人慢性胃炎粘膜内CD3+细胞,S-100+树突状细胞和nNOS表达的意义

采用免疫细胞化学方法探讨胃粘膜内CD3+细胞,S-100+树突状细胞和nNOS的表达与慢性胃炎的关系及意义.检测标本均取自胃窦部活检的胃粘膜组织.结果显示CD3+细胞主要分布于粘膜上皮、腺上皮和固有膜内,而S-100+树突状细胞则主要位于固有膜内,正常组与浅表性胃炎组和萎缩性胃炎组,浅表性胃炎组与萎缩性胃炎组细胞数量有显著性差异( P<0.01),nNOS阳性反应主要位于粘膜上皮和腺上皮的基底部,但各组之间nNOS的表达程度不同,特别是萎缩性胃炎与浅表性胃炎有显著性差异(P<0.01),我们认为,对CD3+ 细胞,S-100 +树突状细胞和nNOS的检测,不仅有助于判断胃炎的病变程度和临床疗效,而且也为胃炎的治疗提供新的启示.     

慢性胃炎粘膜内CD3+细胞,S-100+树突状细胞和nNOS表达与幽门螺旋杆菌感染的关系研究

目的　探讨幽门螺旋杆菌 (Hp)与慢性胃炎粘膜内CD3+ 细胞 ,S 10 0 + 树突状细胞和nNOS表达之间的关系。方法　用免疫细胞化学方法 ,检测Hp+ 和Hp 胃炎患者及正常人的胃窦部活检标本。结果　Hp+ 胃炎组胃窦粘膜内CD3+ 细胞和S 10 0 + 树突状细胞数量明显高于Hp-胃炎和正常组 ,且有显著性差异 (P <0 0 1和P <0 0 5 ) ,而nNOS呈高表达 ,与Hp-胃炎组相比有显著性差异 (P <0 0 1)。结论　Hp+ 胃炎患者胃窦粘膜内CD3+ 细胞和S 10 0 + 树突状细胞的增加是机体对Hp的免疫应答 ,而nNOS在Hp+ 胃炎组的高表达可能与Hp感染有密切关系。     

人慢性胃炎胃粘膜内胃泌素及其mRNA的表达和意义

目的　探讨慢性胃炎胃粘膜对细胞内胃泌素 (G)及其mRNA的表达和意义。方法　标本均来自活检的胃窦部粘膜组织 ,用免疫细胞化学和原位杂交技术。结果　对照组G细胞及GmRNA阳性细胞数 ,与浅表性胃炎组比较无显著性差异 (P >0 0 5 ) ,而与萎缩性胃炎组比较则有显著性差异 (P <0 0 1) ,不论对照组还是胃炎组的G细胞数量与GmRNA阳性细胞均有平行关系。结论　证明浅表性和萎缩性胃炎的胃泌素基因的转录和蛋白表达功能保存完好 ,因此检测G细胞的数量及GmRNA不仅可以帮助了解胃炎的萎缩程度 ,而且能判断临床疗效。     

幽门螺杆菌感染者CD4~+ CD25~+调节性T细胞的检测及其相关性分析

目的检测幽门螺杆菌(Helicobacter pylori,H.pylori)感染阳性的胃部疾病患者外周血中CD4+CD25+调节性T细胞(Treg细胞)的百分含量及转化生长因子-β1(transforming growth factor-β1,TGF-β1)的水平,探讨CD4+CD25+调节性T细胞在H.pylori感染中的免疫调节作用及意义。方法采用流式细胞术检测H.pylori感染的慢性浅表性胃炎、胃癌前病变和胃癌患者外周血中CD4+CD25+调节性T细胞的含量、CD4+CD25+T细胞中表达FOXP3的细胞比例;并采用ELISA方法检测H.pylori感染者血清中TGF-β1的含量,无H.pylori感染的患者作为阴性对照。结果 H.pylori感染的患者外周血中CD4+CD25+调节性T细胞的百分含量及TGF-β1的水平较不伴有H.pylori感染的患者显著升高(P<0.05);H.pylori感染的浅表性胃炎、胃癌前病变及胃癌患者外周血中CD4+CD25+T淋巴细胞的百分含量及CD4+CD25+T细胞中表达FOXP3的细胞比例随病变严重程度的进展逐渐升高,差异有统计学意义(P<0.05);H.pylori感染的患者血清中TGF-β1水平也随病变严重程度的进展逐渐升高,差异有统计学意义(P<0.05)。结论 H.pylori感染可增加CD4+CD25+调节性T细胞的含量和TGF-β1的水平;随着病变严重程度的进展,CD4+CD25+调节性T细胞的含量和TGF-β1的水平逐渐升高,CD4+CD25+调节性T细胞百分含量和TGF-β1水平可作为临床判断病情进展的指标。     

