Cytotoxic effect of anti-lymphocytic globulin on human lymphoid cells

Summary The cytotoxic activity of two anti-lymphocytic globulin (ALG) preparations was investigated by measurement of⁵¹Cr release from human peripheral blood mononuclear cells and tonsil lymphocytes incubated with ALG in the presence and absence of complement. Both types of ALG had a greater complement-dependent cytotoxic effect on tonsil lymphocytes than on peripheral blood mononuclear cells. The ALG preparations were more cytotoxic for a population of tonsil lymphocytes enriched in non-T cells than for one enriched in T cells. This finding does not support the view that ALG has a selective action on T lymphocytes.     

聚合酶链反应方法检测外周血淋巴细胞HLA-A mRNA方法学的建立及在肾移植中的初步应用

目的通过建立检测宿主外周血淋巴细胞(PBL)表面HLA-AmRNA表达程度的实时荧光定量逆转录聚合酶链反应(RT-FQ-PCR)方法,检测肾移植患者移植后外周血淋巴细胞表面HLA-AmRNA的表达程度,探讨其在诊断移植后早期排斥反应的发生的价值及机体免疫状态的关系。方法以6-磷酸葡萄糖脱氢酶(G6PDH)作内参照,用Taqman探针荧光定量技术,建立检测PBLHLA-AmRNA的RT-FQ-PCR方法;并检测35例随访移植患者的不同时间的60个标本和60名正常人的阈循环数(Ct),然后用REST软件分析PBLHLA-AmRNA的表达水平。结果正常人和51个移植标本的PBLHLA-AmRNA对G6PDH的相对表达量分别为0·036±0·012和0·026±0·015(双侧t=1·981,P<0·05),提示服用免疫抑制药(目前临床用药剂量均偏大)后患者PBLHLA-AmRNA表达减弱,机体免疫状态低,无排斥反应发生但易导致感染、肿瘤等并发症的产生;其中9个移植标本的PBLHLA-AmRNA对G6PDH的相对表达量为0·042±0·017(双侧t=2·002,P<0·05),提示较其他51例移植标本患者PBLHLA-AmRNA表达增强的患者,经病例回访查询发现该5例移植患者在这期间有排斥反应(根据临床表现、肾功能检查、影像学及核医学检查的临床排斥反应期)的发生,其中1例在移植后6个不同时间的标本显示2次有增高,每次经治疗后均降低。结论建立的宿主PBLHLA-AmRNART-FQ-PCR检测方法简便、特异、重复性好、可信度高,可用于肾移植后PBLHLA-AmRNA表达水平的检测,评估机体的免疫状态,预测排斥反应的发生。     

人外周血淋巴细胞和粒细胞褪黑素受体亚型mt1 的检测

目的 检测人外周血淋巴细胞和粒细胞的褪黑素受体(MR)亚型mt1和其细胞内定位。方法 分离出健康成年人外周血淋巴细胞和粒细胞，应用RT-PCR技术分别检测其亚型mt1的mRNA，阳性产物用自动测序仪测序；同时将所分离的淋巴细胞和粒细胞制成细胞涂片，应用免疫组化染色检测mt1在淋巴细胞和粒细胞上的分布。结果 RT-PCR产物电泳显示：人外周血淋巴细胞和粒细胞均存在mt1阳性条带，片段长度约370bp，测序结果显示扩增产物与GenBank的人mt1受体亚型的基因序列相吻合；免疫组化结果显示：人外周血淋巴细胞和粒细胞均存在mt1受体蛋白，分布于细胞膜、细胞浆和细胞核，以细胞膜和细胞浆为主。结论 人外周血淋巴细胞和粒细胞中均存在MR的亚型mt1,外周血淋巴细胞和粒细胞为褪黑素作用的外周靶器官，提示褪黑素在生理及病理情况下可能对免疫系统有调节作用。     

