重症中暑患者CD14单核细胞HLA-DR表达的意义

目的:研究重症中暑患者CD14单核细胞HLA-DR表达变化规律及其对临床预后判断的作用.方法:40例重症中暑患者分为死亡组和存活组,用流式细胞仪检测其CD14单核细胞HLA-DR水平,并进行受试者工作特征曲线(ROC曲线)分析,比较两组间CD14+单核细胞HLA-DR表达率的差异,评价其在感染监测中的价值.结果:死亡组CD14+单核细胞HLA-DR表达率为(52.9±10.8)％,比存活组(73.7±11.0)％明显减少(P＜ 0.001).CD14+/HLA-DR表达率对死亡判断的ROC曲线下总面积为0.967,在最佳诊断界点为54.83％时敏感性和特异性分别为96.7％和100.0％.结论:CD14单核细胞HLA-DR表达率是判断重症中暑临床预后的良好指标；CD14+/HLA-DR≤54.83％可作为预后死亡的预警指标.     

CD14+/HLA-DR+单核细胞对脓毒症患者预后判断的应用研究

目的 探讨CD14+/人类白细胞抗原-DR+(HLA-DR+)单核细胞在脓毒症发展进程中的变化情况及对预后判断的临床意义.方法 用流式细胞术(flow cytometry, FCM)检测脓毒症患者(预后良好组和预后不良组)及正常对照组全血中的单核细胞表面CD14+/HLA-DR+的表达情况.对10例脓毒症患者CD14+/HLA-DR+单核细胞进行连续监测. 结果 103例脓毒症患者CD14+/HLA-DR+单核细胞明显低于正常对照组(t=8.147,P<0.05);脓毒症患者预后良好组CD14+/HLA-DR+单核细胞与预后不良组比较差异有统计学意义(χ2=14.9,P<0.05);对10例脓毒症患者CD14+/HLA-DR+单核细胞进行连续监测,发现该指标与疾病的发展变化及预后转归有一定的相关性.结论 CD14+/人类白细胞抗原-DR+单核细胞的检测及动态连续监测对脓毒症患者的预后判断有重要意义,对该病的临床治疗有积极的指导作用.     

CFSE标记结合流式细胞术检测单核细胞Siglec-1刺激CD4~+/CD8~+T细胞增殖

目的建立CFSE标记结合流式细胞术检测T淋巴细胞增殖的方法,并探讨其在研究单核细胞唾液酸黏附素(Si-glec-1)对CD4+/CD8+T细胞增殖中的应用。方法免疫磁珠分离冠状动脉综合征(ACS)患者和健康对照者外周血CD14+单核细胞,与经CFSE标记的第三方健康献血者CD4+/CD8+T细胞共培养5 d,流式细胞术检测T淋巴细胞增殖情况。结果 ACS患者CD14+单核细胞具有比健康人单核细胞更强的刺激CD4+/CD8+T淋巴细胞增殖能力;当用干扰素-α(INF-α)刺激增强正常单核细胞Siglec-1表达后,其刺激T细胞增殖能力增强;用单抗阻断单核细胞Siglec-1后,其刺激T细胞增殖能力减弱。结论 ACS患者增强的CD4+/CD8+T淋巴细胞增殖能力部分是由于激活的单核细胞高表达Siglec-1引起。     

早期目标导向治疗对严重脓毒症患者免疫功能的影响

目的评价早期目标导向治疗对严重脓毒症患者免疫功能的影响。方法前瞻性分析50例严重脓毒症患者,随机分为治疗组及对照组,对照组实行常规补液治疗,治疗组实行早期目标导向治疗,治疗组30例,对照组20例,观察两组治疗前和治疗后第6小时、第12小时、第1天、第3天免疫指标情况。结果严重脓毒症患者早期目标导向治疗对细胞免疫影响的分析,治疗组CD4+T淋巴细胞比对照组明显升高,两组差异有统计学意义(P<0.05),治疗组CD14+单核细胞人类白细胞抗原-DR(HLA-DR)水平明显升高,两组差异有统计学意义(P<0.05);严重脓毒症患者早期目标导向治疗对炎症/抗炎因子影响的分析,治疗组TNF-α降低比对照组明显,两组比较差异有统计学意义(P<0.05),治疗组IL-10升高比对照组明显,两组比较差异有统计学意义(P<0.05)。住院期间,对照组死亡8例,治疗组死亡4例,两组比较差异有统计学意义(P<0.05)。结论严重脓毒症患者早期目标导向治疗可以升高细胞免疫中CD4+T淋巴细胞数及CD14+单核细胞HLA-DR水平,明显降低促炎因子TNF-α水平,升高抗炎因子IL-10水平,并改善严重脓毒症患者的住院生存率。     

