CHOP方案治疗前后外周T细胞淋巴瘤患者外周血CD45RA~+ T和CD45RO~+ T的变化特点

目的:探讨CHOP方案对外周T细胞淋巴瘤(PTCL)患者外周血中初始和记忆T细胞水平的变化及其临床意义。方法:采用流式细胞术检测20例PTCL患者CHOP方案化疗前后外周血中CD4+CD45RA+、CD4+CD45RO+、CD8+CD45RA+和CD8+CD45RO+T细胞的比例,分析疗效与T细胞亚群的关系。结果:PTCL患者化疗前外周血中CD4+T细胞、CD4+CD45RO+细胞的比例明显降低,CD4+CD45RA+、CD8+、CD8+CD45RO+和CD8+CD45RA+T细胞的比例均明显升高(P<0.05),而化疗后PTCL患者CD4+、CD4+CD45RO+细胞的比例较治疗前升高,CD4+CD45RA+、CD8+、CD8+CD45RO+和CD8+CD45RA+T细胞比例则较治疗前稍下降(P<0.05)。治疗前后化疗有效组的CD4+CD45RA+均明显高于化疗无效组(P<0.05)。结论:CHOP治疗对PTCL患者胸腺输出功能有一定的影响,伴有较高胸腺输出功能者对化疗效果更好。     

慢性乙型肝炎患者外周血CD8+/CD28+淋巴细胞及其亚型的研究

目的 探讨慢性乙型肝炎患者外周血CD8+/CD28+淋巴细胞及其亚型与其临床状态和HBV复制的关系.方法 采用流式细胞技术多色荧光分析法,检测研究对象的外周血CD8+淋巴细胞、CD45RO、CD45RA以及CD28的表达及HBV病毒载量.结果 ①慢性HBV携带组CD8+淋巴细胞CD28的表达率(10.64±5.09%)明显低于慢性乙肝组和正常对照组;而CD45RA和CD45RO的表达差异无统计学意义;②慢性乙肝组CD8+/CD45RO+/CD28+的表达(10.99±7.33%)明显高于正常对照组,而慢性乙肝组和慢性HBV携带组CD8+/CD45RA+/CD28+表达率均明显低于正常对照组;③HBeAg阴性慢性乙肝CD8+/CD45RO+/CD28+表达率明显高于阳性组,而CD8+/CD45RA+/CD28+表达率两组无差别.结论 慢性HBV携带者处于病毒携带状态可能与CD8细胞上协同分子CD28的表达低下有关;CD8+/CD45RO+/CD28+亚型淋巴细胞数的升高与慢性乙肝的病情进展及血清HBeAg转换有关.     

结直肠癌患者与健康受试者外周血CD45RA+/CD45RO+淋巴细胞表型分析

目的 分析结直肠癌患者与健康受试者外周血CD45RA+/CD45RO+系列T淋巴细胞表达的差异.方法 运用流式细胞术(FCM)检测2010年1月至2013年12月解放军总医院收治的109例结直肠癌患者(试验组)与64例健康受试者(对照组)外周血CD45RA+、CD45RO+、CD4+ CD45RA+、CD4+ CD45RO+T淋巴细胞亚群表达情况,统计分析试验组和对照组性别和年龄的分布是否存在差异,然后进一步分析CD45 RA等T淋巴细胞亚群与结直肠癌临床分期的关系.结果Ⅰ+Ⅱ期、Ⅲ期和Ⅳ期结直肠癌患者外周血CD45 RA+细胞百分率[三者分别为(56.23±7.75)％、(58.86±7.66)％和(59.02±9.71)％]明显高于对照组[(48.94±12.66)％],差异具有统计学意义(F=11.128,P＜0.001);Ⅲ期和Ⅳ期患者的CD45RO+细胞百分率[分别为(47.19±8.30)％和(45.41±10.45)％]则明显低于对照组[(53.43±11.75)％],差异具有统计学意义(F=5.817,P=0.00083);Ⅲ期和Ⅳ期患者CD45RA+/CD45RO+的比值(分别为1.32 ±0.46和1.43±0.63)明显高于对照组(1.00±0.47),差异具有统计学意义(F=6.986,P=0.000185);Ⅰ+Ⅱ期患者CD4+ CD45 RO+细胞百分率[(31.37±6.39)％]明显高于对照组[(27.49±7.19)％],差异具有统计学意义(F=2.368,P=0.009);Ⅳ期患者CD4+ CD45RA+/CD4+ CD45RO+的比值(0.66±0.39)明显高于Ⅰ+Ⅱ期的患者(0.49±0.23),差异具有统计学意义(F=1.812,P=0.029);各组之间CD4+ CD45RA+细胞百分率无明显统计学差异(F=0.637,P=0.592).结论 随着临床分期的增加,结直肠癌患者外周血CD45RA+细胞逐渐增加而CD45RO+细胞逐渐减少,反映出结直肠癌患者随着肿瘤的进展其免疫功能逐渐抑制、逐渐降低的动态过程;CD4+ CD45RA+细胞和CD4+ CD45RO+细胞在反映结直肠癌患者机体免疫功能方面不如CD45RA+细胞和CD45RO+细胞敏感.     

