淋巴细胞免疫治疗对小鼠自然流产模型母胎界面CD80~+表达及妊娠结局的影响

目的:评价CBA/J×DBA/2小鼠配对组合作为反复自然流产模型的生殖力特点,及其与母胎交界CD80表达间的关系,并研究淋巴细胞免疫治疗(lymphocyte immunotherapy,LIT)对CD80表达水平的影响。方法:对CBA/J×DBA/2小鼠的生殖力特点进行为期120d的观察,并与生殖力正常的4种对照组进行比较。另计算15对CBA/J×DBA/2小鼠孕13d的胚胎吸收率,并用CD80-FITC和CD45-PE双色流式细胞术检测CD80细胞在母胎交界面的构成比。为了明确CD80~+细胞的身份,检测了CD3、DX5(NK细胞)和MHC-Ⅱ在CD80细胞群中的表达水平。此外,检测LIT组与未治疗组CBA/J×DBA/2小鼠胚胎吸收率和CD80细胞的阳性率。结果:CBA/J×DBA/2小鼠的流产特点是为孕10d左右的反复流产。CBA/J×DBA/2小鼠孕13d的胚胎吸收率显著高于BALB/c×DBA/2小鼠(30.8％±16.6％vs.7.7％±6.7％,P相似文献   

阻断协同刺激分子对MMP-9/TIMP-3的表达及妊娠结局的影响

目的：探讨阻断CD86协同刺激分子对自然流产模型孕鼠母胎界面MMP-9和TIMP-3的表达及妊娠结局的影响。方法：实验组于妊娠d 4．5腹腔注射大鼠抗小鼠CD86单抗,对照组注射大鼠同型IgG2b,而正常组不作任何处理。于妊娠d 13．5计算胚胎吸收率,并用免疫组化测定MMP- 9和TIMP-3的表达。结果：实验组的胚胎吸收率和MMP-9均显著低于对照组(P＜0．05),与正常对照组间均无差异；TIMP-3的表达与对照组和正常组比较均无显著差异(P＞0．05)。结论：在妊娠早期阻断CD86协同刺激分子能够通过某些机制诱导自然流产鼠MMP-9/TIMP-3的比值降低并且降低自然流产模型的胚胎吸收率,使模型组的胚胎吸收率恢复至正常妊娠水平。     

干预CD86协同刺激信号对母胎界面Th1/Th2型细胞因子转录调控及妊娠结局的影响

目的探讨干预CD86协同刺激信号对母胎界面Th1/Th2型细胞因子转录调控及妊娠结局的影响。方法:将正常妊娠模型(CBA/J×BALB/c)和自然流产模型(CBA/J×DBA/2J)CBA孕鼠均分为两组:对照组(各10只)于孕d 4、d 6、d 8腹腔注射大鼠IgG;干预组(各10只)于孕d 4、d 6、d 8腹腔注射大鼠抗小鼠CD86 mAb。孕d 9竞争性半定量RT-PCR测定各组母胎界面组织中Th1型(IL-12、IFN-γ)/Th2型(IL-4、IL-10)细胞因子转录水平;孕d 12比较两种模型各组的胚胎吸收率。结果:正常妊娠模型中,干预CD86协同刺激信号对母胎界面Th1/Th2型细胞因子转录水平及妊娠预后均无显著影响(P>0.05)。自然流产模型中,干预CD86协同刺激信号能够升调节母胎界面局部Th2型而降调节Th1型细胞因子转录水平,并显著改善其妊娠预后(P<0.05)。结论:于孕早期干预CD86协同刺激信号能够调控母胎界面局部Th1/Th2型细胞因子转录,形成维持正常妊娠所需的Th2型免疫偏倚,诱导母胎免疫耐受。     

