外周血T淋巴细胞亚群检测在胃癌病情监测及预后评价中的意义

目的：探讨外周血T淋巴细胞亚群检测在胃癌病情监测及预后评价中的价值。方法选择100例胃癌患者作为胃癌组,患者采用mFOLFOX6方案,2周为1个疗程,每位患者化疗4个疗程以上。另选择同期在医院体检的健康人群80例作为对照组,采用流式细胞仪技术检测两组外周血T淋巴细胞亚群水平。结果化疗前胃癌组外周血CD3+、CD4+、CD4+/CD8+显著低于对照组,CD8+显著高于对照组,差异有统计学意义（P﹤0.01）。Ⅰ～Ⅱ期胃癌患者外周血CD3+、CD4+、CD4+/CD8+显著高于Ⅲ～Ⅳ期,CD8+显著低于Ⅲ～Ⅳ期,差异有统计学意义（P﹤0.01）。不同疗效的胃癌患者化疗前外周血T淋巴细胞亚群水平比较差异无统计学意义（P﹥0.05）,化疗后CR+PR组、SD组、PD组外周血CD3+、CD4+、CD4+/CD8+显著降低,CD8+显著上升,差异有统计学意义（P﹤0.01）,且CR+PR组外周血CD3+、CD4+、CD4+/CD8+显著高于SD组和PD组,CD8+显著低于SD组和PD组,差异有统计学意义（P﹤0.01）。结论通过检测血清外周血T淋巴细胞亚群水平能够有助于评估胃癌的病情和预后。     

Differential depression of lymphocyte subsets according to stage in stomach cancer

Lymphocyte surface markers were determined in the peripheral blood lymphocytes (PBL) in 31 stomach cancer patients (15 males and 16 females) and 47 controls (20 males and 27 females) using an indirect immunofluorescence technique. The monoclonal antibodies used were Leu 2a (CD8, suppressor/cytotoxic T cells), Leu 3a (CD4, inducer/helper T cells), Leu 4 (CD3, pan T reagent), Leu 11 (CD16, natural killer cells) and Leu 12 (CD19, B cells). The numbers of PBL, CD3+, CD4+, CD8+, CD16+ and CD19+ cells significantly decreased and the CD4:CD8 value increased in patients with stomach cancer compared to those in healthy volunteers. In stage I, PBL, none of the PBL subsets nor the CD4:CD8 value were significantly different from those of the controls. In stage II, the numbers of PBL, CD3+, CD4+ and CD8+ cells decreased. In stage III, the CD19+ cells decreased in addition to the decreased subsets in stage II. In stage IV, PBL and all subsets measured decreased. The CD4:CD8 value showed significant increases in stages II, III and IV, because the CD8+ cells decreased to a greater extent than did the CD4+ cells. Changes in the subsets were analyzed with regard to age, sex, performance status and smoking history, no significant relation being observed between these factors and lymphocyte subsets. From the present study, we have demonstrated that lymphocyte subsets were differentially depressed in the order of T cells, B cells and natural killer cells, with progression of the stage of disease.     

食管胃交界部腺癌根治性手术对细胞免疫的影响

目的:探讨进展期食管胃交界部腺癌(AEGJ)患者围手术期外周血中淋巴细胞亚群及CD4+CD25+T细胞和程序性细胞死亡1（PD-1）在T淋巴细胞中表达的变化,了解根治性手术创伤对机体细胞免疫功能的影响及机制。 方法:126例进展期食管胃交界部腺癌行开腹根治性R0切除+D2淋巴结清除术,同期体检志愿者87例为对照组。应用流式细胞仪检测对照组和治疗组术前、术后1天、术后3天、术后7天和术后9天外周血CD3+、CD3+CD4+、CD3+CD8+、CD4+CD25+T细胞计数和PD-1在T细胞中的表达水平。结果:治疗组术前和对照组相比较,CD3+、CD3+CD4+T细胞均显著降低（P<0.05）,而CD4+CD25+T细胞和PD-1在CD4+、CD8+T细胞的表达比例均显著增高（P<0.05）。CD4+CD25+T细胞的比例在术后1天迅速下降至谷底,随后逐渐升高,于术后7天达顶峰,然后再次下降。CD4+PD-1+T细胞的比例在术后显著增加:术后第1天达到最大值,然后逐渐下降,但在术后第7天仍显著高于术前。 CD8+PD-1+T细胞的比例则在术后逐渐增加,到术后7天达高峰。结论:食管胃交界部腺癌手术创伤可能是通过各种细胞因子导致CD4+PD-1+T细胞比例改变,进而调节T淋巴细胞亚群比例,抑制细胞免疫能力。     