不同化疗方案治疗T细胞淋巴瘤的疗效比较

目的 探讨不同化疗方案治疗外周T淋巴细胞瘤(peripheral t-cell lymphoma,PTCL)的疗效.方法 将2010年8月至2015年1月医院收治的初诊PTCL患者40例,根据用药方案分为2组,CHOP(环磷酰胺+表柔比星+长春新碱+泼尼松)/CHOP样方案作为CC组24例,Hyper CVAD(大剂量环磷酰胺、表柔比星、长春新碱、地塞米松)/MA(大剂量甲氨蝶呤、阿糖胞苷)方案作为HM组16例,记录近期疗效、生存情况、化疗毒副反应,比较化疗前后T淋巴细胞亚群、CD4+和CD8+T细胞中CD45RA+、CD45RO+T细胞分布情况.结果 HM组缓解率为87.5%高于CC组的54.17%(P<0.05),两组Ⅰ~Ⅱ期化疗缓解率高于Ⅲ~Ⅳ期.两组化疗后CD4+、CD4+/CD8+、CD4+CD45RO+均较治疗前升高,且HM组升高幅度较CC组明显(P<0.05);化疗后CD8+、CD4+CD45RA+、CD8+CD45RA+、CD8+CD45RO+较治疗前下降(P<0.05),其中HM组CD8+、CD4+CD45RA+、CD4+CD45RO+下降幅度较CC组明显.HM组中位无进展生存期为25个月长于CC组的12个月(P<0.05),1年无进展生存期几率为81.25%高于CC组的45.83%(P<0.05);两组2年、3年无进展生存期及1年、2年、3年总生存期比较差异无统计学意义(P>0.05).HM组中性粒细胞减少、血小板减少发生率分别为68.75%、62.5%高于CC组的20.83%、16.67%(P<0.05).结论 Hyper CVAD/MA治疗初诊PTCL近期疗效好,可延长中位PFS,调节机体免疫功能,但中性粒细胞减少、血小板减少发生率较高,应给予高度重视.     

HIV-1 RNA被有效抑制下T细胞与外周血中HIV-1前病毒DNA

目的:考察HIV-1 RNA被有效抑制下T细胞与外周血中HIV-1前病毒DNA的相关性。方法:对云南地区90例确诊为AIDS,HAART持续治疗时间超过6个月,血浆HIV RNA<50 Copies/ml的患者进行研究,采用Spearmans秩和相关回顾性分析CD4+、CD3+、CD8+、CD4+CD28+、CD4+CD45RA+、CD4+CD45RO+、CD38+、CD8+CD38+T细胞的绝对计数和相对计数与外周血中HIV-1前病毒DNA的相关性,同时考察HIV-1前病毒DNA拷贝数与HAART持续治疗时间的相关性。将90例患者根据HAART后CD4+T细胞绝对计数分为A(CD4+<200 cells/μl)、B(200≤CD4+≤349 cells/μl)、C(CD4+≥350 cells/μl)3组,考察3组间T细胞亚群的差别。结果:HIV-1前病毒DNA的log拷贝数与CD4+CD45RA+T细胞绝对计数呈负相关(r=-0.231,P<0.05),与CD4+CD45RA+T细胞相对计数呈明显负相关(r=-0.270,P<0.01);与CD38+T细胞绝对计数呈正相关(r=0.250,P<0.05);与HAART持续治疗时间无相关性。B、C组的CD3+T细胞绝对计数明显高于A组(P<0.01);C组的CD8+T细胞绝对计数高于B组的(P<0.05);B、C组的CD4+CD28+T细胞绝对计数和相对计数明显高于A组(P<0.01),C组的CD4+CD28+T细胞绝对计数高于B组(P<0.05);B、C组的CD4+CD45RA+T细胞、CD4+CD45RO+T细胞绝对计数明显高于A组(P<0.01),C组的CD4+CD45RA+T细胞、CD4+CD45RO+T细胞绝对计数高于B组(P<0.05),B、C组的CD4+CD45RO+T细胞相对计数高于A组(P<0.05);B、C组的CD8+CD38+T细胞绝对计数低于A组(P<0.05),B组的CD8+CD38+T细胞相对计数低于A组(P<0.05);C组的CD38+T细胞绝对计数高于A组(P<0.05)。A、B、C组的HIV-1前病毒DNA的log拷贝数分别为3.561±0.297 9,3.605±0.277 6,3.434±0.289 1(copies/ml),3组间无显著性差异(P>0.05)。结论:经过HAART治疗后HIV-1前病毒DNA可能处于稳定状态,HIV-1前病毒DNA拷贝数只是某些T细胞亚群数量改变的原因之一。     

