Moroccan lymphocyte subsets reference ranges: age,gender, ethnicity,and socio-economic factors dependent differences

ABSTRACT

Lymphocyte subsets reference ranges are helpful for a precise diagnosis and therapy of various diseases. We attempted in the current study to establish Moroccan lymphocyte reference range and reveal age, gender, ethnicity, income, and instructional levels dependent differences. Lymphocyte subsets percentage and absolute count were determined by 4-color flow cytometry in a population study of 145 adults Moroccan healthy volunteers. Analysis showed significant age-dependent changes. Age was associated with a decrease of naïve CD4+ and CD8+ T cells and an increase of memory CD4+ or CD8+ T cells. Activated CD4+ CD38+ and CD8+ CD38+ T cells, Treg as well as NK cell showed age-dependent alterations. In contrast, B cells remained unchanged. A higher percentage of CD3+ and CD4+ T cells was observed in females while CD8+, B and NK cells count were higher in men. Ethnicity, instructional levels, and personal income seem to not influence lymphocyte subsets reference values. This study provides reference ranges for lymphocyte subsets of healthy Moroccan adults. These results can be used for other North African (Maghrebian) countries considering their geographic, ethnic, economic, and cultural similarities.     

应用四色荧光标记流式细胞术检测外周血淋巴细胞亚群的变化

目的 探讨四色荧光标记流式细胞术(Flow cytometry,FCM)检测鼻咽癌患者服用云芝丹参胶囊后外周血淋巴细胞亚群:辅助性T淋巴细胞(CD4 ),抑制性和细胞毒T淋巴细胞(CD8 ),B淋巴细胞(CD19 ),NK(CD16 CD56 ) 细胞的绝对数和百分率等的变化.方法 收集鼻咽癌患者27例,设立中药组和安慰剂对照组,采用四色荧光标记流式细胞术对服用云芝丹参胶囊后的鼻咽癌患者外周抗凝全血的淋巴细胞亚群CD4 细胞、CD8 细胞、B细胞、NK细胞进行绝对计数和相对计数,并对两组结果进行比较分析.结果 四色荧光标记流式细胞术结果显示:接受放射治疗的鼻咽癌患者在服用云芝丹参胶囊16周后,外周血T淋巴细胞的绝对数和百分率,以及Ts、Th细胞绝对数的下降均明显低于安慰剂组(P＜0.05);中药组和安慰剂组NK细胞百分率的变化均显著增高(P＜0.05); 但两组NK细胞绝对值的变化不存在明显差异(P＞0.05).结论 云芝丹参能明显减轻放射治疗对鼻咽癌患者的淋巴毒性.     

Heart rate variability in healthy subjects is related to age and gender

The effects of age and gender on heart rate variability as measured by spectral and time domain analysis of 24 h ECG recordings were evaluated in 101 healthy subjects, 49 men and 52 women (20-69 years of age). In the frequency domain, total power, very low-frequency power, low-frequency power and high-frequency power were negatively correlated to age (P < 0.001 for all variables). Total power decreased by 30% between 20-29 and 60-69 years of age. In the time domain, SDNN-index, the mean of the standard deviations of all normal R-R intervals for all 5 min segments of a 24 h ECG recording, was negatively correlated to age (P <0.001). Total power, very low-frequency power, low-frequency power and the low-frequency/high-frequency ratio were lower in women (P < 0.05, P < 0.05, P < 0.01 and P < 0.01), although the absolute differences were much smaller than for age. There was a pronounced circadian variation; at night total power increased in all age groups (P < 0.01). The results show that age, and to a lesser degree gender, are important determinants of heart rate variability in healthy subjects. Heart rate variability is a valuable tool for risk stratification in cardiovascular disease, but the physiological effects of ageing, with diminishing heart rate variability in older age groups, must also be taken into account.     