细胞免疫功能测定在危重症患者中的临床意义

目的探讨细胞免疫功能与危重症患者病情严重程度和预后的关系。方法选择资料完整的危重症患者43例,根据28d生存情况,将患者分为存活组(21例)及死亡组(22例),并通过急性病生理学和长期健康评价(APACHE)Ⅱ评分计算出分值,采用流式细胞仪对T淋巴细胞亚群及自然杀伤(NK)细胞进行检测。结果与存活组比较,死亡组外周血T淋巴细胞CD3+、CD4+及CD8+细胞计数低于存活组,差异均具有统计学意义([270.32±187.58)、(189.27±156.99)、(88.73±56.74),(489.05±109.18)、(342.05±116.41)、(170.33±68.69)个/μl,P均0.05];死亡组外周血T淋巴细胞CD3+、CD4+、CD8+及NK细胞百分数低于存活组,差异均具有统计学意义([43.07±10.35)%、(21.88±8.04)%、(13.71±6.27)%、(7.35±4.46)%,(52.91±13.24)%、(29.76±6.02)%、(24.45±10.78)%、(14.35±5.03)%,P均0.05]。存活组和死亡组患者APACHEⅡ评分与CD3+、CD4+、CD8+细胞计数均呈负相关(P均0.05),而与CD3+、CD4+、CD8+及NK细胞百分数均无相关性(P均0.05)。结论细胞免疫功能水平高低与危重症患者病情严重程度有关。细胞免疫功能水平可作为评估危重症患者病情和预后的重要检测指标。     

肝移植术后CD14+单核细胞人白细胞DR抗原表达率的变化及其意义

目的:探讨肝移植术后CD14+单核细胞人白细胞DR抗原(HLA-DR)表达率的变化及其在感染监测和判断临床预后方面的作用.方法:65例肝移植术后患者分为非感染组、感染组,用流式细胞仪检测其CD14+单核细胞HLA-DR水平,比较两组间CD14+单核细胞HLA-DR表达率的差别.同时按CD14+单核细胞HLA-DR表达率分布将患者分为HLA-DR≥40%(A组)、21%～39%(B组)、HLA-DR≤20%(C组),比较3组间临床预后的差别.结果:感染组CD14+单核细胞HLA-DR表达率为(25.6±9.2)%,比非感染组的(61.3±18.2)%明显减少(P＜0.05).B、C两组在ICU停留时间、术后至出院时间与A组比较明显延长(P＜0.05),以C组延长更为显著(P＜0.01),B、C两组比较差异则无显著性.A、B、C 3组在感染率和病死率之间比较差异亦有显著性(P＜0.05).结论:CD14+单核细胞HLA-DR表达率是监测肝移植术后感染及判断临床预后的良好指标,特别是针对高危或可疑感染的术后患者,动态的CD14+单核细胞HLA-DR监测对病情判断和治疗调整均有指导意义.     

