[4] Steinborn A, Saran G, Schneider A, et al. The presence of gestational diabetes is associated with increased detection of anti-HLA-class II antibodies in the maternal circulation. Am J Reprod Immunol, 2006, 56: 124 ~ 134
[5] Terasaki PI. Humoral theory of transplantation. Am J Transplant, 2003, 3: 665 ~ 673
[6] Takemoto SK, Zeevi A, Feng S, et al. National conference to assess antibody-mediated rejection in solid organ transplantation. Am J Transplant, 2004, 4: 1033 ~ 1041
[7]吴长有.免疫记忆与疫苗研究开发.中国免疫学杂志, 2005, 21: 4 ~ 7
[8] Zhang L, Wang Y, Liao AH. Quantitative abnormalities of fetal trophoblast cells in maternal circulation in preeclampsia. Prenat Diagn, 2008, 28: 1160 ~ 1166
[9] Liao AH, Liu LP, Ding WP, Zhang L. Functional changes of human peripheral B-lymphocytes in pre-eclampsia. Am J Reprod Immunol, 2009, 61: 313 ~ 321
[10] Karlsson RK, Jennes W, Page-Shafer K, et al. Poorly soluble peptides can mimicauthentic EILSPOT responses. J Immunol Methods, 2004, 285: 89 ~ 92
[11] Pahar B, Li J, Rourke T, et al. Detection of antigen-specific T cell interferonγexpression by ELISPOT and cytokine flow cytometry assays in rhesus macaques. J Immunlo Methods, 2003, 282: 103 ~ 115
[12] Janetzki S, Schaed S, Blachere NE, et al. Evaluation of Elispot assays: influence of method and operator on variability of results. J Immunol Methods, 2004, 291: 175 ~ 183
[13] Lehmann PV. Image analysis and data management of ELISPOT assay results. Methods Mol Biol, 2005, 302: 117 ~ 132
[14] Cox JH, Ferrari G, Kalams SA, et al. Results of an ELISPOT proficiency panel conducted in 11 laboratories participating in international human immunodeficiency virus type 1 vaccine trials. AIDS Res Hum Retroviruses, 2005, 21: 68 ~ 81
[15] Janetzki S, Cox JH, Oden N, et al. Standardization and validation issues of the ELISPOT assay. Methods Mol Biol, 2005, 302: 51 ~ 86
[16]廖爱华,VERA Rebmann,HANS Grosse-Wilde. ELISPOT:一种新的检测人类HLA-Ⅰ类抗体的方法.中国免疫学杂志, 2005, 21: 52 ~ 59
[17]廖爱华,VERA Rebmann,HANS Grosse-Wilde等.体外诱导人类B细胞分泌HLA抗体的实验研究.细胞与分子免疫学杂志, 2007, 23: 641 ~ 644
[18] Sasaki Y, Darmochwal-Kolarz D, Suzuki D, et al. Proportion of peripheral blood and decidual CD4+CD25bright regulatory T cells in pre-eclampsia. Clin Exp Immunol, 2007, 149: 139 ~ 145
[19] Darmochwal-Kolarz D, Saito S, Rolinski J, et al. Activated T lymphocytes in pre-eclampsia. Am J Reprod Immunol, 2007, 58: 39 ~ 45
[20] Lim HW, Hillsamer P, Banham AH, et al. Cutting edge: direct suppression of B cells by CD4+ CD25+ regulatory T cells. J Immunol, 2005, 175: 4180 ~ 4183
[1] Redman CW, Sargent IL. Latest advances in understanding preeclampsia. Science, 2005, 308: 1592 ~ 1594
[2] Sibai B, Dekker G, Kupferminc M. Pre-eclampsia. Lancet, 2005, 365: 785 ~ 799
[3] Khan KS, Wojdyla D, Say L, et al. WHO analysis of causes of maternal death: a systematic review. Lancet, 2006, 367: 1066 ~ 1074
[4] Walker JJ. Pre-eclampsia. Lancet, 2000, 356: 1260 ~ 1265