GD患者外周血CD4+CD28-T细胞亚群的表型特征及临床意义

检测Graves病(GD)患者外周血CD4~+CD28-T细胞水平及其表面CD45RO/CD45RA及ICOS的表达,探讨CD4~+ CD28-T细胞亚群在GD免疫致病机制中的作用。采用三色荧光抗体染色及流式细胞术检测了42例初发GD患者和30例健康者外周血中CD4~+CD28-T细胞的百分率及其表面CD45RO/CD45RA和ICOS表达水平,同时检测其甲状腺功能并进行相关性分析。结果GD患者外周血中CD4~+CD28-T细胞百分率明显高于健康对照组,并高表达ICOS分子,与FT3水平显著正相关;与健康对照组相比,GD患者CD4~+CD28~-CD45RO~+T细胞百分率也显著增高,而CD4~+CD28~-CD45RA~+T细胞呈下降趋势,FT3、FT4水平与CD4~+CD28-T细胞表面CD45RO的表达率呈正相关,而FT3水平与CD45RA表达呈负相关。结论GD患者外周血CD4~+CD28~-T细胞异常增高,表面高表达ICOS分子,具有记忆性细胞的表型特征,与甲状腺功能异常有一定的相关性,CD4~+CD28-T细胞可能是参与GD免疫病理反应的自身反应性T细胞。     

人CK8&18阳性胸腺上皮细胞对脐血CD34~+细胞向T细胞分化的影响

目的:通过CK8/18阳性胸腺上皮细胞与人脐血单个核细胞在Transwell板的共培养,探讨CK8/18阳性TEC对T细胞增殖分化的影响。方法:用胶原酶消化法分离纯化胸腺上皮细胞、免疫组化鉴定分离纯化的TEC,采用密度梯度离心法分离获得脐血单个核细胞,并采用免疫磁珠分选CD34+细胞,将TEC种植培养于Transwell双层培养板上层,使其均匀平铺于上层板底部,再将分选后的细胞加入上层小室,经过48小时的共培养,流式细胞术检测进入下室细胞的表型变化。结果:分离纯化的TEC经免疫组化鉴定CK8/18阳性。TEC与脐血单个核细胞共培养后,CD3+CD4-CD8-双阴性、CD3+CD4+CD8-单阳性细胞显著增加,CD3+CD4+CD8+双阳性细胞亚群细胞减少,CD8单阳性细胞增加不明显;CD45RA阳性细胞比率无显著变化,CD45RO阳性细胞比率显著增加。结论:CK8/18阳性TEC能选择促进脐血单个核细胞CD4+T细胞和CD45RO+细胞增殖。     