非霍奇金淋巴瘤患者外周血中CD4+CD25+ 调节性T 细胞 亚群变化初探

目的:检测非霍奇金淋巴瘤(non-Hodgkin’s lymphoma,NHL)患者外周血中CD4+CD25+调节性T细胞(CD4+CD25+regulatoryT cell,Treg)的改变,探讨Treg与NHL的相关性。方法:病例组(n=60)为本院收治的初诊NHL患者,对照组(n=60)为本院健康体检者,用流式细胞技术联合标记CD4、CD25检测对照组及病例组化疗前、化疗后的外周血中CD4+CD25+调节性T细胞的分布特点。结果:(1)病例组化疗前外周血中CD4+细胞比例显著低于对照组(P<0.05),CD4+CD25+调节性T细胞比例显著高于对照组(P<0.05);(2)病例组化疗后,CD4+细胞比例明显高于化疗前(P<0.05),CD4+CD25+调节性T细胞比例明显低于化疗前(P<0.05);(3)病例组化疗后CD4+细胞比例与对照组无显著差异(P>0.05),而CD4+CD25+调节性T细胞比例显著高于对照组(P<0.05)。结论:非霍奇金淋巴瘤患者外周血中CD4+CD25+调节性T细胞比例升高,存在机体免疫抑制,化疗可降低CD4+CD25+调节性T细胞比例。     

树突状细胞对胃癌前病变的免疫保护分析

摘要 目的：探究树突状细胞(Dendritic cells,DC)对胃癌的免疫保护作用。方法：选择2016年1月至2018年1月于我院接受治疗的145例胃癌、39例慢性萎缩性胃炎、21例不典型增生、27例肠上皮化生以及20例正常对照组患者为研究对象,分别采集其胃粘膜标本进行染色,记录和比较其胃粘膜中S100⁺、CD4⁺和CD8⁺细胞的数量、平均面积以及平均吸光度,并将胃癌患者分为中分化腺癌(49例)、低分化腺癌(53例)和未分化癌(43例)进行对比。结果：(1)胃癌组、慢性萎缩性胃炎组、不典型增生、肠上皮化生组的胃粘膜S100⁺阳性细胞计数明显高于正常对照组（P<0.05）,胃癌组平均吸光度低于对照组,其他3组平均吸光度显著高于对照组,(P<0.05);胃癌组平均面积与正常对照组相比无差异(P>0.05),其他三组平均面积显著高于对照组（P<0.05）;(2)慢性萎缩性胃炎组、肠上皮化生组、不典型增生组患者CD4⁺细胞数均低于对照组(P<0.05);胃癌组、慢性萎缩性胃炎组、肠上皮化生组患者平均面积均低于对照组(P<0.05);胃癌组、慢性萎缩性胃炎组、不典型增生、肠上皮化生组平均吸光度均低于对照组(P<0.05);(3)慢性萎缩性胃炎组、肠上皮化生组、不典型增生组患者CD8⁺细胞数明显高于对照组(P<0.05),胃癌组稍低于对照组(P>0.05);胃癌组患者平均面积低于对照组(P<0.05);胃癌组患者平均吸光值低于对照组,慢性萎缩性胃炎组、肠上皮化生组患者高于对照组(P均<0.05);(4)随着胃癌分化程度的降低,胃癌患者DC细胞数有降低趋势。结论：胃癌前病变患者胃粘膜中DC数量会显著增多,免疫功能加强,DC细胞数量会随胃癌分化程度的降低而减少,分析其原因与DC细胞能够抑制癌前病变有关。     

