乳腺癌异常表达B7-H4对T细胞分泌细胞因子和凋亡的影响

目的:探讨B7-H4在乳腺癌中的表达及其对外周血T细胞分泌细胞因子及增殖、凋亡的影响。方法:应用SP免疫组化法检测B7-H4在乳腺浸润性癌、癌旁组织和纤维腺瘤的表达。流式细胞术分析B7-H4对乳腺癌患者外周血活化T细胞增殖和凋亡的作用。ELISA芯片检测T细胞培养上清液细胞因子的含量。结果:B7-H4在乳腺浸润性癌的阳性表达率为84.62%(44/52),显著高于癌旁组织和纤维腺瘤组织(P0.01,P0.01)。体外淋巴细胞混合培养结果显示,与空白组比较,B7-H4对乳腺癌患者外周血活化T细胞的增殖指数Ki67无明显影响;但B7-H4诱导CD8~+T细胞凋亡的作用强于CD4~+T细胞。B7-H4组FOXP3~+T/CD4~+T也高于空白组(P0.05)。B7-H4组较空白组细胞培养上清液中TGF-β1、IL-17含量均显著增高(P0.05,P0.05)。结论:乳腺浸润性癌异常表达B7-H4。B7-H4在体外能促进CD8~+T细胞凋亡、促进T细胞分泌TGF-β1和IL-17。B7-H4在削弱乳腺癌微环境的抗肿瘤细胞免疫中发挥一定作用。     

B7-H4在人子宫颈癌的表达及其与肿瘤内浸润T细胞亚群的相关性研究

目的:研究共刺激分子B7-H4在人子宫颈癌的表达及其与肿瘤内浸润的FOXP3+、CD4+T、CD8+T细胞数量和分泌细胞因子的关系。方法:采用SP免疫组织化学染色检测B7-H4在30例正常人宫颈组织、30例高级别宫颈上皮内瘤变(Cervical intraepithelial neoplasiaⅡ-Ⅲ,CINⅡ-Ⅲ)和67例子宫颈癌的表达;间接免疫荧光双标(Indirect immunofluorescentdouble-staining)观察肿瘤内浸润的FOXP3+、CD4+T、CD8+T细胞数量及其TGF-β1和IFN-γ分泌情况。结果:B7-H4不表达于正常人宫颈上皮,仅在部分瘤变宫颈上皮微弱表达;子宫颈癌B7-H4的阳性表达率为46%(31/67),显著高于正常人宫颈上皮和瘤变宫颈上皮(P<0.01,P<0.05);子宫颈癌B7-H4阳性组病灶内浸润的CD8+T细胞以及分泌IFN-γ的CD8+T数量显著低于B7-H4阴性组(P<0.001,P<0.035);B7-H4的表达与肿瘤内浸润的FOXP3+细胞、CD4+T细胞以及分泌TGF-β1的CD4+T细胞数量无关(P>0.05,P>0.05)。结论:B7-H4在人子宫颈癌细胞异常高表达,并与肿瘤内浸润的CD8+T细胞数量减少以及分泌IFN-γ减少有关,提示B7-H4在抑制肿瘤微环境的细胞免疫中发挥作用。     