干预CD86协同刺激信号在诱导母胎界面Th2型免疫偏倚中的作用

目的 :探讨干预CD86协同刺激信号在诱导母胎界面局部形成Th2型免疫偏倚中的作用。方法 :将正常妊娠模型 (CBA×BALB/c)和自然流产模型 (CBA×DBA/ 2 )CBA孕鼠均分为两组 ,于孕第 4、6、8天 ,对照组腹腔注射大鼠IgG ,实验组腹腔注射大鼠抗小鼠CD86mAb ;孕第 9天 ,ELISA测定母胎界面组织培养上清中Th1型 (IFN γ、TNF α) /Th2型(IL 4、IL 10 )细胞因子表达水平 ,并计算IL 4 /IFN γ、IL 10 /IFN γ比值 ;孕第 12天比较两种模型各组的胚胎吸收率。结果 :正常妊娠模型中 ,干预CD86协同刺激信号对母胎界面原有的Th2型免疫偏离及妊娠预后均无显著影响。自然流产模型中 ,干预CD86协同刺激信号能够诱导母胎界面局部形成Th2型免疫偏倚并显著改善其妊娠预后。结论 :于孕早期 ,干预CD86协同刺激信号能够改善母胎界面局部细胞因子微环境 ,形成维持正常妊娠所需的Th2型免疫偏倚 ,诱导母胎免疫耐受     

妊娠早期阻断协同刺激分子诱导自然流产模型孕鼠脾脏细胞母-胎免疫耐受的研究

目的　探讨阻断协同刺激分子———CD80 和CD86对自然流产模型孕鼠妊娠结局及孕鼠脾脏免疫细胞对父系抗原免疫耐受状态的影响。方法　将雌性小鼠 (CBA/J)分别与BALB/c及DBA/2两种雄性小鼠合笼交配 ,分别建立正常妊娠模型CBA/J×BALB/c( 2 0只 ,对照组 )和自然流产模型CBA/J×DBA/2 ( 2 0只 ,研究组 )。CBA/J小鼠于妊娠第 4天 (着床期 )腹腔分别注射大鼠同型IgG 0 2mg( 10只 ) ,或大鼠抗小鼠CD80 和CD86单克隆抗体 ( 10只 )。妊娠第 9天 ,采用单向混合淋巴细胞反应 ,分析孕鼠脾脏免疫细胞对父系抗原的增殖能力 ,并测定细胞培养上清液中白细胞介素 2(IL 2 )水平 ,以研究脾脏细胞母 胎免疫耐受状态 ;妊娠第 14天观察两组的胚胎吸收率。结果　 ( 1)研究组中 ,腹腔注射大鼠IgG的孕鼠胚胎吸收率为 2 4 3% ,而注射大鼠抗小鼠CD80 和CD86单克隆抗体的孕鼠胚胎吸收率为 9 8% ,两者比较 ,差异有显著性 (P <0 0 5 )。 ( 2 )应用大鼠抗小鼠CD80 和CD86单克隆抗体 ,使妊娠 9d的孕鼠脾脏免疫细胞对父系抗原的增殖能力及IL 2水平显著下降(P <0 0 5 )。结论　孕早期阻断协同刺激分子 ,可诱导产生孕鼠脾脏免疫细胞对父系抗原的免疫耐受 ,从而使自然流产模型孕鼠的妊娠结局达到正常妊娠水平。     

Blockade of CD80 and CD86 at the time of implantation inhibits maternal rejection to the allogeneic fetus in abortion-prone matings

CD28/CTLA-4 interactions with their specific B7-ligands (CD80 and CD86) play a decisive role in antigenic and allogenic responses. Recently, experimental transplant studies demonstrated that donor-specific tolerance was achieved by blocking these interactions. However, the role of blockade of CD28/B7 costimulatory pathway in the maintenance of materno-fetal tolerance has received little attention. In the present study, abortion-prone CBA/J females mated with DBA/2 males were administered with anti-CD80 and anti-CD86 monoclonal antibodies (mAbs) on day 4 of gestation (time of murine implantation). We demonstrated that the combined use of anti-CD80 and anti-CD86 mAbs induced maternal tolerance to the fetus in the abortion-prone CBA/J mice, and displayed expansion of the maternal CD4(+)CD25+ regulatory T cell population and up-regulated expression of CTLA-4, suggesting an active mechanism of regulatory T cells in suppressing maternal rejection to the fetus. In addition, the anti-CD80/86 mAbs treatment enhanced Th2 and reduced Th1 cytokine production in mice, implying that the development of Th2 cells might contribute to maternal tolerance to her fetus. Together, these findings indicated that blocking CD80 and CD86 enhanced maternal tolerance to her fetus in mice by increasing regulatory T cell function and skewing toward a Th2 response. Our data might provide an enhanced understanding of the maternal-fetal immune relationship and be helpful in clinical trials for immunotherapy of recurrent spontaneous abortion.     

