共刺激分子CD28、CD137在喉癌患者T淋巴细胞中的表达及其与细胞免疫功能失调的关系

目的探讨共刺激分子CD28、CD137与喉癌患者细胞免疫功能状况的关系。方法应用梯度离心法分离并纯化外周血和肿瘤浸润淋巴细胞（tumor infiltrating lymphocyte,TIL）,流式细胞术检测喉癌患者外周血T细胞CD8^＋、CD8^＋CD28^＋、CD8^＋CD28^-、CD28^＋细胞亚群分布和双香豆素（phytohemagglutinin,PHA）刺激前后患者外周血T细胞及TIL中CD137的表达。结果喉癌患者CD8^＋T细胞比率为（32.11±5.07）%,CD8^＋CD28^-T细胞比率为（21.03±5.69）%,均明显高于正常对照组（均P〈0.01）;CD28^＋T细胞比率为（45.53±4.51）%,CD8^＋CD28^＋T细胞比率为（12.70±3.41）%,均明显低于正常对照组（P均〈0.01）。CD137^＋T细胞活化增殖能力与CD28的阳性表达呈明显的正相关,喉癌患者外周血T细胞和TIL在未受PHA刺激时CD137^＋T细胞比率分别为（1.90±0.71）%和（3.52±1.64）%,明显高于正常对照组（P均〈0.01）。但经PHA刺激48小时后,外周血和TIL中CD137^＋-48h/CD137^＋-0h值为分11.48±4.03和10.39±3.02,均明显低于正常对照组,P均〈0.01。结论共刺激分子CD28在CD8^＋T细胞亚群中的表达失调使免疫效应性CD137^＋T细胞活化增殖能力下降,是引起喉癌患者细胞免疫功能紊乱的重要原因之一。 还原     

急性分泌性中耳炎调节性T细胞及T淋巴细胞亚群的分析

目的 检测急性分泌性中耳炎(SOM)患者外周血调节性T细胞及淋巴细胞亚群的变化,综合分析免疫因素在其发病中的作用.方法 以流式细胞术检测38例上感后诱发急性SOM患者,30例健康献血者外周血中CD3+、CD4+、CD8+、NK细胞、B淋巴细胞百分比,CD4 +/CD8+比值,CD4+ CD25+调节性T细胞百分比以及T淋巴细胞亚群的计数.结果 患者组CD3+ CD8+、NK细胞的百分比明显低于正常对照组(P＜0.05);CD4+/CD8+、B淋巴细胞比例明显高于正常对照组(P＜0.05);正常对照组Tregs占CD4 +T淋巴细胞的比例为(6.695±1.09)％,患者组占CD4 +T淋巴细胞的比例为(12.82±4.56)％,与正常对照组比较差异有统计学意义(P＜0.05);两组间CD3+、CD4+细胞比例,差异无统计学意义(P＞0.05);正常对照组及患者组外周血CD3+、CD3+ CD4+、CD3+ CD8+T淋巴细胞计数,两组之间差异无显著性(P＞0.05).结论 急性分泌性中耳炎患者存在着免疫失调,CD4+/CD8+下降,Tregs比例异常增高,说明免疫异常在SOM的发病中起一定的作用.     

