宫颈病变患者外周血T淋巴细胞亚群的相关性研究

目的:探讨HPV阳性不同级别宫颈病变患者T淋巴细胞亚群的变化,为后续免疫治疗提供理论依据。方法:选取高危性HPV阳性患者,其中CINⅡ~Ⅲ53例、宫颈癌31例、宫颈炎和CINI 41例(对照组)。流式细胞仪检测T淋巴细胞亚群的重要指标CD3总T细胞计数、CD4辅助/诱导T细胞计数、CD8抑制/杀伤T细胞计数、CD4/CD8比值的变化。结果:随着宫颈病变程度加重,CD3总T细胞绝对计数、CD4辅助/诱导T细胞绝对计数、CD8抑制/杀伤T细胞计数明显下降,各组间差异有统计学意义(P0.01)。CD4/CD8比值随着宫颈病变加重有下降趋势,但差异无统计学意义(P0.05)。结论:HPV阳性高级别宫颈病变患者的细胞免疫功能均有不同程度的降低,宫颈癌患者降低最显著。检测T淋巴细胞亚群可用于宫颈癌患者的免疫检测,并为预后的评估和临床免疫治疗提供理论依据。     

维吾尔族宫颈癌人乳头瘤病毒感染与CD4+CD25+CD127-调节性T细胞的相关性研究

目的:探讨维吾尔族宫颈癌人乳头瘤病毒(HPV)感染与CD4+CD25+CD127-调节性T细胞(Treg)的相关性.方法:采用HPV核酸扩增分型检测试剂盒及流式细胞术检测66例宫颈癌、35例宫颈上皮内瘤变(CIN)Ⅱ～Ⅲ、45例宫颈CIN Ⅰ维吾尔族患者和40例维吾尔族正常对照组HPV分型和外周血中T细胞亚群及CD4+...     

负载HPV16 E7~(44-62)多肽的脐血DC-CIK细胞抗宫颈癌的免疫效应研究

目的:探讨HPV16 E7~(44-62)多肽对脐血DC-CIK细胞分化和功能调节,以及其对宫颈癌CaSki细胞的体外杀伤效应。方法:采集脐血单个核细胞在体外诱导成DC和CIK细胞,HPV16 E7~(44-62)多肽负载DC与CIK细胞以细胞比1∶5共同培养。流式细胞术(FCM)分析细胞表型。RT-qPCR及Western blot法检测T-bet、GATA-3及Foxp3表达水平。ELISA法检测共培养前后CIK细胞上清液中IFN-γ、IL-12、IL-10水平。CCK-8法检测体外DC-CIK细胞抗宫颈癌Caski细胞的细胞毒杀伤效应。结果:DC及CIK细胞体外诱导成功。共培养6天,HPV16 E7~(44-62)-DC-CIK组的CD3+CD56+、CD3+CD8+、CD3+CD4+细胞比例明显高于DC-CIK组及CIK组(P0.05),DC-CIK组较CIK组高(P0.05);HPV16 E7~(44-62)-DC-CIK组的CD4+CD25+(Treg)阳性率明显低于DC-CIK组及CIK组(P0.05)。HPV16 E7~(44-62)-DC-CIK组的T细胞转录因子T-bet、GATA-3、Foxp3表达与DC-CIK组及CIK组比较,差异有统计学意义(P0.05),其中T-bet表达最高,而GATA-3和Foxp3表达最低。HPV16 E7~(44-62)-DC-CIK组细胞上清液中IFN-γ、IL-12、IL-10与其余两组比较,差异有统计学意义(P0.05),IFN-γ、IL-12表达最高,IL-10表达最低。在各效靶比下,HPV16 E7~(44-62)-DC-CIK组对宫颈癌CaSki细胞杀伤率最高(P0.05)。结论:负载HPV16 E7~(44-62)多肽的DC可增强同源CIK细胞体外抗宫颈癌的免疫作用,本研究为宫颈癌DC-CIK细胞免疫治疗提供新的致敏方法。     