[5] Lynne MM. How useful is uterine artery Doppler ultrasonography in predicting pre-eclampsia and intrauterine growth restriction. CMAJ, 2008, 178: 727 ~ 729
[6] Fernandez A, Prieto B, Escudero A, et al. A monoclonal antibody with potential for aiding non-invasive prenatal diagnosis: utility in screening of pregnant women at risk of preeclampsia. J Histochem Cytochem, 2005, 53: 345 ~ 350
[7] Saito S, Shiozaki A, Nakashima A, et al. The role of the immune system in preeclampsia. Mol Aspects Med, 2007, 28: 192 ~ 209
[8] Billingham RE, Brent L, Medawar PB. Active acquired tolerance of foreign cells. Nature, 1953, 172: 603 ~ 606
[9]王燕,廖爱华.人类白细胞抗原E和F在妊娠中的研究进展.国外医学妇产科学分册. 2007, 48: 77 ~ 86
[10]陈文超,高眉扬.导致母胎免疫耐受机制的研究进展.国外医学妇产科学分册. 2003, 14: 202 ~ 204
[11] Branch DW, Khamashta MA. Antiphospholipid syndrome: obstetric diagnosis, management, and controversies. Obstet Gynecol, 2003, 101: 1333 ~ 1344
[12]陈巧英,李大金,金莉萍等.外周CD4+CD25+T细胞升调参与人类自然流产.生殖与避孕. 2006, 26: 86 ~ 90
[13]赵爱民,林其德.同种免疫型复发性流产的病因与诊治.中国实用妇科与产科杂志, 2007, 23: 901 ~ 904
[14] Trundley A, Moffett A. Human uterine leukocytes and pregnancy. Tissue Antigens, 2004, 63: 1 ~ 12
[15] Moffett-King A. Natural killer cells and pregnancy. Nat Rev Immunol, 2002, 2: 656 ~ 663
[16] Cooper MA, Fehninger TA, Turner SC, et al. Human natural killer cells: a unique innate immunoregulatory role for the CD56bright subset. Blood, 2001, 97: 3146 ~ 3151
[17] Vilches C, Parham P. KIR: diverse, rapidly evolving receptors of innate and adaptive immunity. Annu Rev Immunol, 2002, 20: 217 ~ 251
[18]吴文林.器官移植排斥与耐受的研究进展.泉州师范学院学报, 2003, 21: 77 ~ 82
[19] Saito S, Sakai M, Sasaki Y, et al. Inadequate tolerance induction may induce pre-eclampsia. Journal of Reproductive Immunology, 2007, 76: 30 ~ 39
[20]林其德.子痫前期及子痫的病因学研究进展.中国实用妇科与产科杂志, 2006, 22: 3 ~ 5
[21] Aluvihare VR, Kallikourdis M, Betz AG, et al. Regulatory T cells mediate maternal tolerance to the fetus. Nat Immunol, 2004, 5: 266 ~ 271
[22] Somerset DA, Zheng Y, Kilby MD, et al. Normal human pregnancy is associated with an elevation in the immune suppressive CD25+CD4+regulatory T-cell subset. Immunology, 2004, 112: 38 ~ 43
[23]乐杰主编.妇产科学.第6版.人民卫生出版社, 2004, 9: 97 ~ 105
[24]林秋华主编.疑难妇产科学.科学技术出版社, 2002, 1: 299 ~ 308
[25]吴长有.免疫记忆与疫苗研究开发.中国免疫学杂志, 2005, 21: 4 ~ 7
[26]林其德.妊高征病因学研究的进展.中华妇产科杂志, 1997, 32: 325
[27]林其德.子痫前期/子痫病因及发病机制的研究进展.中国实用妇科与产科杂志, 2004, 20: 577 ~ 579
[28] Tafuri A, Alferink J, Moller P, et al. T cell awareness of paternal alloantigens during pregnancy. Science, 1995, 270: 630 ~ 633
[29] Dekker, G. The partner’s role in the etiology of preeclampsia. J Reprod Immunol, 2002, 57: 203 ~ 215
[30] Koelman CA, Coumans AB, Nijman HW, et al. Correlation between oral sex and a low incidence of preeclampsia: a role for soluble HLA in seminal fluid? J Reprod Immunol, 2000, 46: 155 ~ 166
[31]张展,贾莉婷,王玉萍等. NK细胞、HLA-G和HLA-DRB1等位基因在子痫前期发病机制中的作用研究.河南医学研究, 2007, 16: 1 ~ 6
[32]石永云,凌斌.自然杀伤细胞与妊娠研究进展.国外医学妇产科学分册, 2007, 34: 269 ~ 271