Adoptive transfer of CD4+CD25+ regulatory T cells for prevention and treatment of spontaneous abortion

Objectives

The purpose of this study was to examine whether adoptive transfer with in vitro expanded CD4⁺CD25⁺ regulatory T cells (Tregs) could prevent immune response-mediated spontaneous abortion in mice.

Study design

Female CBA/J mice were mated with male Balb/c as the control with normal pregnancy or with DBA/2J mice as a model of spontaneous abortion. The CBA/J mice mated with DBA/2J were treated intravenously with freshly isolated or in vitro expanded Tregs on day 1 or 4 of pregnancy, respectively. The numbers of surviving and reabsorbed fetuses in the different groups of mice were counted on day 14 of pregnancy, and the concentrations of cytokines in individual sera and the supernatants of cultured Tregs were measured by ELISA.

Results

Adoptive transfer with freshly isolated Tregs only slightly reduced the fetal resorption rate, which was not significantly different from that of the mice without Treg treatment, regardless of treatment at early stage and implementation of pregnancy. In contrast, adoptive transfer with in vitro expanded Tregs significantly reduced the fetal resorption rates, particularly for treatment at early stage of pregnancy (P < 0.05). Furthermore, adoptive transfer with in vitro expanded Tregs at early stage of pregnancy significantly increased the levels of serum IL-10, TGF-β1, and the ratios of IL-10 to IFN-γ.

Conclusions

Our data clearly indicated that adoptive transfer with in vitro expanded Tregs at early stage of pregnancy protected fetuses from spontaneous abortion by re-establishing immune tolerance in mice.     

抗原递呈细胞表面共刺激分子CD80/CD86表达与自然流产的关系

目的 :探讨共刺激分子CD80 /CD86与自然流产的关系。方法 :采用双标记流式细胞分析技术检测自然流产小鼠模型CBA/J×DBA/ 2脾脏及肠系膜淋巴结内 (MLN)抗原递呈细胞MΦ表面CD80 /CD86的表达情况 (n =10 ) ,以正常妊娠小鼠模型CBA/J×BALB /c为对照 (n =5 )。结果 :1、自然流产模型组脾脏内表达CD80MΦ含量为 1.82±0 .4 1% ,与正常妊娠模型组的 1.64%± 0 .61%差异无显著性 (P >0 .0 5 ) ,而表达CD86MΦ含量在自然流产模型组中为 2 .34%± 0 .67% ,明显低于正常妊娠模型组的 5 .98%±2 .4 3% (P <0 .0 5 ) ;2、自然流产模型组MLN内表达CD80MΦ含量为 10 .2 0 %± 5 .4 2 % ,明显高于正常妊娠模型组 1.5 8%± 0 .70 % ,差异有显著性 (P <0 .0 5 ) ,而表达CD86MΦ含量在自然流产模型组中为 1.4 6%± 0 .5 7% ,明显低于正常妊娠模型组 3.96%± 0 .39% (P <0 .0 0 1)。结论 :抗原递呈细胞表面共刺激分子CD80 /CD86的表达异常在自然流产的发病中起重要作用     