协同刺激分子B7-H4在胃癌组织中的表达及定位

探讨负性协同刺激分子B7-H4在胃癌组织及浸润免疫细胞亚群中的表达及定位。应用免疫组织化学、免疫荧光染色及激光共聚焦技术检测B7-H4蛋白在胃癌组织中的表达及定位;流式细胞术检测分析胃癌患者PBMC亚群(CD3+T细胞、CD19+B细胞、CD14+单核细胞及粒细胞)中B7-H4的表达。免疫组织化学检测结果表明,B7-H4在胃癌组织中呈阳性表达。免疫荧光染色及激光共聚焦检测结果显示,B7-H4在胃癌组织中呈阳性表达,并主要定位于胃癌细胞的细胞膜,特别是在Ki67+高度增殖的胃癌细胞中B7-H4呈高表达,且B7-H4在CD34+血管内皮细胞中呈高表达;B7-H4在浸润CD19+B细胞和CD68+巨噬细胞中呈高表达;而在胃癌浸润CD4+T细胞、CD8+T细胞及CD57+NK细胞中呈弱表达。流式细胞术检测结果表明,B7-H4在胃癌患者外周血浸润CD19+B细胞及粒细胞中呈阳性表达,而在浸润CD3+T细胞及CD14+单核细胞中呈弱表达。本研究结果提示B7-H4在胃癌组织中可表达于肿瘤细胞和部分浸润免疫细胞,并可通过其介导的信号途径参与胃癌的发生与发展。检测B7-H4的表达可作为潜在的胃癌诊断、治疗及预后判断的新靶点。     

B7-H3单抗交联作用对Mo-DC体外生物学功能的影响

探讨B7-H3分子对人外周血单核细胞来源树突状细胞(Mo-DC)体外成熟和生物学功能的影响。采用常规方法从健康人外周血单核细胞诱导DC,在诱导过程中,加入B7-H3单抗21D4共培养,经流式细胞术检测Mo-DC上B7-H3分子和其他共刺激分子的表达,ELISA试剂盒检测培养上清中细胞因子IL-10和IFN-γ的分泌量,并采用3H-TdR掺入法测定T细胞的增殖。结果:B7-H3分子在未成熟和成熟Mo-DC上均有高水平表达,抗人B7-H3单抗21D4能上调Mo-DC表面CD80、CD86和CD83的表达,提高Mo-DC的共刺激能力,促进T细胞的体外增殖,并能显著促进T细胞分泌IL-10。由此表明,B7-H3单抗21D4交联作用可以促进Mo-DC体外成熟,上调其共刺激T细胞的能力。     

负性共刺激分子B7-H4在结直肠癌及其微环境中的表达及临床意义

负性共刺激分子B7-H4是新近发现的B7家族的新成员。为了检测人结直肠癌组织中肿瘤细胞及肿瘤浸润淋巴细胞(tumor-infiltrating lymphocyte,TIL)上B7-H4的表达以及分析巨噬细胞、T淋巴细胞及其亚群的浸润,进而探讨其相关临床意义,本课题采用免疫组化SP法检测98例人结直肠癌组织中肿瘤细胞及浸润淋巴细胞上B7-H4的表达以及巨噬细胞、T细胞及其亚群的浸润,统计学分析B7-H4的表达与患者临床病理参数、预后、肿瘤相关巨噬细胞(tumor-associated mac-rophage,TAM)以及CD3+T细胞、CD8+T细胞浸润程度的相关性。结果显示,结直肠癌组织中高表达B7-H4分子,而正常组织中基本不表达,并且在肿瘤细胞及浸润淋巴细胞中均检测到B7-H4的表达。统计学分析表明,浸润淋巴细胞中B7-H4的表达与患者年龄、有无淋巴结转移、是否为粘液腺癌有关(P<0.05),与患者总体生存率呈负相关(P<0.05),且B7-H4高表达组织较B7-H4低表达组织CD3+T细胞高度浸润例数显著增加(P<0.05);肠癌细胞中B7-H4的表达与患者肿瘤分期、淋巴结转移、有无远处转移以及Dukes分期有关(P<0.05)。综上所述,人结直肠癌患者的肠癌细胞及浸润淋巴细胞较正常细胞均异常表达B7-H4分子,CD3+T细胞的浸润程度与浸润淋巴细胞中B7-H4的表达呈显著相关而与肿瘤细胞上B7-H4的表达无关,而肿瘤细胞表达B7-H4与肿瘤分期、淋巴结转移、有无远处转移以及Dukes分期有关。上述结果提示肿瘤细胞表达B7-H4与浸润淋巴细胞表达B7-H4可能分别以不同机制参与了肿瘤的发生和进展,进一步分析肿瘤组织中B7-H4在不同细胞表达的生物学和临床意义具有重要的价值。     

