慢性乙型肝炎患者CD14~(high)CD16~+单核细胞表达特点研究

目的探讨慢性乙型肝炎(CHB)患者外周血中CD14~(high)CD16~+单核细胞频率、功能特点以及与疾病进程之间的关系。方法利用荧光抗体标记结合多色流式技术,检测CHB患者外周血CD14~(high)CD16~+单核细胞频率及表面TLR2分子的表达,用LPS刺激及细胞内染色方法检测CHB患者外周血CD14~(high)CD16~+单核细胞产生细胞因子的能力。结果 CHB患者外周血的CD14~(high)CD16~+单核细胞频率明显高于健康对照;CHB患者CD14~(high)CD16~+单核细胞频率与ALT呈负相关(R=0.739,P0.01),与HBV DNA载量成正相关(R=-0.283,P=0.008);CHB患者外周血CD14~(high)CD16~+单核细胞上TLR2的表达高于其他两个亚群;免疫活化CHB患者外周血在LPS刺激后,CD14~(high)CD16~+单核细胞亚群分泌IL-6的能力均高于CD14highCD16-和CD14lowCD16+单核细胞亚群。结论 CD14~(high)CD16~+单核细胞可能在清除肝炎病毒及肝脏免疫损伤中起重要作用。     

慢性乙型肝炎患者不同免疫状态下外周血自然杀伤细胞亚群的特点及其临床意义

背景:外周血自然杀伤细胞(NK细胞)亚群在各种病毒感染性疾病中有不同程度的改变,其在HBV感染不同免疫状态下的变化尚不明确。目的:研究慢性乙型肝炎(CHB)患者不同免疫状态下外周血NK细胞亚群的变化特点及其临床意义。方法:采用流式细胞术检测46例CHB患者(免疫耐受期25例,免疫清除期21例)和10例健康志愿者的外周血NK细胞亚群比例,采用实时荧光定量PCR技术检测血清HBV DNA载量,同时检测血清ALT水平。结果:免疫清除期CHB患者外周血CD56~(bright)NK细胞比例明显高于免疫耐受期和健康对照组,差异均有统计学意义(0.877±0.493对0.647±0.294和0.546±0.173,P0.05);三组间CD56~(dim)NK细胞比例差异无统计学意义。外周血CD56~(bright)NK细胞比例与血清HBV DNA载量在免疫耐受期呈负相关(r=-0.575,P0.05),在免疫清除期则无相关性。各组不同免疫状态下外周血NK细胞亚群比例与血清ALT水平均无相关性。结论:外周血CD56~(bright)NK细胞比例增高可能是反映CHB患者进入免疫清除期的潜在指标之一,NK细胞及其亚群在抗HBV的适应性免疫中发挥重要调节作用。     

多色流式细胞术检测CHB患者外周血多重细胞因子分泌的Th细胞亚群

目的建立多色流式胞内细胞因子检测方法,明确慢性乙型肝炎(chronic hepatitis B,CHB)免疫清除期患者外周血Th淋巴细胞亚群细胞因子的分泌特点,探讨CHB患者多重细胞因子的表达模式。方法选取2017年3月至2017年4月就诊于首都医科大学附属北京地坛医院的CHB免疫清除期患者10例及健康对照者10例为研究对象。根据多色流式荧光搭配原则,确立外周血T淋巴细胞CD3、CD4、CD8及细胞因子GM-CSF、TNF-α、IFN-γ的六色流式细胞配色方案。利用健康对照者外周血单个核细胞(peripheral blood mo-nonuclear cell,PBMC)调节最适检测电压及荧光补偿,明确PMAIonomycin的体外刺激方案。分离10例CHB患者外周血PBMC细胞,PMA-Ionomycin体外刺激5小时后,行多重流式细胞因子染色,利用多色流式细胞仪LSR Fortessa获取细胞,Flowjo10.0流式细胞软件分析健康对照者及CHB患者外周血Th细胞亚群胞内细胞因子共表达水平。采用Graph Pad Prism 7.0对所得数据进行统计分析。结果建立了多重细胞因子的胞内染色方案,明确了GM-CSF、TNF-α、IFN-γ在Th细胞上的共表达分析策略,确认存在分泌双重细胞因子GM-CSF~+TNF-α~+、GM-CSF~+IFN-γ~+的Th细胞亚群。通过检测CHB患者外周血样本,发现CHB患者分泌GM-CSF、GM-CSF~+TNF-α~+、GMCSF~+IFN-γ~+的Th细胞亚群百分率均显著高于健康对照组,差异具有统计学意义(t=2.576、4.208、2.671,P均0.05)。结论建立的多色流式细胞因子胞内染色方案可用于明确CHB患者外周血分泌双重细胞因子的Th细胞亚群的分布模式;双重细胞因子分泌的Th细胞亚群的检测可能为临床上CHB患者免疫状态的评估提供可靠依据。     