Increase of regulatory T cells in metastatic stage and CTLA-4 over expression in lymphocytes of patients with non-small cell lung cancer (NSCLC)

We hypothesized that the increased percentages of Regulatory T (Treg) cells, as well as over expression of Cytotoxic T Lymphocyte Antigen-4 (CTLA-4) by lymphocyte subsets might be associated with lung cancer. Accordingly, peripheral blood of 23 new cases with non-small cell lung cancer (NSCLC) and 16 healthy volunteers were investigated, by follow cytometry, for the prevalence of CD4+CD25+FoxP3+ Treg cells as well as surface (sur-) and intracellular (In-) expression of CTLA-4 by the main lymphocyte subsets (CD4+, CD8+ and CD19+). Results indicated that NSCLC patients had an increased percentage of Treg cells than controls (7.9±4.1 versus 3.8±1.8, P=0.001). The proportion of Treg cells was observed to be increased by stage increase in patients (stage II=5.2±2.4, stage III=7.9±4.4, stage IV=12.0±2.2), and also significantly higher in metastatic than non-metastatic stages (12.0±2.2 versus 6.8±3.9, P=0.023). Increase of SurCTLA-4- as well as InCTLA-4-expressing lymphocytes in patients were observed in nearly all investigated subsets, but significant differences between patients and controls were observed about InCTLA-4+CD4+ lymphocytes (8.6±7.1 and 3.8±5.3 respectively, P=0.006) as well as SurCTLA-4+CD8+ lymphocytes (0.3±0.2 and 0.2±0.1 respectively, P=0.047). In conclusion, the results suggest that immunotherapy regimen targeting CTLA-4 and Treg cells might be beneficial in lung cancer patients.     

肺癌患者外周血 T- 淋巴细胞亚群 NK 细胞的基线数值及其与预后的关系*

目的:对比肺癌患者与健康者之间淋巴细胞亚群差异,评估肺癌患者外周血T 淋巴细胞亚群及自然杀伤细胞（NK）之基线值与预后的关系。方法:收集2006年2 月至2013年3 月就诊于天津医科大学肿瘤医院病例资料完整的肺癌患者105 例,其中非小细胞肺癌（NSCLC ）86例、小细胞肺癌（SCLC）19例,另选健康对照35例,对比接受治疗前肺癌患者和健康对照者外周血中的CD3+T 细胞、CD4+T 细胞、CD8+T 细胞及NK细胞所占百分比,并回顾性分析86例NSCLC 患者初治时外周血淋巴细胞亚群与预后的关系。结果:肺癌患者外周血CD3+T 细胞、CD4+T 细胞、NK细胞及CD4/CD 8 比值均明显低于健康对照组（P = 0.011,P = 0.007,P <0.001,P=0.025）,而CD8+T 细胞比例高于健康对照组（P = 0.013）。 当CD8+T ≥ 31.8% 及CD4/CD 8< 1.28时NSCLC 患者可以获得一个更长的OS（中位OS分别为36.2 m vs . 20.0 m ,P = 0.010;30.8 m vs . 20.0 m ,P = 0.035）。 而CD3+T 细胞、CD4+T 细胞及NK细胞百分比对NSCLC 患者预后无显著影响。结论:外周血CD8+T 细胞基线水平较高的NSCLC 患者生存较长,此基线水平可能对NSCLC 患者预后有指示作用。      

Influence of perioperative whole blood transfusions on lymphocyte subpopulations in patients with stage II breast cancer

