Isolation and phenotypic characterization of mucosal nasal lymphocytes by direct ex vivo analysis

Cellular inflammation of the nasal mucosa demonstrates a local immune response which plays an important role in allergic rhinitis. The aim of the present study was to characterize nasal mucosal lymphocytes regarding their activation and differentiation state by direct ex vivo flowcytometric analysis. Lymphocytes from the inferior turbinates were isolated by a mechanical method of preparation and, for comparison, from peripheral blood by Ficoll gradient centrifugation. Patients suffering from rhinitis or difficulty in nasal breathing were divided into an allergic (pollen-allergy, n = 13) and non-allergic group (n = 24). Expression of different T- and B-cell markers was determined by flowcytometric analysis. CD4+ T-cells from the nasal mucosa exhibited a memory phenotype (CD45RO+, 97%), were highly activated (CD69+, 43–73%), and showed low expression of the cutaneous lymphocyte antigen (CLA+, 5%). Nasal CD20+ B-lymphocytes expressed significantly higher levels of mIgE and lower levels of CD23 and CD80 than peripheral B-cells. Subsets of CD80+ (4%) and CD86+ (6%) CD20+ B-lymphocytes were identified in the nasal mucosa. No significant differences between allergic and non-allergic individuals were determined. As expected, the data show profound phenotypical differences between circulating peripheral blood and nasal mucosal lymphocytes. Activated memory lymphocytes are present in the nasal mucosa from allergic, but also non-allergic patients and may indicate to a significant role of a local inflammatory state without systemic criteria for allergy. In our study, we show that direct ex vivo isolation of lymphocytes is practicable method and offers a new technique to examine the local nasal allergic immune response using a multiparametric phenotypical analysis. Christin Wolfram and Claudia Rasche contributed equally to this work.     

Lymphocyte measures in treatment-naïve 13-15-year old adolescents with alcohol use disorders

Many adolescents have chronic exposure to hazardous levels of alcohol. This is likely to be a significant predictor of health outcomes, including those related to immunity. We assessed substance use and biochemical immunological parameters in heavy drinking adolescents (meeting DSM-IV criteria for alcohol dependence) and light/nondrinking control adolescents in Cape Town. Lifetime alcohol dose, measured in standard units of alcohol, was orders of magnitude higher in alcohol-dependent (AD) participants than controls. All adolescent AD had a “weekends-only” style of alcohol consumption. The AD group was chosen to represent relatively “pure” AD, with minimal other drug use and no psychiatric diagnoses. With these narrow parameters in place, we found that AD adolescents were lymphopenic compared with controls, with significantly lower mean numbers of absolute circulating CD3+, CD4+, and CD8+ T-lymphocytes. On conclusion, we found that adolescent AD individuals with excessive alcohol intake, in a weekend binge-drinking style but without comorbid drug or psychiatric disorders, may be at increased risk of lymphopenia. This alcohol misuse may increase infectious disease susceptibility (including TB and HIV) by reducing immune system capabilities. Complex interactions of alcohol with other documented high-risk activities may further compound health risks.     

维生素D与慢性荨麻疹的研究进展

慢性荨麻疹患者的平均维生素D与对照组相比明显偏低,且与病情严重程度及病程存在负相关.尽管尚不明确其中的因果关系,两者间的潜在联系可能与维生素D所起的免疫调节作用有关.有研究表明,维生素D可通过结合维生素D受体影响下游信号通路及核内转录水平,从而对肥大细胞、淋巴细胞以及嗜酸性粒细胞等多种免疫细胞的功能产生调节效应.维生素D可抑制肥大细胞释放活性介质,还可影响T淋巴细胞亚群动态平衡和迁移,引起干扰素γ水平下降和白细胞介素10水平升高等变化,可能是维生素D影响荨麻疹发病的重要机制.同时,临床药物试验显示,补充维生素D对慢性荨麻疹的治疗具有潜在的辅助作用.因此,若能继续明确维生素D与慢性荨麻疹发病的因果关系,从细胞及基因层面研究维生素D对肥大细胞等主要效应细胞的具体影响,将有助于为慢性荨麻疹的诊断和治疗找到新的理论依据.     

鼻咽癌组织VEGF表达及淋巴细胞浸润对患者预后的影响

目的：研究鼻咽癌活检组织中血管内皮生长因子（VEGF）的表达及瘤巢中淋巴细胞的浸润程度对鼻咽癌患者远期预后的影响。方法：以免疫组化S-P法检测42例初诊无远处转移鼻咽癌患者活检组织中VEGF的表达情况，在HE切片上观察瘤巢中淋巴细胞的浸润程度，对其与患者临床病理特征及远期生存的关系进行分析。结果：VEGF低表达组与高表达组5年生存率分别为86．67％和37．04％（P=0．002），瘤巢低淋巴细胞浸润组与中、高组5年生存率分别为23．5％和76．0％（P=0．0014）；VEGF低表达组与高表达组、瘤巢低淋巴细胞浸润组与中、高组的生存曲线分布差异有统计学意义。VEGF表达水平与N分期正相关，瘤巢淋巴细胞浸润程度与患者年龄呈负相关。结论：VEGF高表达、瘤巢低淋巴细胞浸润的鼻咽癌患者5年生存率降低，VEGF表达水平、瘤巢淋巴细胞浸润情况可以作为评价鼻咽癌患者预后的参考指标。     