外周血CD8^(+)T淋巴细胞亚群检测对过敏性鼻炎临床疗效的评估价值CSCD

目的探讨外周血CD8^(+)T淋巴细胞亚群检测对过敏性鼻炎(AR)临床疗效的评估价值。方法前瞻性选取2021年3月~2022年2月温州市中西医结合医院耳鼻咽喉科门诊治疗的120例AR患者和40例健康志愿者,分别设为研究组和对照组,比较两组外周血CD8^(+)T细胞、CD8^(+)CD28^(+)T细胞、CD8^(+)CD28-T细胞比率;研究组接受鼻用激素联合口服抗组胺药物治疗方案,根据治疗效果分为有效组(n=107)和无效组(n=13),比较有效组和无效组治疗前CD8^(+)T细胞、CD8^(+)CD28^(+)T细胞、CD8^(+)CD28-T细胞比率;Pearson相关性分析法分析外周血CD8^(+)T淋巴细胞亚群水平与鼻部症状总评分(totalnose symptomscore,TNSS)的相关性;以受试者工作特征(ROC)曲线分析治疗前CD8^(+)T淋巴细胞亚群对AR临床疗效的预测价值。结果研究组治疗前外周血CD8^(+)CD28^(+)T细胞比率高于对照组,CD8^(+)CD28-T细胞比率低于对照组(P<0.05)。研究组总有效率为89.17%;有效组治疗前CD8^(+)CD28^(+)T细胞比率低于无效组,CD8^(+)CD28-T细胞比率高于无效组(P<0.05);Pearson相关性分析显示,CD8^(+)CD28^(+)T细胞比率与TNSS评分呈正相关(r=0.324,P=0.006),CD8^(+)CD28-T细胞比率与TNSS评分呈负相关(r=-0.541,P=0.000)。ROC曲线结果显示,CD8^(+)T细胞、CD8^(+)CD28^(+)T细胞、CD8^(+)CD28-T细胞比率评估AR治疗无效的曲线下面积(AUC)(95%CI)分别为0.647(0.555~0.732)、0.683(0.592~0.765)、0.911(0.845~0.955),其中CD8^(+)CD28-T细胞的评估效能优于CD8^(+)T细胞、CD8^(+)CD28^(+)T细胞比率(P<0.05)。结论AR患者外周血CD8^(+)CD28^(+)T细胞比率升高,CD8^(+)CD28-T细胞比率降低,CD8^(+)T细胞比率相对稳定,治疗前CD8^(+)CD28-T细胞比率可作为预测AR患者的临床疗效的参考指标。     

树突状细胞表型抗原在喉咽癌颈部淋巴结中的表达

目的：探讨喉咽癌颈部淋巴结中树突状细胞（Dc）表型抗原的表达与喉咽癌转移及预后的关系。方法：对37例喉咽癌144枚淋巴结DC的表型抗原S-100、CD1a、CD83的表达用Envision HIS的方法检测,并观察CD45RO^＋T细胞与DC的关系。结果：S-100^＋DC、CD1a^＋DC、CD83^＋DC在转移组淋巴结中的数目较非转移组淋巴结中减少,差异有统计学意义。CD83^＋DC在转移淋巴结的癌巢中的数目较癌旁中的少,差异有统计学意义。在生存组淋巴结中的浸润数目较死亡组多,差异有统计学意义。CD45RO^＋T细胞主要分布在CD83^＋DC的周围。结论：表达不同表型抗原的DC具有不同的功能,CD83^＋DCs被认为是成熟DC分布在癌旁与其激活T细胞的抗原提呈功能有关,对于肿瘤免疫防御机制的建立起着重要作用。DC在淋巴结中的浸润程度及表型抗原的表达情况是反映宿主肿瘤免疫状况的重要指标,也是预测喉咽癌转移和预后的一项重要指标。     

获得性免疫缺陷综合征合并慢性扁桃体炎患者Th1及Treg应答水平及意义

目的　探讨获得性免疫缺陷综合征（acquired immunodeficiency syndrome,AIDS）合并慢性扁桃体炎患者外周血和扁桃体组织中Th1/Treg应答情况。方法  采用RT-PCR检测了外周血和扁桃体T-bet /Foxp3的表达;同时采用免疫组织化学法检测T-bet/Foxp3以及CD4/CD8细胞的数量;采用全血胞内细胞因子染色方法,分析CD4+IFN-γ+T/CD4+CD25+ Foxp3+细胞表达水平。结果  FACS检测AIDS伴慢性扁桃体炎患者CD4+IFN-γ+T细胞表达（20.65±7.14）%显著低于健康对照（39.13±13.30）%和慢性扁桃体炎患者（36.89±10.05）%;CD4+CD25+Foxp3+细胞表达（7.38±2.63）%则明显高于健康对照（4.31±2.14）%和慢性扁桃体炎患者（4.79±2.23）%;CD4 细胞计数（104.5±60.96）显著低于健康对照（663.9±170.3）和慢性扁桃体炎患者（546.4±151.9）;CD8细胞计数（580.5±130.7）显著高于健康对照（364.7±69.85）和慢性扁桃体炎患者（443.0±150.5）;RT-PCR检测AIDS伴慢性扁桃体炎患者T-bet表达（4.831±2.61）显著低于健康对照（20.12±11.31）和慢性扁桃体炎患者（9.83±3.54）;Foxp3表达（102.5±47.42）显著高于健康对照（57.28±25.19）;免疫组化检测AIDS伴慢性扁桃体炎患者扁桃体组织中T-bet表达（80.20±2.20）显著高于慢性扁桃体炎患者（31.9±2.82）;Foxp3表达和慢性扁桃体炎患者差异无统计学意义;CD4细胞 (42.09±4.09）显著低于慢性扁桃体炎患 者（80.93±3.42）;CD8细胞（80.45±2.81）显著高于慢性扁桃体炎患者（37.57±3.11）。结论　患者外周血和扁桃体组织中存在明显不同的Th1/Treg免疫反应,Th1/Treg失衡在AIDS伴慢性扁桃体炎患者致病机制中起着重要作用。     

