肝脏间质树突状细胞在多器官功能障碍综合征中的变化与意义

目的探讨肝脏间质树突状细胞在多器官功能障碍综合征（MODS）免疫紊乱机制中的影响与作用。方法150只C57BL／6小鼠经腹腔注射酵母多糖复制MODS模型，随机分为正常对照组和致伤后3～6h、12～48h、5～7d及10～12d组。观察各组小鼠肝脏间质树突状细胞的形态学变化及其表面标记物CD11c、CD205、CD80和I—A^B的表达水平；用流式细胞术检测各组外周血CD4^＋与CD8^＋的T细胞数量及CD4’／CD8’比值。结果急性损伤期（12～48h）肝脏间质树突状细胞大量增生，CD11c、CD205、CD80和I—A^b表达均较正常对照组显著上升（P均〈0．01），外周血T细胞CD4^＋／CD8^＋比值则明显下降（P〈O．01）；功能衰竭期（10～12d）肝脏间质树突状细胞继续增生，但CD205、CD80和I—A^b表达较急性损伤期显著减少（P〈0．05或P〈0．01），外周血T细胞CD4^-／CD8^＋比值下降至最低。结论肝脏间质树突状细胞参与并影响了MODS中肝脏局部与全身免疫失衡及免疫抑制过程。     

肾脏间质树突状细胞在多器官功能障碍综合征中的变化与作用

目的探讨肾脏间质树突状细胞在多器官功能障碍综合征免疫紊乱中的影响与作用。方法腹腔注射酵母多糖制造C57BL/6小鼠MODS模型,分为正常、3～6h、12～48h、5—7d和10-12d组。观察各组小鼠肾脏间质树突状细胞的病理学变化及其表面标记物CD11c、CD205、CD80和I—A^b在MODS各期肾脏中的表达水平;流式细胞术检测外周血CD4^＋与CD8^＋的T细胞数量与比值。结果在急性损伤期,肾脏间质树突状细胞大量增生,CD80和I-A^b表达较正常组差异具有统计学意义（P〈0．01和P〈0．05）,外周血T细胞CD4^＋／CD8^＋比值则明显下降（P〈0．01）。功能衰竭期,肾脏间质树突状细胞增生至最高点,但其CD80和I-A^b表达较急性损伤期显著减少（P〈0．01）,外周血T细胞CD4^＋／CD8^＋比值下降至最低（P〈0．01）。结论肾脏间质树突状细胞参与并影响了MODS中肾脏局部与全身的免疫失衡及免疫抑制过程。     

Flt3配体对多器官功能障碍综合征小鼠脾脏树突状细胞及细胞免疫功能的修复作用

目的探讨Flt3配体（FL）对多器官功能障碍综合征（MODS）晚期免疫失衡的修复作用与意义。方法按随机数字表法将90只实验鼠分为正常对照组、MODS组和FL治疗组，每组30只。用流式细胞技术检测各组脾脏树突状细胞（DC）、外周血单个核细胞主要组织相容性复合物Ⅱ类分子（I—A^b）表达变化及T细胞亚群变化，光、电镜下观察脾脏组织结构和DC变化。结果MODS组脾脏中未成熟DC明显增加（P〈0．05）；外周血单个核细胞I—A^b表达及CD4／CD8比值明显下降（P〈0．01和P〈0．05）；脾脏白髓消散，脾小体明显减少，DC多呈凋亡与退变崩解改变，周围淋巴细胞大量凋亡；活杀前动物死亡率为18％。治疗组小鼠脾脏中成熟DC数量明显增加（P〈0．05）；外周血单个核细胞I—A^b表达接近正常水平，CD4／CD8比值较MODS组明显上调；脾脏组织形态与DC的病理改变较MODS组明显减轻；实验鼠死亡率（7％）也明显降低。结论FL可以通过促进DC增生和活化，有效改善MODS晚期细胞免疫功能，进而缓解MODS的进程。     