Preliminary reports have suggested an adverse relationship between blood transfusion and survival after surgery in patients with solid tumour. One might postulate that from these studies, perioperative blood transfusions alter host immune defences. We therefore examined the influence of homologous whole blood transfusion on circulating lymphocyte subpopulations in transfused patients compared with non-transfused patients. Fifty-one women with Stage II breast cancer who underwent surgical procedures were studied. Patients were classified into two groups on the basis of whether or not they had received blood transfusion. The lymphocyte subpopulations were analyzed by flow cytometry before cancer surgery and three weeks after the operation. CD3+, CD4+, CD8+, and CD20+ cells as the lymphocyte subsets were quantitated using appropriate monoclonal antibodies. No significant differences between pre- and postoperative lymphocyte subset levels were seen in non-transfused patients. However, there was a statistically significant increase in the CD8+ cell count; decreasing CD4+ cell count and decreased CD3+ cells levels were observed in the transfused group (P < 0.05). Although these early results of the study suggest that the blood transfusions could be associated with alterations in lymphocyte populations, additional studies are needed to elucidate the possible mechanism of the transfusion-induced immunological modulations.     

Impact of topotecan-based chemotherapy on the immune system of advanced ovarian cancer patients: an immunophenotypic study

The effects of topotecan-based chemotherapy (CT) on peripheral blood lymphocyte (PBL) subsets were evaluated in ovarian cancer patients. Fourteen patients with epithelial ovarian cancer, at the diagnosis or relapsed after platinum-based CT, were treated with: a) topotecan in association with carboplatin and taxanes as first line CT; b) topotecan alone or c) topotecan in association with carboplatin both as second line of treatment after platinum. The phenotype of PBL was determined before starting treatment and immediately before each CT course by flow cytometric analysis. Before starting CT, the absolute number of lymphocytes and the CD2+, CD3+, CD4+ subsets were significantly lower in pre-treated patients and not significantly altered in CT-naive patients with respect to a cohort of 20 healthy donors utilized as control. Lymphocytes co-expressing CD4+/CD8+ were significantly higher in both subgroups of patients than in normal donors. CD4+/CD45RA+ and CD4+/CD45RO+ subsets were significantly decreased in pre-treated patients and normal in CT-naive patients. CD3+/HLA-DR+ T cell population significantly increased in CT-naive patients at baseline. During CT and after its discontinuation, no relevant changes were recorded for both subgroups of patients with respect to the baseline in lymphocyte absolute count, CD2+, CD3+, CD4+, CD4+/CD45RO+ subsets, while CD4+/CD45RA+ subpopulation was significantly decreased in CT-naive patients. CD8+, CD19+, CD20+, CD16+, CD56+, CD2+/CD25+ subsets did not differ statistically comparing to normal donors both at baseline and during CT. The treatment was well tolerated and no patient developed non-neutropenic infection. Topotecan-based therapy does not have a negative impact on PBL in either CT-naive or in pretreated ovarian cancer patients. This information should be considered when utilizing topotecan with other anticancer drugs in the adjuvant setting as well as when dose-intensification of topotecan with stem cell support is planned.     

肿瘤患者外周血免疫细胞亚群和有关细胞因子含量的相关性研究

目的:评价血液中CD3+、CD4+、CD8+、CD19+、自然杀伤（NK）细胞含量和细胞因子肿瘤坏死因子（TNF）、白介素-2（IL-2）、IL-6、IL-10的相关性。方法:对80例不同肿瘤患者和20例健康对照者进行细胞亚群和细胞因子检测。使用流式细胞仪测定样本外周血CD3+、CD4+、CD8+、NK、CD19+细胞比例,同时使用酶联免疫吸附试验测定外周血中TNF、IL-2、IL-6、IL-10水平。应用配对资料t检验分析肿瘤患者和正常对照者的细胞亚群和细胞因子有无差异,应用直线相关性t检验分析各细胞亚群和细胞因子之间有无线性相关。结果:肿瘤患者CD8+细胞含量高于正常对照者,NK细胞含量低于正常对照者,血液中CD3+细胞含量和细胞因子TNF、IL-2、IL-6、IL-10均无相关性,CD4+、CD8+细胞含量和IL-2线性相关,NK细胞含量和IL-2、IL-10线性相关,CD19+细胞含量和IL-6线性相关（但差异无统计学意义）。结论:肿瘤患者细胞免疫状况和正常人有统计学差异,细胞因子未发现统计学差异;免疫细胞亚群和细胞因子有一定线性相关。     