炎性关节炎患者调节性T细胞亚群的变化及其对TNF-α拮抗剂治疗的反应

人外周血CD4+Foxp3+调节性T细胞(regulatory T cells,Treg)的表型和功能存在异质性。本文用Foxp3和CD45RO共同标记Treg,研究强直性脊柱炎(AS)和类风湿性关节炎(RA)患者外周血Treg亚群数量以及接受TNF-α拮抗剂治疗对其亚群的改变。分别采集未经治疗的21例活动性AS患者和27例活动性RA患者的外周血。两种疾病各有14名患者接受TNF-α拮抗剂治疗40周。应用流式细胞术检测外周血Treg细胞数量。在CD4+T细胞中,活动性AS和RA患者其表型为CD45RO+Foxp3hi的效应型Treg(AS与正常对照相比,P=0.020;RA与正常对照相比,P0.001)和表型为CD45RO-Foxp3low的幼稚型Treg(AS与正常对照相比,P=0.009;RA与正常对照相比,P=0.001)细胞数量均显著下降。与正常对照相比,AS和RA患者的CD4+Foxp3+T细胞中非Treg亚群细胞(表型为CD45RO+Foxp3low)的比例显著增加(AS与正常对照相比,P=0.027;RA与正常对照相比,P0.001)。TNF-α拮抗剂治疗后,AS患者外周血中效应型Treg(P=0.025)和幼稚型Treg(P=0.005)细胞数量均增加,RA患者中仅效应型Treg细胞数量增加(P=0.003)。因此,不仅分析CD4+Foxp3+T细胞总数,而且分析其中不同表型的亚群细胞数量更有助于明确炎性关节炎的发病机制。     

妊娠17～37周正常和感染胎儿的T细胞免疫

目的探讨妊娠17-37周正常和宫内感染胎儿的T细胞免疫。方法超声引导下行脐带穿刺术收集129例胎儿外周血,其中97例为正常对照组胎儿血,32例为宫内感染组胎儿血(单纯疱疹病毒感染、弓形虫感染、风疹病毒感染),采用双色免疫荧光标记流式细胞仪技术测定胎儿外周血T细胞亚群百分率,3H-TdR掺入法检测T细胞增殖能力,分析生理状态下胎儿的T细胞亚群和T细胞增殖情况、宫内感染时胎儿T细胞亚群和增殖反应的变化。结果生理状态下胎儿CD3 、CD4 、CD8 T细胞百分率随孕龄增长而增加(P<0.01),相关系数为:r(CD3 )=0.63、r(CD4 )=0.45、r(CD8 )=0.27。CD4 /CD8 比值,CD8 HLA-DR 、CD3 CD45RO 、CD3 CD45RA T细胞百分率,T细胞增殖能力与孕龄无相关。CD3 CD45RA T细胞百分率为(96±2)%。宫内感染时,胎儿CD3 、CD4 、CD8 HLA-DR T细胞百分率减少,CD8 T细胞百分率增多,CD4 /CD8 比值降低,T细胞增殖能力下降,与正常对照组相比差异显著。CD3 CD45RO 、CD3 CD45RA T细胞百分率无明显变化。结论妊娠17-37周胎儿各T细胞亚群百分率随孕龄增长情况不同,但多数仍处于“初始状态”;宫内感染时,胎儿容易产生T细胞免疫抑制。     

人外周血CD4~+ IL-21~+记忆T细胞的特征

目的:探讨人外周血中白细胞介素21(IL-21)的产生细胞及其特征。方法:分离人外周血单个核细胞(PBMC),分为不刺激或anti-CD3(OKT3)、OKT3+anti-CD28、PMA+ionomycin刺激四个组,流式细胞术(FCM)检测产生IL-21的细胞亚群。PMA+ionomycin刺激PBMC、纯化CD4+、CD4+CD45RA-、CD4+CD45RA+细胞、脐带血单个核细胞(CB-MC),FCM分析产生IL-21细胞的表型特征和IL-21与Th1、Th2、Th17和Th22细胞因子之间的关系。结果:与OKT3、OKT3+anti-CD28相比,PMA+ionomycin能诱导最高量的IL-21产生。产生IL-21的主要细胞为CD4+T细胞,少数CD8+T细胞。CD4+IL-21+T细胞表达CD45RO,不表达CD45RA,其中部分细胞表达CCR6、CCR7或CXCR5。CD4+CD45RA-细胞表达IL-21远高于CD4+CD45RA+细胞。进一步研究表明,PBMC产生IL-21,而CBMC不产生。此外,大约24%的CD4+IL-21+细胞表达IFN-γ,小于10%CD4+IL-21+细胞表达IL-4、IL-17或IL-22。结论:人PBMC在多克隆刺激的条件下,可以诱导IL-21的产生。产生IL-21的主要细胞亚群具有记忆CD4+T细胞的表型。其中一部分CD4+IL-21+T细胞的表型独立于Th1、Th2、Th17和Th22细胞亚群。     