消瘤汤治疗鼻内翻性乳头状瘤的疗效及其作用机理研究

目的 观察消瘤汤对鼻内翻性乳头状瘤术后复发与恶变的抑制作用及对宿主细胞免疫功能的影响.方法 鼻内翻性乳头状瘤患者60例,术后随机分为治疗组和对照组,每组各30例.治疗组术后服消瘤汤半年,对照组不用药物治疗,均随访8个月以上,观察病变复发和恶变情况.分别在术前及术后半年,以流式细胞仪检测患者外周血T淋巴细胞亚群及NK细胞活性,观察消瘤汤对宿主细胞免疫功能的影响.结果 治疗组复发2例（2/30,6.7%）,对照组复发8例（8/28,28.6%,其中3例存在轻至中度不典型增生）,差异具有统计学意义（P＜0.05）.与术前检测结果比较,治疗组的CD3^＋、CD4^＋T细胞及NK细胞比例升高,CD8^＋T细胞比例降低,对照组的CD3^＋、CD4^＋T细胞及NK细胞比例均降低,而CD8^＋T细胞比例升高.结论 消瘤汤可降低鼻内翻性乳头状瘤患者的术后复发率,并通过改善宿主细胞免疫功能而起到抑制其复发与恶变的作用.     

螨变应原特异性免疫治疗1年后免疫学变化

目的探讨标准化尘螨变应原疫苗进行集群免疫治疗过程中的免疫学机制。方法将60例螨过敏变应性鼻炎（AR）患者分为集群免疫治疗组（n=30）和药物治疗组（n=30）。检测基线和治疗1年后受试者血清中总IgE、螨特异性IgE、特异性IgG4水平,通过流式细胞术检测外周血单核细胞中Th1、Th2、天然调节性T细胞（CD4^＋CD25^＋Foxp3^＋ T细胞）和I型调节性T细胞（Tr1细胞,CD4^＋IL-10^＋IL-4^-T细胞）在CD4^＋T细胞中百分比。结果经过1年治疗后,总IgE和螨特异性IgE水平未见明显改变,但免疫治疗组患者血清中螨特异性IgG4水平明显升高（P〈0．0001）。Th1、Th2型细胞和天然调节性T细胞（CD4^＋CD25^＋Foxp3^＋T细胞）占CD4^＋T细胞的百分比在治疗前后均无明显变化（P=ns）,而Tr1细胞在免疫治疗1年后明显增高（P〈0．001）。结论特异性IgG4和Tr1细胞增高可作为产生免疫耐受的免疫学指标。     

复发性鼻息肉中T淋巴细胞亚群的表达

目的 :检测复发性鼻息肉组织中T淋巴细胞亚群的表达 ,探讨T淋巴细胞介导的免疫反应在鼻息肉复发中所起的作用。方法 :应用荧光免疫流式细胞术检测 17例复发性鼻息肉患者鼻息肉组织、外周血T淋巴细胞亚群CD4 、CD8 、CD4 5RO 的表达 ,并与正常人下鼻甲黏膜及外周血的相应指标进行比较。 结果 :复发性鼻息肉组织中有大量T淋巴细胞浸润 [(39.6 5±2 .0 8) % ],而在健康下鼻甲黏膜中几乎未见CD3 细胞。在复发性鼻息肉中 ,CD3 CD4 细胞[(6 4.4 46± 5 .2 97) % ]多于CD3 CD8 细胞 [(35 .5 5 4± 5 .2 97) % ](P <0 .0 5 ) ,CD3 CD4 5RO 细胞 [(2 2 .6 49± 2 .789) % ]也显著多于正常人外周血CD3 CD4 5RO 细胞 [(3.896± 0 .384 ) % ](P<0 .0 5 ) ,CD3 CD4 /CD3 CD8 比值为 1.95 6± 0 .0 93,复发性鼻息肉患者外周血比值为 2 .36 7±0 .12 8,正常人外周血比值为 1.6 0 6± 0 .0 96 ,其差异均有统计学意义 (均P <0 .0 5 )。结论 :复发性鼻息肉组织中有大量T淋巴细胞表达 ,且T细胞亚群比例失调 ,显示细胞免疫功能的紊乱在鼻息肉的形成与复发中起重要作用     