TACE及MWA治疗对肝癌患者免疫功能的影响

目的:探讨肝动脉灌注化疗栓塞（TACE）及微波消融(MWA)对肝癌患者免疫系统的影响及临床意义。方法:搜集我院116例肝癌患者完整资料,据治疗方式不同分组:单纯接受MWA治疗40例为研究组,单纯接受TACE治疗76例为对照组;两组分别于术前2天、术后3天取外周血,测定T细胞亚群（CD3+、CD4+、CD8+、CD4+/CD8+、NK细胞、CD19+）及体液免疫（IgG、IgM、IgA、C3、C4）水平。结果:MWA组治疗前后CD3+、CD4+、CD56+细胞比例上升,CD8+、CD19+细胞比例下降,差异均无统计学意义（P＞0.05）,CD4+/CD8+比值上升,差异有统计学意义（P＜0.05）;TACE组治疗前后CD3+、CD8+细胞比例上升,CD4+、CD56+细胞比例和CD4+/CD8+比值下降,差异有统计学意义（P＜0.05）,CD19+细胞比例下降,差异无统计学意义（P＞0.05）。两组手术治疗前后IgG、IgM、IgA、C3、C4定量均下降,但MWA组治疗前后差异无统计学意义（P＞0.05）,TACE组治疗前后差异有统计学意义（P＜0.05）。结论:TACE明显抑制肝癌患者免疫功能,MWA能增强肝癌患者主要抗肿瘤免疫;TACE联合MWA同时治疗基础上辅以免疫治疗可能是治疗中晚期肝癌患者疗效更明显的手段之一。     

肺癌患者化疗前后免疫细胞格局的改变及其临床意义

目的〖HT5"SS〗：研究肺癌患者化疗前后免疫细胞数量、亚群比例、细胞表型及功能的改变,并探讨其临床意义。〖HT5W〗方法：〖HT5"SS〗 对23例次肺癌患者化疗前后的白细胞、淋巴细胞和中性粒细胞计数,以流式细胞术对CD3+、CD4+及CD8+T细胞亚群比例以及记忆样表型T细胞进行检测和分析;体外以PHA刺激外周血淋巴细胞,培养48 h后检测对K562靶细胞的杀伤效应。〖HT5W〗结果： 〖HT5"SS〗化疗后外周淋巴细胞数量迅速减少,1周左右达最低水平,此后逐渐恢复至化疗前水平;淋巴细胞恢复过程中, CD3+、CD8+T细胞亚群比例升高,记忆样表型T细胞（CD44high,CD62Llow）比例增加;增生期淋巴细胞在体外以PHA刺激后对K562细胞的杀伤能力没有下降,个别患者的杀伤能力还有所增强。〖HT5W〗结论： 〖HT5"SS〗肺癌患者化疗后不仅没有降低免疫功能,反而可增强其免疫效应,该结果为临床肿瘤患者化疗后开展免疫生物治疗提供了实验依据。     

Decreased absolute counts of T lymphocyte subsets and their relation to disease in squamous cell carcinoma of the head and neck.

PURPOSE: Apoptosis of circulating CD8+ T cells seen in patients with squamous cell carcinoma of the head and neck [SCCHN (Hoffmann T, et al. Clin Cancer Res 2002;8:2553-62)] suggested a possibility of lymphocyte imbalance. Therefore, absolute numbers and percentages of lymphocyte subsets were examined in the peripheral blood of SCCHN patients and controls. EXPERIMENTAL DESIGN: Venous blood was obtained from 146 patients with SCCHN and 54 normal volunteers. Absolute numbers of CD3+, CD4+, and CD8+ T lymphocytes were determined using fluorobeads in a flow cytometry-based technique. Percentages of T lymphocyte subsets were also evaluated by flow cytometry. The patients were grouped at the time of blood draw [active versus no evidence of disease (NED), type of therapy administered, and the length of follow-up]. RESULTS: Patients with SCCHN had significantly lower absolute numbers of CD3+ CD4+, and CD8+ T cells than normal controls. However, no differences in the percentages of T-cell subsets between patients and normal controls were observed. Patients with active disease had significantly lower CD3+ and CD4+ T-cell counts than those with NED. Patients who had NED after surgery and radiotherapy had the lowest T-cell counts among the NED cohort. Patients who had NED for >2 years did not recover their T-cell counts, and the T-cell imbalance was evident many years after curative surgery. The tumor-node-metastasis (TNM) stage or site of the disease was not related to the absolute T-cell count. Patients with recurrent disease at the time of blood draw tended to have the lowest CD4+ T-cell counts. CONCLUSIONS: Patients with SCCHN have altered lymphocyte homeostasis, which persists for months or years after curative therapies.     