慢性乙型肝炎患者外周血CD45RA+、CD45RO+T淋巴细胞亚群的特点及临床意义

目的：探讨慢性乙型肝炎患者外周血CD4^＋CD45RA^＋、CD4^＋CD45RO^＋、CD8^＋CD45RA^＋和CD8^＋CD45RO^＋T淋巴细胞亚群的特点及其与肝病病情的关系。方法：采集46例轻中度慢性乙型肝炎患者、58例重度慢性乙型肝炎患者和30例健康人的外周抗凝血，应用流式细胞技术三色荧光分析法对其外周血中CD4^＋CD45RA^＋、CD4^＋CD45RO^＋、CD8^＋CD45RA^＋和CD8^＋CD45RO＋T淋巴细胞亚群进行检测。结果：轻中、重度慢性乙型肝炎患者与正常人相比，其外周血中CD4^＋、CD8^＋T细胞均无明显改变；CD8^＋CD45RA^＋T细胞均明显降低，CD8^＋CD45RO^＋T细胞均明显增高，而CD4^＋CD45RA^＋、CD4^＋CD45RO^＋T细胞均无明显改变；重度慢性乙型肝炎患者与轻中度慢性乙型肝炎患者相比，CD8^＋CD45RA^＋T细胞明显降低（P〈0.05），CD8^＋CD45RO^＋T细胞明显升高（P〈0.05）。结论：乙型肝炎慢性化过程中，CD8^＋CD45RO^＋T细胞起重要作用且与慢性乙型肝炎患者病情的进展呈正相关；检测CD4^＋CD45RA^＋、CD4^＋CD45RO^＋、CD8^＋CD45RA^＋和CD8^＋CD45RO^＋T淋巴细胞亚群比检测CD4^＋和CD8^＋T细胞亚群更能正确、充分、全面地了解慢性乙型肝炎的发病机制和预后，从而有效地指导临床治疗。     

蕈样肉芽肿患者外周血单一核细胞CD45RA及CD45RO表达的研究

目的探讨蕈样肉芽肿（Mycosis fungoides,MF）患者外周血单个核细胞CD45RA及CD45RO的表达及其与MF发病的关系。方法应用双荧光抗体标记、流式细胞仪检测15例MF患者外周血单个核细胞CD45RA及CD45RO的表达。结果（1）MF患者外周血CD3^＋、CD3^＋CD8^＋细胞与正常对照比较差异不显著（P〉0.05）。（2）MF患者外周血CD4^＋细胞低于正常对照,差异非常显著（P〈0.001）。（3）MF患者外周血T细胞CD3^＋CD4^＋/CD3^＋CD8^＋比值低于正常对照,差异显著（P〈0.001）。（4）MF患者外周血CD45RA^＋细胞低于正常对照,差异非常显著（P〈0.001）,CD45RO^＋细胞高于正常对照,差异非常显著（P〈0.001）。（5）MF患者外周血CD45RO^＋/CD45RA^＋比值高于正常对照,差异非常显著（P〈0.001）。（6）MF患者外周血CD4^＋CD45RA^＋细胞低于正常对照,差异非常显著（P〈0.001）。（7）MF患者外周血CD4^＋CD45RO^＋细胞及CD8^＋CD45RO^＋细胞均高于正常对照,差异非常显著（均P〈0.001）。（8）MF患者外周血CD4^＋CD45RO^＋/CD4^＋CD45RA^＋比值及CD8^＋CD45RO^＋/CD8^＋CD45RA^＋比值均高于正常对照,差异非常显著（P〈0.01及P〈0.001）。结论MF患者外周血中,不仅存在CD4^＋亚群失调和CD4^＋/CD8^＋比值降低,而且在CD4^＋和CD8^＋亚群中也存在CD45RA^＋、CD45RO^＋亚群失调和CD45RO^＋/CD45RA^＋比值升高,从而导致的机体免疫功能紊乱,可能与MF的发病或病情加剧有关。     

Distribution of Alloreactivity Amongst the CD45 Isoforms of Circulating CD4 and CD8 T Lymphocytes