变应性鼻炎小鼠外周调节性T细胞的表达特征

目的运用流式细胞仪检测小鼠变应性鼻炎模型的外周血单个核细胞（peripheral blood mononuclear cell,PBMC）和脾脏单个核细胞中Foxp3^＋、CD4^＋、CD25^＋的表达情况,比较与正常对照组间是否存在差异。方法选取8周龄成熟雌性BALB/c小鼠30只,随机分为实验组和对照组各15只。卵清蛋白致敏建立BALB/c小鼠变应性鼻炎模型。21天实验结束取鼻腔HE染色观察形态学特点及差异;分别取脾脏和外周血分离获得单个核细胞,流式细胞仪检测其Foxp3^＋、CD4^＋、CD25^＋的表达情况。结果卵清蛋白造模成功,实验组小鼠均出现喷嚏、搔鼻、鼻周红肿等症状;镜下可见鼻黏膜下较多嗜酸性粒细胞、浆细胞及肥大细胞浸润。实验组PBMC中CD4^＋CD25^＋T细胞和Foxp3^＋CD4^＋CD25^＋T细胞明显低于对照组,均具有极显著性差异（P〈0.01）。小鼠脾脏单个核细胞中CD4^＋CD25^＋T细胞和Foxp3^＋CD4^＋CD25^＋T细胞均略低于对照组,但统计学显示无显著性差异（P〉0.05）。结论调节性T细胞（regulatory T cell,Treg）在变应性鼻炎中作用机制尚未完全明晰,本实验从分子水平分析变应性鼻炎中CD4^＋细胞的表达特征,旨在为Treg的深入研究提供又一实验依据。     

扁桃体淋巴细胞亚群及其活性

本文对32例扁桃体炎患者切除的扁桃体标本应用双染色流式细胞仪(FCM)测定淋巴细胞亚群及其活性。患者分两组,21岁以上16例(成人组),15岁以下16例(儿童组)。肥大型扁桃体炎14例,复发型扁桃体炎18例。淋巴细胞与用异硫氰基荧光素(FITC)或藻红蛋白(PE)标记的相应特异单克喹抗体结合后进行 FCM 测定。结果发现扁桃体组织中辅助 T 细胞(CD_4~+Leu8~-)较外周血显著增高(P<0.005),而诱导 T 细胞(CD_4~+Leu8~+)、抑制 T 细胞(CD8~+CD11~+)及细胞毒性 T 细胞(CD8~+CD11~-)较外周血少(P<0.005),尤其抑制细胞在扁桃体组织中非常少见。成人患者扁桃体组织中四种 T 细胞所占比例均不同程度地较儿童患者高。肥大型组的细胞毒性 T 细胞所占比例明显高于复发型组(P<0.05),而复发型组的诱导 T 细胞和抑制 T 细胞所占比例又明显高于肥大型组(P<0.005)。外周血活性 CD4~+或 CD8~+细胞不到总淋巴细胞的1%,     

Double immunoenzymatic and two-color flow cytometric analyses of tonsillar T-cell subsets]

Distribution of various T-cell subsets in the palatine tonsil was investigated by two-color flow cytometry and double immunoenzymatic stain. Tonsillar lymphocytes contained many (about 20%) helper T (CD4+ Leu8-) cells and few (only 1%) suppressor T (CD8+ CD11b+) cells. In interfollicular area, each T-cell subset, i.e., helper, helper-inducer (CD4+ CD29+), suppressor-inducer (CD4+ CD45RA+), cytotoxic (CD8+ CD11b-), and suppressor, was identified by double immunoenzymatic labeling technique using alkaline phosphatase and peroxidase. On the other hand, only two T-cell subsets, helper, and cytotoxic T-cell subpopulations, were recognized in germial center. These results indicate that double immunoenzymatic stain as well as two-color flow cytometry gives us useful informations in terms of tonsillar T-cell subsets.     