CD14+单核细胞人类白细胞DR抗原及C反应蛋白评估脓毒症患者预后的临床价值

目的:评价外周血CD14+单核细胞人类白细胞DR抗原(HLA-DR/CD14+)和C-反应蛋白(CRP)对脓毒症患者预后的判断价值和临床意义。方法:选择入住ICU的脓毒症患者,治疗前测定其外周血HLA-DR/CD14+及CRP、进行APACHEⅡ评分和Marshall评分,观察追踪28天,根据其预后将患者分为存活组和死亡组,分析2组的参数差异,评估参数与患者预后的关系。结果:本研究共入选脓毒症患者21例,12例存活,9例死亡。存活组患者年龄为(65.67&#177;17.15)岁明显低于死亡组的(71.78&#177;8.87)岁(P&lt;0.05);存活组患者HLA-DR/CD14+水平为(28.9&#177;16.1)%明显高于死亡组的(18.7&#177;12.7)%(P&lt;0.05);存活组患者CRP水平为(56.1&#177;18.7)mg/L明显低于死亡组的(91.1&#177;26.3)mg/L(P&lt;0.05)。结论:脓毒症患者的免疫状态与其预后有关,HLA-DR/CD14+和CRP水平对预后判定具有一定的临床价值。     

严重多发伤患者血糖和细胞免疫功能变化及与预后的关系

目的探讨严重多发伤患者血糖及细胞免疫功能变化特点及与预后的关系。方法检测55例严重多发伤患者空腹血糖及细胞免疫功能指标,并与同期60例轻度多发伤患者(对照组)进行比较,并分析血糖与细胞免疫功能的相关性。根据预后将55例严重多发伤患者分为死亡组和存活组,比较2组的血糖和细胞免疫功能指标,分析其与预后的相关性。结果严重多发伤组血糖显著高于对照组(P<0.05),严重多发伤组CD3+、CD4+、CD4+/CD8+显著低于对照组(P<0.05),血糖与CD3+、CD4+、CD4+/CD8+呈负相关(P<0.05)。死亡组血糖显著高于存活组(P<0.05),CD4+/CD8+显著低于存活组(P<0.05)。结论严重多发伤患者应激血糖水平往往更高,且存在一定程度的细胞免疫功能缺陷,细胞免疫功能与血糖具有一定的相关性,两者均与预后有关。     

脓毒症患者CD4+CD25+调节性T细胞和Th17细胞检测的临床意义

目的 探讨脓毒症患者外周血辅助性T细胞(T Helper,Th)17及CD4+CD25+调节性T细胞(Treg细胞)表达的变化和检测的临床意义.方法 将2008-01～2010-11在我医院ICU住院的脓毒症患者40例按照疾病严重程度分为三组:脓毒症组14例、严重脓毒症组15例和脓毒症休克组11例.再将脓毒症患者根据28 d预后分为死亡组(D,n=11)和存活组(S,n=29).所有入选的40例脓毒症患者在入选当天行流式细胞术检测血中Th17细胞及CD4+CD25+调节性T细胞的比例.比较脓毒症组、严重脓毒症组和脓毒症休克组患者血中Th17细胞及CD4+CD25+调节性T细胞的比例变化,以及生存组和死亡组上述指标的差异.分析CD4+CD25+调节性T细胞和Th17细胞免疫平衡紊乱与病情严重程度的关系.同时选取20例健康人为对照组.结果 健康对照组Th17细胞表达率为(0.91±0.38)%,CD4+CD25+调节性T细胞表达率为(0.39±0.23)%;脓毒症患者较正常对照组这两项指标明显升高,差异有统计学意义(P＜0.05).其中脓毒症组分别为(2.09±0.53)%和(1.72±0.59)%;严重脓毒症组为(3.90±0.80)%和(2.72±0.22)%;脓毒性休克组为(1.85±0.35)%和(3.55±0.51)%.Th17表达率严重脓毒症组最高,与其他两组比较差异有统计学意义(P<0.05).而脓毒症休克组与脓毒症组比较差异无统计学意义(P＞0.05).CD4+CD25+调节性T细胞表达率脓毒症休克组最高,严重脓毒症组次之,脓毒症组最低,三组间两两比较差异有统计学意义(P<0.05).死亡组血中CD4+CD25+调节性T细胞表达率明显高于存活组[(3.03±0.91)%和(2.43±0.79)%,P<0.05],Th17细胞表达率明显低于存活组[(2.01±0.66)%和(2.97±1.15)%,P<0.05].结论 Th17细胞和CD4+CD25+调节性T细胞在脓毒症的免疫发病机制中可能起着重要作用.检测脓毒症患者外周血Th17细胞和CD4+CD25+调节性T细胞能够反映机体的细胞免疫状态,对评估患者预后有重要临床价值.     