海拔4500m地区藏、汉人群外周血淋巴细胞亚群检测

目的探讨高原缺氧对机体免疫功能及其免疫调节功能的影响。方法采用流式细胞术对高海拔地区(4 500 m)52名健康汉族人群和35名健康藏族人群进行了淋巴细胞亚群CD4+、CD8+、CD3-CD19+(B细胞)、CD3-CD56+(NK细胞)测定,并与中度海拔(西宁)地区45名健康汉族人群进行了比较。结果 2个海拔的CD4+、CD8+、CD3-CD19+、CD3-CD56+各项中,4 500 m人群CD8+高于西宁组,CD4+明显低于西宁组,P<0.05;CD19+(B细胞)西宁组高于高海拔组,P<0.05;海拔4 500 m汉族、世居藏族人群的CD4+、CD8+无统计学意义比较,P>0.05;CD3-CD56+(NK)藏族低于汉族,P<0.05;CD3-CD19+(B细胞)藏族高于汉族,P<0.05。结论高原低氧环境影响机体的免疫调节功能。     

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。     

Cell-type specific response of peripheral blood lymphocytes to methotrexate in the treatment of rheumatoid arthritis

The mode of action of methotrexate in the treatment of rheumatoid arthritis is still questionable. Although in vitro results suggest an immunosuppressive effect of methotrexate, several clinical studies have failed to confirm these effects in patients treated with oral low-dose methotrexate. With respect to the highly variable bioavailability of methotrexate, we investigated the effects of an intravenous administration of 15 mg methotrexate per week on peripheral blood lymphocyte subsets in eight patients with rheumatoid arthritis. Methotrexate after 12 weeks significantly (P<0.01) reduced total peripheral blood lymphocytes and led to a pronounced redistribution of lymphocyte subsets with a preferred reductive effect on B-lymphocytes (P<0.005) and T-lymphocytes (P<0.05). Natural killer cells and killer cell-like T cells, on the other hand, were unaffected by the treatment. Our results suggest a cell-type specific effect of intravenously administered low-dose methotrexate on peripheral blood lymphocytes. This effect, in our opinion, may contribute to the mode of action of methotrexate as an immunosuppressive drug in the treatment of rheumatoid arthritis.Abbreviations NSAID nonsteroidal anti-inflammatory drug - DMARD disease-modifying antirheumatic drug - MTX methotrexate - RF rheumatoid factor - CRP C-reactive protein - BSR blood sedimentation rate - NK cells natural killer cells - VAS visual analog scale Correspondence to: T.C. Wascher     

Lymphocyte subsets in the peripheral blood of patients with multiple myeloma and benign monoclonal gammopathy

Summary A panel of monoclonal antibodies that identify various antigens present on T and B cells was used to characterize circulating blood lymphocyte subsets in multiple myeloma [(MM) - 13 patients] and benign monoclonal gammopathy [(BMG) - 5 patients]. In MM and BMG an increase in B cell proportions (BA 1 positive cells) was observed, whereas T cells (Lyt 3 positive cells) were reduced compared to normal controls. However, with respect to the T cell subset distribution, a marked diversity between MM and BMG was noted. This may help to differentiate BMG from MM. In MM a relative decrease in inducer/helper T cells (OKT 4 positive cells) and increase in suppressor/cytotoxic T cells (OKT 8 positive cells) as well as in NK/K T cells (Leu 7 positive cells) was observed. On the other hand, in BMG the relative T cell subset distribution was comparable to those seen in normal subjects.Supported in parts by the Deutsche Forschungsgemeinschaft (Ru 215/2)     

Dysregulation of blood lymphocyte subsets and natural killer cells in schistosomal liver cirrhosis and hepatocellular carcinoma