夏至草提取物对失血性休克大鼠转归时淋巴循环的影响

目的：探讨夏至草提取物对失血性休克大鼠转归时淋巴循环的作用。方法：Wistar雄性大鼠16只，分成实验组和对照组两组，采用淋巴学研究方法，观察夏至草提取物对失血性休克大鼠肠淋巴循环的干预作用。结果：在休克输血输液治疗时，夏至草治疗组的肠淋巴流量、淋巴细胞输出量显著高于对照组（P〈0．01，P〈0．05）。此外，在休克输血输液治疗时，夏至草可明显降低淋巴液表观黏度，增加淋巴液中淋巴细胞和单核细胞数目，其作用明显强于对照治疗组（P〈0．01，P〈0．05）。结论：在失血性休克转归时，夏至草能通过增强淋巴管转运功能，降低淋巴液黏度来增加淋巴细胞的数目以改善失血性休克时的淋巴循障碍。     

周期联合甲氨蝶呤 环磷酰胺对类风湿关节炎患者外周血淋巴细胞周期及凋亡的影响

目的探讨甲氨蝶呤(MTX)和环磷酰胺(CTX)及两者联合治疗对类风湿关节炎(RA)患者外周血淋巴细胞(PBL)周期及凋亡的影响。方法入选活动期RA患者30例,随机分为3组(MTX组、CTX组及MTX+CTX联合组),应用流式细胞术检测MTX、CTX及联合治疗前后PBL周期及凋亡的变化,同时对10名健康志愿者PBL的周期及凋亡情况进行检测。结果活动期RA患者基线PBL各期细胞比例与正常人相比,G0/G1期细胞比例低于正常对照组,S期细胞比例高于正常对照组,差异均有统计学意义;基线早期凋亡率与正常人相比差异无统计学意义。MTX组患者用药后较用药前G0/G1期细胞比例增加,S期细胞比例减少,PBL早期凋亡率明显增加,差异均有统计学意义。CTX组患者用药前后PBL各期细胞比例差异无统计学意义;用药后较用药前PBL早期凋亡率明显增加,差异有统计学意义。MTX+CTX联合组用药后较用药前PBLG0/G1期细胞比例增加,S期细胞比例减少,早期凋亡率较MTX组、CTX组明显增加,差异均有统计学意义。MTX组及MTX+CTX联合组用药前后外周血淋巴细胞G0/G1期细胞比例变化与早期凋亡率变化之间呈显著正相关。结论MTX+CTX联合治疗能更加有效地诱导阻滞于G0/G1期的PBL发生凋亡。     

Defective B-cell and regulatory T-cell function in Wiskott-Aldrich syndrome

We report two Chinese boys with Wiskott-Aldrich syndrome presenting with gastro-intestinal bleeding, eczema and recurrent infection. They had thrombocytopenia and the mean platelet volume was small. Serum IgG and IgA were elevated and lymphocyte proliferation in response to phytohaemagglutinin, concanavalin A and pokeweed mitogen was defective. Despite documented herpes simplex virus type 1 and cytomegalovirus infection in one patient, he did not mount any humoral response. The generation of antibody-secreting cells in response to pokeweed mitogen was markedly defective in a plaque-forming cell assay. Both patients' regulatory T-cell and B-cell functions were defective in this assay. The genetic defect in Wiskott-Aldrich syndrome therefore affects T-cells, B-cells and platelets.     

Clinical aspects and distribution of immunologically active cells in the nasal mucosa of patients with nasal polyps after endoscopic sinus surgery and treatment with topical corticosteroids

Summary Clinical parameters of 72 patients who were operated upon for nasal polyps were evaluated as well as biopsy specimens of the mucosa of the middle and inferior turbinates of 41 of these patients. Biopsies were taken at the time of endoscopic sinus surgery (ESS), after 6 months and after 1 year in 23 patients. During the follow-up period the patients were treated with topical corticosteroids (budesonide). At the time of ESS significantly more CD8+ (suppressor/cytotoxic) cells than CD4+ (helper/inducer) cells were found in the middle and inferior turbinates. At 6 months significantly more CD4+ cells were found than at the time of ESS, whereas at 1 year the number of CD4+ cells had decreased and was lower than at 6 months. These data support the theory that the occurrence of nasal polyps is associated with T-cell-dependent disturbances. Clinical evaluation revealed that most of the patients with chronic airway obstruction had better pulmonary functions postoperatively or required less medication for lung disease. These findings show that ESS combined with topical corticosteroids has a positive effect on upper and lower respiratory tract pathology. Correspondence to: A. E. Stoop     

乳腺癌前哨淋巴结活检技术

目的 探讨不同方法检测乳腺癌前哨淋巴结（sentinel lymph node,SLN)的可行性及其临床意义。方法 71例经细针穿刺或术中冰冻切片诊断为乳腺癌而临床无腋窝淋巴结肿大的女性患者,随机分为4组,其中亚甲蓝淋巴示踪24例,活性染料异硫蓝示踪29例,核素^99m锝－硫胶体示踪8例,染料与核素联合示踪10例,确认并切除前哨淋巴结,将其单独送病理,随后行乳腺癌手术,包括传统的腋窝淋巴结清扫术。结果 71例患者中有60例成功确定前哨淋巴结,检出率84．5％,其中亚甲蓝组75．0％（18／24）,异硫蓝组86．2％（25／29）,核素组检出7例,联合组全部检出;该技术总的敏感性83．3％,4组分别为70．0％、90．0％、100％、100％;假阴性率各组分别为30．0％、10．0％、0、0;总的准确率93．3％,各组分别为83％、96．0％、100％、100％。各组寻找前哨淋巴结平均花费时间分别为29、22、7、6min。结论 前哨淋巴结活检技术在临床是可行的。绝大多数前哨淋巴结可以比较准确地反映其余腋窝淋巴结的组织学特点。活性染料与核素联合示踪既直观又准确,是最佳的选择方法。