幽门螺旋杆菌感染的慢性胃炎胃窦粘膜内NF-κB和TGF-α表达及其意义

目的　探讨幽门螺旋杆菌 (Hp)与慢性胃炎患者胃窦粘膜内NF κB和TGF α表达之间的关系。方法　用免疫细胞化学方法 ,检测Hp+ 和Hp-胃炎患者及正常人的胃窦部活检标本。结果　NF κB和TGF α在正常组胃粘膜内弱表达。在胃炎病人 ,NF κB和TGF α表达增强。特别是在Hp+ 组 ,NF κB和TGF α呈高表达 ,与Hp-组和正常组比较均有显著性差异 (P <0 0 5和P <0 0 1)。此外在G细胞和部分腺上皮的细胞核内也呈高表达。TGF α常以颗粒状的特征位于腺上皮细胞核上区而且高表达。结论　NF κB和TGF α的表达在胃炎组增强 ,而且Hp+ 组远高于其他两组。NF κB和TGF α二者的表达平行且成呈正相关。     

人和大鼠胃窦部神经内分泌细胞分布和形态学的比较研究

目的探讨人和大鼠胃窦部神经内分泌细胞的分布和形态学特征。方法采用免疫细胞化学方法,检测人和大鼠胃窦部粘膜内生长抑素细胞(D细胞)、胃泌素细胞(G细胞)、5-羟色胺细胞(5-HT细胞)、嗜铬粒素A细胞(CgA细胞)的分布。结果人和大鼠的G、D细胞的特征是都具有细胞突起,但是在细胞密度及其分布上是不同的;5-HT细胞的分布在两组稍有不同,在大鼠胃窦部,大多数5-HT细胞位于腺体基部,而人的5-HT细胞主要在间质,少数位于腺上皮内;在两组中,CgA细胞几乎布满整个胃粘膜,其数量也高于其他神经内分泌细胞,CgA细胞形态多样,胞质内充满细小颗粒。结论1)人与大鼠的G、D细胞通常都伴有突起,但其分布不同。2)5-HT细胞形态多样,分布于间质和腺上皮内。3)CgA细胞特征是胞质内充满细小颗粒,细胞形态多样,几乎布满整个粘膜。     

棕色田鼠胃肠道5-羟色胺细胞的免疫组织化学定位

应用ABC免疫组织化学方法,对棕色田鼠胃肠道内5-羟色胺细胞进行了免疫组织化学定位和形态学观察.结果 表明:5-HTIR细胞在棕色田鼠胃肠道的各段均有分布,其中以十二指肠和直肠段分布密度最高,胃底、胃幽门部、空肠、回肠、盲肠段其次,胃贲门部、胃体、结肠段分布密度最低.5-羟色胺细胞位于胃腺上皮、肠粘膜上皮、肠腺上皮及固有膜,有圆形、椭圆形、梭形、楔形和不规则形,有的还具有胞突.对棕色田鼠胃肠道5-HTIR细胞的分布、形态与功能相适应的特点进行了讨论.     