炎症性肠病患者外周血CD4^＋CD25^＋Treg细胞的表达及意义

[目的]探讨CD4^＋CD25^＋Treg细胞在炎症性肠病（IBD）发病中的作用及其与疾病活动性的关系。[方法]采用三色流式细胞术检测40例IBD患者，其它肠病患者30例和健康对照者30例。IBD患者中活动期患者25例，缓解期患者15例，使用和未使用激素和／或免疫抑制剂活动期IBD患者分别为16例和9例。对以上各组外周血中CD4^＋CD25^＋T细胞亚群的百分率进行测定。[结果]疾病活动期IBD患者外周血CD4^＋CD25^＋Treg细胞比例明显低于其他肠病和正常对照组（P〈0．01），疾病活动期IBD患者外周血CD4^＋CD25^＋Treg细胞比例明显低于疾病稳定期患者（P〈0．01）。活动期IBD患者中使用激素和／或免疫抑制剂与未使用激素和／或免疫抑制剂结果差异有统计学意义。IBD患者外周血CD4^＋CD25^＋Treg细胞表达率与疾病活动指数评分呈负相关性。[结论]IBD患者外周血CD4^＋CD25^＋Treg细胞异常表达，可能参与疾病的发生发展，与疾病的活动性密切相关。     

罗红霉素对慢性阻塞性肺疾病急性加重期患者细胞免疫功能的影响

目的：观察慢性阻塞性疾病（COPD）急性加重期患者外周血CD4^＋、CD8^＋T淋巴细胞及CD4^＋／CD8^＋变化，探讨COPD急性加重期患者的细胞免疫功能及罗红霉素对其的影响。方法：15位健康体检者（对照组，A组）、27例COPD急性加重期患者常规治疗组（B组）和27例COPD急性加重期患者常规加罗红霉素治疗组（C组）于发病初及治疗8W后检测肺功能，测定外周血中CD4^＋和CD8^＋T淋巴细胞占总淋巴细胞的百分率。结果与正常体检者及C组治疗后比较，COPD急性加重期患者外周血CD8叶。T淋巴细胞升高，CD4^＋／CD8^＋比值降低，且CD8^＋T淋巴细胞与CD4^＋／CD8^＋比值与FVC％pred、FEV1％pred and FEV1／FVC呈显著相关性。结论：COPD急性加重期患者存在细胞免疫功能异常，CD8^＋T淋巴细胞免疫反应增强是COPD急性加重期患者气流受限的可能发病机制之一，罗红霉素通过调节COPD患者急性加重期细胞免疫反应而减轻气流受限，改善肺功能。     

淋巴细胞亚群不同凋亡在狼疮性肾炎患者发病中的意义

目的探讨狼疮性肾炎（LN）患者不同时期外周血单个核细胞凋亡率及CD3^＋CD4^＋、CD3^＋CD8^＋、CD3^＋CU4^＋Fas、CD3^＋CD8^＋Fas、sFas表达的意义。方法56例经肾活检证实的LN患者中，活动期38例，静止期18例，均采用酶联免疫吸附试验（ELISA）检测sFas和流式细胞仪检测PBMC凋亡率及CD3^＋CD4^＋、CD3^＋CD8^＋、CD3^＋CD4^＋Fas、CD3^＋CD8^＋Fas等指标。结果LN静止期sFas较健康对照组增高，活动期PBMC凋亡率、sFas明显升高（P〈0．01），CD3^＋CD4^＋细胞较健康对照组明显减少（P〈0．01）、CD3^＋CD3^＋细胞比健康对照组增多（P〈0．01）；PBMC凋亡率与sFas呈正相关。结论LN患者外周血T淋巴细胞的不同凋亡，特别是CD3^＋CD4^＋／CD3^＋CD8^＋比例失调，是导致LN病情发展的重要机制之一。sFas可能抑制肾小球系膜细胞凋亡，而且可能是LN进展中的一个因素。     

外周血CD4^＋CD25^high调节性T细胞比例变化在肾移植受者移植后免疫监测中的应用

背景：相关实验表明调节性T细胞在移植物免疫耐受中起重要作用。 目的：观察外周血CD4^＋CD25^high调节性T细胞比例变化与肾移植受者移植后免疫变化的相关性。 设计、时间及地点：回顾性病例分析,于2007-09／2008-07在广东省第二人民医院器官移植中心及其实验室完成。 参试者：52例病情稳定的维持性血液透析患者行同种异体肾移植治疗。 方法：肾移植后患者均服用三联免疫抑制剂。所有患者移植后发生急性移植肾排斥反应以及感染均按照相应指南诊断及治疗。分别于移植前和移植后1,2,4,8,12周以及发生排斥反应和感染时抽血检测外周血CD4^＋CD25^＋调节性T细胞。按其免疫力恢复情况分为正常组26例,排斥反应组17例,感染组9例。 主要观察指标：用流式细胞仪检测外周血CD4^＋CD25^high调节性T细胞的比例,所得结果进行相关分析。 结果：与正常组比较：排斥反应组CD4^＋CD25^highFoxP3／CD4^＋的比值降低（P〈0．05）,而感染组显著增高（P〈0．01）。与感染组比较：排斥反应组CD4^＋CD25^highFoxP3／CD4^＋的比值显著降低（P〈0．01）。 结论：肾移植后受者外周血CD4^＋CD25^high调节性T细胞比例与受者免疫状态密切相关。CD4^＋CD25^high调节性T细胞比例的变化可以反应机体的免疫状态的变化,其升高或降低可以作为预测肾移植受者移植后发生感染或排斥反应的指标之一。     