初诊肺癌患者多次化疗外周血免疫相关指标动态变化的研究

  目的  探索免疫治疗与化疗相结合的有利切入点,为临床免疫治疗介入提供试验依据。  方法  收集2015年11月至2016年12月新乡医学院第一附属医院初诊的23例肺癌患者,全部患者均完成连续5个周期的化疗,采用流式细胞术方法检测化疗过程中患者外周血免疫相关指标如CD4+T细胞、CD8+T细胞、CD19+B细胞和CD16+、CD56+、NK细胞比率,T细胞表面共信号分子PD-1、PD-L1、CD137、CTLA-4、CCR-4、LAG-3以及细胞因子表达情况。分析多周期过程中,上述指标动态变化趋势。  结果  在肺癌患者多个周期的化疗过程中,机体CD8+T淋巴细胞、CD19+B淋巴细胞和CD16+、CD56+、NK细胞水平下调,CD4+T淋巴细胞数量增加,差异具有统计学意义（P＜0.05）;T细胞表面共抑制分子PD-1、CTLA-4和CCR-4随治疗进行表达下调,差异具有统计学意义（P＜0.05）。  结论  肺癌患者在多个周期的化疗中,实施淋巴细胞亚群数量及T细胞表面共抑制分子监测具有重要意义,化疗中后期针对免疫检查点高表达患者,联合应用PD-1抗体、PD-L1抗体、CTLA-4抗体或CCR-4抗体可能具有更优的治疗效果。      

老年肾癌患者血清T淋巴细胞亚群变化对其预后及生存的影响

目的:探讨分析老年肾癌患者血清T淋巴细胞亚群变化对其预后及生存的影响。方法:选择2013年4月至2015年3月我院收治的78例老年肾癌患者，再选择同期来我院体检的健康人群50例作为对照组，术前以及术后2周采用流式细胞技术检测肾癌患者外周血T淋巴细胞亚群水平变化情况，并与对照组比较。对患者进行随访分析，采用Kaplan-Meier生存分析术后不同T淋巴细胞亚群水平对患者生存预后的影响。结果:肾癌组患者术前、术后2周外周血CD3+、CD4+以及CD4+/CD8+水平均显著低于对照组（P＜0.05）。肾癌组术后2周外周血CD3+、CD4+以及CD4+/CD8+水平较术前有显著升高（P＜0.05）。CD3+>48%肾癌患者总体生存情况显著优于CD3+≤48%患者（P＜0.05）。CD4+>30%肾癌患者总体生存情况显著优于CD4+≤30%患者（P＜0.05）。CD8+>25%肾癌患者总体生存情况与CD8+≤25%患者生存情况无差异（P＞0.05）。CD4+/CD8+>1.0肾癌患者总体生存情况显著优于CD4+/CD8+≤1.0患者（P＜0.05）。结论:老年肾癌患者存在广泛的细胞免疫功能紊乱现象，手术治疗后能够有效去除瘤负荷而从一定程度上逆转免疫功能，术后免疫功能指标可反映患者预后情况，免疫功能状态越差的患生存情况越差。     