IL-4与IL-10联合免疫对趋化因子受体CCR3、CCR5、CXCR3的选择性诱导对自然流产模型小鼠胚胎丢失率的影响

目的:观察IL-4与IL-10对趋化因子受体CCR3、CCR5和CXCR3的选择性诱导对自然流产模型小鼠胚胎丢失率的影响,探讨趋化因子受体CCR3、CCR5、CXCR3在诱导妊娠免疫耐受中的作用。方法:建立自然流产小鼠模型与正常妊娠小鼠模型,观察细胞因子IL-4与IL-10对CCR3、CCR5、CXCR3的选择性诱导作用,用双标记流式细胞分析技术,检测正常妊娠模型组孕鼠(CBA/J×BALB/c)、自然流产模型无干预组孕鼠(CBA/J×DBA/2)、自然流产模型IL-4免疫组孕鼠、自然流产模型IL-4+IL-10联合免疫组孕鼠和自然流产模型-生理盐水(NS)免疫组孕鼠中外周血CD4+T细胞CCR3、CCR5、CXCR3等3类趋化因子受体的表达率,并观察各组孕鼠胚胎丢失率。结果:(1)流产模型无干预组胚胎丢失率显著高于正常妊娠模型组,差异有统计学意义(P<0.01),IL-4免疫组、IL-4+IL-10联合免疫组胚胎丢失率皆明显低于NS组与流产模型无干预组(P<0.01,P<0.01);(2)自然流产模无干预组外周血CD4+T细胞CCR3表达水平明显低于正常妊娠模型组(P<0.01),而CCR5、CXCR3表达率明显高于正常妊娠模型组(P<0.01);转输IL-4、IL-4+IL-10后CCR3表达率明显上调、CXCR3表达率明显下降,均与流产模型无干预组差异有显著性(P<0.01)。此外,IL-4+IL-10联合免疫组外周血CD4+T细胞上CCR5表达率也明显低于流产模型无干预组(P<0.05),但IL-4免疫组与流产模型无干预组无明显差异(P>0.05)。结论:CD4+T细胞上CCR3、CCR5、CXCR3表达异常可能在自然流产发病中起重要作用,细胞因子IL-4与IL-10联合作用可能通过诱导CCR3高表达,抑制CCR5与CXCR3表达来诱导妊娠免疫耐受,降低胚胎丢失率。     

协同刺激分子CD86在母-胎免疫调节中的作用

目的　探讨协同刺激分子CD86在母 胎免疫调节中的作用。方法　建立自然流产模型(CBA×DBA/ 2 )及正常妊娠模型 (CBA×BALB/c)。两种模型分别再分为 3个组 :(1)以大鼠的IgG为对照的对照组 ;(2 )于妊娠第 4、6、8、10天 ,分别给CBA孕鼠腹腔注射大鼠抗小鼠CD86单克隆抗体的多次干预组 ;(3)仅于妊娠第 4天 ,给CBA孕鼠单次腹腔注射抗CD86单克隆抗体的单次干预组。每次腹腔注射的抗体剂量均为 10 0 μg。于妊娠第 14天计算各组胚胎吸收率。 结果　 (1)自然流产模型与正常妊娠模型的对照组胚胎吸收率分别为 2 7 78%和 8 42 %。 (2 )于妊娠第 4、6、8、10天腹腔注射抗CD86单克隆抗体后 ,自然流产模型的多次干预组胚胎吸收率下降至 9 6 8% (P <0 0 5 ) ;而正常妊娠模型的多次干预组胚胎吸收率上升至 13 5 4% (P >0 0 5 )。 (3)妊娠第 4天单次腹腔注射抗CD86单克隆抗体 ,自然流产模型单次干预组的胚胎吸收率下降至 7 14% (P <0 0 0 1) ;而正常妊娠模型单次干预组的胚胎吸收率上升至 11 39% (P >0 0 5 )。结论　妊娠早期 ,尤其在胚胎对母体的致敏阶段 (着床期 ) ,阻断母 胎界面的CD86协同刺激信号 ,将诱导母体对胚胎的免疫耐受 ,从而减少胚胎吸收 ,提高妊娠成功率     