[33] Hanna J, Goldman-Wohl D, Hamani Y, et al. Decidual NK cells regulate key developmental processes at the human fetal-maternal interface. Nat Med, 2006, 12: 1065 ~ 1074
[34]姚凤球,凌斌.母胎界面免疫微环境的研究进展.国外医学妇产科学分册, 2005, 32: 281 ~ 284
[35]全松,陈雷宁.胚胎着床的子宫内膜免疫学研究现状与展望.广东医学, 2007, 28: 1553 ~ 1555
[36] Croy BA, He H, Esadeg S, et al. Uterine natural killer cells: insights into their cellular and molecular biology from mouse modeling. Reproduction, 2003, 126: 149 ~ 160
[37] Koopman LA, Kopcow HD, Rybalov B, et al. Human decidual natural killer cells are a unique NK cell subset with immunomodulatory potential. J Exp Med, 2003, 198: 1201 ~ 1212
[38] Loke YW, King A. Immunology of human placental implantation: clinical implications of our current understanding. Mol Med Today, 1999, 3: 153 ~ 159
[39] Koopman LA, Kopcow HD, RybaIov B, el a1. Human decidual natural killer cells are a unique NK cell subset with immunomodulatory potential. J Exp Med, 2003, 198: 1201 ~ 1212
[40] Moffett-King A. Natural killer cells and pregnancy. NaI Rev Immunol, 2002, 2: 656 ~663
[41] de Luca BI, Battini L, Simonelli M, et al. Increased HLA-DR homozygosity associated with preeclampsia. Hum Reprod, 2000, 15: 1807 ~ 1812
[42] Shevach EM, McHugh RS, Piccirillo CA et al. Control of T-cell activation by CD4+CD25+ suppressor T cells. Immunol Rev, 2001, 182: 59 ~ 67
[43] Saito S, Sasaki Y, SakaiM. CD4+CD25high regulatory T cells in human p pregnancy. J Reprod Immunol, 2005, 65: 111 ~ 120
[44] Heikkinen J, Mottonen M, Alanen A, et al. Phenotypic characterization of regulatory T cells in the human decidua. Clin Exp Immunol, 2004, 136: 373 ~ 378
[45]贺银燕,李大金. CD4+CD25+调节性T细胞在母胎免疫耐受中的作用.中国免疫学杂志, 2007, 23: 374 ~ 376
[46] Sasaki Y, Sakai M, Miyazaki S, et al. Decidual and peripheral blood CD4+CD25+ regulatory T cells in early p pregnancy subjects and spontaneous abortion cases. Mol Hum Reprod, 2004, 10: 347 ~ 353
[47] Somerset DA, Zheng Y, Kilby MD, et al. Normal human p pregnancy is associated with an elevation in the immune suppressive CD25+CD4+ regulatory T-cell subset. Immunology, 2004, 112: 38 ~ 43
[48] Zenclussen AC, Gerlof K, Zenclussen ML, et al. Abnormal T-cell reactivity against paternal antigens in spontaneous abortion: adoptive transfer of pregnancy-induced CD4+CD25+ T regulatory cells prevents fetal rejection in a murine abortion model. Am J Pathol, 2005, 166: 811 ~ 822
[49] Tebar F, Villalonga P, Sorkina T, et al. Calmodulin regulates intracellular trafficking of epidermal growth factor receptor and the MAPK signaling pathway. Mol Biol Cell, 2002, 13: 2057 ~ 2068
[50] Thienel T, Chwalisz K, Winterhager E. Expression of MAPkinases (Erk1/2) during decidualization in the rat: regulation by progesterone and nitric oxide. Mol Hum Reprod, 2002, 8: 465 ~ 474
[51] Strakova Z, Srisuparp S, Fazleabas AT. IL-1βduring in vitro decidualization in primate. J Reprod Immunol, 2002, 55: 35 ~ 47