脓毒症患者外周血CD14+单核细胞HLA-DR、CD86和T淋巴细胞亚群表达水平的变化及其协同表达研究

目的探讨脓毒症患者外周血CD14~+单核细胞HLA-DR、CD86和T淋巴细胞亚群表达水平的变化及其协同表达的意义。方法选取2020年1-7月铜陵市人民医院重症医学科收治且确诊的脓毒症患者(脓毒症组)作为研究对象,并以同期入院体检健康者作为对照组。脓毒症组患者入院第1天、第3天及第5天抽取外周静脉血,用流式细胞术分析CD14~+单核细胞HLA-DR、CD86~+CD14~+和T淋巴细胞亚群表达水平,同时检测对照组入院体检当日上述各指标水平并对两组各指标进行分析比较。结果脓毒症组入院第1天患者外周血CD14~+单核细胞HLA-DR、CD86~+CD14~+、总T淋巴细胞、CD4~+T淋巴细胞表达水平明显低于对照组,差异均有统计学意义(P0.05);脓毒症组入院第3天、第5天患者外周血CD14~+单核细胞HLA-DR、CD86~+CD14~+、CD4~+T淋巴细胞表达水平明显低于对照组,差异均有统计学意义(P0.05)。结论脓毒症患者体内存在免疫麻痹,CD14~+单核细胞HLA-DR及CD86均可以评估脓毒症患者免疫状况。     

严重烧伤患者外周血单核细胞表面人白细胞DR抗原变化的初步研究

目的 :探讨烧伤后外周血单核细胞表面人白细胞 DR抗原受体 (HL A DR)的动态变化规律及意义。方法 :选取临床烧伤患者 30例 ,依据病程长短选取病程中 1～ 5个时间点静脉采血 ,以流式细胞仪测定外周血单核细胞 HL A DR的表达率 ,并根据烧伤程度分组进行分析。结果 :伤后患者外周血单核细胞 HL A DR的表达率明显降低 ,降低程度及持续时间与伤情有关 ,特重烧伤患者与中度烧伤患者〔(4.30± 1.5 0 ) %比(13.86 %± 2 .4 0 ) %〕、中度烧伤患者与轻度烧伤患者〔(13.86± 2 .4 0 ) %比 (5 8.80± 5 .6 0 ) %〕比较差异均显著(P均 <0 .0 1)。结论 :单核细胞 HL A DR的表达率是反映免疫功能的简单实用的指标。重症烧伤后免疫麻痹可持续较长时间 ,必要的免疫加强治疗有重要意义。     

丙酮酸乙酯对烫伤延迟复苏大鼠脾淋巴细胞增殖及凋亡影响

目的观察丙酮酸乙酯对烫伤延迟复苏大鼠脾淋巴细胞增殖及凋亡的影响,并对其机制进行初步探讨。方法72只Wistar大鼠随机分为假烫伤组(n=24)、烫伤组(n=24)和丙酮酸乙酯(3.23mg/ml)治疗组(n=24),分别于伤后1、3和5d活杀大鼠,检测脾淋巴细胞增殖能力及凋亡情况。结果烫伤延迟复苏后1～5d,脾淋巴细胞对丝裂原刺激的增殖反应明显受抑制(P均<0.05);在烫伤后第1d,脾CD3+CD4+T细胞凋亡显著增加(P<0.05)。应用丙酮酸乙酯治疗能够明显恢复烫伤后1～5d脾淋巴细胞的增殖反应(P<0.05),同时烫伤后第1d的凋亡明显下降(P<0.05)。结论丙酮酸乙酯能够有效恢复烫伤延迟复苏后脾淋巴细胞的增殖能力,并减轻脾淋巴细胞凋亡。     

Molecular or pharmacologic inhibition of the CD14 signaling pathway protects against burn-related myocardial inflammation and dysfunction