联合检测外周血T淋巴细胞AgNOR活性及T淋巴细胞亚群评估食管癌围手术期细胞免疫变化规律

背景与目的食管癌患者存在着细胞免疫功能抑制,测定T淋巴细胞核仁区的AgNOR活性及T淋巴细胞亚群,可以了解T淋巴细胞的免疫活性状态,对研究肿瘤的生物学行为具有重要意义。本研究旨在通过动态观察食管癌患者围手术期外周血T淋巴细胞AgNOR的活性及T淋巴细胞亚群的变化规律,研究细胞免疫机能状态及手术对食管癌患者细胞免疫功能的影响。方法分别抽取75例健康人外周血和70例食管癌患者术前、术后第3天、术后第9天的外周血,利用KL型肿瘤免疫图像分析系统检测AgNOR活性,流式细胞术分析T淋巴细胞亚群。结果食管癌患者术前AgNOR活性[(5.5±0.8)%]明显低于健康人[(7.1±0.8)%](P<0.00),CD3 、CD3 CD4 无明显变化(P>0.05),CD4 CD25 、CD4 /CD8 明显上升(P<0.05),CD3 CD8 、CD8 CD25 亚群明显下降(P<0.00)。术后第3天外周血T淋巴细胞AgNOR、CD3 、CD3 CD4 与CD4 /CD8 最低,术后第9天最高;CD3 CD8 亚群、CD4 CD25 亚群、CD8 CD25 亚群则从第3天开始逐渐恢复,术后第9天最高。食管癌患者术后第9天CD3 、CD3 CD4 与健康人相比无显著性差异(P>0.05);外周血T淋巴细胞AgNOR、CD3 CD8 与CD8 CD25 仍未恢复到健康人水平(P<0.05);CD4 CD25 仍明显高于健康人(P<0.00)。结论手术创伤引起食管癌患者暂时性细胞免疫抑制,但从第3天开始可观察到细胞免疫功能恢复,术后第9天继续恢复,但仍未恢复到正常水平。     

晚期肺腺癌患者一线化疗后T淋巴细胞亚群变化及临床意义

背景与目的机体的免疫功能异常与恶性肿瘤的发生、发展、转移及预后密切相关。T淋巴细胞亚群是反映细胞免疫功能的重要指标之一。本实验研究晚期肺腺癌患者一线化疗后外周血T淋巴细胞数量的动态变化,探讨化疗后机体免疫状态变化的动态过程,为制定化疗联合免疫治疗方案提供实验依据。方法 49例经病理学确诊的IIIb期-IV期肺腺癌患者,与33例正常人比较外周血T淋巴细胞数量。然后患者随机进入2个实验组,分别采用培美曲塞、顺铂联合方案及多西他赛、顺铂联合方案化疗,应用流式细胞仪检测化疗前后不同时间点淋巴细胞的组成。结果肺癌患者的CD3+、CD3+CD4+、CD4+CD25+等T细胞的数量与健康对照组比较存在差异,P值分别为0.012,0.034和0.006;化疗后第4天及第7-10天CD3+、CD3+CD4+比例升高,至第21天逐渐恢复至治疗前水平;2个化疗组比较CD3+细胞比例均升高,而培美曲塞组第4天CD3+、CD3+CD4+、CD4+/CD8+升高,CD3+CD8+及CD8+CD28-比例降低,差异均具有统计学意义。部分缓解患者较早期疾病进展患者的第4天及第7-10天CD3+CD4+细胞升高;第4天CD3+CD8+、CD8+CD28-细胞降低。结论晚期肺腺癌患者免疫功能处于抑制状态。化疗后第4天免疫功能得到一定程度恢复,至第21天恢复至治疗前水平。培美曲塞似乎对化疗后短期内免疫功能的改善有更明显的作用。化疗后免疫格局的改变可能与预后有关。     

细胞因子诱导的杀伤细胞对恶性肿瘤患者细胞表型的影响

目的:探讨细胞因子诱导的杀伤细胞(cytokine induced killer cells,CIK cells)治疗对恶性肿瘤患者淋巴细胞亚群的影响,对比接受CIK细胞免疫治疗前后,培养细胞表型及患者淋巴亚群的变化.方法:选取辽宁省肿瘤医院手术、放化疗治疗结束1个月以上或无法进行以上治疗,单纯接受4个周期CIK细胞免疫治疗的恶性肿瘤患者67名入组.应用流式细胞术检测分析接受4周期CIK细胞免疫治疗前后患者淋巴细胞CD3+CD4+、CD3+CD8+、CD4+/CD8+、CD3+CD56+、CD3-CD56+、CD3+PD1+亚群的变化情况.同时对各次输注细胞表型(CD3+CD4+、CD3+CD8+、CD3+CD56+)进行检测,观察其变化.结果:患者接受4周期CIK细胞免疫治疗后,与治疗前相比,淋巴细胞CD3+CD4+、CD3+CD8+、CD4+/CD8+、CD3+CD56+亚群略有升高,但无统计学差异.CD3-CD56+、CD3+PD1+亚群无明显变化.值得注意的是,输注细胞表型与第一次培养后相比,第二次、第三次的CIK细胞CD3+CD8+、CD3+CD56+表型呈阶梯型上升,结果有统计学差异.第三次与第四次无明显变化.结论:CIK细胞免疫治疗能稳定肿瘤患者免疫状态.接受过CIK细胞回输的患者,再次抽取外周血进行培养时,可增加体内CD3+CD56+前体细胞增殖和定向分化的能力,有望得到远期获益.     