某退役铀矿山周围居民淋巴细胞微核率分析

目的 调查某退役铀矿山周围居民外周血淋巴细胞微核情况,初步探讨环境因素对微核的影响,为综合治理退役铀矿周围环境提供科学依据。方法 采集100名退役铀矿山周围居民外周静脉血,用常规淋巴细胞微核培养法,检查淋巴细胞微核,进行统计分析。结果 在100名居民中,微核检出率为88.0%,细胞微核率（9.46±6.62）‰、微核细胞率为（8.77±5.92）‰;按性别和年龄分别分组分析得出,细胞微核率和微核细胞率整体随着年龄的增加而增加,但男性和女性增加的速度和幅度在各年龄段稍有不同,男性各组间无明显差异（P>0.05）,女性微核细胞率各组间无明显差异（P>0.05）,细胞微核率各组间差异具有统计学意义（P<0.05）。结论 该退役铀矿山周围居民外周血淋巴细胞微核检出率、淋巴细胞微核率和微核细胞率远超过国内外报道的本底范围,应对退役铀矿周围环境采取更具针对性的综合治理措施,以确保退役铀矿山周围居民的健康。     

内毒素血症大鼠淋巴细胞中隆钙素基因相关肽（CGRP）释放及mRNA表达的变化

目的　测定大鼠内毒素血症期间淋巴细胞释放和合成降钙素基因相关肽 (CGRP)的变化。方法　采用逆转录聚合酶链反应 (RT- PCR)测定大鼠淋巴细胞 CGRP m RNA水平。用放射性标记法测定血浆 CGRP水平。结果　注射内毒素大鼠血浆和淋巴细胞释放 CGRP均增高 ,淋巴细胞中 CGRP m RNA表达也明显增高。结论　提示内毒素不仅刺激淋巴细胞释放 CGRP,而且能通过某些机制激活淋巴细胞 CGRP m RNA的转录 ,使 CGRP合成增加 ,以作为 CGRP大量释放的重要补充来源。     

Effects of Ethanol on Cellular Immunity to Facultative Intracellular Bacteria

Alcohol abuse has been associated with an increase in infectious diseases caused by pathogenic and opportunistic microorganisms. Study results obtained from this laboratory and other laboratories have shown that consumption of large amounts of ethanol is associated with numerous changes in the immune system. The purpose of this article is to report findings obtained from this laboratory, as well as review those obtained from other laboratories, from experiments designed to evaluate the effects of ethanol on various components of antimicrobial host-defense mechanisms. The effects of ethanol on various aspects of immunity obtained with the use of in vivo and in vitro model systems are reviewed as they pertain to antimicrobial defenses. All current data would support the suggestion that ethanol affects both the development of an antigen-specific immune response and the effector mechanisms of the cellular immune response. Findings obtained from animal models show that ethanol prevents the formation of granulomas in infected tissues, perhaps by inhibiting the response of macrophages to T-cell cytokines. Data obtained from this laboratory also support the suggestion that the inability of the immune system to control the intracellular growth of microorganisms results in an exaggerated inflammatory response that is responsible for at least a part of the tissue damage.     

吡喹酮对血吸虫卵肉芽肿与肝纤维化作用的研究Ⅱ吡喹酮对小鼠肺部血吸虫卵肉芽肿内细胞的影响

目的研究应用吡喹酮后小鼠肺部血吸虫卵肉芽肿内细胞的变化。方法48只小鼠均分4组。A组:虫卵致敏组,先于小鼠腹壁皮下注入新鲜日本血吸虫卵,10d后由尾静脉注入虫卵;B组:吡喹酮短期应用组,致敏方法同A组,于尾静脉注入虫卵前1d起灌服吡喹酮300mg/(kg.d)连续3d;C组:吡喹酮持续应用组,致敏方法同A组,于尾静脉注入虫卵前1d起灌服吡喹酮75mg/kg,1日2次,连续5d,停药2d后反复用药至剖检;D组:第29天起应用吡喹酮组,服法同C组。各组均在尾静脉注射虫卵后7、14、28、56d分别剖杀3只,取肺组织切片,HE染色,在每组肺连续切片内选含毛蚴的虫卵肉芽肿25～30个,分类计数每个肉芽肿内的中性粒细胞、嗜酸性粒细胞、淋巴细胞、纤维母细胞、巨噬细胞,计算其平均值并作比较。结果与A组比较,应用吡喹酮的3个实验组,中性粒细胞、嗜酸性粒细胞、巨噬细胞均显著减少,纤维母细胞增高的幅度也明显为低,差异均有统计学意义,而以C组最为显著,第56天,中性粒细胞、嗜酸性粒细胞、巨噬细胞分别减少了54.4%、87.0%和23.1%,纤维母细胞增高的幅度减少了59.4%。各组淋巴细胞数目变化不大。结论吡喹酮能明显抑制虫卵肉芽肿内炎细胞的聚集、纤维母细胞的增生。     