Subsets of tonsillar lymphocytes and activated cells in each subset analysed by two-color flow cytometry

We analysed subsets of tonsillar lymphocytes and activated cells in each subset by two-color flow cytometry. There were many helper T cells (CD 4+ leu 8-) and few inducer T cells (CD 4+ leu 8+) in the tonsil. The situation was the reverse in the peripheral blood. The tonsil had few suppressor T cells (CD 8+ CD 11+). The proportion of activated cells (HLA-DR+) was low in the peripheral blood and high in the tonsil. In the tonsil, the ratio of cells with transferrin receptors to total lymphocytes was higher in the B than in the T subset, and higher in the subset of CD 4+ than in that of CD 8+ cells. Activated and proliferating lymphocytes were more abundant in the tonsils of children than in those of adults and more abundant in patients with hyperplastic tonsillitis than in those with recurrent tonsillitis.     

Is immune system influenced by adenotonsillectomy in children?

OBJECTIVE: Tonsils and adenoids are lymphoid tissues that are located in the pharynx and play an important role against invading antigens of the upper respiratory tract. The present study analyses serum immunoglobulin levels and peripheral blood (PB) lymphocyte subsets in children, 24-48 h prior to and 4-6 weeks after adenotonsillectomy, in order to determine early effects of adenotonsillectomy on the immune system. METHODS: The study population consists of 15 children (aged 4-10 years) who underwent adenotonsillectomy because of adenoidal hypertrophy and chronic tonsillitis and 15 age-matched healthy children without a history of adenotonsillectomy. Serum IgG, IgA and IgM levels were measured by nephelometry. PB lymphocyte subsets were analysed by using monoclonal antibodies and flow cytometry. RESULTS: Children with chronic tonsillitis have increased levels of CD19+ B lymphocytes compared to healthy controls in the pre-operative period. The percentage of B lymphocytes bearing CD23 was found to be significantly higher in patients, most likely representing in vivo B lymphocyte activation due to chronic antigenic stimulation. After the adenotonsillectomy, despite ongoing B lymphocyte activation, CD8+ T lymphocyte levels increased and B cell levels returned to normal. A slight decrease in serum IgG, IgA and IgM levels was detected in the post-operative period compared to prior levels. CONCLUSION: Adenotonsillectomy performed in children leads to alterations that may reflect a compensatory response of the developing immune system after the removal of the lymphoid tissue in the setting of chronic antigenic stimulation. However, these changes do not cause significant immune deficiency.     

Correlation between immunohistochemical indices of adenoid vegetations and health of ailing children

We studied correlations between clinical characteristics of ailing children and their immunohistochemical parameters. Clinical status was assessed by the degree of adenoid vegetations, duration of the disease, the presence of chronic tonsillitis or its combination with adenoiditis. We used the following immunohistochemical parameters: the level of CD3+ in histological slices and the presence or absence of IgA-producing cells. Correlation was found between the presence or absence of IgA-producing cells and the degree of adenoid vegetations as well as the presence only of chronic tonsillitis or chronic tonsillitis plus adenoiditis. We found also important but statistically insignificant differences: in a group of patients with the disease duration up to 2 years CD3+ cells were absent, while in children with the disease duration more than 5 years these cells were present. Thus, immunohistochemical parameters are rather informative in interpretation of a clinical picture in ailing children.     

Lymphocyte subsets of maxillary mucosa in chronic inflammation

Subsets of infiltrating lymphocytes within maxillary sinus mucosae of patients with chronic sinusitis were investigated by immunoperoxidase staining of frozen sections with the use of monoclonal antibodies. The most commonly observed infiltrating cell type was suppressor/cytotoxic T cells (CD8+) and smaller subpopulations of lymphocytes were helper/inducer T cells (CD4+) and B cells (CD20+). Variable numbers of HLA-DR+ cells were commonly observed in the lamina propria. The fibrous type of chronic sinusitis was found to have more suppressor/cytotoxic T cells (CD8+) and lower CD4/CD8 ratio than the other histopathological types.     