FOLFOX方案化疗对结直肠癌患者免疫细胞数的影响

背景与目的：化疗在杀伤肿瘤的同时,也损害了机体的正常免疫。很多文献曾报道晚期肠癌的化疗,常使细胞免疫功能抑制加重。本研究探讨FOLFOX方案即奥沙利铂联合亚叶酸钙及氟尿嘧啶联合化疗对结直肠癌患者免疫细胞数的影响及与健康人的差异。方法：80例结直肠癌患者行FOLFOX方案化疗（奥沙利铂85mg/m2,静脉滴注,第1天;亚叶酸钙200mg/m2,静脉滴注,第1天;氟尿嘧啶400mg/m2,第1天,2400mg/m2,持续静脉滴注46h）,2周1次,2次为1个疗程,采用流式细胞仪测定化疗前及化疗后2周、4周外周血T淋巴细胞亚群和NK细胞的活性,比较化疗前后的变化,同时根据临床分期进行亚组分析比较,以健康人作对照分析。结果：本组结直肠癌患者第1天、第2周及第4周外周血CD3＋、CD4＋、CD8＋、CD4＋/CD8＋及NK细胞活性比较均无显著下降（P〉0.05）,但患者外周血CD3＋、CD4＋、NK细胞数量及CD4＋/CD8＋比值与健康人相比下降,而CD8＋细胞比例显著升高,差异有显著性（P〈0.05）,提示免疫功能较健康人下降,而且外周血T淋巴细胞亚群和NK细胞数量的改变与结直肠癌临床病理分期有关,分期越晚,CD3＋、CD4＋及NK细胞数量CD4＋/CD8＋细胞比值越低,CD8＋细胞比例越高;Ⅰ、Ⅱ期结直肠癌患者与Ⅲ、Ⅳ期患者之间差异有显著性（P〈0.05）。结论：FOLFOX方案治疗结直肠癌疗效肯定,可改善患者生存质量,而且对机体免疫力影响小。检测淋巴细胞亚群对判断患者的免疫功能、预测肿瘤患者的预后以及指导临床是否需要应用免疫增强剂均有重要意义。     

外周调节性细胞在接受非小细胞肺癌放疗患者中的运用

  目的  探讨外周调节性T细胞、NK细胞及T淋巴细胞亚群在接受放疗的非小细胞肺癌(non-small cell lung cancer, NSCLC)患者中的预测价值。  方法  选择2016年1月至2017年12月九江市第三人民医院105例NSCLC患者及同期体格检查的45例健康受试者进行研究, 其中62例为初诊患者, 43例为复发患者。抽取患者空腹外周静脉血对其淋巴细胞亚群进行检查, 并采用单因素与多因素分析NSCLC无进展生存的影响因素。  结果  与健康对照组相比, NSCLC患者CD4+CD25+Foxp3+调节性T细胞的比例显著升高(P < 0.05); NSCLC患者复发时CD4+CD25+Foxp3+调节性T细胞高表达与初诊患者相比得到进一步验证, CD19+B细胞、CD4+T细胞和CD4/CD8比值显著下降, NSCLC患者CD8+T细胞和CD8+CD28-T细胞明显增加(P < 0.05), 而NK、NKT、γδT、CD3+、CD8+ CD28+T细胞在患者与对照组之间无显著性差异(P > 0.05)。40例复发患者中, 转移患者与复发患者相比, CD4+CD25+Foxp3+调节性T细胞增多; CD4+T细胞比例、CD4/CD8比值下降, 差异具有统计学意义(P < 0.05)。CD4+CD25+Foxp3+调节性T细胞比例高与无进展生存期相关(HR=2.55, 95%CI:1.07~6.11, P=0.019)。其他淋巴细胞亚群与无进展生存期之间无显著性关联。除淋巴细胞亚群外, 其中性粒细胞/淋巴细胞比率与无进展生存期呈负相关(HR=2.66, 95%CI:1.01~7.05, P=0.033), 淋巴细胞与无进展生存期呈正相关(HR=0.42, 95%CI:0.17~1.03, P=0.042)。根据临床病理特点, NSCLC患者的肿瘤分化程度、T分期、N分期与临床预后相关(P < 0.05)。多因素分析结果显示, 无进展生存的独立性影响因素为CD4+CD25+Foxp3+调节性T细胞及N分期(P < 0.05)。疾病进展患者调节性T细胞高表达比例显著高于疾病稳定、部分缓解及完全缓解的患者(P < 0.05)。通过受试者工作特征曲线(ROC)对外周调节性T细胞预测NSCLC无进展生存的诊断效能进行分析, AUC为0.915, 当最佳阈值取2.85时, 外周调节性T细胞预测NSCLC患者无进展生存的预测敏感性为87.7%, 特异性为71.4%。  结论  NSCLC放疗患者外周血CD4+CD25+Foxp3+调节性T细胞增多, 是NSCLC进展的独立性影响因素, 具有预测NSCLC患者预后的作用。      