B7-H3在人肝癌细胞株HepG2对外周血CD8+T细胞活化、周期及分泌IL-17调节中的作用

目的 研究B7-H3在人肝癌细胞株HepG2对人外周血CD8+T细胞活化、周期及IL-17分泌等调节中的作用.方法 RT-PCR及FCM检测B7-H3在HepG2细胞上的表达;应用脂质体法将PGPU6/GFP/neo-B7-H3shRNA质粒转入肝癌细胞株HepG2,阻断B7-H3的表达;免疫磁珠分选健康人外周血CD8+T细胞;FCM分析B7-H3分子在HepG2细胞对PHA刺激下CD8+T细胞活化、周期及PMA刺激下CD8+T细胞分泌IL-17调节中的作用.结果 肝癌细胞株HepG2高表达B7-H3分子,PGPU6/GFP/neo-B7-H3 shRNA质粒能有效阻断B7-H3在HepG2细胞上的表达;FCM分析结果显示,肝癌细胞株HepG2对CD8+T细胞活化及周期均有抑制作用;阻断B7-H3的表达后,明显减弱HepG2细胞对CD8+T细胞早期活化表型CD69表达的抑制作用,且能够通过下调CD8+T细胞Go/G1期细胞数量,上调S期细胞数量逆转HepG2细胞对CD8+T细胞周期的阻滞作用;在HepG2存在条件下,CD8+T细胞对IL-17的分泌明显增加,阻断B7-H3的表达后,IL-17的分泌被进一步上调.结论 HepG2细胞高表达B7-H3分子;B7-H3能够协同HepG2细胞对CD8+T细胞活化表型CD69的表达及细胞周期的抑制作用;HepG2细胞上调CD8+T细胞对IL-17的分泌作用,但B7-H3可抑制该上调作用.     

原发性肝癌组织中B7-H3表达与浸润的CD8~+ T细胞数负相关

目的探讨原发性肝癌组织中B7-H3的表达与CD8+T细胞浸润之间的关系。方法采用免疫组织化学技术分析原发性肝癌组织和正常肝组织中B7-H3的表达、CD8+T细胞的浸润数量,对B7-H3的表达与CD8+T细胞浸润数量的相关性进行统计学分析。结果 63例肝癌患者中B7-H3高表达26例(41.3%),低表达37例(58.7%),总阳性率为90.5%,其中正常肝组织基本不表达B7-H3。B7-H3高表达组CD8+T细胞浸润数量明显低于B7-H3低表达组,CD8+T细胞浸润数量与肝癌组织中B7-H3的表达呈负相关。肝癌组织中B7-H3的表达和CD8+T细胞数量在肝癌患者的临床病理分期、是否有局部淋巴结转移以及远隔器官转移等方面具有显著性差异。结论原发性肝癌组织中高表达B7-H3,与肿瘤组织中CD8+T细胞浸润呈负相关。     

人B7-H4基因转染细胞的构建及其对T细胞的共刺激效应的初步研究

目的 构建含有人B7-H4基因的重组逆转录病毒载体，获得稳定表达B7-H4基因的1929细胞并初步研究其对T细胞的共信号作用及可能机制。方法从含人B7-H4基因的cDNA序列FLJ22418中，采用PCR扩增出B7-H4全长基因，双酶切装入逆转录病毒载体pEGZ-HA-Term，与辅助病毒载体脂质体法共转染包装细胞293T，用其培养上清感染1929细胞72h后，经Zeocin筛选出稳定表达B7-H4蛋白的L929细胞株；采用免疫荧光标记和流式细胞仪分析免疫分子的表达、^3H-TdR掺入检测细胞增殖以及ELISA测定细胞因子的水平。结果构建了含人B7-H4基因的重组逆转录病毒载体和获得含有B7-H4基因的重组病毒，筛选获得能稳定高表达人B7-H4蛋白的I_929转基因细胞；该转基因细胞对T细胞体外具有抑制增殖、活化和细胞因子分泌的作用。结论构建了稳定表达人B7-H4蛋白的细胞株，B7-H4分子在体外通过抑制T细胞分泌IL-2和促进其凋亡作用可显著地下调T细胞功能的效应。     

