狼疮性肾炎患者血清NETs、TWEAK、外周血CD4+T/CD8+T比例与疾病活动度及肾脏预后的关系

摘要 目的：探讨狼疮性肾炎（LN）患者血清中性粒细胞胞外诱捕网（NETs）、肿瘤坏死因子样凋亡微弱诱导剂（TWEAK）、外周血分化簇（CD）4⁺T/CD8⁺T比例与疾病活动度及肾脏预后的关系。方法：选取2021年8月～2022年8月川北医学院附属医院肾内科收治的LN患者137例（LN组）,根据系统性红斑狼疮疾病活动指数（SLEDAI）-2000评分分为轻度活动组（52例）、中度活动组（45例）、重度活动组（40例）。随访1年,根据肾脏相关终点事件发生情况分为预后不良组（43例）和预后良好组（94例）,另选取同期76名体检健康志愿者（对照组）。采用酶联免疫吸附法检测血清NETs、TWEAK水平,流式细胞术检测外周血CD4⁺T/CD8⁺T比例。Spearman相关性分析LN患者血清NETs、TWEAK和外周血CD4⁺T/CD8⁺T与SLEDAI-2000评分的相关性,多因素Logistic回归分析LN患者预后不良的因素,受试者工作特征曲线分析血清NETs、TWEAK和外周血CD4⁺T/CD8⁺T对LN患者预后不良的预测价值。结果：与对照组比较,LN组血清NETs、TWEAK水平升高,外周血CD4⁺T/CD8⁺T降低（P＜0.05）。轻度活动组、中度活动组、重度活动组血清NETs、TWEAK依次升高,外周血CD4⁺T/CD8⁺T依次降低（P＜0.05）。LN患者SLEDAI-2000评分与血清NETs、TWEAK呈正相关,与外周血CD4⁺T/CD8⁺T呈负相关（P＜0.05）。慢性肾脏病分期4期、SLEDAI-2000评分升高、NETs升高、TWEAK升高为LN患者预后不良的独立危险因素,估算肾小球滤过率升高、CD4⁺T/CD8⁺T升高为独立保护因素（P＜0.05）。血清NETs、TWEAK和外周血CD4⁺T/CD8⁺T联合预测LN患者预后不良的曲线下面积为0.943,大于血清NETs、TWEAK和外周血CD4⁺T/CD8⁺T单独预测的0.790、0.788、0.799（P＜0.05）。结论：LN患者血清NETs、TWEAK水平升高,外周血CD4⁺T/CD8⁺T降低,与疾病活动度及肾脏预后不良密切相关,血清NETs、TWEAK联合外周血CD4⁺T/CD8⁺T预测LN患者肾脏预后的价值较高。     

组蛋白去乙酰化酶3对外周CD4+ T细胞分化的影响

目的 探讨组蛋白去乙酰化酶3（HDAC3）对外周CD4⁺ T细胞分化及功能的调控作用。方法 采用CD4cre酶介导Hdac3杂合基因缺失小鼠（Hdac3^fl/flCD4^cre+/-）及其野生型正常对照小鼠（Hdac3^fl/fl，WT），流式细胞术检测HDAC3缺失对外周CD4⁺和CD8⁺ T细胞比例和数量的影响；在体外佛波酯（PMA）和离子霉素（Ionomycin）刺激条件下，流式细胞术检测HDAC3缺失对CD4⁺ T细胞中IFN-γ、IL-4和IL-17A的表达以及Tfh细胞产生的影响；采用ELISA检测HDAC3缺失对小鼠血清IFN-γ、IL-4和IL-17表达的影响；分选Hdac3^fl/flCD4^cre+/-和WT小鼠外周初始CD4⁺ T细胞，分别在Th1和Th2分化条件下培养，细胞内染色检测HDAC3缺失对Th1、Th2以及Th17相关细胞因子及其特异转录因子表达的影响；采用Microarray检测HDAC3缺失对CD4⁺ T细胞分化亚群相关基因表达的影响；采用链脲佐菌素（STZ）处理小鼠构建I型糖尿病（TIDM）疾病模型，检测HDAC3缺失对T1DM发病的影响。结果 与WT小鼠相比，Hdac3^fl/flCD4^cre+/-小鼠外周CD4⁺和CD8⁺ T细胞的比例和数量显著降低。Hdac3^fl/flCD4^cre+/-小鼠CD4⁺ T细胞及血清中IFN-γ的表达显著降低，而IL-4和IL-17A的表达显著增加，Tfh细胞比例也显著增加；HDAC3缺失抑制体外培养CD4⁺ T细胞向Th1分化但促进其向Th2分化；Microarray检测发现HDAC3缺失导致Th1型细胞谱系基因表达降低，而Th2、Th17以及Tfh细胞谱系基因表达增加；在STZ诱导条件下，HDAC3缺失抑制小鼠T1DM的发生和CD4⁺ T细胞向Th1分化。结论 HDAC3促进外周CD4⁺ T细胞向Th1细胞分化并加重T1DM的发生。     