幽门螺杆菌感染对慢性胃炎胃窦部EC细胞和CgA细胞的影响

目的探讨幽门螺杆菌（Helicobacter pylori Hp）感染与慢性胃炎（CG）患者胃窦粘膜内肠嗜铬细胞（Enterochromaffin,EC细胞）、嗜铬粒素A细胞（CgA细胞）变化的关系。方法采用免疫细胞化学方法,检测Hp相关性慢性胃炎胃窦部粘膜内EC细胞、CgA细胞的分布。结果1．Hp^＋组CgA^＋细胞数低,与Hp^＋组和正常组比较差异显著（P〈0．01）;2.EC细胞数随病理类型加重呈下降趋势,三组间的差异性极显著（P〈0．01）,慢性萎缩性胃炎组（CAG）CgA^＋细胞数与慢性非萎缩性胃炎（CNAG）组和正常组比较,差异极显著（P〈0．01）。结论Hp感染和慢性胃炎胃窦部EC细胞和CgA细胞之间存在密切关系。     

雷公藤甲素对哮喘豚鼠外周血CD_4~+和CD_8~+淋巴细胞的影响

探讨雷公藤甲素在治疗哮喘中对外周血 T淋巴细胞的影响机制 ,采用免疫细胞化学方法检测 30例豚鼠外周血淋巴细胞 CD+ 4 、 CD+ 8的表达。实验动物分为对照组、哮喘组和雷公藤甲素治疗组 (治疗组 ) ,每组各 1 0只。结果表明 ,治疗组CD+ 4 淋巴细胞表达阳性率及表达强度明显低于哮喘组 (P<0 .0 1 ) ,CD+ 8阳性率高于哮喘组 (P<0 .0 5 ) ,与对照组比较差异无显著性。本研究认为 ,雷公藤甲素可能通过增高哮喘豚鼠 CD+ 8淋巴细胞 ,降低 CD+ 4 淋巴细胞来发挥抗哮喘气道炎症作用。     

Isolation and Purification of Colon Lamina Propria Dendritic Cells from Mice with Colitis

Dendritic cells are prime antigen presenting cells for stimulation of T cell immune responses. These cells are present in trace amounts in normal tissue. At sites of disease the increased frequency of these cells interacting with T cells may provide the basis for the release of pro-inflammatory mediators and contribute to localised cell and tissue damage. Studies on dendritic cells in the colon lamina propria of inflammatory bowel disease (IBD) mice have been limited due to the difficulties encountered in the isolation and purification of sufficient numbers of these cells. This is the first detailed, reproducible method provided in the literature for the isolation of colon lamina propria dendritic cells from mice with colitis, yielding optimum purity of cells and sufficient numbers to advance the study of dendritic cell function in the colons of mice. The most frequently used identification marker of murine DC is the CD11c surface antigen. We have adapted, combined, and improved procedures developed for the isolation of other cell types, to develop an efficient procedure for the isolation of dendritic cells from colon tissue. This protocol describes a step-by-step method for optimising the purity and recovery of lamina propria CD11c+ dendritic cells from mice colons.     

慢性胃炎患者粘膜内EGF和TFF1的表达及G细胞的相关研究

目的探讨EGF和TFF1在慢性胃炎患者胃窦粘膜内的表达及G细胞相关研究.方法免疫细胞化学.结果 EGF和TFF1在慢性胃炎粘膜内的表达与正常对照组相比均有减弱或消失,而且有显著性差异(P<0.01).在萎缩性胃炎的肠化区EGF和TFF1均为阴性,但与其相邻的腺体内TFF1仍为阳性反应.G细胞数目在Hp 和Hp-组之间无显著性差异,与正常对照组比较亦无差异(P>0.05);但萎缩性胃炎的G细胞显著减少,与正常对照组有差异(P<0.01).结论慢性胃炎粘膜内的EGF和TFF1表达减弱或消失与Hp或其它致病原有关.肠化区内的TFF1消失,可能与细胞恶变有关.Hp 组患者的G细胞无增加.     