Maternal lymphocyte subpopulations (CD45RA+ and CD45RO+) in preeclampsia

OBJECTIVE: The maternal syndrome of preeclampsia has been attributed to a systemic intravascular inflammatory response and endothelial cell dysfunction. The stimulus responsible for intravascular inflammation in preeclampsia has not been determined. The expression of CD45 isoforms on the surface of human T cells has been used to classify CD4(+) T lymphocytes into na?ve cells (CD45RA+) and memory T cells (CD45RO+). An increased percentage of CD45RO+ cells has been interpreted as consistent with previous exposure to microbial products or other antigens. The purpose of this study was to determine whether preeclampsia is associated with a change in the proportion of CD45RA+ and CD45RO+. STUDY DESIGN: A prospective study was conducted in patients with preeclampsia (n = 24) and normal pregnancy (n = 75). The percentage of CD45RA+ and CD45RO+ on CD4(+) T lymphocytes in peripheral blood was determined using flow cytometry and monoclonal antibodies. Results were reported as a percentage of CD4(+) lymphocytes. Parametric statistics were used for analysis. A probability value of <.05 was considered statistically significant. RESULTS: Patients with preeclampsia had a significantly higher percentage of CD45RO+ than normal pregnant women (P <.01). A significantly lower percentage of CD45RA+ was found in patients with preeclampsia than in normal pregnant women (P <.01). CONCLUSION: Preeclampsia is associated with an increase in the percentage of CD45RO+ and a decrease in the CD45RA+ lymphocyte subpopulation. Therefore, patients with preeclampsia have evidence of previous antigenic exposure, the nature of which remains to be established.     

The therapeutic potential of the recombinant antigen from Dirofilaria immitis (rDiAg) for immune-mediated pregnancy loss

The mammalian fetuses are semi-allograft for mothers. Therefore the failure of immunological tolerance often causes pregnancy loss. Recently, the effects of helminthes therapy for immune mediated diseases have been reported. In the present study we employed the murine model to examine the therapeutic potential of the recombinant antigen from a nematoda parasite, Dirofilaria immitis for immune mediated pregnancy loss. Recombinant D. immitis polyproteins (rDiAg) had been cloned and selected by us for the strongest immuno-regulatory activities in parasite antigens. Female CBA/J mice were injected with sterilized rDiAg or PBS solution using micro-osmotic pumps before mating. Pregnant CBA/J mice were sacrificed on day 13.5 for scoring the number of resorbed and viable fetuses for histological and immunological analysis. The serum cytokine concentrations were measured using suspension array system. The resorption rate of mock-treated mice was 42.9% (resorbed fetus 12/total fetus 28). The resorption rate was decreased to 11.1% (resorbed fetus 3/total fetus 27) with rDiAg treatments. The IL-4, IL-23 and TNF-α concentrations in serum were significantly lower in rDiAg-treated mice than mock-treated mice. The serum IL-17 level was also reduced in rDiAg-treated mice but the difference was not significant. The rDiAg treatment reduced the resorption rates of CBA/J×DBA/2J mouse model, which mimic human pregnancy failures with allo-immune backgrounds. Our observations suggest as the first time of therapeutic potentials of the rDiAg for pregnancy loss.     

Expression of intercellular adhesion molecule-1 (ICAM-1) on the surface of peripheral blood lymphocytes of pregnant women with pregnancy-induced hypertension