Signaling through toll-like receptor 4 (TLR4) plays an obligate role in burn-related myocardial dysfunction. We hypothesized that signaling through CD14, a cellular receptor for endotoxin that lacks a transmembrane domain but is coupled to TLR4, also plays a role in postburn myocardial inflammation and dysfunction. Burn covering 40% total body surface area (or sham burn for controls) was produced in wild-type (WT) and CD14 knockout (KO) as well as vehicle-treated and geldanamycin-treated WT mice (1 microg/g body weight) to inhibit CD14 signaling. Groups included (1) WT shams, (2) CD14 KO sham, (3) WT burns, (4) CD14 KO burns, (5) vehicle-treated WT shams, (6) geldanamycin-treated WT shams, (7) vehicle-treated WT burns, and (8) geldanamycin-treated WT burns. Twenty-four hours after burn, cardiac function (Langendorff) and cardiomyocyte secretion of inflammatory cytokines TNF-alpha, IL-1 beta, and IL-6 (in pg/mL; 5 x 10(4) myocytes) were studied in all groups. Relative to sham WT controls, burn trauma in increased cardiac myocyte secretion of inflammatory cytokines (TNF-alpha, IL-1 beta, and IL-6 rose from 59 +/- 10 to 171 +/- 8; 6 +/- 0.2 to 78 +/- 1; and 88 +/- 3 to 170 +/- 12 pg/mL, respectively; P < 0.05) and produced robust cardiac contractile dysfunction (left ventricular pressure and +dP/dt fell from 105 +/- 4 to 73 +/- 5 mmHg and 2,400 +/- 73 to 1,803 +/- 90 mmHg/s; P < 0.05). Inability to signal through the CD14/TLR4 pathway (induced by CD14/KO or inhibition of CD14 expression by administration of geldanamycin) attenuated TNF-alpha, IL-1 beta, and IL-6 production in response to burn injury and improved postburn myocardial contractile function. Our data suggest that signaling through the CD14 pathway plays an obligate role in cardiac inflammation/dysfunction which occurs after major burn injury.     

严重烧伤患者促炎和抗炎细胞因子的变化及其意义

目的检测严重烧伤患者血清中促炎和抗炎细胞因子水平,探讨严重烧伤患者体内免疫状态变化的特点。方法选取烧伤总体表面积（TBSA）〉30%的患者64例,按烧伤面积将严重烧伤患者分为2组：Ⅰ组39例（TBSA 30%~50%）,Ⅱ组25例（TBSA〉50%）;同时选取年龄和性别相匹配的20例健康人作为对照组。分别收集烧伤患者伤后1、3、7和21d的血清,采用酶联免疫吸附试验法（ELISA）检测血清中促炎细胞因子IL-6、IL-8、TNF-α和抗炎细胞因子IL-4、IL-10、G-CSF的浓度。结果 （1）与对照组相比,两组烧伤患者血清中促炎细胞因子IL-6、IL-8、TNF-α和抗炎细胞因子IL-4、IL-10、G-CSF在烧伤后不同时间点均显著增高（P〈0.01）。（2）烧伤Ⅱ组IL-6水平在伤后7d明显高于烧伤Ⅰ组（P〈0.05或P〈0.01）,并持续较长时间。烧伤Ⅱ组血清中IL-8和TNF-α水平在伤后1d就显著高于烧Ⅰ组（P〈0.01）。（3）烧伤II组IL-4、G-CSF水平在伤后3d明显高于烧伤Ⅰ组（P〈0.05或P〈0.01）,并持续较长时间。烧伤Ⅱ组血清中IL-10水平在伤后1d就显著高于烧伤Ⅰ组（P〈0.01）。结论严重烧伤患者体内促炎抗炎细胞因子处于失衡状态,并且不同烧伤面积的烧伤患者促炎和抗炎细胞因子的表达存在明显差异,早期监测严重烧伤患者的促炎和抗炎细胞因子水平对疾病的治疗具有重要意义。     

Changes in blood lymphocyte populations after multiple trauma: association with posttraumatic complications.