急性白血病儿童外周血CD4^＋CD25^＋调节性T细胞和NK细胞的变化及其在白血病免疫中的意义

本研究观察急性白血病患儿外周血CD4^＋CCD25^＋调节性T细胞（CD4^＋CD25^＋Treg）及自然杀伤细胞（nature killer cell,NK）在不同病程阶段的变化,了解白血病患儿的免疫状态,以探讨CD4^＋CD25^＋Treg细胞及NK细胞在急性白血病肿瘤免疫中的意义。建立流式细胞术检测外周血CD4^＋CD25^＋Treg细胞和NK细胞的方法：检测急性白血病初诊患儿25例、完全缓解患儿28例及20例正常健康对照者外周血CD4^＋CD25^＋Treg细胞及NK细胞的数量及比例。结果表明：初诊组、完全缓解组及对照组外周血CD4^＋CD25^＋CD127^＋占CD4^＋T细胞的比例分别为（9．55±2．41）％,（8．54±2．51）％和（6．25±0．85）％,在初诊患儿组和缓解患儿组高于正常对照组,且在初诊患儿组高于完全缓解组（P〈0．05）;同时,与正常对照组比较,急性白血病患儿的NK细胞数量减少,完全缓解后患儿组NK细胞数量仍低于正常对照（4．11±3．87％和10．41±7．20％w14．06±5．95％,P〈0．05）。结论：联合应用CD4、CD25及cDl27检测Treg细胞简便可行、重复性好、检测结果可靠、准确,CD4^＋CD25^＋CD127^＋T细胞可较好地反映CD4^＋CD25^＋Treg细胞的比例。急性白血病患儿外周血中Treg细胞数量升高,NK细胞数量降低,表明急性白血病患儿NK细胞免疫功能处于抑制状态。Treg细胞可能在白血病的发生、发展中起一定作用,参与NK细胞的调节可能是Treg细胞在白血病免疫中的一个环节。     

肾综合征出血热患者免疫发病机制的探讨

目的 研究肾综合征出血热患者的细胞免疫功能异常情况。方法 应用流式细胞仪检测T细胞亚群CD4^＋T淋巴细胞、CD8^＋T淋巴细胞。结果 肾综合征出血热患者早期、后期与正常人比较，均有CD4^＋淋巴细胞计数降低（P=0.000，P=0．000），CD8^＋T淋巴细胞计数明显升高（P=0．000，P=0．002），CD4^＋／CD8^＋明显降低（P=0．000，P=0．000）。CD4^＋T淋巴细胞计数病程早期与病程后期无明显差异（P=0．985），CD8^＋1淋巴细胞计数病程早期明显高于病程后期（P=0．000），CD4^＋／CD8^＋病程早期明显低于病程后期（P=0．000）。结论 肾综合征出血热患者发病早期存在明显细胞免疫功能异常．在发病后期逐渐恢复。     

登革热患者外周血CD4^＋、CD8^＋T细胞及IL-10的检测分析

目的：观察登革热患者外周血CD4^＋、CD8^＋T细胞及白介素-10（IL-10）的变化，探讨其在登革热疾病的发生和发展中的作用。方法：12例健康人和18例登革热患者于治疗前后采静脉血。采用流式细胞仪检测CD4^＋、CD8^＋T细胞并计算CD4^＋／CD8^＋比值，ELISA法检测血清IL-10水平，同时作外周血白细胞、血小板计数。结果：与正常健康人和治疗后相比，登革热患者治疗前外周血CD4^＋细胞百分比、CD4^＋／CD8^＋比值明显降低（P〈0．01）。CD8^＋细胞百分比和血清IL-10显著增高（P〈0．01），白细胞、血小板计数均明显下降（P〈0．01）；治疗后患者以上指标均与健康人无明显差异。结论：CD4^＋、CD8^＋ T细胞在登革热病毒感染后异常激活，CD4^＋／CD8^＋比值明显降低，IL-10分泌增高，可能在登革热的发病中起重要作用。     