OBJECTIVES: Intercellular adhesion molecule-1 (ICAM-1, CD54) is involved in process of leukocytes adhesion to endothelium as well as in their migration to surrounding tissues. There is much evidence that pregnancy-induced hypertension (PIH) presents a state of endothelial destruction mediated partially by increased ICAM-1 expression on endothelial cells and neutrofils. DESIGN: The aim of this study was to evaluate the expression of ICAM-1 (CD54) molecule on the peripheral blood lymphocytes of pregnant women with PIH studied "in vitro". MATERIALS AND METHODS: Preeclampsia (PE) and transient hypertension (TH) were defined according to USA National Health Institute criteria. The study group consisted of 16 women with preeclampsia (PE), 12 women with transient hypertension (TH) and 9 women with physiological pregnancy. The group of 8 nonpregnant women served as controls. Exclusion criteria were: uterine contractions, infection and steroid therapy before blood sampling. Peripheral blood was obtained by venipuncture. Lymphocytes were isolated and cultured by using standard procedures. Mitogenic doses of phytohaemaglutynin (PHA) were added to each culture. Immunofluorescent marking techniques with anty-CD54 one-step monoclonal antibodies were performed. Analysis was made with FACSCalibur flow-cytometer with 488 nm argon laser using CellQuest programme. The results were described as the percentage of CD54+ lymphocytes and MFI index corresponding density of CD54 molecules on the lymphocyte surface. Statistical analysis was performed using t-Student and U-Mann-Whitney tests. The work was sponsored by KBN 4 P05E 118,15 grant. RESULTS: The percentage of CD54+ lymphocytes in physiological pregnancy compared to nonpregnant women did not differ significantly (56.9 +/- 20.8% vs. 57.2 +/- 14.0%, p = 0.97). The MFI value was increased in pregnant women but in comparison with nonpregnant women did not reach statistical significance (34.7 +/- 35.7 vs. 17.8 +/- 4.3, p = 0.20). The percentage of CD54+ lymphocytes in TH group compared to normal pregnant women did not differ significantly (52.2 +/- 18.6% vs. 56.9 +/- 20.8%, p = 0.58) but MFI value was significantly increased (100.6 +/- 81.5 vs. 34.7 +/- 35.7). In PE group compared to normal pregnant women the percentage of CD54+ lymphocytes as well as MFI value were significantly increased (CD54+: 70.8 +/- 12.9% vs. 56.9 +/- 20.8%, p < 0.05; MFI: 170.8 +/- 91.7 vs. 34.7 +/- 35.7, p < 0.0005). CONCLUSIONS: 1/expression of ICAM-1 molecule on peripheral blood lymphocytes studied "in vitro" during normal pregnancy is not different in comparison to the nonpregnant state, but 2/ in pregnancy complicated with PIH is significantly increased, especially in PE, 3/described changes are a sign of the lymphocyte activation and may be responsible for endothelial destruction observed in PIH.     

低剂量米非司酮对植入窗口期子宫内膜中子宫自然杀伤细胞数量及其亚型含量的影响

目的 探讨低剂量米非司酮对植入窗口期子宫内膜中子宫自然杀伤(uNK)细胞数量及其亚型含量的影响.方法 收集正常妇女植入窗口期的子宫内膜14份,将每份内膜组织平均分为3部分,分别用浓度为65 nmol/L米非司酮(A组)、200 nmol/L米非司酮(B组)和0 nmol/L米非司酮(对照组)进行体外培养,运用免疫组化和流式细胞技术检测uNK细胞数量及CD-3CD+56CD-16亚型、CD-3CD+56CD+16亚型的百分含量.结果 (1)A、B及对照组子宫内膜中CD+56uNK细胞数量分别为(148±11)、(150±12)和(121±7)个,A、B组分别与对照组比较,差异均有统计学意义(P＜0.05);A、B组之间比较,差异无统计学意义(P＞0.05).(2)A组和B组子宫内膜中CD-3CD+56亚型百分含量分别为(44±5)%和(48±4)%,高于对照组[(35±3)%],差异有统计学意义(P＜0.05),A、B组间比较,差异无统计学意义(P＞0.05);A组和B组子宫内膜CD-3 CD+56CD-16亚型百分含量分别为(42±5)%和(45±5)%,高于对照组[(33±3)%],差异有统计学意义(P＜0.05),A、B组间比较,差异无统计学意义(P＞0.05);CD-3CD+56CD+16亚型百分含量分别为(2.70±0.24)%、(3.26±0.37)%和(2.33±0.29)%,3组间比较,差异无统计学意义(P＞0.05).结论 低剂量米非司酮可通过增加uNK细胞数量及CD-3CD+56CD-16亚型百分含量,使植入窗口期子宫内膜局部免疫微环境失平衡,从而可能导致胚胎植入的失败.     