替比夫定对乙肝孕妇T淋巴细胞亚群的影响

目的:分析孕晚期应用替比夫定(Ld T)对乙肝孕妇细胞免疫功能的影响,为降低乙肝高病毒血症孕产妇死亡率提供临床依据。方法:收集2013年3月至12月我院收治的乙肝表面抗原阳性(HBs Ag)阳性、乙肝e抗原(HBe Ag)阳性、HBV DNA≥106IU/ml的59例孕妇,根据孕晚期是否服用Ld T,分为Ld T治疗组和未治疗组。同时收集同期HBs Ag(+)、HBe Ag(-)、HBV DNA106IU/ml的孕妇30例为低病毒组,HBs Ag(-)的孕妇30例为正常对照组。运用流式细胞术分析Ld T对乙肝孕妇外周血CD3~+T细胞、CD4~+T细胞、CD8~+T细胞及CD4+CD25+Treg细胞(Tregs)的影响。结果:正常组、低病毒组、高病毒未治疗组和高病毒Ld T治疗组的CD3~+T细胞比例无明显差异;与正常组比较,高病毒和低病毒组中CD4~+T细胞比例升高、CD8~+T细胞降低、CD4~+/CD8~+比值进一步升高,Tregs比例显著升高。经Ld T治疗后,T细胞亚群基本处于正常水平。与高病毒未治疗组相比,Ld T治疗组的CD4~+T细胞比例下降,CD8~+T细胞比例也有上升趋势,CD4~+/CD8~+比值显著下降,Tregs比例显著降低。结论:乙肝高病毒血症孕妇体内细胞免疫功能紊乱。Ld T治疗可能解除Tregs对乙肝孕妇机体细胞免疫功能的抑制作用,恢复CD4~+T细胞与CD8~+T细胞之间的平衡状态。Ld T治疗可能通过免疫调节在有效阻断母婴垂直传播的同时,还有可能降低重型肝炎发生率,从而降低孕产妇死亡率。     

顺铂对小鼠宫颈癌局部CD8~+T细胞的影响及可能机制

目的:探讨顺铂对小鼠宫颈癌局部CD8~+T细胞的影响及可能机制。方法:建立TC-1肿瘤细胞C57BL/6小鼠模型,分为对照组(4只)和顺铂组(4只),对照组给予磷酸盐缓冲溶液(PBS)100μl,顺铂组给予顺铂5 mg/kg,两组均腹腔注射,3天1次,共2次。免疫组化方法检测两组肿瘤组织中CD8阳性T细胞(CD8~+T)、程序性凋亡配体1(PD-L1)的表达,并对小鼠宫颈癌组织中CD8~+T细胞与PD-L1表达的相关性进行分析,同时采用流式细胞仪检测并比较两组脾单核淋巴细胞中CD4~+CD25~+T调节细胞(Treg)数量比例。结果:①顺铂组模型小鼠宫颈癌组织中CD8~+T表达的MOD值为3.24±0.39明显低于对照组4.70±0.28,差异有统计学意义(P0.05);顺铂组PD-L1表达的MOD值为7.65±1.82高于对照组2.89±0.70,差异有统计学意义(P0.05)。②小鼠肿瘤组织中CD8~+T细胞与PD-L1表达呈负相关(r=-0.61,P0.05)。③顺铂组小鼠脾脏内单核淋巴细胞中Treg细胞数量(71.50±4.04)%低于对照组(87.00±6.06)%,差异有统计学意义(P0.05)。结论:顺铂可增加宫颈癌组织PD-L1表达,减少宫颈癌组织局部CD8~+T细胞数量,抑制机体免疫内环境,并且顺铂对机体免疫的抑制作用可能并不来源于对Treg细胞的影响。应用PD-L1抗体阻断顺铂诱发的PD-L1表达可能可促进顺铂的化疗效果。     

卵巢早衰患者外周血CD4~+CD25~+Treg细胞的变化及意义

目的:探讨CD4+CD25+调节性T细胞(即CD4+CD25+Treg细胞)在卵巢早衰发病机制中的作用。方法:流式细胞仪定量检测卵巢早衰(premature ovarian failure,POF)患者、卵巢储备功能下降(diminished ovarian reserve,DOR)患者及健康对照组外周血CD4+T、CD8+T细胞及CD4+CD25+Treg细胞数量,应用3H-thymidine掺入法测定POF患者及对照组外周血CD4+CD25+Treg细胞对效应性T细胞的增殖抑制功能。结果:与对照组相比,POF患者及DOR患者CD4+CD25+Treg细胞比例降低(P<0.01)、POF患者CD4+T/CD8+T细胞比值增高(P<0.05),DOR患者CD4+T/CD8+T细胞比值无明显变化(P>0.05);POF患者免疫抑制功能无明显降低(P>0.05)。结论:CD4+CD25+Treg细胞比例降低与T细胞亚群失衡可能是POF的发病机制。     