传染性单核细胞增多症患儿NLR、CD4+/CD8+比值、ADA与EBV-DNA载量的相关性分析及对肝损害的影响

摘要 目的：探讨传染性单核细胞增多症（IM）患儿外周血中性粒细胞/淋巴细胞比值（NLR）、CD4⁺/CD8⁺比值、腺苷脱氨酶（ADA）与EB病毒（EBV）-脱氧核糖核酸（DNA）载量的相关性,分析其对IM患儿肝损害的影响。方法：选择2019年1月至2022年4月我院儿科收治的102例IM患儿（IM组）,另选择同期我科收治的95例EB病毒检测阴性的发热患儿（非IM组）和体检健康的73例健康儿童（对照组）。根据是否发生肝损害将IM患儿分为肝损害组（61例）和非肝损害组（41例）。比较外周血NLR、CD4⁺/CD8⁺比值、ADA与EBV-DNA载量,Pearson法分析NLR、CD4⁺/CD8⁺比值、ADA与EBV-DNA载量的相关性。多因素Logistic回归分析IM患儿发生肝损害的影响因素。结果：IM组ADA高于非IM组和对照组（P＜0.05）,且非IM组高于对照组（P＜0.05）,NLR、CD4⁺/CD8⁺比值低于非IM组和对照组（P＜0.05）,且非IM组低于对照组（P＜0.05）,IM组EBV-DNA载量高于非IM组（P＜0.05）。IM患儿ADA与EBV-DNA载量呈正相关（r=0.493,P＜0.05）,NLR、CD4⁺/CD8⁺比值与EBV-DNA载量呈负相关（r=-0.419、-472,P＜0.05）。肝损害组ADA、EBV-DNA载量高于非肝损害组（P＜0.05）,NLR、CD4⁺/CD8+⁺比值低于非肝损害组（P＜0.05）。肝脏肿大、高EBV-DNA载量、高ADA是IM患儿肝损害的危险因素（P＜0.05）,高NLR、高CD4⁺/ CD8⁺比值是保护因素（P＜0.05）。结论：IM患儿ADA增高,NLR、CD4⁺/CD8⁺比值降低,与EBV-DNA载量增加以及肝损害有关。     

YKL-40、CD4+/CD8+比值与腺病毒肺炎患儿炎性因子和并发喘息的关系研究

摘要 目的：探讨血清壳多糖酶3样蛋白1（YKL-40）、外周血CD4⁺/CD8⁺比值与腺病毒肺炎（AP）患儿炎性因子和并发喘息的关系。方法：选取2019年10月～2022年10月湖北省妇幼保健院收治的97例AP患儿为AP组,根据是否并发喘息分别为喘息组和无喘息组,另选取同期50例体检健康儿童为对照组。收集AP患儿的临床资料,采用酶联免疫吸附法检测血清YKL-40和炎性因子[白细胞介素（IL）-6、IL-8、肿瘤坏死因子-α（TNFα）]水平,流式细胞术检测外周血CD4⁺、CD8⁺比例并计算CD4⁺/CD8⁺比值。采用Spearman相关性分析AP患儿血清YKL-40、CD4⁺/CD8⁺比值与炎性因子水平的相关性,多因素Logistic回归分析AP患儿并发喘息的影响因素。结果：与对照组比较,AP组血清YKL-40、外周血CD8⁺比例升高,CD4⁺比例、CD4⁺/CD8⁺比值降低（P＜0.05）。AP组血清IL-6、IL-8、TNF-α水平高于对照组（P＜0.05）。Spearman相关性分析显示,AP患儿血清YKL-40与IL-6、IL-8、TNF-α水平呈正相关,外周血CD4⁺/CD8⁺比值与IL-6、IL-8、TNF-α水平呈负相关（P＜0.05）。97例AP患儿住院期间喘息发生率为50.52%（49/97）。多因素Logistic回归分析显示,呼吸衰竭、小气道病变、特应性体质和血清IL-6、IL-8、TNF-α、YKL-40升高为AP患儿并发喘息的独立危险因素,外周血CD4⁺/CD8⁺比值升高为独立保护因素（P＜0.05）。结论：AP患儿血清YKL-40水平升高和外周血CD4⁺/CD8⁺比值降低,与炎性因子水平升高和并发喘息密切相关。     