The expression of different isoforms of the CD45 surface molecule allows lymphocytes to be divided into two nonoverlapping categories, CD45RA and CD45RO. Previous studies of CD4 T cells have shown that responses to soluble antigens are present predominantly in the RO subset and to mitogens in the RA, alloreactivity being present in both subsets. Responses of CD8 T cells have not been investigated in such detail, nor have responses been compared to CD4 cells. Here we report the alloreactive responses of both CD45RA and RO subsets amongst both CD4 and CD8 T lymphocytes. Following isolation of CD4 and CD8 cells with immunomagnetic beads, CD45 subsets were separated by negative depletion using specific monoclonal antibodies. CD45RA populations were greater than 97% pure and CD45RO cells greater than 91%. One-way primary mixed lymphocyte reactions were established using the purified responder cells with irradiated allogeneic peripheral blood mononuclear cells as stimulators; experiments were all repeated at least three times. In assays of CD4⁺ RA and RO subsets, reactivity was present in both isoforms, being consistently, but not significantly, greater amongst the RO subset. With CD8⁺ T cells, reactivity was also present in both isoforms, but was significantly greater in the CD45RA subset, with mean proliferation 2.5–3-fold that of the CD45RO cells ( P  < 0.05).     

大肠癌患者与正常健康人淋巴细胞表型差异分析

目的:分析大肠癌患者与正常健康人20项淋巴细胞亚群变化情况。方法:流式细胞术(FCM)检测168例大肠癌患者与102例正常健康人外周血20项淋巴细胞亚群,SPSS17.0软件进行统计学处理。结果:与正常健康人相比,大肠癌患者CD29+、45RA+、CD4+CD45RO+和CD8+CD28-细胞百分比例明显升高,而NKT、45RO+、CD4+CD45RA+、CD28+和CD8+CD28+细胞百分比例则明显降低(P<0.05)。Ⅳ期患者组CD8+CD28-细胞百分比例明显高于正常健康人组、Ⅰ+Ⅱ期患者组及Ⅲ期患者组(P<0.05)。结论:大肠癌患者的细胞免疫功能降低,体液免疫功能有所增强;传统指标难以准确评价患者的免疫功能,包括CD8+CD28+、CD8+CD28-在内的多项细胞亚群分析体系也许能更好的反映大肠癌患者的免疫状况。     

Abnormalities in the T and NK lymphocyte phenotype in patients with Nijmegen breakage syndrome

Nijmegen breakage syndrome (NBS) is a rare autosomal recessive disorder characterized by spontaneous chromosomal instability with predisposition to immunodeficiency and cancer. In order to assess the cellular basis of the compromised immune response of NBS patients, the distribution of functionally distinct lymphocyte subsets in peripheral blood was evaluated by means of double-colour flow cytometry. The study involved the 36 lymphopenic patients with a total lymphocyte count < or =1500 microl (group A) and seven patients (group B) having the absolute lymphocyte count comparable with the age-matched controls (> or =3000 microl). Regardless of the total lymphocyte count the NBS patients showed: (1) profound deficiency of CD4+ and CD3/CD8+ T cell subsets and up to fourfold increase in natural killer (NK) cells, almost lack of naive CD4+ T cells expressing CD45RA isoform, unchanged percentage of naive CD8+ cell subset (CD8/CD45RA+) but bearing the CD8 receptor of low density (CD8low); (2) normal expression of CD45RA isoform in the CD56+ lymphocyte subset, profound decrease in alpha beta but up to threefold increase in gamma delta-T cell-receptor (TCR)-positive T cells; (3) shift towards the memory phenotype in both CD4+ and CD8+ lymphocyte subpopulations expressing CD45RO isoform (over-expression of CD45RO in terms of both the fluorescence intensity for CD45RO isoform and the number of positive cells); and (4) an increase in fluorescence intensity for the CD45RA isoform in NK cells population. These results indicate either a failure in T cell regeneration in the thymic pathway (deficiency of naive CD4+ cells) and/or more dominant contribution of non-thymic pathways in lymphocyte renewal reflected by an increase in the population of CD4+ and CD8+ memory cells, gamma delta-TCR positive T as well as NK cell subsets.     