肺癌患者淋巴细胞亚群在外周血中的表达及其与预后的关系

背景与目的肺癌是最常见的恶性肿瘤之一,本研究旨在探讨肺癌患者外周血中淋巴细胞亚群的表达及与预后的关系。方法采用流式细胞仪检测221例原发性肺癌首诊患者外周血淋巴细胞亚群CD3+、CD4+、CD8+、CD4+/CD8+、CD19+、CD25+、CD44+及NK细胞所占比例,并与96例健康人的血标本对比,结合临床及随访资料进行统计分析。结果与健康对照组对比,肺癌患者淋巴细胞亚群8项指标中CD3+及CD8+明显低于健康对照组,CD4+/CD8+、CD19+、CD25+、CD44+及NK细胞明显高于健康对照组(P<0.05)。与非小细胞肺癌(non-small cell lung cancer,NSCLC)相比,小细胞肺癌(small cell lung cancer,SCLC)的CD8+明显升高而CD4+和CD4+/CD8+明显下降(P<0.05)。化疗后与化疗前相比CD3+明显上升,NK细胞、CD19+及CD44+明显下降(P<0.05),其中CD44+在化疗后表达不升高者有生存优势(P=0.021),而其余3项指标与患者预期生存无关。结论肺癌患者外周血淋巴细胞亚群普遍发生改变,CD44+在化疗后的改变可能与预后相关。     

自体外周血造血干细胞移植过程中淋巴细胞亚群和CD34+细胞变化的动态研究

目的动态观察动员、采集过程中外周血及造血干细胞中的淋巴细胞亚群和造血干细胞含量变化,及时指导临床选择最佳采集时机。方法 采用流式细胞术(FCM)测定大剂量化疗(HDC)联合重组人粒细胞集落刺激因子(rhG-CSF)对47例恶性实体瘤患者自体外周血造血干细胞移植(APBSCT)过程中,外周血和外周血造血干细胞(PBPC)中的淋巴细胞亚群CD3、CD4、CD8、NK、CD19和造血干细胞CD34+及其亚群CD34+CD38-、CD34+ Thy1+、AC133+细胞含量的变化,同时用体外集落培养的方法评价干细胞克隆形成能力。结果 动员后外周血淋巴细胞亚群CD3、CD4、CD8、NK和CD19细胞含量均低于动员前,其中CD3、CD8含量明显低于基础状态(P=0.007,P=0.016),而动员后外周血中的CD34及其亚群含量均明显高于动员前(P<0.05)。动员后中位时间第16天(15～17 d)外周血中的CD34含量达到最高峰,且第1次采集物中的造血干细胞含量最高。PBPC中除CD4、CD4/CD8明显低于外周血外(P<0.000 5),其他淋巴细胞亚群含量与外周血比较无明显改变。外周血单个核细胞中CD34+细胞含量与PBPC中CD34+细胞、CD34+CD38-及粒/单系集落形成单位(CFU-GM)、红系爆式集落形成单位(BFU—E)均存在显著相关性(P<0.05),而与采集的单个核细胞(MNC)总数、CD34+Thy1+、AC133+细胞含量间无相关关