慢性乙肝患者肝组织内CD4+、CD8+和CD25+T淋巴细胞的表达和研究

探讨肝内免疫活性细胞与乙肝病毒（HBV）慢性感染的关系。运用免疫组织化学法分析54例CHB患者肝内CD25^＋T细胞以及部分患者肝内CD4^＋T细胞、CD8^＋T细胞的表达,并与9例正常肝组织进行比较。CD25^＋和CD8^＋T细胞在HBV感染组和非HBV感染组中的表达差异均存在显著性（P〈0．05和P〈0．01）,CIM^＋T细胞在HBV感染组的表达虽有增多的趋势,但统计学上无显著意义（P〉0．05）。HBV感染组CD25^＋T细胞表达与I临床病理炎症分级和ALT的变化相关,随着炎症分级的加重和ALT的升高,其表达逐渐增强（P〈0．05和P〈0．01）。CD4^＋、CD8^＋T细胞表达与临床病理炎症分级和ALT的变化无关（P〉0．05）。CHB患者肝内CD25^＋T细胞表达增强与机体对HBV产生免疫耐受有关,CHB患者肝内CIM^＋、CD8^＋T淋巴细胞的表达异常,功能性免疫活性细胞浸润量不足,不能有效清除HBV。     

四色流式分析冠心病患者外周血单核细胞亚群及单核细胞-血小板聚集体的变化

目的:建立基于四色的流式细胞术检测外周血单核细胞亚群及各亚群单核细胞-血小板聚集体(MPA)的方法,并观察冠心病患者外周血单核细胞亚群及各亚群MPA的变化特点。方法:本研究纳入经冠状动脉造影证实的冠心病患者和健康对照者各25例,记录所有研究对象的一般临床资料,并采集外周静脉血,依据单核细胞和血小板的标志性分子CD86、CD14、CD16和CD41对单核细胞亚群和MPA进行四色的流式细胞术(FCM)分析。利用抗CD86-PE-Cy5圈出总的单核细胞(总Mon)群,利用抗CD14-FITC和抗CD16-PE将总Mon分为:经典型(CD14++CD16-,Mon1)、中间型(CD14++CD16+,Mon2)和非经典型(CD14+CD16++,Mon3)3个亚群,最后应用CD41-PE-Cy7分析总Mon和各个亚群的MPA,并应用Flow-count TM荧光微球对各个亚群及MPA的绝对量进行计数,比较两组研究对象外周血单核细胞各亚群及各亚群MPA的差异。结果:冠心病组患者外周血Mon2和Mon3所占百分比和绝对计数均显著高于对照组(P<0.05),而Mon1和总Mon两组间比较差异无统计学意义;与对照组相比,冠心病患者总MPA和Mon2相关MPA所占百分比和绝对计数亦显著增高(P<0.05),而Mon1相关MPA和Mon3相关MPA的百分比及绝对计数两组间比较差异无统计学意义。结论:基于CD14、CD16、CD86和CD41的四色流式分析能够有效定量分析外周血单核细胞各亚群及各亚群MPA。中间型单核细胞及其形成的MPA水平显著升高,为临床上冠心病患者的诊治策略提供了新思路。     