[52] Scherle PA, Ma W, Lim H, et al. Regulation of cyclooxygenase-2 induction in the mouse uterus during decidualization. An event of early pregnancy. J Biol Chem, 2000, 275: 37086 ~ 37092
[53] Fallarino F, Grohmann U, Hwang KW, et al. Modulation of tryptophan catabolism by regulatory T cells. Nat Immunol, 2003, 4: 1206 ~ 1212
[54] Liao AH, Liu LP, Ding WP, Zhang L. Functional changes of human peripheral B-lymphocytes in pre-eclampsia. Am J Reprod Immunol, 2009, 61: 313 ~ 321
[55] Sasaki Y, Darmochwal-Kolarz D, Suzuki D, et al. Proportion of peripheral blood and decidual CD4+CD25bright regulatory T cells in pre-eclampsia. Clin Exp Immunol, 2007, 149: 139 ~ 145
[56] Darmochwal-Kolarz D, Saito S, Rolinski J, et al. Activated T lymphocytes in pre-eclampsia. Am J Reprod Immunol, 2007, 58: 39 ~ 45
[57] Lim HW, Hillsamer P, Banham AH, et al. Cutting edge: direct suppression of B cells by CD4+CD25+ regulatory T cells. J Immunol, 2005, 175: 4180~4183
[58]林其德.复发性流产与自身抗体关系探讨.中华妇产科杂志, 1993, 28: 674 ~ 677
[59] Chamley LW, Duncalf AM, Mitchell MD, et al. Action of anticardiolipin and antibodies to beta2-glycoprotein-I on trophoblast proliferation as a mechanism for fetal death. Lancet, 1998, 352: 1037 ~ 1038
[60] Clark DA. Is there any evidence for immunologically mediated or immunologically modifiable early pregnancy failure? J Assisted Reprod Genet, 2003, 20: 63 ~ 72
[61] Zhang L, Wang Y, Liao AH: Quantitative abnormalities of fetal trophoblast cells in maternal circulation in preeclampsia. Prenat Diagn, 2008, 28: 1160 ~ 1166
[62] Tangye SG, Avery DT, Hodgkin PD. A division-linked mechanism for the rapid generation of Ig-secreting cells from human memory B cells. J Immunol, 2003, 170: 261 ~ 269
[63] Rebmann V, Busemann A, Lindemann M, et al. Detection of HLA-G5 secreting cells. Hum Immunol, 2003, 64: 1017 ~ 1024
[64] Karlsson RK, Jennes W, Page-Shafer K, et al. Poorly soluble peptides can mimic authentic EILSPOT responses. J Immunlo Methods, 2004, 285: 89 ~ 92
[65] Pahar B, Li J, Rourke T, et al. Detection of antigen-specific T cell interferonγexpression by ELISPOT and cytokine flow cytometry assays in rhesus macaques. J Immunol Methods, 2003, 282: 103 ~ 115
[66] Cox JH, Ferrari G, Kalams SA, et al. Results if an ELISPOT proficiency panel conducted in 11 laboratories participating in international human immunodeficiency virus type 1 vaccine trials. AIDS Res Hum Retroviruses, 2005, 21: 68 ~ 81
[67] Janetzki S, Schaed S, Blachere NE, et al. Evaluation of elispot assay: influence of method and operator on variability of results. J Immunlo Methods, 2004, 291: 175 ~ 183
[68] Janetzki S, Cox JH, Oden N, et al. Standardization and validation issues of the ELISPOT assay. Methods Mol Biol, 2005, 302: 51 ~ 86
[69]廖爱华,VERA Rebmann,HANS Grosse-Wilde. ELISPOT:一种新的检测人类HLA-Ⅰ类抗体的方法.中国免疫学杂志, 2005, 21: 52 ~ 59
[70]朱薏,高眉扬. HLA-DM基因夫妇共享率与子痫前期相关性.实用医学杂志, 2007, 23(17): 2645 ~ 2647