原发性肝癌组织中B7-H3表达与浸润的CD8^+ T细胞数负相关北大核心CSCD

目的探讨原发性肝癌组织中B7-H3的表达与CD8+T细胞浸润之间的关系。方法采用免疫组织化学技术分析原发性肝癌组织和正常肝组织中B7-H3的表达、CD8+T细胞的浸润数量,对B7-H3的表达与CD8+T细胞浸润数量的相关性进行统计学分析。结果 63例肝癌患者中B7-H3高表达26例(41.3%),低表达37例(58.7%),总阳性率为90.5%,其中正常肝组织基本不表达B7-H3。B7-H3高表达组CD8+T细胞浸润数量明显低于B7-H3低表达组,CD8+T细胞浸润数量与肝癌组织中B7-H3的表达呈负相关。肝癌组织中B7-H3的表达和CD8+T细胞数量在肝癌患者的临床病理分期、是否有局部淋巴结转移以及远隔器官转移等方面具有显著性差异。结论原发性肝癌组织中高表达B7-H3,与肿瘤组织中CD8+T细胞浸润呈负相关。     

乳腺癌中HCCR-1的表达及干扰HCCR-1对MCF-7细胞生物学行为的影响

目的 探讨HCCR-1在乳腺浸润性导管癌中的表达及其对乳腺癌细胞体外增殖、凋亡、迁移的影响。方法 采用免疫组化法检测HCCR-1在乳腺浸润性导管癌中的表达，分析其与乳腺癌临床病理特征的关系。应用siRNA干扰乳腺癌细胞株MCF-7中HCCR-1表达，采用qRT-PCR和Western blot法验证siRNA的干扰效率。应用CCK-8实验检测细胞增殖；细胞凋亡实验检测细胞凋亡；Transwell实验检测细胞迁移能力。结果 HCCR-1在乳腺浸润性导管癌中高表达(66.9%,107/160)。HCCR-1表达与临床分期、组织学分级、淋巴结转移、ER表达、PR表达相关(P<0.05)。HCCR-1-siRNA可显著下调MCF-7中HCCR-1的表达(P<0.001)。CCK8实验显示：干扰HCCR-1表达可抑制MCF-7的增殖(P<0.05)。细胞凋亡实验显示：干扰HCCR-1表达可促进MCF-7的凋亡(P<0.05)。Transwell实验显示：干扰HCCR-1表达可抑制MCF-7的迁移(P<0.05)。结论 HCCR-1在乳腺浸润性导管癌中高表达，HCCR...     

B7-H4 overexpression impairs the immune response of T cells in human cervical carcinomas

Objective

To investigate B7-H4 expression and its correlation with the number of infiltrating T lymphocytes and cytokine production by those lymphocytes in human cervical cancer and to determine the effect of recombinant B7-H4 on the active peripheral blood T cells of the patients in vitro.

Methods

B7-H4 expression was detected in 67 cases of cervical cancer using immunohistochemical staining. Tumor-infiltrating CD8⁺T, CD4⁺T, and FOXP3⁺ (Forkhead Box P3) T lymphocytes and their levels of IFN-γ and TGF-β₁ production were determined by immunofluorescent double-staining. After the peripheral blood T lymphocytes of patients were co-cultured with B7-H4, proliferation, apoptosis, and cell subtypes were analyzed using flow cytometry. Cytokines in the supernatant were detected by ELISA.