Abstract. Immunological factors are important in the pathogenesis of a wide spectrum of hepatobiliary diseases. Using flow cytometry, we determined the changes in lymphocyte subsets and natural killer cells in 123 individuals (81 patients with liver disease and 42 healthy volunteers). The liver diseases included periportal fibrosis (PPF, 10 patients), liver cirrhosis (LC, 31 patients), and hepatocellular carcinoma (HCC, 40 patients). Schistosomiasis and viral hepatitis B and C were the putative etiological agents of liver diseases. Immunophenotyping by indirect immunofluorescence was conducted using monoclonal antibodies to CD3 (T-lymphocytes), CD4 (helper/inducer T-cells), CD8 (suppressor/cytotoxic T-cells), and CD57 (natural killer cells) cell surface markers. Immunophenotyping of PPF patients showed no significant changes in all markers compared with the healthy controls. However, there was a significant decrease (P<0.01) in CD3 and CD4 T-cells, and a highly significant increase (P<0.001) in CD57 T-cells in patients with LC or HCC. In addition, LC and HCC patients showed no significant change in CD8 T-cells compared with controls. In conclusion, the progression of liver diseases is associated with a dysregulation of cellular immune responses. T-lymphocytes and natural killer cells may play a role in the immunopathogenesis of liver cirrhosis and HCC.     

乙型流感患者外周血白细胞和淋巴细胞亚群变化分析

目的 探讨乙型流感患者外周血白细胞和淋巴细胞亚群的变化特点,为乙型流感的诊断、治疗和预后判断提供实验室依据.方法 采用全血细胞分析和流式细胞分析法分别检测47例乙型流感患者急性期和恢复期的外周血白细胞及淋巴细胞亚群,并与38名健康人比较.结果 乙型流感轻症患者外周血白细胞总数在急性期显著下降,恢复期迅速上升,而重症患者外周血白细胞总数、中性粒细胞百分比及绝对值在急性期显著上升,恢复期迅速下降;所有乙型流感患者淋巴细胞总数、CD3、CD4、CD8、CD19在急性期显著下降,恢复期迅速上升;轻症患者的NK细胞绝对值在急性期与恢复期都与健康对照组无显著差异,而重症患者NK细胞绝对值在急性期显著下降,恢复期迅速上升.结论 乙型流感患者在急性期出现白细胞总数及中性粒细胞的显著上升,而NK细胞绝对值的大幅降低可以提示病情重症化倾向.     

AFP-L3阳性肝细胞癌患者肿瘤分期和外周血淋巴细胞亚群的研究

目的: 分析小扁豆凝素结合型甲胎蛋白(AFP-L3)阳性的肝细胞癌患者肿瘤分期及外周血淋巴细胞的改变,探讨AFP-L3水平与肿瘤分期及免疫状态的关系。方法:采集60例肝细胞癌患者外周血,检测AFP及AFP-L3含量。根据外周血AFP-L3比例将60例原发性肝细胞癌患者分为2组:AFP-L3(+)组和AFP-L3(-)组。流式细胞术检测外周血中各淋巴细胞亚群(NK、CD4⁺、CD8⁺、CD4/CD8、Treg)的比例。结果:AFP-L3(+)组患者UICC分期Ⅲ+Ⅳ期为29例,AFP-L3(-)组为7例;AFP-L3(+)组患者BCLC分期C+D期为20例,AFP-L3(-)组为3例。AFP-L3(+)组患者发生血管侵犯为17例,远高于AFP-L3(-)组(1例)。AFP-L3(+)组患者外周血中NK细胞比例及CD4/CD8比值下降,CD8⁺T和Treg细胞显著比例上升。结论:外周血AFP-L3阳性的肝癌患者容易发生血管侵犯,且分期较差,可能与淋巴细胞亚群发生变化、免疫系统受到抑制有关。     

类风湿性关节炎患者2H4+T淋巴细胞亚群的流式细胞仪分析

了解类风湿性关节炎患者免疫功能的异常变化，探讨其发病机制。方法，使用间接免疫荧光和直接免疫荧光方法对１１例ＲＡ患者和１２例健康人的外周血Ｔ淋巴细胞进行了研究。结果：ＲＡ患者与对照组比较ＣＤ３＾＋Ｔ细胞和ＣＤ＾＋４Ｔ细胞地明显变化，ＣＤ８＾＋Ｔ细胞明显减少，     