不同年龄、性别的健康成年人21项淋巴细胞亚群分析

建立健康成年人外周血21项淋巴细胞亚群正常参考值,并分析不同年龄、性别健康成年人淋巴细胞亚群之间的差异.用三标荧光抗体对102例健康成年人外周血进行标记,流式细胞仪检测并分析结果,SPSS 10.0软件进行统计学处理.结果显示:21～89岁健康成年人外周血中CD3+、CD3+CD4+、CD3+CD8+、CD4+/CD8...     

Selective recruitment of lymphocyte subsets to the inflamed appendix.

Total lymphocyte counts and the distribution of lymphocyte subsets were determined in peripheral venous blood and appendiceal mononuclear cells from 60 patients who underwent appendicectomy for the clinical diagnosis of appendicitis. A significant peripheral lymphopenia was observed in the 46 patients with histologically confirmed acute appendicitis which was accompanied by an increase in the appendiceal lymphocyte concentration. There was an even greater depletion of CD45RO+ (memory) T lymphocytes in peripheral blood and an increase in the inflamed appendix. Reciprocal changes were observed in the CD45RA+ (naive) T lymphocyte subset. These changes were reflected in the local arterial and venous CD45RA and CD45RO T lymphocyte subsets. Proliferation studies showed an expanded functional repertoire of T lymphocytes in the inflamed appendix. Selective recruitment of memory T lymphocytes from the peripheral blood to the inflamed appendix was demonstrated.     

Age-related modifications of the human alphabeta T cell repertoire due to different clonal expansions in the CD4+ and CD8+ subsets

We have studied the effects of a life-long antigen stimulation on the clonal heterogeneity of human peripheral T cell subsets, as defined by their CD45 isoform expression. CD4+ or CD8+ T cells were obtained from healthy donors ranging in age from 20 to 100 years, and sorted into CD45RA+ and CD45RO+ populations. A modified PCR-heteroduplex analysis was then used to directly compare the TCR Vbeta clonal make up of either compartment pair. We find that the CD4+ T cell repertoire remains largely polyclonal throughout life, since CD4+ expanded clones are rare and accumulate predominantly in the CD45RO+ compartment of exceptionally old donors (100 years old). In contrast, the CD8+ T cell subset contains expanded clones which are already detectable in young adults and become very frequent in 70- to 75-year-old donors in both CD45RA+ and CD45RO+ compartments analyzed. Interestingly, some expanded clones are detectable in the CD45RA+ or in both CD45RA+ and CD45RO+ compartments of either CD4+ or CD8+ T cells. These results indicate that the age-dependent accumulation of expanded clones starts earlier and is more pronounced in the CD8+ than in the CD4+ T cell subset, reinforcing the concept that clonal expansion in the two subsets is controlled by substantially different mechanisms. Furthermore, whereas the finding of expanded CD45RO+ T cell clones is explained by antigen- driven proliferation, the detection of expanded clones in the CD45RA+ or in both CD45RA+ and CD45RO+ compartments would support the hypothesis of reversion from the CD45RO+ to the CD45RA+ phenotype after antigen encounter.      

CD45 isoform expression during T cell development in the thymus.

Various isoforms of leukocyte common antigen, or CD45, are expressed differentially on T cells at different stages of development and activation. We report studies on CD45 isoform expression on various subsets of human T cells using two- and three-color flow cytometry and cell depletion. Bone marrow cells that were depleted of CD3+ and HLA-DR+ cells were CD45RA-RO-. The earliest CD3-CD4-CD8-CD19- thymocytes were CD45RO- with 20%-30% CD45RA+ cells. The most prominent population of CD4+CD8+ double-positive thymocytes were CD45RA-RO+. Even the CD4+CD8+ blasts were greater than 90% CD45RO+. About 80% of single-positive thymocytes (CD4+CD8- or CD4-CD8+) were also CD45RO+. Only 4.3% of CD4+ and 18% of CD8+ single-positive thymocytes were CD45RA+. In contrast, cord blood T cells which represent the stage that immediately follows single-positive thymocytes, contained 90% CD45RA+ cells. Thus, in terms of CD45 isoform expression, single-positive thymocytes are more like double-positive cells than cord blood T cells. These results suggest the following sequence of CD45 isoform switching during T cell development: CD45RA-RO- or RA+RO- (double-negative thymocytes)----RA-RO+ (double-positive and most single-positive thymocytes)----RA+RO- (cord blood T cells), the last switch from CD45RO to CD45RA occurring as a final step of maturation in the thymus.