Myeloid CD11c+ S100+ dendritic cells express indoleamine 2,3-dioxygenase at the inflammatory border to invasive lower lip squamous cell carcinoma

The prevalence of squamous cell carcinoma of the lower lip (SCC-LL) is increasing worlwide. The expression of the enzyme indoleamine 2,3-dioxygenase (IDO) by antigen-presenting cells and/or tumor cells leads to tumor escape by inhibiting T cell-mediated rejection responses. The aim of this study was to determine the expression of IDO in SCC-LL. IDO-expression was analyzed in 47 SCC-LL, together with the expression of markers of T-cells (CD3), myeloid DCs (S100, CD11c), macrophages (CD68, CD11c), Langerhans cells (CD1a, Langerin (CD207)), plasmacytoid DCs (CD123), and regulatory T cells (Foxp3) by immunohistochemistry and immunofluorescence analysis. Twelve specimens out of 47 LL-SCCs contained cells that expressed IDO. IDO-positivity was strongly associated with the intensity of the cancer-associated infiltrate (P=0.0007). IDO-positive cells are located right along the border between the developing tumor and the inflammatory infiltrate. Immunofluorescence stainings showed that CD11c+S100+CD68- dendritic cells (DCs) express IDO in SCC-LL. IDO expression in LL-SCC may aid immune escape and chronic inflammation to promote cancer progression. Inhibition of IDO might be a therapeutic strategy to increase the anti-tumor immune response in SCC-LL.     

Carboxylated glycans mediate colitis through activation of NF-kappa B

The role of carbohydrate modifications of glycoproteins in leukocyte trafficking is well established, but less is known concerning how glycans influence pathogenesis of inflammation. We previously identified a carboxylate modification of N-linked glycans that is recognized by S100A8, S100A9, and S100A12. The glycans are expressed on macrophages and dendritic cells of normal colonic lamina propria, and in inflammatory infiltrates in colon tissues from Crohn's disease patients. We assessed the contribution of these glycans to the development of colitis induced by CD4(+)CD45RB(high) T cell transfer to Rag1(-/-) mice. Administration of an anti-carboxylate glycan Ab markedly reduced clinical and histological disease in preventive and early therapeutic protocols. Ab treatment reduced accumulation of CD4(+) T cells in colon. This was accompanied by reduction in inflammatory cells, reduced expression of proinflammatory cytokines and of S100A8, S100A9, and receptor for advanced glycation end products. In vitro, the Ab inhibited expression of LPS-elicited cytokines and induced apoptosis of activated macrophages. It specifically blocked activation of NF-kappaB p65 in lamina propria cells of colitic mice and in activated macrophages. These results indicate that carboxylate-glycan-dependent pathways contribute to the early onset of colitis.     

CD4 positive Leu-8 negative helper-inducer T cells predominate in the human intestinal lamina propria

The regulatory function of peripheral blood CD4 T cells correlates with the presence or absence of the membrane glycoprotein recognized by anti-Leu-8 antibody; CD4,Leu8- T cells help Ig synthesis and CD4,Leu-8+ T cells suppress Ig synthesis. In contrast to CD4 T cells from the peripheral blood and organized gut-associated lymphoid tissues, intestinal lamina propria CD4 T cells were found to have diminished expression of the Leu-8 Ag. Therefore, studies were performed to determine whether the decreased expression of the Leu-8 Ag on lamina propria CD4 T cells correlates with a difference in the ability of peripheral blood and lamina propria CD4 T cells to regulate PWM-stimulated Ig synthesis. At high T cell to non-T cell ratios, the helper function of lamina propria CD4 T cells was significantly higher than that of peripheral blood CD4 T cells. When CD4 T cells were incubated with anti-Leu-8 antibody, the suppressor function of peripheral blood CD4 T cells was increased, but lamina propria CD4 T cells did not suppress Ig synthesis. No difference was found between the helper function of CD4,Leu-8- T cells and the suppressor function of CD4, Leu-8+ T cells isolated from either the peripheral blood or the lamina propria. Thus, the difference in the regulatory function of CD4 T cells from the peripheral blood and the lamina propria is due to the quantitative difference in CD4,Leu-8+ T cells in these sites. Consequently, the intestinal lamina propria is a site enriched in CD4,Leu-8- T cells which predominantly mediate help for Ig synthesis.     

gamma delta T cells in the human intestine express surface markers of activation and are preferentially located in the epithelium