Vaccination against spontaneous abortion in mice

We report here that the high rate of spontaneous resorption observed in CBA/J female mice mated with DBA/2 J males can be dramatically reduced by vaccination with Balb/c male spleen cells, but not by CBA/J or DBA/2 J male spleen cells. This effect correlates with the differential ability of Balb/c spleen cells to induce MLR suppressor activity in CBA/J female mice, and should lead to a better understanding of the immunology of the materno-fetal relationship.     

原因不明复发性流产患者外周血及蜕膜组织中CD+4CD+25调节性T细胞比例的变化

目的 探讨原因不明复发性流产(URSA)患者外周血及蜕膜组织中CD+4CD+25调节性T(Tr)细胞比例的变化.方法 采用双荧光标记流式细胞分析技术检测25例URSA患者(流产组)、34例正常早孕妇女(正常妊娠组)和22例正常非孕妇女(正常非孕组)外周血及蜕膜组织中CD+4CD+25Tr细胞的比例.结果 (1)流产组和正常妊娠组妇女外周血CD+4 CDbright25T细胞的比例[分别为(1.55±0.77)%、(2.65±1.10)%]均显著高于正常非孕组妇女[(0.39±0.14)%],分别比较,差异均有统计学意义(P<0.05);流产组妇女外周血CD+4 CDbright25T细胞的比例显著低于正常妊娠组妇女,两组比较,差异有统计学意义(P<0.05).(2)流产组妇女蜕膜组织中CD+4 CDbright25T细胞比例[(0.59±0.23)%]显著低于正常妊娠组妇女[(1.24±0.55)%],两组比较,差异有统计学意义(P<0.01).流产组妇女蜕膜组织中CD+4CDdim25T细胞比例[(4.23±1.52)%]与正常妊娠组[(3.75±1.88)%]比较,差异无统计学意义(P>0.05).(3)正常妊娠组妇女蜕膜组织中CD+4CDbright25T细胞占CD+4T细胞的比例(CD+4CDbright25/CD+4)为(13.10±10.25)%,显著高于外周血[(5.59±2.62)%],两者比较,差异有统计学意义(P<0.05);流产组患者蜕膜组织中CD+4CDbright25/CD+4比例[(5.16±2.83)%]与外周血[(4.64±2.07)%]比较,差异无统计学意义(P>0.05).结论 CD+4CD+25Tr细胞数量在早孕期显著升高,参与了正常妊娠的维持,有可能是调控母-胎界面局部免疫耐受形成的一个重要因素;CD+4 CD+25Tr细胞数量的减少可能与URSA的发生有关.     

复发性流产患者外周血T淋巴细胞亚群及CD4^＋辅助性T淋巴细胞亚型的变化

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DC-sign+ CD163+ macrophages expressing hyaluronan receptor LYVE-1 are located within chorion villi of the placenta

The purpose of this study was to investigate with immunohistochemical methods antigen presenting cells and their relationship to blood and lymphatic vessels in human term placenta. Fetal placental antigen presenting cells, historically also known as Hofbauer cells, were located in the chorionic villi below the syncytiotrophoblast and in the vicinity of fetal capillaries. DC-SIGN/CD209 expression was observed on CD163+, CD68+, CD45+, HLA-A,B,C+, DC-LAMP/CD208-, CD86-, Langerin/CD207-, FXIIIa-, CD1a- cells consistent with the macrophage nature of these cells. These fetal DC-SIGN+ cells lack HLA-DR, -DP, -DQ expression. Moreover, we show for the first time that they co-express the hyaluronan receptor LYVE-1. In contrast, no LYVE-1+ vessel structures, i.e. lymphatic vessels, were detected. Human term decidua hosted a variety of CD45+ cells, further phenotyped as CD163+, DC-SIGN+, CD68+, HLA-DR+, HLA-A,B,C+. Mature dendritic cells were never observed in human term placenta. In summary, human term placenta is an immunoprivileged organ without lymphatic drainage and with numerous DC-SIGN+ macrophages within the chorionic villi. We hypothesize that these cells may fulfil a function in innate responses against pathogens as well as be involved in the homeostasis of hyaluronan metabolism in the rapidly differentiating placenta.     