2型糖尿病并发肺结核患者PCT、HMGB1、CD4+/CD8+比值与继发肺部感染的关系

摘要 目的：探讨2型糖尿病并发肺结核患者降钙素原（PCT）、高迁移率族蛋白1（HMGB1）、CD4⁺/CD8⁺比值与继发肺部感染的关系。方法：选择2019年1月至2022年6月四川大学华西医院呼吸与危重症医学科收治的97例2型糖尿病并发肺结核患者,根据入院治疗时是否继发肺部感染分为肺部感染组（53例）及非肺部感染组（44例）。检测两组血清PCT、HMGB1水平以及外周血CD4⁺/CD8⁺比值。单因素和多因素Logistic回归分析2型糖尿病并发肺结核患者继发肺部感染的因素。受试者工作特征（ROC）曲线分析PCT、HMGB1和CD4⁺/CD8⁺比值预测2型糖尿病并发肺结核患者继发肺部感染的价值。结果：肺部感染组血清PCT、HMGB1水平高于非肺部感染组（P＜0.05）,外周血CD4⁺/CD8⁺比值低于非肺部感染组（P＜0.05）。糖化血红蛋白及血清PCT、HMGB1水平升高是2型糖尿病并发肺结核患者继发肺部感染的危险因素（P＜0.05）,高CD4⁺/CD8⁺比值是保护因素（P＜0.05）。PCT、HMGB1、CD4⁺/CD8⁺比值预测2型糖尿病并发肺结核患者继发肺部感染的曲线下面积为0.719、0.761、0.738,联合PCT、HMGB1和CD4⁺/CD8⁺比值预测的曲线下面积为0.878,高于各指标单独预测。结论：2型糖尿病并发肺结核患者血清PCT、HMGB1水平增高,外周血CD4⁺/CD8⁺比值降低,均与继发肺部感染有关,PCT、HMGB1联合CD4⁺/CD8⁺比值可辅助预测2型糖尿病并发肺结核患者继发肺部感染的风险。     

消疬膏对脾胃虚弱证颈部淋巴结核患者CD4+CD25highFoxP3+调节性T淋巴细胞和胃肠激素的影响

摘要 目的：研究消疬膏对脾胃虚弱证颈部淋巴结核患者CD4⁺CD25^highFoxP3⁺调节性T淋巴细胞和胃肠激素的影响。方法：选择南京市中西医结合医院于2020年3月~2022年1月期间收治的颈部淋巴结核患者120例。按照随机数字表法分为对照组（60例）和研究组（60例）。对照组接受常规抗结核方案，研究组接受消疬膏联合常规抗结核方案，两组疗程均为6个月。观察两组治疗前和治疗6个月后的中医证候积分、CD4⁺CD25^highFoxP3⁺调节性T淋巴细胞相关指标和胃肠激素的变化，记录两组治疗期间用药安全性。结果：两组治疗6个月后肿疡大小、肿疡皮色、面色、纳呆、倦怠乏力、便溏、舌质、脉象评分均下降，且研究组低于对照组（P<0.05）。研究组治疗6个月后CD4⁺CD25^highFoxP3⁺调节性T淋巴细胞、白介素-10（IL-10）水平低于对照组，γ-干扰素（IFN-γ）水平高于对照组（P<0.05）。研究组治疗6个月后胆囊收缩素（CCK）、胃泌素（GAS）水平低于对照组，胃动素（MTL）水平高于对照组（P<0.05）。研究组的不良反应总发生率低于对照组（P<0.05）。结论：消疬膏治疗脾胃虚弱证颈部淋巴结核患者，可有效改善患者临床症状，可能与调节CD4⁺CD25^highFoxP3⁺调节性T淋巴细胞和胃肠激素水平有关。     