甲型H1N1流感患者外周血白细胞和淋巴细胞亚群变化分析

目的 探讨甲型H1N1流感患者白细胞和淋巴细胞亚群的变化特点,为甲型H1N1流感的诊断、治疗和预后判断提供实验室依据.方法 采用全血细胞分析和流式细胞分析法分别检测59例甲型H1N1流感患者急性期和恢复期的外周血白细胞及淋巴细胞亚群,与43名健康人和24例普通甲型流感患者比较.结果 甲型H1N1流感轻症患者白细胞总数在急性期显著下降,重症患者白细胞总数下降不显著,而中性粒细胞显著升高;与普通甲型流感相似,所有H1N1流感患者淋巴细胞总数、CD3、CD4、CD8百分比和绝对值在急性期大幅度下降,恢复期迅速回升;而重症患者NK和NKT细胞绝对值在急性期下降幅度超过20%.结论 甲型H1N1流感患者白细胞和淋巴细胞亚群变化与普通甲流相似;急性期NK、NKT细胞绝对值的大幅降低可以提示病情重症化倾向.     

HAART对人类免疫缺陷病毒感染者淋巴细胞亚群的影响

应用流式细胞分析法通过分析检测41例AIDS患者与30例健康对照之间,以及21例AIDS患者在接受HAART治疗前后外周血淋巴细胞亚群的表达情况,研究感染HIV病毒以及HAART治疗对淋巴细胞亚群的影响。结果显示AIDS患者与健康对照相比,外周血CD4~+CD45RA~+、CD4~+CD28~+、CD4~+CD45RO~+、CD4~+CD95~+细胞比例明显低于正常对照,CD8~+CD95~+、CD8~+CD38~+、CD56~+细胞比例明显高于正常对照;经HAART治疗后,CD4~+CD45RA~+,CD4~+CD28~+细胞比例比治疗前升高,CD56~+细胞比例比治疗前显著下降。结果表明CD4~+T淋巴细胞表面CD45RA、CD28表达,以及CD56~+细胞能够预测AIDS患者HAART治疗效果。     

G-CSF对外周血树突状细胞亚群比例的影响

目的 :了解G CSF对小鼠外周血树突状细胞 (DC)亚群比例的影响。方法 :以不同剂量 (0、5 .0、10 .0或 15 .0 μg)的G CSF ,分别皮下注射于BALB/c小鼠 ,连续 6d ,每天 1次。第 6天分离外周血DC ,用流式细胞仪检测DC1(CD11c+ CD8a-)和DC2 (CD11c+ CD8a+ )的比例并计算其绝对值。结果 :经不同剂量的G CSF刺激后 ,外周血DC1的绝对值无显著变化 ,而DC2的绝对值增加 5倍 (9.6× 10 6/Lvs 5 5 .1× 10 6/L ,P <0 .0 1) ,DC1/DC2比值降低 (4.2vs 0 .7,P <0 .0 1)。结论 :G CSF在体内可提高外周血DC2的绝对数 ,对DC1的数无明显影响 ,从而使DC1/DC2的比例倒置     

性别、年龄和季节对血清25-羟维生素D水平的影响

目的探讨性别、年龄和季节对血清25-羟维生素D(25OHD)的影响。方法调取2014年12月至2018年10月北京协和医院血清25OHD检测结果,共计34062例。采用罗氏e601自动分析仪及其配套试剂检测血清25OHD。应用回归分析评估性别、年龄和季节对血清25OHD水平的影响。结果男性血清25OHD水平明显高于女性(P<0.001)。血清25OHD在8月份达到高峰为22.1 ng/mL。夏季对25OHD水平影响最大(标准化系数=0.273)。结论性别和季节显著影响血清25OHD水平。     