OBJECTIVE: To study the frequency of several lymphocyte subsets, circulating cytokines, and prostaglandin plasma values at their time course over a period of 14 days in severely injured trauma patients in relation to the development of sepsis and multiple organ failure (MOF). DESIGN: Prospective study. SETTING: An operative intensive care unit (ICU) of a university hospital. PATIENTS: Sixty-eight consecutive severely injured trauma patients. INTERVENTIONS: Patients were separated into patients without sepsis and MOF (group 1, n = 51), and patients who developed sepsis and MOF (group 2, n = 17) during their stay in the ICU. Therapy was adjusted to the standards of modern intensive care management by physicians who were not involved in the study. MEASUREMENTS AND MAIN RESULTS: In arterial blood samples, the profile of lymphocyte subset frequencies was performed by flow cytometry and, together with interleukin (IL)-1, IL-10, tumor necrosis factor (TNF)-alpha soluble TNF-alpha receptor 1 (sTNF-alpha r1 [p55]), and prostaglandin E2 (PGE2alpha)-alpha, serially measured after arrival in the ICU (baseline value) and during the next 14 days. Mean plasma IL-1 (29.3 +/- 5.8 [SD] pg/mL), TNF-alpha (138.5 +/- 22.4 pg/mL), and soluble TNF-alpha r1 (6.1 +/- 0.3 ng/mL) values were significantly higher in group 2 patients before clinical evidence of sepsis and MOF. With the onset of severe infections in group 2 patients, IL-1, TNF-alpha, and sTNF-alpha r1 values decreased, while immunosuppressive IL-10 (191.7 +/- 29.1 pg/mL) and PGE2alpha (87.7 +/- 20.4 pg/mL) values further increased and remained elevated during the time course. Analysis of lymphocyte subsets revealed a fall in total lymphocyte levels, in CD4+ T lymphocytes, and natural killer (NK) cells, but no change in CD8+ T lymphocyte subset. Despite a marked change in the T helper (CD4+) to T suppressor (CD8+) ratio (from 1:1.72 to 1:1.10), patients without MOF (group 1) had no significant difference in any of the markers tested compared with baseline values. In addition to the inverse CD4+/CD8+ T cell ratio (from 1:1.75 to 1:0.91) and increased activated T cells, each of these markers was significantly elevated and peaked before the onset of MOF in group 2 patients. CONCLUSIONS: A severely depressed cellular immune response associated with increased suppressive mediators might be closely related to the development of severe sepsis and MOF in trauma patients. Therefore, an in-depth understanding of the deficits in host defense following multiple trauma will provide the basis for therapeutic interventions.     

p53基因修饰树突细胞诱导特异性抗肿瘤免疫应答的实验研究

目的　探讨含p5 3基因的质粒pC5 3 SN3转染的树突细胞 (DC)体外诱导特异性抗肿瘤细胞毒性T淋巴细胞 (CTL)。方法　应用脂质体介导将质粒pC5 3 SN3转染肺癌患者单个核细胞 [HLA A2 + ]衍生的DC ,然后与未经纯化的自体T细胞混合培养以诱导CTL(T pC5 3 SN3) ,并通过乳酸脱氢酶(LDH)释放实验测定其对p5 3基因突变的HLA A2 + 人肺癌细胞系Calu 6的杀伤活性。结果　用质粒pC5 3 SN3转染DC后 ,发现CD1a、CD83显著升高 ,转染前表达率分别为 (5 .4 5± 0 .89) %、(3.2 6± 0 .4 7) % ,转染后分别为 (5 2 .15± 11.5 6 ) %、(2 5 .78± 12 .35 ) %。但CD86 、CD4 0 、HLA DR等DC相关标志与转染前无明显改变。LDH释放实验显示 ,以Calu 6作为靶细胞 ,T pC5 3 SN3诱导的杀伤作用显著高于T IL 2(IL 2 10 0U ml刺激外周血单个核细胞产生的CTL) ,效靶比为 10∶1时其杀伤活性分别为 (5 6 .79±15 .6 7) %和 (39.33± 9.88) %。同时细胞表面标记检测可见T pC5 3 SN3细胞中以CD8+ 细胞为主 ,且CD6 9、CD4 5RO CD8表达均显著升高。结论　p5 3基因修饰的DC对p5 3突变的人肺癌细胞系可有效诱导HLA A2限制的特异性CTL。     