泼尼松治疗原发性肾病综合征对患儿CD4+CD25+调节性T细胞的影响分析

目的　观察原发性肾病综合征（PNS）患儿泼尼松治疗前后CD4+CD25+ 调节性T细胞（CD4+CD25+ Tr）的变化,阐明肾上腺糖皮质激素治疗儿童PNS的免疫机制。方法　选择2004—2007年在深圳市儿童医院住院治疗的初发PNS患儿42例,其中激素敏感型32例,激素耐药型10例。同期25例健康体检儿童作为对照组。流式细胞术检测泼尼松治疗前后外周血CD3+CD4+CD8-、CD3+CD4-CD8+、CD4+CD25+Tr的比例,荧光定量聚合酶链反应（Real-time PCR）检测泼尼松治疗前后外周血单个核细胞（PBMC）叉头型基因P3（Foxp3）、细胞毒性T淋巴细胞相关抗原4（CTLA-4）和糖皮质激素诱导的肿瘤坏死因子受体 （GITR）基因mRNA的表达。结果 　与对照组比较,PNS患儿外周血CD3+CD4+CD8- T细胞、CD3+CD4-CD8+ T细胞、CD4+CD25+ Tr比例无明显改变（P > 0.05）。激素敏感型PNS患儿CD4+CD25+Tr比例在泼尼松治疗后明显增高,差异有统计学意义（P < 0.01）;激素耐药型PNS患儿CD4+CD25+Tr比例在泼尼松治疗前后无明显改变（P > 0.05）。激素敏感型PNS患儿在泼尼松治疗后PBMC细胞Foxp3、CTLA-4和GITR基因mRNA的表达明显增高;而激素耐药型PNS患儿泼尼松治疗前后Foxp3、CTLA-4基因表达无明显改变,仅GITR表达明显增高。 结论 泼尼松等肾上腺糖皮质激素类药物可通过上调激素敏感型PNS患儿CD4+CD25+调节性T细胞的表达发挥免疫治疗作用。     

Immunohistochemical analysis of CD4+ and CD8+ T-cell subsets in high risk human papillomavirus-associated pre-malignant and malignant lesions of the uterine cervix

OBJECTIVES: Humoral and cellular immune responses are likely to play a key role for the clearance or persistence and progression of high risk (HR) HPV-associated cervical lesions. Although there are many studies describing the systemic T-cell responses to HPV16 and 18 proteins, few data are available regarding the cellular mucosal immune responses. We used immunohistochemistry to characterize populations of T-immune cells (CD4+, CD8+, CD45RO+) in HR-HPV-infected precancerous and cancerous lesions of the uterine cervix. METHODS: Four biopsies from normal cervix, 9 CIN1 which have regressed (rCIN), 5 CIN1 which have progressed (pCIN) to high grade lesions, 13 CIN3 and 11 invasive carcinomas were included. All dysplasias and carcinomas were HR-HPV-positive and low-risk-HPV-negative. They were stained with monoclonal antibodies specific for CD4, CD8 and CD45RO and examined by microscopy. STATISTICAL ANALYSIS: The Kruskal-Wallis test and the Siegel's and Castelan's method were used. RESULTS.: CD4+ cells predominated in regressing CIN1 both within the stroma and the epithelium with the highest CD4+/CD8+ ratio compared with pCIN1, CIN3 and invasive carcinoma. At the exception of CD45RO+ cells, T cells were detected with similar frequencies in both pCIN1 and CIN3. However, in 7 out of 10 CIN3, CD4+ and CD8+ cells were visible as organized lymphoid follicle structure. The CD8+ and CD45RO+ cells far exceeded the CD4+ cells in invasive cancers. CONCLUSIONS: Density and distribution of immune T cells depend on the malignant potential of HR-HPV lesions. These results suggest that the studied lymphocyte subsets have an important role to fulfil during the natural history of HR-HPV-associated lesions.