Results

B7-H4 was expressed in 46% (31/67) of the cases of cervical cancer. The number of infiltrating CD8⁺T lymphocytes and their IFN-γ production in positive B7-H4 expression cervical cancers was significantly lower than in negative B7-H4 cases (P < 0.01, P < 0.05), but there was no significant difference between cases positive and negative for B7-H4 with respect to infiltrating FOXP3⁺T and CD4⁺T cells or TGF-β1 production. After co-culture with B7-H4 for 48 h, the patients’ activated T lymphocytes were arrested at G1/G2 phase. The Ki67 positive rates of CD4⁺T and CD8⁺T cells were 2.13 ± 0.13% and 1.03 ± 1.33%, and they were lower than in the blank group. The proportion of CD4⁺T and CD8⁺T cells decreased, but CD4⁺T/CD8⁺T and the proportion of CD4⁺CD25⁺Foxp3⁺T cells increased. In addition, concentrations of IL-10 and TGF-β1 in the supernatant of co-cultured T cells increased significantly (P < 0.05, P < 0.05), but that of IFN-γ decreased. B7-H4 had no significant effect on apoptosis of the T cells.

Conclusion

B7-H4 is overexpressed in human cervical cancers, and it is associated with lower numbers of tumor-infiltrating CD8⁺T lymphocytes and therefore less IFN-γ production. In vitro, B7-H4 inhibits the proliferation of CD4⁺T and CD8⁺T but promotes the proliferation of Tregs and the secretion of IL-10 and TGF-β1. B7-H4 plays an important role in depressing the anti-tumor immunity of CD8⁺T cell in microenvironments of cervical cancer.     

CD95 ligand expression as a mechanism of immune escape in breast cancer

Interaction of CD95 (Apo-1/Fas) and its ligand (CD95L) plays an important role in the regulation of the immune response, since CD95+ lymphocytes may be killed after engagement of the CD95 receptor. Studying the CD95/CD95L system in 40 cases of breast cancer, the malignant cells expressed CD95L, but lost CD95 expression, when compared with non-malignant mammary tissue. Jurkat T cells incubated on breast cancer sections underwent CD95L-specific apoptosis. The rate of apoptosis correlated with the CD95L mRNA levels of the tissue samples. In four breast cancer cell lines, CD95L expression was increased by interferon-gamma (IFN-gamma), which resulted in higher levels of CD95L-specific apoptosis in co-cultured Jurkat T cells. Since IFN-gamma is mainly secreted by activated T cells, up-regulation of CD95L in breast cancer cells in response to IFN-gamma may thus counterselect activated tumour-infiltrating T cells and favour the immune escape of breast cancer. As demonstrated by inhibition of matrix metalloproteinases, CD95L expressed on breast cancer cells can also be shed from the cell membrane into the culture supernatant. Supernatants derived from cultured breast cancer cells induced apoptosis in Jurkat T cells via CD95L. In breast cancer patients, depletion of CD4+ and CD8+ peripheral blood lymphocytes was significantly correlated with CD95L expression in the tumours. This might be suggestive for a relationship between CD95L expression by breast cancer and systemic immunosuppression.     

急性淋巴细胞白血病患者外周血CD4+CD25+T调节细胞与白细胞介素2、白细胞介素10、转化生长因子β的检测*

背景：目前急性淋巴细胞白血病的病因尚未完全清楚,而CD4+CD25+T调节细胞对机体的免疫反应可能起着一定的调节作用。 目的：观察急性淋巴细胞白血病患者外周血CD4+CD25+T调节细胞分泌白细胞介素2、白细胞介素10及转化生长因子β的情况。 方法：采用密度梯度离心法分离22例B细胞系列的急性淋巴细胞白血病(B-ALL)患者、13例T细胞系列的急性淋巴细胞白血病(T-ALL)患者和18名正常人外周血单个核细胞。采用免疫磁珠法分离出CD4+CD25+T调节细胞进行培养,同时用双抗体夹心酶联免疫吸附试验检测细胞培养上清中白细胞介素2、白细胞介素10及转化生长因子β的水平。 结果与结论：B-ALL患者和T-ALL患者外周血CD4+CD25+T调节细胞培养上清中白细胞介素10和转化生长因子β水平显著高于正常人水平,而白细胞介素2水平显著低于正常人水平(P < 0.05);B-ALL和T-ALL患者的CD4+CD25+T调节细胞培养上清中3种因子表达差异无显著性意义(P > 0.05)。提示CD4+CD25+T调节细胞可能通过增加分泌白细胞介素10和转化生长因子β、减少分泌白细胞介素2等干扰抗肿瘤的免疫反应。      