丙型肝炎病毒感染者外周血淋巴细胞亚群的变化及意义

目的调查丙型肝炎患者外周血淋巴细胞亚群变化及影响因素。方法利用流式细胞术检测241例丙型肝炎患者和117例健康人群外周血淋巴细胞亚群数值,通过回顾性分析,调查我国丙型肝炎人群淋巴细胞亚群变化及其相关的影响因素。结果在健康人群中,CD3＋CD4＋T细胞、CD3＋CD16＋CD56＋NKT细胞频率以及CD4/CD8比值与年龄呈显著正相关;而在慢性丙型肝炎人群中,CD3＋CD4＋T细胞与年龄呈显著正相关,CD3-CD19＋B细胞则与年龄呈显著负相关;在肝硬化人群中,只有CD3-CD16＋CD56＋NK细胞频率与年龄呈显著正相关;在15～49岁的健康人及慢性肝炎人群中,女性CD3＋CD4＋T细胞亚群频率高于男性,而在肝硬化患者中女性CD3-CD19＋B细胞频率低于男性;同时,在HCV感染的不同阶段,CD3-CD16＋CD56＋NK细胞亚群频率均较正常人显著降低,而CD3＋CD8＋T细胞频率则显著升高,在15岁以上人群,CD3-CD19＋B细胞在健康人、慢性肝炎、肝硬化人群中呈持续升高的现象。结论通过回顾性分析健康人群和HCV感染不同阶段人群淋巴细胞亚群的分布特点及其与HCV疾病进展、年龄、性别的关系,为临床科学评价丙型肝炎人群免疫状态提供重要的免疫指标。     

慢性乙型肝炎患者不同免疫状态及抗病毒治疗早期外周血NK细胞亚群变化特点

目的观察慢性乙型肝炎(chronic hepatitis B,CHB)患者不同免疫状态下外周血NK细胞亚群特点,比较聚乙二醇干扰素及核苷(酸)类似物抗病毒治疗早期外周血NK细胞亚群变化差异。方法收集门诊不同免疫状态下的CHB患者81例,其中免疫耐受期(immune-tolerance phase,IT)患者22例,免疫活化期(immune-activation phase,IA)患者52例,非活动携带组(inactive carry phase,IC)7例,同时纳入健康志愿者(healthy controls,HC)39例作为对照。对27例符合抗病毒治疗标准的患者进行抗病毒治疗,并完成随访。其中22例采用核苷(酸)类似物治疗,5例采用聚乙二醇干扰素治疗。流式细胞术检测患者抗病毒治疗0周、4周、8周、12周及24周外周血NK细胞亚群的比例。结果免疫耐受组、非活动携带组NK细胞总数较健康对照组明显上升(P0.05)。非活动携带组CD56~(dim)NK细胞比例较健康对照组升高(P=0.02)。免疫耐受组及免疫活化组患者外周血CD56~(bright)NK细胞以及CD56~(bright)/CD56~(dim)NK比值与健康对照组相比均明显升高(P0.05);比较CHB患者聚乙二醇干扰素及核苷(酸)类似物抗病毒治疗各时间点外周血NK亚群比例发现,CD56~(dim)NK细胞比例无明显差异。聚乙二醇干扰素组CD56~(bright)NK细胞比例在8周、12周、24周时明显高于核苷(酸)类似物组,差异有统计学意义(P0.05)。结论外周血NK细胞亚群可能与慢性HBV持续感染相关。在抗HBV治疗过程中,聚乙二醇干扰素可能通过上调CD56brightNK细胞发挥免疫调节作用。     

单核细胞表型与弓形体基因型关系概述

单核细胞在抗寄生虫免疫过程中发挥重要作用。人体外周血单核细胞由CD14++CD16-和CD14+CD16+2个亚群组成。弓形体基因型可分为Ⅰ、Ⅱ和Ⅲ型,不同基因型的弓形体能引起不同单核细胞的活化并能诱导单核细胞表型改变及细胞因子的释放。本文就单核细胞表型与弓形虫基因型之间可能的关系做一简要综述。     

小细胞肺癌患者外周血淋巴细胞亚群分析

目的观察小细胞肺癌(SCLC)患者外周血中T淋巴细胞亚群的水平,探讨SCLC免疫变化及其意义。方法采用流式细胞技术,根据白细胞表面抗原分化群(CD)检测33例SCLC患者外周血中的CD+3、CD+4、CD+8、CD+19及CD+16CD+56、活化T细胞比例,以30例健康人做对照,结合临床病理及随访资料,比较其差异。结果 SCLC患者外周血CD+3、CD+19细胞较对照组低;活化T细胞两组间无明显统计学意义。结论SCLC患者机体的细胞免疫异常,与病情发展、临床分期密切相关。检测T淋巴细胞亚群有助于观察患者的免疫状态。     