外周血T细胞CD38和HLA-DR的表达水平在儿童传染性单核细胞增多症中的临床意义

摘要 目的：探讨传染性单核细胞增多症（IM）患儿外周血T细胞活化分子CD38和人类白细胞抗原DR（HLA-DR）表达水平的临床意义。方法：采用流式细胞术分别检测45例IM患儿急性期和恢复期的活化分子CD38和HLA-DR在T细胞的表达水平,并与30例健康体检儿童进行对比。分析IM患儿急性期CD38和HLA-DR在T细胞的表达水平与EB病毒载量、肝功能指标、外周血异型淋巴细胞比例、淋巴细胞计数的相关性,并采用ROC曲线分析CD8⁺CD38⁺T和CD8⁺HLA-DR+T细胞百分比的诊断效能。结果：与对照组比较,IM急性期患儿的CD38和HLA-DR在T细胞的表达水平显著升高（P＜0.05）。CD8⁺CD38⁺T、CD8⁺HLA-DR+T细胞百分比分别与EBV-DNA、ALT、AST、LDH、异型淋巴细胞百分比、淋巴细胞计数呈正相关（P＜0.05）,与白蛋白（ALB）呈负相关（P＜0.05）;CD4⁺CD38⁺T、CD4⁺HLA-DR+T细胞百分比与上述指标无显著相关性（P均＞0.05）。IM恢复期CD38和HLA-DR在T细胞的表达水平较急性期明显降低（P＜0.05）。ROC曲线分析CD8⁺CD38⁺T、CD8⁺HLA-DR+T细胞百分比显示诊断儿童IM的AUC值分别为0.931和0.993,特异度均为100%,灵敏度分别为88.89 %和93.33 %。结论：流式法检测CD38和HLA-DR在T细胞的变化有助于判断病情变化。外周血CD8⁺CD38⁺T、CD8⁺HLA-DR+T细胞百分比不仅能反映出IM急性期肝功能损伤严重程度,还可作为儿童IM的流式诊断指标。     

五灵胶囊联合富马酸丙酚替诺福韦片对慢性乙型肝炎患者氧化应激、CD4+CD25+调节性T细胞和血清MMP-1、MMP-2、TIMP-1的影响

摘要 目的：探讨五灵胶囊联合富马酸丙酚替诺福韦片对慢性乙型肝炎（CHB）患者氧化应激、CD4⁺CD25⁺调节性T细胞和血清基质金属蛋白酶-1（MMP-1）、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶组织抑制因子-1（TIMP-1）的影响。方法：纳入苏州大学附属传染病医院2021年6月~2022年12月期间收治的CHB患者122例,采用随机数字表法分为对照组（n=61,富马酸丙酚替诺福韦片）和研究组（n=61,五灵胶囊联合富马酸丙酚替诺福韦片）。对比两组疗效、氧化应激指标、CD4⁺CD25⁺调节性T细胞、肝功能指标[总胆红素（TBIL）、谷丙转氨酶（ALT）、谷氨酰转肽酶（GGT）]、血清MMP-1、MMP-2、TIMP-1水平和不良反应发生情况。结果：研究组的临床总有效率高于对照组（P<0.05）。两组治疗后丙二醛（MDA）下降,且研究组低于对照组同时间点;超氧化物歧化酶（SOD）升高,且研究组高于对照组同时间点（P<0.05）。两组治疗后CD4⁺CD25⁺调节性T细胞下降,且研究组低于对照组同时间点（P<0.05）。两组治疗后MMP-1、MMP-2、TIMP-1下降,且研究组低于对照组同时间点（P<0.05）。两组治疗后TBIL、ALT、GGT下降,且研究组低于对照组同时间点（P<0.05）。两组不良反应总发生率组间对比未见差异（P>0.05）。结论：五灵胶囊联合富马酸丙酚替诺福韦片治疗CHB患者,可有效减轻机体氧化应激,调节CD4⁺CD25⁺调节性T细胞和血清MMP-1、MMP-2、TIMP-1水平。     