演习人员外周血淋巴细胞热休克蛋白70的表达及相关因素分析

目的观察军事演习与热休克蛋白70表达的关系,并分析影响演习人员热休克蛋白（heat shock protein 70,HSP70）表达的因素。方法某次演习期间,选取参演官兵137人为演习组,观摩演习官兵50名为对照组。问卷调查一般情况和心理应激状况,进行身体测量和血乳酸测定,采用Western blotting法检测外周血淋巴细胞中HSP70的表达水平。组间比较采用t检验,采用多元线性回归分析考察相关影响因素。结果与对照组比较,演习组外周血淋巴细胞HSP70水平显著增加（0.39±0.06vs0.60±0.09）（P〈0.01）,这种表达与年龄、受教育程度、家庭人口、吸烟、血乳酸和心理应激状况自评得分相关（P〈0.05,P〈0.01）,与体质量指数（body mass index,BMI）、收缩压、白细胞计数（WBC）、红细胞计数（RBC）、血红蛋白（HGB）等无明显相关（P均〉0.05）;多元线性回归分析显示血乳酸、心理应激状况和受教育程度是影响演习人员HSP70表达的最显著的因素。结论演习人员外周血淋巴细胞HSP70表达水平升高,这种升高与血乳酸、心理应激状况自评得分和受教育程度等相关。     

Impact of perfluorooctanesulfonate and perfluorooctanoic acid on human peripheral leukocytes

Perfluorinated compounds (PFCs), such as perfluorooctanesulfonate (PFOS) and perfluorooctanoic acid (PFOA), are xenobiotics that can be detected worldwide in the environment, wildlife, and humans. So far, the immunotoxicity of PFCs has only been investigated in rodents, but not in humans. In this study, we explore the impact of PFOS and PFOA on selected functions of human leukocytes in vitro. PFOS induced a significant decrease of natural killer-cell activity and reduced the release of the pro-inflammatory cytokine TNF-α following lipopolysaccharide (LPS)-stimulation. Furthermore, the plasma PFOS concentrations (2.09-8.98 ng/ml) found in our study subjects correlated positively with the LPS-stimulated IL-6 release. PFOA augmented significantly calcitriol-induced monocytic differentiation of the HL-60 cell line. Additionally, there was a significant linear relationship between LPS-stimulated TNF-α and IL-6 release, and the plasma PFOA (1.20-6.92 ng/ml) concentrations of the study subjects. In conclusion, the investigated PFCs affect human immune cells mainly with regard to natural killer-cell cytotoxicity and the pro-inflammatory cytokine release by stimulated macrophages.     

Raf／MEK／ERK信号通路对CXCL16诱导的CIA淋巴细胞增殖的影响

目的探讨细胞外信号调节激酶（ERK）信号通路在CXCL16诱导的CIA小鼠淋巴细胞增殖中的调控作用。方法CCK8法检测CXCL16对CIA小鼠淋巴细胞增殖的影响,ELISA法检测细胞上清中TNF-α及RANKL表达,WesternBlot检测p-Raf-1、p-ERK1/2蛋白的表达,同时观察MEK抑制剂对CXCLl6诱导的上述变化的影响。结果CXCL16对正常小鼠淋巴细胞增殖的刺激作用不明显,但CIA＋CXCLl6组淋巴细胞增殖能力明显高于CIA空白组及CIA＋CXCL16＋抑制剂组（P〈0．05）。CIA＋CXCL16组TNF—α水平明显高于其他各组（P〈0．01）。RANKL水平在CIA＋CXCL16组明显高于CIA空白组（P〈0．01）,但与CIA＋CXCL16＋抑制剂组相比无明显差别。p-Raf-1／β—actin及p-ERK1/2／β—actin在CIA＋CXCL16组明显高于CIA空白组（P〈0．05）。结论CXCL16可能通过激活Raf／MEK／ERK信号通路,诱导CIA小鼠淋巴细胞增殖活化。