老年原发性高血压患者外周血淋巴细胞亚群及活化状态的研究

目的：探讨老年原发性高血压患者外周血淋巴细胞亚群活化状态变化。方法采用流式细胞术检测65岁以上老年原发性高血压患者（观察组）170例和体检健康者（对照组）50例外周血淋巴细胞亚群以及活化状态。结果观察组B淋巴细胞表达的 CD3-CD19＋高于对照组,T 淋巴细胞活化表达的 CD3＋CD8＋CD38＋高于对照组,CD3＋HLA- DR＋, CD3＋CD8＋HLA- DR＋以及NK细胞表达的CD16＋CD56＋高于对照组（均P＜0.05）。T淋巴细胞表达的CD3＋低于对照组（均P＜0.05）。T细胞亚群表达的CD3＋CD4＋、CD3＋CD8＋比例下降,调节性T细胞CD3＋CD4＋CD25＋比例上升,但与对照组比较差异无统计学意义（均P＞0.05）。结论老年原发性高血压患者B淋巴细胞、NK淋巴细胞和CD8＋T细胞的活化能力显著增加。     

中国乙型肝炎患者外周血淋巴细胞亚群频率参考值范围

目的 调查中国乙型肝炎患者外周血淋巴细胞亚群频率参考值范围.方法 利用流式细胞术检测2846例乙型肝炎患者和117例健康人群外周血淋巴细胞亚群数值,调查我国健康人群和乙型肝炎人群的参考值范围.结果 调查了16～60岁健康人群和HBV感染相关的急性肝炎、慢性肝炎、重型肝炎和肝硬化人群外周血CD3+T淋巴细胞、CD3+CD...     

乙型肝炎慢加急性肝功能衰竭患者免疫功能抑制与疾病严重程度的研究

目的 探讨乙型肝炎慢加急性肝功能衰竭(ACLF)患者体内免疫功能抑制与疾病严重程度的关系.方法 收集上海公共卫生临床中心2009年8月至2010年4月住院治疗的乙型肝炎ACLF患者27例(ACLF组),活动性慢性乙型肝炎患者28例(CHB组)和健康志愿者8名(对照组)的临床资料及外周血标本.以APACHEⅢ评分及肝性脑病程度作为反映疾病严重程度的量化指标.应用流式细胞术检测患者外周血中T淋巴细胞亚群的绝对计数和单核细胞表面人类白细胞抗原(HLA)-DR的表达量.采用CBA试剂盒中检测患者血浆中促炎性细胞因子及抑炎性细胞因子[干扰素(IFN)-γ、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-2、IL-4、IL-10]的水平.采用SPSS 16.0软件进行数据统计.结果 与CHB组和对照组相比,ACLF组患者抑炎性细胞因子IL-10含量显著增加(Z=-4.279,U=124,P＜0.01;Z=-3.871,U=9.5,P=0.0001),促炎性因子IFN-γ、TNF-α、IL-2、IL-4则处于检测值下限;外周血中单核细胞表面HLA-DR表达量显著下调(Z=-4.714,U=98,P＜0.01;Z=-4.086,U=4,P＜0.01),HLA-DR表达量和APACHEⅢ评分存在显著负相关(R2=0.2667,P=0.0167);适应性免疫细胞CD4+T淋巴细胞绝对数量减少(Z=-4.411,U=116,P＜0.01;Z=-3.575,U=17,P=0.004).结论 乙型肝炎ACLF患者的免疫系统存在单核细胞功能抑制、CD4+T细胞耗竭及高含量抑炎性细胞因子的免疫紊乱状态,单核细胞HLA-DR表达量持续下降是反映疾病严重程度的指标.