多器官功能障碍综合征52例尸检脾及脾树突状细胞的病理学观察

目的探讨多器官功能障碍综合征（MODS）脾及其树突状细胞的病变特点与作用。方法收集52例MODS死亡病例与25例正常脾标本,应用光镜、电镜和免疫组化（S-100、CD1a、CD80及HLA-DR）方法观察脾及其树突状细胞的病理变化。结果镜下见脾小体消失,白髓消散伴淋巴细胞大量凋亡,树突状细胞数目增加而活性减退,CD4（＋）/CD8（＋）T淋巴细胞比例显著下降。结论MODS终末期外周免疫器官脾及其树突状细胞严重损伤和功能耗竭,而且不同诱因所致MODS病例的病变基本一致。提示脾树突状细胞的病变与免疫抑制及MODS形成有重要关系。     

Constitutive presentation of a natural tissue autoantigen exclusively by dendritic cells in the draining lymph node

The major histocompatibility complex (MHC)-dependent presentation of processed tissue-specific self-antigens can contribute to either peripheral (extrathymic) tolerance or the differentiation of autoreactive T cells. Here, we have studied the MHC class II molecule presentation of gastric parietal cell (PC)-specific H(+)/K(+)-ATPase, which induces a destructive autoimmune gastritis in BALB/c mice lacking CD4(+) CD25(+) regulatory T cells. Immunofluorescence microscopy showed physical association of CD11c(+) dendritic cells (DCs) with PCs in the gastric mucosa. H(+)/K(+)-ATPase protein was found within vesicular compartments of a few CD11c(+) DCs only in the draining gastric lymph node (LN) and these antigen-containing DCs increased markedly in number with the onset of tissue destruction in autoimmune animals. Both CD8alpha(hi) and CD8alpha(lo) gastric DCs, but not peripheral or mesenteric DCs, showed evidence of constitutive in vivo processing and presentation of H(+)/K(+)-ATPase. These data provide direct support for a widely held model of local tissue antigen uptake and trafficking by DCs in normal animals and demonstrate that DCs in the draining LN can present a tissue-specific self-antigen under noninflammatory conditions without fully deleting autoreactive T cells or inducing active autoimmunity.     

T-lymphocyte subset distribution in human spleen

BACKGROUND: When analyzing human cellular immune responses, most focus is placed on the peripheral blood (PB) and, to a lesser extent, the lymph nodes. To date the spleen has not been analyzed with regard to its role in adaptive cellular immunity and more notably not with respect to T-cell immune responses. MATERIALS AND METHODS: We analyzed the splenic lymphocyte compartment in comparison with the PB lymphocyte compartment regarding the number of NK cells, B cells, CD4(+), CD8(+) T cells and CMV-specific CD8(+) T cells. Furthermore, we analyzed the distribution of naive, memory and effector subsets of CD4(+) and CD8(+) T cells in these compartments. RESULTS: The spleen contains proportionally more B cells and less CD4(+) and CD8(+) T cells than PB. The percentage of CD8(+) T cells is greater in the spleen, leading to an inverse CD4/CD8 ratio. Both splenic CD4(+) and CD8(+) T-cell populations show a greater number of activated cells, and splenic CD8(+) T cells show a more differentiated cytotoxic CD27(-)CD45RA(+) memory phenotype. CONCLUSIONS: Our findings show that the distribution of the different lymphocyte subsets is markedly different between the spleen and the PB, thus inferring an important and distinct role for the spleen in CD4(+) and CD8(+) T-cell activation.     

MODS大鼠胸腺树突状细胞病理改变及作用的研究

目的：探讨胸腺树突状细胞在多脏器功能障碍综合征（MODS）发病机制中的作用,为严重创伤后脓毒症和MODS的基础研究与临床防治提供新的思路。方法：运用光镜、电镜观察与免疫组织化学（CD1a与S－100）及原位末端标记法（TUNEL）研究MODS大鼠胸腺中树突状细胞的变化及其与细胞凋亡的关系。结果：复制小鼠酵母多糖MODS模型,伤后12小时（MODS早期）观察到胸腺树突状细胞增生和过高反应,伴有淋巴细胞大量凋亡;伤后2－3日（MODS进展期）,树突状细胞数量显著减少、形态萎缩,淋巴细胞凋亡减轻。结论：树突状细胞的病理改变不仅可能是MODS的早发病变和启动因素之一,且可能是影响全身炎症反应综合征（SIRS）或代偿性抗炎反应综合征（CARS）形成和转归,使病程走向MODS的一种重要细胞病变。     