视频脑电图联合PEWS评分、NSE、CD4+/CD8+比值对病毒性脑炎患儿病情评估及预后预测的临床价值

摘要 目的：探讨视频脑电图（VEEG）联合儿童早期预警评分（PEWS）、神经元特异性烯醇化酶（NSE）、CD4⁺/CD8⁺比值对病毒性脑炎患儿病情评估及预后预测的价值。方法：选择2020年3月至2022年3月南京医科大学附属儿童医院收治的152例病毒性脑炎患儿，根据病情严重程度将患儿分为重症组（67例）和轻症组（85例），另选择72例无神经系统损伤住院患儿为对照组。治疗2周后，根据儿童格拉斯哥预后量表（CGOS）将其分为预后良好组（4~5级，89例）与预后不良组（1~3级，63例）。所有研究对象均接受PEWS测评和VEEG检查，检测血清NSE水平，计算CD4⁺/CD8⁺比值。采用单因素和多因素Logistic回归分析影响病毒性脑炎患儿预后的因素。采用受试者工作特征（ROC）曲线分析PEWS、VEEG 及血清NSE、CD4⁺/CD8⁺比值预测病毒性脑炎患儿预后的价值。结果：重症组PEWS、VEEG重度异常比例及血清NSE水平高于轻症组和对照组，CD4⁺/CD8⁺比值低于轻症组和对照组（P＜0.05）；轻症组PEWS、VEEG重度异常比例及血清NSE水平高于对照组，CD4⁺/CD8⁺比值低于对照组（P＜0.05）。预后不良组PEWS、VEEG重度异常比例、血清NSE水平高于预后良好组，CD4⁺/CD8⁺比值低于预后良好组（P＜0.05）。多因素Logistic回归分析结果显示，持续惊厥、高PEWS、VEEG重度异常、血清NSE水平升高是病毒性脑炎患儿预后不良的危险因素，CD4⁺/CD8⁺比值升高是保护因素（P＜0.05）。联合PEWS、VEEG 及血清NSE、CD4⁺/CD8⁺比值预测病毒性脑炎患儿预后曲线下面积为0.859，高于各指标单独预测。结论：病毒性脑炎患儿高PEWS、VEEG重度异常、血清NSE水平升高、CD4⁺/CD8⁺比值降低，与病情加重和预后不良有关，联合以上四项指标辅助预测病毒性脑炎患儿预后的价值较高。     

脓毒症患者外周血T淋巴细胞PD-1表达变化及胸腺肽α-1的免疫调理作用研究

摘要 目的：探讨脓毒症患者外周血T淋巴细胞程序性细胞死亡受体1（PD-1）表达特点,分析胸腺肽?琢-1治疗对患者免疫功能的影响。方法：选择2018年3月至2020年6月我院重症医学科收治的140例脓毒症患者（脓毒症组）和同期于我院进行体检的95例健康志愿者（对照组）,根据急性生理与慢性健康评估Ⅱ（APACHE Ⅱ）、序贯器官衰竭评估（SOFA）评分结果将脓毒症患者分为APACHE Ⅱ 0~10分组（51例）、11~20分组（62例）和＞20分组（27例）;SOFA评分0~5分组（48例）、6~10分组（60例）和＞10分组（32例）。检测外周血CD4⁺T细胞上PD-1表达、CD8⁺T细胞上PD-1表达,比较组间差异性。Pearson秩相关性分析外周血CD4⁺T细胞上PD-1表达、CD8⁺T细胞上PD-1表达与APACHE Ⅱ、SOFA评分相关性。根据治疗方法将脓毒症患者分为A组（60例）和B组（80例）,A组给予常规综合治疗和乌司他丁治疗,B组在A组的基础上联合胸腺肽α-1治疗,比较两组治疗前后外周血T淋巴细胞（CD3⁺、CD4⁺、CD8⁺）、NK细胞（CD3-CD16⁺CD56⁺）差异。结果：脓毒症组外周血CD4⁺T细胞上PD-1表达、CD8⁺T细胞上PD-1表达高于对照组（P＜0.001）,外周血CD4⁺T细胞上PD-1表达、CD8⁺T细胞上PD-1表达随APACHE Ⅱ、SOFA评分的增加而增高,各组间差异显著（P＜0.05）。Pearson秩相关分析结果显示外周血CD4⁺T细胞上PD-1表达、CD8+T细胞上PD-1表达与APACHE Ⅱ评分、SOFA评分呈正相关（r=0.569、0.475;0.653、0.509,P均＜0.05）。B组治疗后CD3⁺、CD4⁺、CD3-CD16⁺CD56⁺高于A组（P＜0.05）,CD8⁺低于A组（P＜0.05）。结论：脓毒症患者外周血CD4⁺、CD8⁺T细胞上PD-1表达均增高,其表达与病情严重程度密切相关。给予胸腺肽α-1治疗可改善患者免疫功能。     