Abstract：

Objective To explore the association of immune suppression with the severity of HBV-related acute-on-chronic liver failure(ACLF) patients.Methods From August 2009 to April 2010 in Shanghai Public Health Clinical Center, the peripheral blood samples and clinical data of 27 HBV-related ACLF patients (ACLF group), 28 patients wit h chronic active hepatitis B (CHB group)and 8 healthy individuals (Control group) were collected.APACHE Ⅲ score and the grade of hepatic encephalopathy were as quantitative index to evaluate the severity degree of the disease.The absolute counts of the subsets of T lymphocytes and human leukocyte antigen (HLA)-DR expression on the surface of monocyte in patients' peripheral blood were examined by flow cytometry, the proinflammatory cytokines and anti-inflammatory cytokines(IFN-γ, TNF-α, IL-2, IL-4, IL-10) in patients' plasma were detected by cytometric bead array (CBA) kit.The data was analyzed with SPSS 16.0 software.Results Compared with CHB group and control group, the level of anti-inflammatory eytokire IL-10 markedly increased ir HBV-related ACLF patients (Z= -4.279 ,U= 124, P＜0.01;Z= - 3.871, U= 9.5, P= 0.0001 ), however the level of pro-inflammatory cytokines IFN-γ、 TFN-α、IL-2 、 IL-4 in plasma were at low limit of detectable value.Meanwhile the expression quantity of HLA-DR on the peripheral blood monocytes significantly down-regulated (Z= -4.714, U= 98, P＜0.01;7= - 4.086, U = 4, P＜ 0.01), and there was negative correlation between HLA-DR expression quantity and APACHE Ⅲ score (R2 =0.2667, P=0.0167).In addition, the absolute counts of CD4+T lymphocytes in adaptive immune cells significantly decreased (Z= -4.411, U= 116, P＜0.01; Z=-3.575, U= 17, P= 0.0004).Conclusions The immune system of HBV-related ACLF patients displays immune dysfunction like monocyte function inhibition; CD4+ T lymphocytes depletion and high level of anti-inflammatory eytokines, the persistent down-regulation of the HLA-DR expression on monocyte is an indicator for the severity of disease.     

Incidence of HIV infection in monocyte subpopulations characterized by CD4 and HLA-DR surface density.

OBJECTIVE: The purpose of this study was to determine any correlation between the expression of CD4 antigen on the surface of monocytes, and the frequency with which these cells are infected with HIV. DESIGN: CD4 surface expression on monocytes is significantly less than that expressed on CD4+ lymphocytes. Nevertheless, all monocytes express the HIV CD4 receptor and infected individuals have a significant decrease in the number of monocytes that express a higher density of CD4 surface fluorescence. METHODS: Three-color flow cytometric analysis was used to characterize monocyte-enriched peripheral blood mononuclear cells (PBMC) in terms of surface expression of CD4, CD14 (macrophage antigen), and class II major histocompatibility antigen (HLA-DR). HLA-DR+ monocytes from HIV-positive individuals were sorted into two subpopulations based on either 'bright' or 'dim' CD4 surface expression. A polymerase chain reaction (PCR) assay was used to detect the presence of proviral HIV sequences within the DNA from 10(5) cells from each sorted population. RESULTS: Post-sort analysis revealed that the dim CD4+ monocyte subset expressed dim HLA-DR surface antigen, while the bright CD4+ monocyte subset contained both bright and dim HLA-DR+ cells. PCR results showed that four out of eight dim CD4+ monocyte subsets contained proviral HIV DNA, compared with one out of eight bright CD4+ monocyte subsets.     

原发性肝癌和乙型肝炎肝硬化患者外周血淋巴细胞凋亡率、T细胞亚群和NK细胞的水平变化和临床意义

目的通过检测乙型肝炎肝硬化和合并乙型肝炎病毒感染的原发性肝细胞癌患者的外周血的粒细胞、单核细胞、NK细胞、T细胞及其亚群和淋巴细胞及其凋亡率,探讨两者在细胞免疫方面的差异.方法用Ficoll Hypaque离心法分离外周血单个核细胞(PBMNC),流式细胞仪检测外周血T细胞及其亚群、NK细胞和淋巴细胞、单核细胞和粒细胞,AnnexinV/FITCKit检测凋亡细胞.结果乙型肝炎肝硬化患者的外周血单核细胞、粒细胞、淋巴细胞、CD3-CD16 CD56 NK细胞、CD3 T细胞、CD3 CD4 T细胞、CD3 CD4 T细胞和CD4 CD8 T细胞比值,均与正常对照组无显著性差异(P＞0.05);但淋巴细胞凋亡率明显低于对照组(P＜0.01).原发性肝癌外周血CD3-CD16 CD56 NK细胞、单核细胞和CD4 /CD8 T细胞比值与肝硬化组和正常对照组比较,均无显著性差异(P＞0.05),而粒细胞明显升高(P＜0.05);CD3 T细胞、CD3 CD4 T细胞和CD3 CD8 T细胞均较另两组明显减少(P＜0.05),淋巴细胞及其凋亡率均明显低于另两组(P＜0.01).结论乙型肝炎肝硬化患者的外周血细胞免疫只发生不显著的变化,但淋巴细胞的凋亡率明显降低.原发性肝癌外周血的细胞免疫和淋巴细胞凋亡率均明显低下.     