负载宫颈癌肿瘤裂解物的树突状细胞诱导的CTL杀伤效应

目的:利用树突状细胞(DC)递呈肿瘤抗原的生物学特性,提高细胞毒性T淋巴细胞(CTL)对宫颈癌细胞的杀伤活性。方法:联合应用重组人粒巨噬细胞集落刺激因子(GMCSF)、白细胞介素4(IL4)、肿瘤坏死因子α(TNFα)诱导培养宫颈癌患者外周血DC,以宫颈癌组织肿瘤相关抗原(TAA)激活DC,诱导异体T细胞增殖分化为CTL,以MTT法测定CTL增殖活性及其对宫颈癌Hela细胞株的特异性杀伤活性。结果:宫颈癌患者外周血来源DC负载宫颈癌肿瘤裂解物后,可促进CTL增殖,诱导产生的CTL对宫颈癌细胞具有高效而特异的杀伤率,且显著高于异种肿瘤细胞(P<0.05)。结论:DC能递呈宫颈癌肿瘤抗原,诱导产生抗原特异和高效的CTL,提示负载肿瘤裂解物的DC,具有为宫颈癌患者进行特异性免疫治疗的临床应用前景。     

HPV16E7肽疫苗对人免疫重建荷人宫颈癌细胞株SiHa细胞SCID鼠免疫功能的影响

目的:观察HPV16E7多肽疫苗对人免疫重建荷人宫颈癌细胞株SiHa细胞严重联合免疫缺陷(SCID)鼠移植瘤的影响。方法:对SCID鼠腹腔注射人外周血淋巴细胞(PBL)后24小时,皮下接种HPV16阳性的人宫颈癌细胞株SiHa细胞,建立人免疫重建荷SiHa细胞的SCID鼠模型,当移植瘤最小体积达100mm3时,三次背部皮下接种疫苗,每次接种间隔二周,观察荷瘤鼠一般生物学特性,检测血浆中人IgG含量、外周血中人CD3+、CD4+、CD8+T细胞、脾重、肿瘤浸润淋巴细胞(TIL)以及脾细胞毒杀伤功能。实验设立空白组,仅为腹腔内注射PBL,疫苗成份仅为不完全佐剂(IFA);随机多肽组,为腹腔内注射PBL,皮下接种SiHa细胞,疫苗成份仅为随机多肽+T辅助多肽+IFA;T辅助多肽组,为腹腔内注射PBL,皮下接种SiHa细胞,疫苗成份仅为T辅助多肽+IFA;多肽治疗组,为腹腔内注射PBL,皮下接种SiHa细胞,疫苗成份为HPV16E711-20(YMLDLQ-PETT),HPV16E786-93(TLGIVCPI)+T辅助多肽+IFA。结果:各组SCID鼠血浆中人IgG水平,8周内随重建时间延长而升高(P<0.05),多肽治疗组达2957.85μg/m l,但各组间无显著性差异;外周血中均可检测到人CD3+、CD4+、CD8+T细胞,但各组间无显著性差异(P>0.05);与空白组比较,其他三组SCID鼠脾重均显著增加(P<0.05)。组织学观察到移植瘤局部TIL浸润及明显的出血和坏死,但经免疫组化未检测到人CD8+T细胞。与空白组比较,多肽治疗组、随机多肽组、辅助多肽组的脾细胞毒杀伤功能均显著降低(P<0.05)。结论:人免疫功能重建的荷SiHa细胞的SCID鼠体内重建的人免疫功能随荷瘤时间的延长受到抑制,多肽疫苗在一定程度上能减轻免疫抑制状态。     