白细胞介素-2和10 mRNA表达差异在CD4+CD25+T细胞诱导大鼠肝移植免疫耐受中的意义

目的 探讨白细胞介素-2(IL-2)mRNA及IL-10 mRNA表达水平对大鼠肝移植耐受的影响.方法 将实验大鼠随机分3组:Ⅰ组为急性排斥组; Ⅱ组为CD4+CD25+T细胞处理组,供体Wistar大鼠,受体SD大鼠;Ⅲ组为移植对照组,供体、受体均为SD大鼠.每组12对.肝移植前7 d,Ⅱ组受体大鼠经阴茎背静脉注射含供体脾淋巴细胞的培养液,Ⅰ组、Ⅲ组注射等量生理盐水;术后7 d随机取6只大鼠用逆转录-聚合酶链反应(RT-PCR)测定肝组织中细胞因子IL-2 mRNA及IL-10 mRNA的表达,用流式细胞仪检测各组移植肝内分离出的淋巴细胞含量,同时检测肝脏病理学的变化;另6只观察移植大鼠的生存期.结果 IL-2mRNA在Ⅰ组大鼠移植物内出现高表达,Ⅱ组仅有微弱表达,Ⅲ组则未见表达,IL-10 mRNA仅在Ⅱ组中表达,且表达程度较强.Ⅱ组和Ⅲ组大鼠存活期均超过30 d,与Ⅰ组(8～11 d)比较差异有统计学意义(P均＜0.01).Ⅰ组大鼠移植后肝脏有大量淋巴细胞浸润,数量明显高于Ⅱ组和Ⅲ组[(14.31±3.41)×106 /g比(5.04±1.13)×106 /g和(1.55±0.40)×106 /g,P均＜0.01],且显示中度排斥反应.Ⅱ组大鼠有中等量淋巴细胞浸润,病理为无排斥或不确定性排斥,且淋巴细胞中CD4+百分比[(43.31±8.07)%]和CD4+CD25+百分比C(11.39±1.92)%]均显著高于Ⅰ组[(33.65±7.25)%,(3.05±0.62)%]和Ⅲ组[(31.18±6.52)%,(3.37±0.72)%],差异有统计学意义(P＜0.05或P＜0.01).Ⅲ组未见明显淋巴细胞浸润,病理为无排斥反应.结论 IL-2参与了移植排斥的发生,而IL-10在CD4+CD25+T细胞诱导免疫耐受中具有非常重要的作用.     

Increased circulating immunosuppressive CD14(+)HLA-DR(-/low) cells correlate with clinical cancer stage and pathological grade in patients with bladder carcinoma

This study investigated CD14(+)HLA-DR(-/low) cells in peripheral blood mononuclear cells (PBMCs) from 64 patients with bladder carcinoma (BC) and 14 healthy controls. Cell phenotypes were determined and CD14(+)HLA-DR(-/low) cells, CD14(+)HLA-DR(+) cells and PBMCs depleted of CD14(+)HLA-DR(-/low) cells were isolated. Proliferation of stimulated PBMCs and interferon-γ (IFN-γ) production after addition of CD14(+)HLA-DR(-/low) and CD14(+)HLA-DR(+) cells at different ratios were measured. IFN-γ production was also measured after addition of L-arginine and/or antitransforming growth factor-β (TGF-β) neutralizing monoclonal antibody, and in PBMCs depleted of CD14(+)HLA-DR(-/low) cells. The proportion of CD14(+)HLA-DR(-/low) cells in BC patients was significantly higher than in controls. CD14(+)HLA-DR(-/low) cells significantly decreased T-cell proliferation and IFN-γ production in a dose-dependent manner. This suppressive activity was partially reversed by L-arginine or anti-TGF-β. Enhanced IFN-γ secretion was also seen in PBMCs depleted of CD14(+)HLA-DR(-/low) cells. The level of CD14(+)HLA-DR(-/low) cells was associated with gender, tumour size, number of tumours, cancer pathological grade and clinical stage. CD14(+)HLA-DR(-/low) cells may represent a subpopulation of myeloid-derived suppressor cells in BC patients.     