We have investigated the expression of the alpha beta and the gamma delta T cell receptor (TCR) in the human intestine. By immunohistology we found that 39% of CD3+ intraepithelial lymphocytes (IEL) expressed the gamma delta TCR compared to 3% of CD3+ lamina propria lymphocytes (LPL). Cytofluorometric analysis of isolated cells revealed a significantly higher proportion of gamma delta T cells among CD3+ IEL compared to LPL and peripheral blood lymphocytes. This was due to an increase in both CD8+ (low density) and CD4-CD8- gamma delta T cells in IEL. Most alpha beta IEL expressed high-density CD8. About 30% of both IEL and LPL expressed CD25 (alpha-chain of the IL-2 receptor). HML-1 expression was detected on nearly all IEL and on 27% of LPL. CD25 and HML-1 were preferentially expressed on intestinal alpha beta and gamma delta T cells, respectively. Thus, human gamma delta T cells are located preferentially in the gut epithelium and are phenotypically different from alpha beta T cells, which constitute the majority of both LPL and IEL.     

Changes in mitotic rate and GFAP expression in the primary olfactory axis of Streptozotocin-induced diabetic rats

Many diabetic individuals develop anosmia but the mechanism(s) causing the dysfunction in the olfactory system is (are) unknown. Glial fibrillary acidic protein expression is reduced in diabetic retinopathy and is also reduced, with unknown consequences, in other brain regions of diabetic rats. We used immunohistochemistry and immunoblotting from untreated control and streptozotocin-induced type 1 (insulin dependent) diabetic rats to investigate main olfactory epithelial mitotic rate and glial fibrillary acidic protein expression in the lamina propria of the sensory epithelium and in the olfactory bulb. Numbers of bromodeoxyuridine-positive cells were significantly lower in the diabetic sensory epithelium compared to non-diabetic controls. Immunohistochemical observations suggested a qualitative difference in glial fibrillary acidic protein expression in both regions examined especially in the olfactory bulb external plexiform layer and the lamina propria. Immunoblot analysis confirmed that the diabetic olfactory bulb and lamina propria expressed less glial fibrillary acidic protein compared to the non-diabetic control group. The lower expression levels in the olfactory bulb external plexiform layer suggested by immunohistochemistry do not reflect a change in the number of astrocytes since the numbers of S100B(+) cells were not different between the two groups.     

Physiological role of macrophage inflammatory protein-3 alpha induction during maturation of intestinal macrophages

Intestinal macrophages (IMAC) are a central component in the defense of the intestinal mucosa against luminal microbes. In normal mucosa, monocytes differentiate to immunologically tolerant IMAC with a typical phenotype lacking activation markers such as CD14 and TLRs 2 and 4. CD33+ IMAC were isolated from normal intestinal mucosa by immunomagnetic beads. A subtractive hybridization subtracting mRNA from normal IMAC from those of in vitro differentiated macrophages was performed. IMAC differentiation was studied in multicellular spheroids (MCS). Functional assays on migration of CD45R0+ T cells were performed in MCS coculture models. Of 76 clones, 3 obtained by subtractive mRNA hybridization showed >99% homology to mRNA of MIP-3alpha, indicating that this chemokine is induced in IMAC compared with in vitro differentiated macrophages. MIP-3alpha protein expression was confirmed in cryostat sections of normal intestinal mucosa by immunohistochemistry. IMAC in the lamina propria stained positive for MIP-3alpha. FACS of purified IMAC clearly indicated expression of MIP-3alpha in these cells. In the MCS-in vitro differentiation model for IMAC, MIP-3alpha protein expression was absent on day 1 but detectable on day 7 of coculture, demonstrating the induction of MIP-3alpha during differentiation of IMAC. IMAC attracted CD45R0+ T cells to migrate into an MCS coculture model. In human mucosa, a close contact between IMAC and CD45R0+ T cells could be demonstrated. MIP-3alpha is induced during the differentiation of monocytes into IMAC. Our data suggest that MIP-3alpha expression could be involved in the recruitment of CD45R0+ cells into the lamina propria.