Identification and characterization of a novel monocyte subpopulation in human peripheral blood

With the aid of two-color immunofluorescence and flow cytometry, a new subset of cells coexpressing CD14 and CD16 antigens can be identified in human peripheral blood. Using the monoclonal antibody My4, these CD14+/CD16+ cells account for 2.2% of the mononuclear cells and form about 13% of all cells identified by the monocyte-specific CD14 monoclonal antibody. The CD14+/CD16+ cells can be assigned to the monocyte lineage based on typical morphology, on expression of additional monocyte-associated molecules, on the ability to form reactive oxygen intermediates and on the expression of monocyte-specific NaF-sensitive esterase. Light scatter analysis revealed lower forward angle and right angle light scatter for the CD14+/CD16+ cells compared with the regular monocytes, and the average cell size was determined to be 13.8 and 18.4 microns, respectively. Expression of class II antigens on these "small monocytes" was twofold higher compared with the regular monocytes. By contrast, the capacity to perform adherence to plastic surfaces, as well as the ability to phagocytize antibody-coated erythrocytes was clearly reduced in the CD14+/CD16+ monocyte subset as compared with the regular monocytes. Hence the CD14+/CD16+ cells appear to represent a new monocyte subset with a distinct functional repertoire. A survey of various tissues revealed that a large proportion of the alveolar macrophages, but not of the peritoneal macrophages, express the CD14+/CD16+ phenotype.     

Heterogeneity of Human Monocytes: An Optimized Four-Color Flow Cytometry Protocol for Analysis of Monocyte Subsets

Monocytes are central mediators in the development of atherosclerotic plaques. They circulate in blood and eventually migrate into tissue including the vessel wall where they give rise to macrophages and dendritic cells. The existence of monocyte subsets with distinct roles in homeostasis and inflammation suggests specialization of function. These subsets are identified based on expression of the CD14 and CD16 markers. Routinely applicable protocols remain elusive, however. Here, we present an optimized four-color flow cytometry protocol for analysis of human blood monocyte subsets using a specific PE-Cy5–conjugated monoclonal antibody (mAb) to HLA-DR, a PE-Cy7-conjugated mAb to CD14, a FITC-conjugated mAb to CD16, and PE-conjugated mAbs to additional markers relevant to monocyte function. Classical CD14⁺CD16⁻ monocytes (here termed “Mo1” subset) expressed high CCR2, CD36, CD64, and CD62L, but low CX₃CR1, whereas “nonclassical” CD14^loCD16⁺ monocytes (Mo3) essentially showed the inverse expression pattern. CD14⁺CD16⁺ monocytes (Mo2) expressed high HLA-DR, CD36, and CD64. In patients with stable coronary artery disease (n = 13), classical monocytes were decreased, whereas “nonclassical” monocytes were increased 90% compared with healthy subjects with angiographically normal coronary arteries (n = 14). Classical monocytes from CAD patients expressed higher CX₃CR1 and CCR2 than controls. Thus, stable CAD is associated with expansion of the nonclassical monocyte subset and increased expression of inflammatory markers on monocytes. Flow cytometric analysis of monocyte subsets and marker expression may provide valuable information on vascular inflammation. This may translate into the identification of monocyte subsets as selective therapeutic targets, thus avoiding adverse events associated with indiscriminate monocyte inhibition.     

CD16+ monocytes in patients with cancer: spontaneous elevation and pharmacologic induction by recombinant human macrophage colony- stimulating factor