Lymphocyte subpopulations in the surface secretion on the adenoid

The significance and protective efficacy of surface secretions on mucosal membranes in the upper airways are well recognized. The aim of our study was to reveal the role of the adenoids as a source of cellular components in the mucosal secretion. The adenoid removed because of its hypertrophy and the samples of surface secretions taken by "imprint method" described by Ebenfelt et al. from the group of 38 children were examined. By flow cytometry the lymphocyte subsets with following antigens: CD3, CD4, CD8, CD19, CD23, CD16+56, CD45RA, CD45RO, HLA-DR were analyzed. The results were compared between the groups of younger and older children and in the group of concomitant otitis media with effusion (OMS) and pure adenoid hypertrophy. The percentage of lymphocyte subsets in adenoid and secretion were similar, except for lymphocytes Th with memory phenotype which were more numerous in the adenoid and lymphocytes B CD23+ which were more numerous in the secretion. In the adenoid the percentage of T and Th lymphocytes increased with age and the percentage of B lymphocytes decreased. In the secretion the percentage of lymphocytes Th and B CD23+ was higher among older children. In cases of otitis media with effusion higher percentage of lymphocytes B (CD19+ and CD19+CD23+) and lower of lymphocytes Ts and Th with naive phenotype were observed in the secretion. In adenoid however lower percentage of lymphocytes B and higher of lymphocytes Ts was observed in cases of OMS.     

Early stage impacts of tonsillectomy on immune functions of children

OBJECTIVE: The purpose of this study was to investigate the changes in the humoral and cellular immunity of patients with chronic tonsillitis before and 1 month after tonsillectomy. PATIENTS AND METHODS: 37 patients scheduled for tonsillectomy were enrolled in this study. The levels of CD3+, CD4+, CD8+, CD19+, CD25+ and CD16+ + 56+ were measured for cellular immunity, and levels of IgG, IgA, IgM, C3 and C4 were measured for humoral immunity in blood samples taken from these patients before and 1 month after the operation. RESULTS: The levels of CD3+, CD8+ and CD19+ were reduced in post-operative period as compared to pre-operative period but this was not statistically significant (P > 0.05). However it was found that the level of CD4+ was significantly increased while the level of CD25+ was reduced (P < 0.05) in the post-operative period. There were statistically significant differences between pre- and post-operative levels of immunoglobulins, C3 and C4, which were decreased after tonsillectomy (P < 0.05), but these levels were comparable with those of the control group. CONCLUSION: The results from the present study indicate that humoral and cellular immunity of patients undergoing tonsillectomy were decreased in the early period and came to normal later. The cellular and humoral immune responses are stimulated in patients with tonsillitis, and tonsillectomy removes this stimulus without negatively affecting the patient's immune functions.     

The role and expression of CD27/CD70 in tonsil

We studied the expression and function of CD27 and CD70, which are reported to play an important role in the T-B cell interaction in tonsillar lymphocytes of patients with chronic tonsillitis. In comparison of CD27 and CD70 in peripheral lymphocytes of each patients, no significant difference was found. Furthermore, CD27 and CD70 in T-B cell interaction and antibody production in the tonsil was studied. Our findings was confirmed that CD27 and CD70 are involved in interactions between T/T, T/B and B/B cells. A comparative study of CD27- CD4+ cells and CD70+ CD4 cells, so-called memory cells in the T cell subset, was made in tonsillar lymphocytes and peripheral lymphocytes. No significant difference was found in the presence of CD27+ CD4+ cells, but the CD27- CD4+ cells increased in tonsillar lymphocytes. This suggests recurrent antigenic stimulation and the presence of memory cells in tonsillar lymphocytes. Localization of CD27 and CD70 in tonsillar tissue was also investigate.     

慢性扁桃体炎患者外周血T淋巴细胞亚群测定

目的：探讨慢性扁桃体炎患者免疫状态。方法：对６０例慢性扁桃体炎患者进行外周血Ｔ淋巴细胞亚群测定。结果：慢性扁桃体炎患者的ＣＤ４,ＣＤ４／ＣＤ８较正常对照组降低（Ｐ〈０．０１）。结论：慢性扁桃体炎患者细胞免疫功能受抑制,免疫系统失衡。