宫颈癌患者外周血CD4+CD25+调节性T细胞及其Foxp3表达的临床意义

目的评价宫颈癌患者外周血CD4+CD25+调节性T细胞(Regulatery T cell,Treg)及其Foxp3表达的临床意义。方法收集中国医科大学附属盛京医院自2012年1月至2013年6月手术治疗的早期宫颈癌患者(FIGO分期≤Ⅱ期)34例,高级别宫颈上皮内瘤变(CIN2/3)患者30例以及20例健康女性作为正常对照。流式细胞技术检测各研究组手术前、后及对照组外周血Treg及Foxp3+Treg细胞占CD4+T细胞比例,应用t检验和单因素方差法进行统计学分析。结果比较宫颈癌、CIN2/3及对照组外周血Treg及Foxp3+Treg细胞所占CD4+T细胞比例,各组间差异有统计学意义(P0.05);术后早期(7d),宫颈癌及CIN2/3患者外周血Foxp3+Treg细胞所占比例较术前显著下降(P0.05),但Treg细胞所占比例无显著性变化(P0.05);术后恢复期(1个月),宫颈癌及CIN2/3患者外周血Foxp3+Treg细胞所占比例较术后早期无显著变化,但Treg细胞所占比例较术后早期显著性降低(P0.05)。结论宫颈癌患者外周血Treg细胞及其Foxp3的表达均与病变程度相关;Treg细胞Foxp3的表达具有不稳定性;宫颈癌肿瘤微环境可能是维持Treg细胞分化增殖及Foxp3稳定表达的外在客观条件。     

妊娠中晚期外周血T淋巴细胞亚群和NK细胞的观察

目的 :检测孕妇外周血T淋巴细胞亚群和NK细胞的变化 ,探讨正常妊娠时母体的细胞免疫状态。方法 :健康孕妇 92例按孕周分为 3组 :中孕组 (孕周 13～ 2 7+ 6周 )、晚孕未足月组 (孕周 2 8～ 36 + 6周 )和足月组 (孕周 37～ 4 1+ 6周 ) ;另取同期健康未孕生育年龄妇女 2 0例作对照组。用流式细胞仪检测其外周血T淋巴细胞亚群和NK细胞的相对数 ,结合外周血白细胞计数计算其绝对数。结果 :正常孕妇外周血白细胞总数显著增加 ,其中粒细胞百分数和绝对数均显著增加 ,单核细胞绝对数增加 ,淋巴细胞百分数和绝对数均显著减少 ;CD3+ 细胞百分数显著增加 ,CD4 + 细胞百分数和绝对数均显著减少 ,CD4 + /CD8+ 比值显著下降 ,CD8+ 细胞差异无显著性 ;NK细胞百分数和绝对数均显著减少。随着孕周进展 ,CD4 + 细胞百分数和绝对数均逐渐减少 ,CD4 + /CD8+ 比值逐渐下降 ,中孕组与晚孕未足月组比较 ,差异有显著性 (P <0 .0 5 )。结论 :妊娠期母体细胞免疫功能处于免疫抑制状态 ;随着妊娠进展 ,这种抑制有一定程度的下降。     

子宫颈上皮内病变及子宫颈癌患者外周血及局部免疫功能分析#br#

Objective?To investigate the distribution of T cell subsets in peripheral blood of different cervical lesions and the level of cytokines secreted, and it’s difference in the location and density of immune cell distribution in the epithelium and stroma of cervical lesion tissues. Methods?We used CD series cell detection slide quality control kit to detect the distribution of T cell subsets in peripheral blood of 55 patients with different cervical lesions, analyzed the changes of peripheral blood cytokines, and analyzed the localization and density distribution of T lymphocytes (CD4+T, CD8 +T) in the tissue microenvironment of 64 patients with different cervical lesions by immunohistochemistry. Results?The level of CD4+T and CD8+T in peripheral blood of patients with low grade lesion (LSIL) were lower, with 479.13±229.65 unit and 378.00±231.74 unit respectively. The level of CD4+T and CD8+T in high grade cervical squamous intraepithelial lesion(HSIL)was 816.30±284.65 unit and 668.75±268.92 unit respectively, and the level of CD4+T and CD8+T in carcinoma was 824.80±330.65 unit and 564.00±58.31 unit. Meanwhile the concentration of IL-17 in the serum of LSIL patients was higher than that of HSIL (P<0.05). CD8+T cells in epithelial and stroma tissues of cervical cancer were higher than those in control, LSIL and HSIL tissues, with statistical significance (P<0.05). However, CD4+T cells in the stroma of cancer tissues were higher than those of control, LSIL and HSIL tissues, with statistical significance (P<0.001). Conclusion?The patients with cervical lesion have abnormal immune function in the systemic immunity and local microenvironment of the lesion, both CD8+T cells and CD4+T cells play a key role in eliminating HPV-infected cervical epithelial cells. The development of cervical lesions is related to the cellular inflammatory factors.     