血清IL-6、IL-8、IgM抗体及T细胞亚群水平对新生儿先天性梅毒的诊断价值

目的:探讨血清白介素-6(IL-6)、白介素-8(IL-8)、IgM抗体及T细胞亚群对先天性梅毒新生儿的诊断价值。方法:选择2015年5月至2017年5月在我院进行临床治疗的先天性梅毒新生儿81例为观察组,另选同期来我院进行健康体检81例新生儿为对照组。比较两组患者血清IL-6、IL-8、T细胞亚群中CD~(3+)、CD~(4+)、CD~(8+)、CD~(4+)/CD~(8+)细胞及IgM抗体的阳性率。结果:治疗后,观察组血清IL-6、IL-8水平均明显高于对照组(P0.05),T细胞亚群中CD~(3+)、CD~(4+)、CD~(4+)/CD~(8+)明显低于对照组,而CD~(8+)T细胞比例高于对照组(P0.05)。19S-IgM-TP ELISA法检测出IgM的阳性率92.59%,明显高于TRUST法(74.07%)及TP-ELSA法(70.37%)(P0.05)。ROC曲线中,血清IL-8特异度为88.34%明显高于血清IL-6特异度81.48%、IgM抗体特异度60.13%、T细胞亚群特异度65.34%;IgM抗体的曲线面积88.91 cm~2明显大于IL-6的曲线面积45.09 cm~2、IL-8的曲线面积76.19 cm~2、T细胞亚群的曲线面积77.35 cm~2;T细胞亚群准备性67.89%明显高于IL-6准确性60.39%、IL-8准确性51.09%、IgM抗体准确性50.12;IgM抗体的灵敏度60.13%高于IL-6灵敏度59.19%、IL-8灵敏度42.35%、T细胞亚群灵敏度59.37%。具有比较意义(P0.05)。结论:血清IL-6、IL-8水平、T细胞亚群中CD~(3+)、CD~(4+)、CD~(8+)、CD~(4+)/CD~(8+)及IgM抗体阳性率是诊断先天性梅毒新生儿的重要指标。     

Rapid turnover of the CD8(+)CD28(-) T-cell subset of effector cells in the circulation of patients with head and neck cancer

CD8⁺ T cells in the circulation of patients with head and neck cancer (HNC) were previously shown to be significantly more sensitive to, and preferentially targeted for, apoptosis than CD4⁺ T cells (Hoffmann et al., Clin Cancer Res, 8:2553–2562, 2002). To distinguish global from CD8⁺ subset-specific apoptosis, we studied Annexin-binding to naïve, memory, and effector subsets of CD8⁺ cells by multicolor flow cytometry. Age-related changes in naïve and effector CD8⁺ cell subsets were observed in patients and normal controls (NC). The frequencies of naïve (CD28⁺CD45RO^-) CD8⁺ T cells were lower and those of memory (CD28⁺CD45RO⁺) and effector (CD28^-) CD8⁺ T cells significantly higher in the circulation of HNC patients relative to age-matched NC. Among CD8⁺ T cells, the CD28^- effector cell subset contained the highest proportion of Annexin-binding cells, while the naïve CD28⁺CD45RO^- subset contained the lowest. This suggested a high turnover rate of the CD8⁺CD28^- effector cell subset in patients with HNC, which was being compensated by a rapid transition of naïve CD8⁺ T cells to the effector cell pool. Following tumor resection, the frequency of CD8⁺CD28^- T cells normalized in the patients, an indication that the presence of tumor had an influence on the size of CD8⁺CD28^- T-cell pool. Ex vivo, in mixed lymphocyte-tumor cultures (MLTC) with semiallogeneic T cells as responders, CD8⁺CD28^- T cells could be generated from CD8⁺CD28⁺ cells by repeated stimulations with tumor cells. These CD8⁺CD28^- effector cells lysed the tumor, produced IFN- in response to the tumor, and strongly expressed granzyme B. Thus, the high rate of their apoptosis in the circulation of patients with HNC might be expected to contribute to tumor progression. However, the ex vivo generation of this cell subset was suppressed by strong CD28/B7 ligation or by overexpresson of MHC molecules on tumor cells, suggesting that adequate costimulation is necessary for protection from apoptosis. It appears that interactions of immune and tumor cells might determine the fate of this terminally differentiated effector cell subset.Supported in part by NIH grants: PO-1 DE 12321 and RO-1 CA 82016 to Theresa L. Whiteside.     