Soluble B7-H1: differences in production between dendritic cells and T cells

Tumor cells aberrantly express several T cell inhibitory molecules including members of the B7-H co-regulatory family. Presumably tumor-expressed B7-H1 and B7-H3 confer resistance to elimination by the immune system. In addition, elevated levels of soluble B7-H1 (sB7-H1) has been identified in the sera of cancer patients, including renal carcinoma patients and is associated with increased cancer related death. Here we report that sB7-H1 is produced and released by activated mature dendritic cells (mDC). Immature DC, macrophages, monocytes, or T cells are refractory to releasing sB7-H1. Exposure of CD4+ and CD8+ T cells to mDC-derived sB7-H1 molecules induced apoptosis. These data suggest that the immunobiology of B7-H1 is perhaps more complex than previously thought. sB7-H1 molecules may represent an unanticipated contributing factor to immune homeostasis. That both immune and tumor cells can be sources of sB7-H1 suggests that optimization of co-regulatory blockade immunotherapy for solid malignancies of necessity will require impact of targeting tumor and immune-derived B7-H1 molecules.     

B7-h4 expression in a range of breast pathology: correlation with tumor T-cell infiltration.

B7-H4, a member of the B7 family, is involved in the regulation of antigen-specific immune responses. Its expression in a range of breast pathology and correlation to the number of CD3 and CD8 tumor infiltrating T-lymphocytes in invasive carcinomas were explored. The proportion of B7-H4 positive cells, staining pattern, and intensity were evaluated within diagnostic groups (normal and benign lesional, potentially premalignant and in situ carcinoma, and invasive carcinoma) on archival tissue blocks by immunohistochemistry. The proportion and intensity of B7-H4 expression was progressively increased across the major diagnostic groups. There was a significant association between a high proportion of B7-H4 positive cells in invasive ductal carcinomas and decreased number of tumor infiltrating lymphocytes (P=0.002). The cellular distribution of B7-H4 appears altered in the spectrum of normal to malignant breast. Its overexpression may help cancers avoid immune detection.     

组蛋白去乙酰化酶抑制剂罗米地辛对小鼠T细胞表型及功能的影响

目的:通过体外实验观察罗米地辛对效应T细胞和调节性T细胞的作用。方法:取CFSE标记的淋巴细胞、CD4+T细胞和CD8+T细胞作为反应细胞,实验组加入不同浓度梯度(1、3、5μmol/L)的罗米地辛及CD3/CD28单抗进行淋巴细胞培养,以仅加入CD3/CD28单抗作为阳性对照组,另设空白对照组。培养72 h后检测各组细胞的增殖情况。以淋巴细胞作为反应细胞,实验组、阳性对照组及空白对照组设定同上,72 h后检测各组中CD4+Foxp3+T细胞与CD4+T细胞的比例变化。同时采用ELISA检测培养液中相关细胞因子,如TNF-α、IL-10及TGF-β水平的变化。结果:罗米地辛剂量依赖性地抑制CD3/CD28单抗诱导的淋巴细胞、CD4+T细胞和CD8+T细胞的增殖(P0.05)。在CD3/CD28单抗存在的条件下,1μmol/L的罗米地辛不能诱导CD4+Foxp3+T细胞的比例上调(P0.05)。但提高罗米地辛的浓度至3μmol/L和5μmol/L后,CD4+Foxp3+T细胞的比例显著提高(P0.05)。随着罗米地辛浓度的增加,TNF-α和IL-10水平呈剂量依赖性降低,但各实验组明显高于空白对照组而低于阳性对照组(P0.05)。TGF-β在阳性对照组虽稍有升高,但与空白对照组相比无显著差异(P0.05)。而随着罗米地辛浓度的增加,TGF-β水平呈剂量依赖性升高,3实验组间及与空白对照组、阳性对照组间差异显著(P0.05)。结论:体外实验研究显示罗米地辛不仅能够有效抑制效应性T细胞的增殖,而且一定浓度的罗米地辛可上调调节性T细胞的比例,这可能与TGF-β升高有关,而与IL-10变化无关。     