妊娠期糖尿病患者母儿免疫水平变化的临床研究

目的:研究妊娠期糖尿病(GDM)患者母儿免疫球蛋白(Ig)、补体(C)、T淋巴细胞亚群以及NK细胞水平的变化,探讨GDM对母儿体液免疫和细胞免疫的影响。方法:选择58例不同糖耐量水平的GDM患者为研究对象根据是否需要胰岛素治疗进一步分为GDM1组和GDM2组,以30例健康孕妇做对照。免疫比浊法测定外周静脉血与脐血的免疫球蛋白、补体水平;流式细胞技术测定外周静脉血与脐血的T细胞亚群及NK细胞水平。结果:GDM组外周血CD4+、CD4+/CD8+、CD16+CD56+、IgG含量均下降,CD8+、C3、C4、IgE含量升高,差异有统计意义(P<0.05);CD3+、IgA、IgM含量下降,差异无统计学意义(P>0.05)。GDM组脐血中CD3+、CD4+、CD4+/CD8+、CD16+CD56+含量均下降,差异有统计学意义(P<0.05);IgM、IgG含量下降,CD8+、IgE含量升高,差异无统计学意义(P>0.05)。与GDM1组相比,GDM2的指标呈现更明显的变化趋势。结论:妊娠期糖尿病存在母儿体液免疫和细胞免疫的失衡,且病情越重,这种改变越显著。     

Development, characterization and distribution of adoptively transferred peripheral blood lymphocytes primed by human papillomavirus 18 E7--pulsed autologous dendritic cells in a patient with metastatic adenocarcinoma of the uterine cervix

We describe a 27-year-old woman with systemic chemoresistant and radioresistant metastatic disease secondary to a recurrence of human papillomavirus (HPV) 18 infected cervical adenocarcinoma of the uterine cervix who received adoptive transfer of peripheral blood T cells stimulated with HPV 18 E7-pulsed autologous dendritic cells (DC). Extensive in vitro characterization of the DC-activated T cells derived from peripheral blood mononuclear cells (PBMC) included phenotypic analysis, cytotoxicity and intracellular cytokine production. High cytotoxicity activity was observed by CD8+T cells against autologous tumor cells, but not against NK-sensitive K562 cells, autologous Con-A lymphoblasts, or autologous Epstein-Barr virus-transformed lymphoblastoid cells. Blocking studies demonstrated that lytic activity was significantly inhibited by pretreatment of tumor targets with MAb specific for HLA class I as well as that of effector cells with anti-CD8, anti-LFA-1, but not anti CD3 MAb. Two-color flow cytometric analysis of the cytotoxic T cells revealed that a significant proportion of CD8+ cells was also CD56+. These double positive CTLs were thymically derived, as shown by expression of heterodimeric CD8 molecules (alpha/beta CD8) and were endowed with high cytotoxic activity against tumor cells. Analysis of intracellular cytokine expression showed that the striking majority of E7-pulsed DC activated CD8+ T cells strongly expressed IFN-gamma, TNF-alpha and IL-2 but not IL-4. The patient received two infusions of cytotoxic tumor-specific T cells at 2 week intervals, and in vivo distribution of the T cells was followed by 111 oxine labeling and serial gamma camera imaging. Persistent accumulation of radioactivity in the lungs, which harbored extensive metastatic disease, was detected up to 120 hrs after the infusion. Taken together, these results illustrate the potential of E7-specific and tumor-specific CTL-based immunotherapy for the treatment of patients with invasive cervical cancer.     