经阿托伐他汀治疗的急性冠脉综合征患者CD4+T、CD4+CD28+T水平变化及与预后的关系

目的:探讨经阿托伐他汀治疗的急性冠脉综合征(ACS)的CD4~+T、CD4~+CD28~+T水平变化及与预后的关系。方法:选择128例ACS患者,随机分为对照组(64例)和观察组两组(64例),其中对照组患者给予常规治疗,观察组患者在上述基础上外加阿托伐他汀治疗。比较两组患者的细胞因子、CD4~+T及CD4~+CD28~+T水平变化,随访6个月,观察两组患者预后终点事件发生情况。并将观察组患者根据预后是否并发终点事件,将其分为预后良组(未并发终点事件)和预后不良组(并发终点事件)两组,分析不同预后ACS患者CD4~+T、CD4~+CD28~+T水平变化及与预后发生终点事件的相关性。结果:治疗后两组的高敏C反应蛋白(hs-CRP)、干扰素-γ(IFN-γ)水平均明显降低,白细胞介素-10(IL-10)、转化生长因子β1(TGF-β1)水平均明显升高,且观察组改善更为明显,差异均有统计学意义(P0.05)。两组患者治疗后CD4~+T明显升高,CD8~+T、CD4~+CD28~+T明显降低,且观察组上述指标改善更为明显,差异均有统计学意义(P0.05)。随访6月中,对照组患者预后有23例终点事件发生,观察组患者预后有26例终点事件发生,差异无统计学意义(P0.05)。与预后良好组相比,预后不良组患者的CD4~+T降低,CD4~+CD28~+T升高,差异均有统计学意义(P0.05)。经阿托伐他汀治疗的ACS患者预后发生终点事件与CD4~+T水平呈现负相关(r=-0.682,P=0.000),与CD4~+CD28~+T水平呈现正相关(r=0.733,P=0.000)。结论:经阿托伐他汀治疗的ACS患者预后发生终点事件与CD4~+T水平呈现负相关,与CD4~+CD28~+T水平呈现正相关,可为临床ACS患者预后的预测提供参考。     

Apoptosis of CD4<Superscript>+</Superscript>CD25<Superscript>high</Superscript> T cells in response to Sirolimus requires activation of T cell receptor and is modulated by IL-2

Targeted molecular therapies inhibit proliferation and survival of cancer cells but may also affect immune cells. We have evaluated the effects of Sirolimus and Sorafenib on proliferation and survival of lymphoid cell subsets. Both drugs were cytotoxic to CD4⁺CD25^high T cells, and were growth inhibitory for CD4⁺ and CD8⁺ T cells. Cytotoxicity depended on CD3/CD28 stimulation and was detectable within 12 h, with 80–90% of CD4⁺CD25^high cells killed by 72 h. Cell death was due to apoptosis, based on Annexin V and 7AAD staining. Addition of IL-2 prevented the apoptotic response to Sirolimus, potentially accounting for reports that Sirolimus can enhance proliferation of CD4⁺CD25^high cells. These results predict that Sirolimus or Sorafenib would reduce CD4⁺CD25^high cells if administered prior to antigenic stimulation in an immunotherapy protocol. However, administration of IL-2 protects CD4⁺CD25^high T cells from cytotoxic effects of Sirolimus, a response that may be considered in design of therapeutic protocols.     