非霍奇金淋巴瘤患者T细胞亚群、NK细胞检测的临床意义

目的：研究非霍奇金淋巴瘤（NHL）患者外周血T淋巴细胞亚群、NK细胞检测结果的变化与该病的关系及与慢性淋巴腺炎患者细胞免疫功能的不同变化。方法：采用流式细胞仪（FCM）检测非霍奇金淋巴瘤（NHL）患者、慢性淋巴腺炎及正常人外周血T淋巴细胞亚群比例、NK细胞的变化。结果：非雹奇金淋巴瘤患者与正常人比较总的T淋巴细胞、辅助性T淋巴细胞及CD4^＋／CD8^＋比值明显下降（P〈0．05），细胞毒性T淋巴细胞明显升高（P〈0．05），NK细胞则无明显变化（P〉0．05）。非霍奇金淋巴瘤患者与慢性淋巴腺炎患者比较，细胞毒性T淋巴细胞、NK细胞明显升高（P〈0.05），而总的T淋巴细胞、辅助性T淋巴细胞无明显改变（P〉0．05），CD4^＋／CD8^＋比值略有下降但无明显统计学意义。结论：非霍奇金淋巴瘤患者细胞免疫功能明显受到抑制，T细胞亚群及NK细胞的检测对NHL的诊断、治疗、预后判断有一定的临床价值。     

流式细胞术检测肿瘤患者外周血T淋巴细胞亚群研究

目的 探讨肿瘤患者T淋巴细胞亚群的变化及临床意义。方法 采用流式细胞术检测27例肿瘤患者和25例正常对照组的T淋巴细胞亚群。结果 肿瘤患者CD3^＋T细胞、CD4^＋T细胞和CD4^＋／CD8^＋比值明显低于对照组并有统计学意义，而CD8^＋T细胞显著高于对照组（P〈0．01）。结论 肿瘤患者的免疫功能下降。流式细胞术对肿瘤患者的免疫功能的检测具有快速、敏感、准确的特点，对于评价疗效、判断预后具有重要价值。     

The effects of age and gender on gingival tissue and peripheral blood T-lymphocyte subsets: a study in mice

The aim of this study is to investigate the effects of age and gender on relative number of CD4+ and CD8+ T lymphocyte and CD4+/CD8+ ratios in gingival tissue and peripheral blood of periodontally healthy BALB/c mice with flow cytometric analysis. The study was carried out on periodontally healthy 60 BALB/c mice. They were separated into five groups according to the life expectancy: Group I (newborn, 1-10 days old), Group II (age at weaning, 21-28 days old), Group III (age of sexual maturity, puberty, 7-8 weeks old), Group IV (adult, 8 months old), and Group V (the aged, 14 and over). Males and females were equally represented in each group. CD4+ and CD8+ T lymphocytes and CD4+/CD8+ ratio values in gingival tissue and peripheral blood were determined using flow cytometry in the biopsy samples. The relative number of CD8+ T lymphocyte in gingival tissue was higher in puberty (p < 0.05) and the aged (p < 0.05), with more significantly difference in males (p < 0.05). The CD4+/CD8+ ratios in gingival tissue were lower in puberty group and the-aged groups compared to the other groups (p < 0.05). The peripheral blood CD4+/CD8+ ratio increased in puberty and the aged groups. These differences were more significant in females than in males (p < 0.05). There were considerable negative correlations between CD4+/CD8+ T lymphocyte ratio in gingival tissue and CD4+/CD8+ T lymphocyte ratio in peripheral blood in puberty (r = 0.647, p < 0.01) and the aged ( r = 0.599, p < 0.05). In conclusion, our data suggest that CD4+/CD8+ T lymphocyte ratios in peripheral blood increase periodontally healthy mice in puberty and the old groups, while CD4+/CD8+ T lymphocyte ratios in gingival tissue decrease in the same groups. The relative number of CD8+ T lymphocytes increases in gingival tissue in puberty and the old groups when it decreases in peripheral blood. Results from this study indicated that periodontally healthy BALB/c mice may represent important information to determine the character of an immune response in the course of a lifetime.     

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...