反复胚胎种植失败免疫治疗疗效及淋巴细胞免疫表型变化的研究

目的:探讨淋巴细胞主动免疫治疗不明原因反复胚胎种植失败(RIF)患者的疗效及治疗前后外周血淋巴细胞免疫表型的变化。方法:对研究组83例不明原因RIF的患者进行淋巴细胞主动免疫治疗(LIT);对照组96例不明原因RIF患者移植前不给予特殊处理。并按照胚胎移植属性分为鲜胚移植和冻胚移植,所移植胚胎均为卵裂期优质胚胎,接受LIT后新鲜胚胎移植为研究组A(51例);未行LIT接受新鲜胚胎移植为对照组A(57例)。接受LIT后冷冻胚胎移植为研究组B(32例),未行LIT接受冷冻胚胎移植为对照组B(39例)。比较各组的临床妊娠情况,并观察LIT治疗前后外周血淋巴细胞免疫表型变化。结果:(1)研究组A的临床妊娠率(47.06%)高于对照组A(22.81%),研究组B的临床妊娠率(46.88%)高于对照组B(23.77%),差异有统计学意义(P0.05);研究组A的临床妊娠率与研究组B比较,差异无统计学意义(P0.05)。(2)与治疗前相比,LIT后外周血CD3~+CD4~+T细胞占淋巴细胞的比率下降,CD3~+CD8~+T细胞占淋巴细胞的比率上升、CD4~+/CD8~+比值下降,NK细胞(CD3-CD16~+CD56~+)占淋巴细胞的比率下降,差异均有统计学意义(P0.05)。结论:淋巴细胞主动免疫治疗对RIF患者成功妊娠有促进作用,淋巴细胞主动免疫治疗可能促使外周血淋巴细胞免疫表型向增强母胎免疫耐受的方向改变。     

Murine female reproductive tract intraepithelial lymphocytes display selection characteristics distinct from both peripheral and other mucosal T cells

Despite immense effort, the development of vaccines effective at mucosal sites has proceeded at a faltering pace. Efforts concentrating on humoral immunity but neglecting cellular immunity may be misdirected by ignoring many viral mucosal pathogens. Improved understanding of the development and maintenance of lymphocytes populating the reproductive tract (rtIELs) may inform advances in vaccination strategies for sexually transmitted diseases. Recent studies highlight tissue-specific differences in the development of mucosal immunity and suggest that the local milieu may play a role in selection, maintenance and function of resident lymphocytes. Here, we describe MHC class I and thymus dependence of subpopulations of rtIELs. TCRalphabeta+ CD8alphabeta+ T cells in the periphery, intestine, and female reproductive tract are all developmentally dependent on classical class I MHC and the thymus. TCRalphabeta+ CD8alphaalpha+ are absent from the periphery and the rtIELs, but are present and classical MHC class I-independent, in the intestine. In contrast to intestinal TCRgammadelta+ cells, TCRgammadelta+ rtIELs are CD8 negative and thymus dependent. In contrast to peripheral TCRgammadelta+ cells, murine TCRgammadelta+ rtIELs express not a diverse array of Vdelta genes, but rather, a canonical Vdelta1. In summary, lymphocytes isolated from the murine female reproductive tract have characteristics distinct from both peripheral T cells and those found at other mucosal sites. Therefore, for the purpose of vaccination strategies, the female reproductive tract should be regarded neither as peripheral nor mucosal, but rather as a tissue with distinctive immunological characteristics.     

CD8+CD45RA+CD27-CD28-T-cell subset in PBL of cervical cancer patients representing CD8+T-cells being able to recognize cervical cancer associated antigens provided by HPV 16 E7

OBJECTIVE: In response to antigenic stimulation, naive MHC-class I restricted and antigen-specific CD8+CD45RA+CD28+T-cells undergo clonal expansion and differentiate into CD8+CD45RO+ memory T-cells. Upon re- encounter with the nominal antigen, CD45RO+ T-cells are able to convert to CD8+CD45RA+CD28-T-cells displaying potent immune effector functions, including TNF-alpha production. This T-cell subpopulation constitutes a minor population in healthy individuals. In the present study we are currently evaluating whether this particular T-cell subset in PBL represents CD8+T-cells which may be able to recognize cervical cancer associated antigens provided by HPV 16 E7. MATERIAL AND METHODS: Flow-cytometric cell sorted CD8+CD45RA+CD28- and CD8+CD45RA+CD28-T-cells were obtained from patients with cervical cancer and tested for recognition of HLA-A2 restricted peptides derived from the human papillomavirus (HPV)16-E7 gene product using ELISA. HPV DNA in tumor tissue was detected by PCR. RESULTS: We show that the effector CD8+CD45RA+CD28-T-cell subset is expanded in peripheral blood lymphocytes (PBL) from patients with cervical cancer, but also in PBL from patients with an acute mycobacterial infection. CD8+T-cells from 3/6 cancer patients showed a peptide-specific immune response which could be segregated in peptide epitopes which elicited either a strong TNF-alpha production, or GM-CSF and IL-2 secretion. Peptide-reactivity could exclusively be detected in the ex vivo freshly isolated CD8+CD45RA+CD28-T-cell population. A similar situation was found to be true for HLA-A2 presented peptide epitopes derived from M. tuberculosis-associated antigens presented to T-cells obtained from patients with tuberculosis. CONCLUSIONS: The sorting of CD8+CD45RA+CD28-T-cells enables to determine the fine specificity of CD8+ effector T-cells without the need for in vitro manipulation and aids to define the most appropriate target epitopes for novel vaccine designs.     