CD4CD8 thymocytes are induced to cell death by a small dose of puromycin via ER stress

When the CD4⁺CD8⁺ thymic lymphoma cells were treated with puromycin, we found that most of the cells died at 0.3-1 μg/ml of puromycin within 24 h. However, cell death was greatly reduced when the dose of puromycin was increased. Similar dose-pattern of cell death was observed in thymocytes and the sensitivity to puromycin was greater in CD4⁺CD8⁺ thymocytes than CD4⁺CD8⁻ thymocytes. The induction of apoptosis was blocked by the protein synthesis inhibitor cycloheximide, and to some extent by transfection of Bcl-xL or Bcl-2 genes. Expression of GRP78 was up-regulated after treatment with a small dose of puromycin, and the cell death by puromycin was blocked in the presence of caspase 12 inhibitor. These results indicated that the induction of cell death by low-dose puromycin was due to endoplasmic reticulum stress. Furthermore, we found that dexamethasone, a synthetic glucocorticoid, and puromycin worked synergistically to induce cell death in thymocytes.     

4-1BB costimulation enhances HSV-1-specific CD8+ T cell responses by the induction of CD11c+CD8+ T cells

Since 4-1BB plays a predominant role in CD8+ T cell responses, we investigated the effects of 4-1BB triggering on the primary and memory CD8+ T responses to HSV-1 infection. 4-1BB was detected on 10-15% of CD4+ and CD8+ T cells following the infection. 4-1BB-positive T cells were in the proliferative mode and showed the enhanced expression of anti-apoptotic proteins. Agonistic anti-4-1BB treatment exerted preferential expansion of CD8+ T cells and gB/H-2Kb-positive CD8+ T cells, and enhanced cytotoxicity against HSV-1 that was mainly mediated by CD11c+CD8+ T cells. CD11c+CD8+ T cells were re-expanded following re-challenge with HSV-1 at post-infection day 50, indicating that CD11c+CD8+ phenotype was maintained in memory CD8+ T cell pool. Our studies demonstrated that 4-1BB stimulation enhanced both primary and memory anti-HSV-1 CD8+ T cell responses, which was mediated by a massive expansion of antigen-specific CD11c+CD8+ T cells.     

Cadmium‐induced apoptosis and phenotypic changes in mouse thymocytes

At present cadmium (Cd)induced immunotoxicity and the mechanisms involved have not been fully elucidated. The main objective of the present study is to explore the apoptogenic property of Cd in primary cultured mouse thymocytes and its effect on cell surface marker expression and phenotypic changes. Cdinduced thymocyte apoptosis was determined by TdTmediated dUTP nick end labeling (TUNEL) assay, DNA content/cell cycle analysis and DNA gel electrophoresis. The results showed that Cd was able to cause apoptosis in mouse thymocytes in a time and dosedependent manner. Moreover, different subsets of thymocytes possessed different susceptibility to the apoptotic effect of Cd, in the order of CD8⁺ > CD4^–CD8^– (double negative cells, DN) > CD4⁺CD8⁺ (double positive cells, DP) > CD4⁺. Cd treatment also altered thymocyte surface marker expression, leading to evident phenotypic changes. Such changes were characterized by a decline in DP cells and a marked decrease in CD4⁺/CD8⁺ ratio, mainly due to a significant increase in CD8⁺ subsets. These observations help to obtain a better understanding of the immunotoxic and immunomodulatory effects of Cd.     

CD8CD25 T cells reduce atherosclerosis in apoE(−/−) mice

Background

It is increasingly evident that CD8⁺ T cells are involved in atherosclerosis but the specific subtypes have yet to be defined. CD8⁺CD25⁺ T cells exert suppressive effects on immune signaling and modulate experimental autoimmune disorders but their role in atherosclerosis remains to be determined. The phenotype and functional role of CD8⁺CD25⁺ T cells in experimental atherosclerosis were investigated in this study.

Methods and results

CD8⁺CD25⁺ T cells were observed in atherosclerotic plaques of apoE(−/−) mice fed hypercholesterolemic diet. Characterization by flow cytometric analysis and functional evaluation using a CFSE-based proliferation assays revealed a suppressive phenotype and function of splenic CD8⁺CD25⁺ T cells from apoE(−/−) mice. Depletion of CD8⁺CD25⁺ from total CD8⁺ T cells rendered higher cytolytic activity of the remaining CD8⁺CD25⁻ T cells. Adoptive transfer of CD8⁺CD25⁺ T cells into apoE(−/−) mice suppressed the proliferation of splenic CD4⁺ T cells and significantly reduced atherosclerosis in recipient mice.

Conclusions

Our study has identified an athero-protective role for CD8⁺CD25⁺ T cells in experimental atherosclerosis.