Spontaneous release of interleukin 2 by lung T lymphocytes in active pulmonary sarcoidosis is primarily from the Leu3+DR+ T cell subset.

The inflammation within the lower respiratory tract of individuals with pulmonary sarcoidosis is dominated by large numbers of helper T lymphocytes that proliferate and spontaneously release interleukin 2 (IL-2). To identify the lymphocyte subpopulation that releases IL-2 in this disorder, lung lymphocytes recovered by bronchoalveolar lavage were characterized using the monoclonal antibodies Leu4 (T lymphocyte), Leu3 (helper/inducer), Leu2 (suppressor/cytotoxic), and anti-HLA-DR, and separated by panning and flow cytometry. The majority of the IL-2 spontaneously released by T cells in the sarcoid lung was contributed by the Leu3+ cell population (Leu3+65 +/- 23 IL-2 units released/10(6) cells per 24 h; Leu2+ 9 +/- 8, P less than 0.04). Further characterization of the lung Leu3+ T cells in sarcoid demonstrated that 30 +/- 3% were expressing HLA-DR molecules on their surface compared with 6 +/- 1% in normals (P less than 0.01). Importantly, the subpopulation of Leu3+ lung T lymphocytes expressing a high intensity of HLA-DR molecules on their surface was responsible for the majority of the release of IL-2 in the sarcoid lung (Leu3+ high-intensity DR 42 +/- 17 U/10(6) cells per 24 h, Leu3+ low-intensity DR 8 +/- 1 U/10(6) cells per 24 h; P less than 0.01). Thus, the spontaneous release of IL-2 in the lung of sarcoid patients appears to be localized to a subset of Leu3+ high-intensity DR ("activated" lung helper/inducer) T lymphocytes. Because the sarcoid lung is characterized by markedly increased numbers of these cells, it is likely that this compartmentalized T cell population plays a major role in sustaining the exaggerated localized immune processes of this disorder.     

Large-scale immunomagnetic selection of CD14+ monocytes to generate dendritic cells for cancer immunotherapy: a phase I study

Dendritic cells (DC) are professional antigen-presenting cells that are widely used in the experimental immunotherapy of cancer. For clinical use GMP-like protocols for the preparation of functionally active dendritic cells (DC) in large numbers and at high purity are needed. However, the currently available protocols have certain disadvantages. In this study we tested the generation and clinical applicability of DC from monocyte preparations produced by immunomagnetic CD14(+) selection using a semiautomated clinical scale immunomagnetic column. Peripheral blood mononuclear cells (PBMC) of 10 patients with metastatic solid tumors were used. With the immunomagnetic separation, we obtained a cell suspension of high CD14(+) purity (median 97.4%, range 94.9-99.0) with a high monocyte yield (median 82.3%, range 63.9-100.0). Differentiation of CD14(+) cells into mature monocyte-derived DC was induced by incubation with IL-4, GM-CSF, TNF-alpha, PGE(2), IL-1 beta, and IL-6. Mature DC showed a high expression of CD83, HLA-DR, and the co-stimulatory molecules CD80 and CD86. Overall CD83(+) yield was 12.1% (range 4.0-29.4). Allogeneic T stimulatory capacity could be demonstrated for all DC preparations in proliferation assays. No significant differences in marker expression or T cell stimulation was detected between fresh DC and those derived from cryopreserved immature DC. Clinical administration of autologous DC by three different parenteral routes was tolerated by all 10 patients without systemic signs of toxicity. Our results indicate that immunomagnetic isolation of CD14(+) monocytes using the CliniMACS device is a suitable method for clinical-scale generation of functional DC under GMP-grade conditions. The selection can be performed in a closed system. Therefore, immunomagnetic CD14(+) selection can be seen as an alternative way to generate DC for clinical tumor vaccination protocols.