Enhanced immunocompetent cells in chlamydial cervicitis

OBJECTIVE: To investigate changes in cell-mediated immunophenotypes by flow cytometry in endocervical secretions and peripheral blood in women with Chlamydia trachomatis infection. STUDY DESIGN: Fifty women attending the gynaecology outpatient department of Safdarjang Hospital, New Delhi, India, with signs and symptoms of cervicitis were enrolled. All patients underwent endocervical screening for C trachomatis (direct fluorescence antibody test [DFA]), and any coinfection with Candida (Gram stain), bacterial vaginosis (Gram stain), Neisseria gonorrhoeae (Gram stain), Trichomonas vaginalis (wet mount) and HIV (enzyme-linked immunosorbent assay) was ruled out. Flow cytometry was done to investigate changes in immunophenotypes in endocervical secretions and peripheral blood using monoclonal antibodies for surface markers (CD3, CD4, CD8, CD19, CD45 and CD83). Data were analyzed by chi2 test, while means were compared using Student's t test. RESULTS: C trachomatis positivity was found to be 36% (n = 18). Forty-eight patients constituted the study population since 2 patients coinfected with Candida, bacterial vaginosis and T vaginalis were excluded. A statistically significant enhancement in CD4+, CD8+ and dendritic cellular phenotypes was observed in the endocervical secretions of Chlamydia-positive patients, while B cells showed no marked difference. In the parallel study of matched peripheral blood, immunophenotypes did not show statistically significant results. CONCLUSION: Increased influx of CD4+, CD8+ and dendritic cells in the endocervix is an indication of cell-mediated immunity in response to C trachomatis infection. Local immune response in the cervical region is independent of systemic response. The mechanism by which local mucosal and systemic immune cells interact to repel or enhance susceptibility to C trachomatis infection requires further study.     

Local immune response in squamous cell carcinoma of the uterine cervix

The role of Langerhans cells as antigen-presenting cells was examined in cervical carcinomas. Frozen samples were obtained from 34 women with stage Ib and II cervical carcinomas. Langerhans cells (CD1), T lymphocytes (CD4 and CD8), B lymphocytes (CD22), and natural killer (CD57, NK) cells were all quantitatively assessed in cervical carcinomas using immunohistochemical methods. These results were related to the MHC class I and II expression on the tumor cells. The majority of Langerhans cells were distributed among cancer cells and they were positively correlated with CD4+, NK and B cells in cervical carcinomas. This is suggestive of the presence of local immune response. The numbers of Langerhans, CD4+, CD8+ and NK cells did not significantly correlate with age at operation, lymph node metastases or depth of cervical wall invasion. The downregulation of MHC class I expression found in 8 (24%) carcinomas was not associated with the decrease in the number of immunologic cells. The upregulation of MHC class II expression found in 26 (76%) carcinomas was significantly associated with the increase in the number of Langerhans cells (p < 0.007). However, the association between the upregulation of MHC-II expression and CD4+ cells did not reach statistical significance (p < 0.07). This is probably due to a small case in this study. MHC-II-restricted immunity may partly contribute to the local immune response in stages Ib and II squamous cell carcinoma of the uterine cervix.     

原发性痛经患者外周血T淋巴细胞亚群的研究

目的 :探讨原发性痛经患者月经周期中外周血T淋巴细胞亚群的变化 ,分析原发性痛经患者的免疫状况。方法 :原发性痛经患者及正常对照组各 10例 ,分别于月经期、卵泡期、排卵期、黄体期取静脉血 ,应用流式细胞仪测定血浆CD4 + 、CD8+ 百分比值。结果 :原发性痛经患者月经期、卵泡期CD4 + 百分比值及CD4 /CD8的比值显著低于正常对照组 (P <0 0 5 ) ,原发性痛经患者整个月经周期CD8+ 百分值均明显高于正常对照组 (P <0 .0 5 )。结论 :原发性痛经患者存在免疫功能低下