Immune accessory functions of human endothelial cells are modulated by overexpression of B7-H1 (PDL1)

B7-H1 (PDL1) is a B7-related protein that inhibits T-cell responses. Human endothelial cells (EC), which can support polyclonal stimulation (by anti-CD3 or Phytohemagglutinin (PHA)) or direct alloantigen stimulation of T cells, basally express B7-H1 and increase expression in response to IFN-gamma or coculture with allogeneic T cells. Previous studies have suggested that endogenous B7-H1 on EC reduces T-cell responses. We engineered overexpression of B7-H1 in EC (B7H1-EC) to evaluate whether this manipulation could reduce T-cell responses even further. Compared with green fluorescent protein-transduced EC (GFP-EC), B7H1-EC support less anti-CD3 or PHA-induced proliferation of CD4+ memory T cells; naive CD4+ T-cell or CD8+ T-cell responses were less inhibited. The effect of transduced B7H1-EC was more apparent when the EC were fixed prior to coculture, a manipulation that reduces the strength of costimulation and prevents upregulation of the endogenous B7-H1 molecule. T-cell activation markers, including CD25, CD62L, CD152 (CTLA-4), and CD154 (CD40L), were not altered by EC overexpression of B7-H1, whereas there was a reduction in CD69. B7-H1 reduced secretion of IL-2 and IL-10 by memory T cells. B7H1-EC were less able to stimulate allogeneic proliferation of CD4+ memory T cells than control EC. These data suggest that B7-H1 overexpression may be a useful approach for reducing allogeneic CD4+ memory T-cell responses to EC.     

胎盘蛋白14增强调节性T细胞诱导滤泡调节性T细胞比例增加的作用及机制研究

目的：研究胎盘蛋白14（PP14）增强调节性T细胞（Treg细胞）诱导滤泡调节性T细胞（Tfr细胞）增殖的作用及机制。方法：分离健康志愿者外周血中的淋巴细胞；磁珠分选淋巴细胞中的Treg细胞；流式检测CD4⁺CD25⁺Foxp3⁺Treg细胞比例及其表面CTLA-4和HLA-G的表达；ELISA检测上清中IL-10和TGF-β的浓度；采用Transwell小室进行Treg细胞和淋巴细胞的共培养实验；流式检测淋巴细胞中CD4⁺CXCR5⁺Foxp3⁺Tfr细胞的比例。结果：相对于对照组，PP14组Treg细胞的比例增加了1.26倍（t=4.748，P=0.009 0），而其表面CTLA-4和HLA-G的表达与对照组没有显著的统计学差异（t=0.108，P=0.918 6；t=0.439，P=0.682 8）；ELISA的结果显示相对于对照组，PP14组Treg细胞培养上清中IL-10和TGF-β的浓度分别上升了3.66倍和3.86倍（t=6.817，P=0.002 4；t=7.537，P=0.001 7）；共培养实验显示PP14能够增强Treg细胞诱导Tfr细胞比例的增加，相对于共培养组，PP14组Tfr细胞比例增加了2.71倍（t=4.815，P=0.008 6），IL-10和TGF-β的阻断均能部分阻断这种增强作用（t=4.868，P=0.008 2；t=3.670，P=0.021 4）。结论：PP14能够通过分泌IL-10和TGF-β，显著增强Treg细胞诱导Tfr细胞比例的增加，这为不明原因复发性流产的治疗提供了一个新的靶点。