Characterization of resident and migratory dendritic cells in human lymph nodes

Dendritic cells (DCs) initiate adaptive immune responses in lymph nodes (LNs). In mice, LN DCs can be divided into resident and tissue-derived populations, the latter of which migrate from the peripheral tissues. In humans, different subsets of DCs have been identified in the blood, spleen, and skin, but less is known about populations of resident and migratory tissue-derived DCs in LNs. We have analyzed DCs in human LNs and identified two populations of resident DCs that are present in all LNs analyzed, as well as in the spleen and tonsil, and correspond to the two known blood DC subtypes. We also identify three main populations of skin-derived migratory DCs that are present only in skin-draining LNs and correspond to the DC subsets found in the skin. Resident DCs subsets induce both Th1 and Th2 cytokines in naive allogeneic T lymphocytes, whereas the corresponding blood subsets failed to induce efficient Th2 polarization. LN-resident DCs also cross-present antigen without in vitro activation, whereas blood DCs fail to do so. Among migratory DCs, one subset was poor at both CD4(+) and CD8(+) T cell activation, whereas the other subsets induced only Th2 polarization. We conclude that in humans, skin-draining LNs host both resident and migratory DC subsets with distinct functional abilities.     

Flt3 ligand-treated neonatal mice have increased innate immunity against intracellular pathogens and efficiently control virus infections

Flt-3 ligand (FL), a hematopoetic growth factor, increases the number of dendritic cells (DCs), B cells, and natural killer cells in adult mice but the effect in neonates was unknown. We show that FL treatment of newborn mice induced a >100-fold increase in the innate resistance against infection with herpes simplex virus type 1 and Listeria monocytogenes. This resistance required interferon (IFN)-alpha/beta for viral and interleukin (IL)-12 for bacterial infections. Long-term survival after viral but not bacterial infection was increased approximately 100-fold by FL treatment. After treatment, CD11c(+)/major histocompatibility complex type II(+) and CD11c(+)/B220(+) DC lineage cells were the only cell populations increased in the spleen, liver, peritoneum, and skin. DC induction was independent of IFNs, IL-2, -4, -7, -9, -15, and mature T and B cells. The data suggest that FL increases the number of DCs in neonates and possibly in other immune-compromised individuals, which in turn improves IFN-alpha/beta- and IL-12-associated immune responses.     

Compartmentalized production of CCL17 in vivo: strong inducibility in peripheral dendritic cells contrasts selective absence from the spleen

Dendritic cells (DCs)(*) fulfill an important regulatory function at the interface of the innate and adaptive immune system. The thymus and activation-regulated chemokine (TARC/CCL17) is produced by DCs and facilitates the attraction of activated T cells. Using a fluorescence-based in vivo reporter system, we show that CCL17 expression in mice is found in activated Langerhans cells and mature DCs located in various lymphoid and nonlymphoid organs, and is up-regulated after stimulation with Toll-like receptor ligands. DCs expressing CCL17 belong to the CD11b(+)CD8(-)Dec205(+) DC subset, including the myeloid-related DCs located in the subepithelial dome of Peyer's patches. CCL17-deficient mice mount diminished T cell-dependent contact hypersensitivity responses and display a deficiency in rejection of allogeneic organ transplants. In contrast to lymphoid organs located at external barriers of the skin and mucosa, CCL17 is not expressed in the spleen, even after systemic microbial challenge or after in vitro stimulation. These findings indicate that CCL17 production is a hallmark of local DC stimulation in peripheral organs but is absent from the spleen as a filter of blood-borne antigens.     

Correlation between B lymphocyte abnormality and disease activity in patients with idiopathic membranous nephropathy

This study investigated the relationship between peripheral blood B lymphocytes, regulatory T-cells and T lymphocyte subsets, the distribution of B lymphocytes in the kidney, and the pathogenesis of idiopathic membranous nephropathy (IMN). Lymphocyte subsets were measured using flow cytometry in 66 patients with clinically-confirmed IMN and in 40 healthy control subjects. Compared with healthy subjects, the number of peripheral blood B lymphocytes was significantly increased in IMN patients and that of regulatory T-cells was significantly decreased, accompanied by an increased CD4(+)/CD8(+) T-cell ratio. There was no relationship between the number of peripheral blood B lymphocytes and markers of kidney function. Although the number of infiltrating B lymphocytes in the kidney of IMN patients was higher, there was no relationship with the number of peripheral blood B lymphocytes. In conclusion, there was no relationship between peripheral blood B lymphocytes and disease activity, suggesting that peripheral blood B lymphocytes are not a biomarker of disease activity and therapeutic efficacy in IMN.