The small subset of circulating monocytes that express the maturation- associated CD16 antigen has recently been reported to be elevated in patients with bacterial sepsis. We now show that this novel CD16+ monocyte population is also spontaneously expanded in patients with cancer. We studied 14 patients with metastatic gastrointestinal carcinoma enrolled ina clinical trial of recombinant human macrophage colony-stimulating factor (rhMCSF) plus monoclonal antibody D612. We found that before any cytokine treatment, 12 of 14 patients constitutively displayed significant elevations in both the percentage and the absolute number of CD16+ monocytes, as compared with both normal subjects and ill patients with elevated monocyte counts but without malignancy. CD16+ monocytes accounted for 46% +/- 22% of total monocytes in the patients with cancer versus 5% +/- 3% for controls (P < .01). The increase was not attributable to infection or intercurrent illness and appeared to be associated with the underlying malignancy itself. A similar spontaneous elevation of CD16+ monocytes was observed in 35 of 44 additional patients diagnosed with a variety of other solid tumors. When patients with gastrointestinal carcinoma were treated with rhMCSF, there was a marked further increase in the percentage of CD16+ monocytes (to 83% +/- 11%), as well as in the absolute number of CD16+ cells and the level of CD16 antigen expression. In every case, the patients with cancer showed a greater CD16+ monocyte response than the maximal response obtained in normal volunteer subjects treated witha similar regimen of rhMCSF (n = 5, P < .001), suggesting that the presence of malignancy primed patients for enhanced responsiveness to rhMCSF. We hypothesize that spontaneous expansion of the CD16+ monocyte population may represent a novel biologic marker for a widespread and previously unsuspected host immune response to malignancy.     

外周血CD14~+CD16~+细胞与稳定型心绞痛患者冠状动脉硬化病变程度的相关性研究

目的:单核细胞在整个动脉硬化进程中至关重要,根据细胞表面CD14和CD16的差异表达,单核细胞可分为CD14+CD16-和CD14+CD16+两种亚群,本研究目的是探讨外周血单核细胞亚群与稳定型心绞痛患者冠状动脉硬化病变程度的相关性。方法:入选97例进行冠状动脉造影的稳定型心绞痛患者,根据造影结果分为无冠心病组(non-coronary artery disease,non-CAD)、单支病变组(single-vessel disease,SVD)及多支病变组(multiple-vessel disease,MVD),并对病变的严重程度进行Gensini评分。通过流式细胞术测定外周血两种单核细胞亚群数量。结果:外周血单核细胞CD14+CD16+亚群在MVD组的比例高于SVD组和non-CAD组(MVD:15.2±4.6;SVD:12.5±3.7,P0.001;non-CAD:7.5±2.5,P0.001),且与Gensini评分的相关性分析显示,CD14+CD16+亚群与冠状动脉病变严重程度呈正相关。结论:外周血CD14+CD16+细胞与稳定型心绞痛患者冠状动脉硬化病变程度相关。     

Expression and clinical significance of inhibitory receptor Leukocyte-associated immunoglobulin-like receptor-1 on peripheral blood T cells of chronic hepatitis B patients: A cross-sectional study

Leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1) is an inhibitory receptor that is expressed on the surface of multiple immune cells and plays key roles in immune modulation. In patients with chronic hepatitis B (CHB), T cell number and functions are abnormal and the expression of inhibitory receptors is elevated. However, the expression of LAIR-1 on T cells in patients with CHB is still undetermined.We recruited 320 patients with CHB in different disease phases and 17 healthy donors. Serum biochemical and virological examinations were performed for each participant, and their demographic and clinical data were collected. According to the latest American Association for the Study of Liver Disease guidelines, we categorized the patients into 4 groups: immune active, immune tolerant, inactive CHB, and gray zone. Additionally, we tested the expression of LAIR-1 on T cells and T cell subsets using flow cytometry.We observed a significant decrease in LAIR-1 expression on CD3+ T cells and its two subsets (CD4+ and CD8+ T cells) in patients with CHB. LAIR-1 expression on T cells was the lowest in the immune active group. LAIR-1 expression levels on CD4+ and CD8+ T cells showed a significant negative association with hepatitis B virus (HBV) DNA load and were lower in hepatitis B e antigen (HBeAg)-positive patients than in HBeAg-negative patients (P < .05). In addition, LAIR-1 expression levels on CD3+, CD4+, and CD8+ T cells were all negatively associated with liver inflammation and fibrosis parameters, such as alanine aminotransferase and aspartate aminotransferase levels, FibroScan value, and aspartate aminotransferase-to-platelet ratio index score.LAIR-1 expression levels on T cells were associated with HBV DNA load and liver inflammation and fibrosis parameters, indicating that LAIR-1 may play an important regulatory role in HBV-induced T cell immune pathogenesis and may be a therapeutic target for CHB.