Lymphocyte Subsets in the Blood

Removal of the largest single lymphoid organ, the spleen, leads to an increase in severe infections. To prevent this, transplantation of splenic fragments can be performed, which may, however, cause an increase in CD8⁺ lymphocytes in the blood of these patients. This is controversial since in the clinical situation it is often difficult to account for the different age of the patients, the time point after the operation and many other factors known to influence the number of lymphocyte subsets.
Using a well-defined animal model, B, T, CD4⁺, and CD8⁺ lymphocytes were determined preoperatively in adult rats. Then, either sham splenectomy, splenectomy, or splenic autotrans-plantation was performed and the animals were followed up for 15 months after the operation.
The surgical procedure itself, the site of blood sampling and ageing all influenced the number of lymphocyte subsets profoundly. Furthermore, giving the data as relative or absolute numbers leads to different results.
Splenectomy caused lymphocytosis, due to a significant increase in B and CD8⁺ lymphocytes, as did splenic autotransplantation, which indicates that the number of lymphocyte subsets in the blood should not be used to argue in favour of or against splenic autotransplantation.
This study demonstrates that the number of lymphocyte subsets in the blood is influenced by many factors and therefore should be determined in a highly standardized fashion.     

T淋巴细胞在不同类型白血病患者外周血中的变化

目的通过对不同类型白血病患者外周血T淋巴细胞亚群(CD3+、CD4+、CD8+)的检测以评价不同类型白血病患者的细胞免疫功能。方法采用流式细胞术检测200例不同类型白血病患者和52例健康志愿者外周血T细胞亚群的水平。结果与正常对照组相比,急性白血病和慢性白血病患者的CD4+/CD8+比值都是下调的(P<0.05);急性白血病患者(包括急淋和急粒)的CD3+、CD8+细胞水平是上升的,且急淋患者还伴随着CD4+细胞的降低(P<0.05);慢性白血病患者的CD3+、CD4+细胞水平是下调的,其中慢淋患者的CD3+细胞下调更为显著,而慢粒患者则是以CD8+细胞增高为主(P<0.05)。结论不同类型白血病患者外周血T淋巴细胞均存在免疫异常,且具有不同异常方式,对指导治疗及疗效观察有一定的临床意义。     

Cytochemical Analysis of Human Peripheral Blood Lymphocyte Subsets Defined by Monoclonal Antibodies

Human blood lymphocyte subpopulaiions, revealed by a panel of commercially available monoclonal antibodies by means of a resetting technique, were submitted to direct cyto-chemicat analysis and were shown to have distinguishing characteristics. T cells reactive with OKT3 antibody (T3⁺) displayed higher beta-glucuronidase and dot-like alpha-napnthyl acetate acid esterase (ANAE) activity than T3⁻ cells. Helper/inducer cells (T4⁺) were characterized by a high level of dot-like ANAE activity, whereas cytotoxic/suppressor cells (T8⁺) displayed selective naphthol-ASD-chloroacetate esterase activity. These results provide evidence for association of some cytoplasmic enzyme activities with the expression of membrane differentiation markers defined by monoclonal antibodies.     

Peripheral Blood Lymphocyte Subsets in Adolescents: a Longitudinal Analysis from the REACH Project

Flow cytometry analysis of lymphocyte subset markers was performed for a group of sexually active, human immunodeficiency virus (HIV)-negative adolescents over a 2-year period to establish normative data. Data were collected in the REACH Project (Reaching for Excellence in Adolescent Care and Health), a multicenter, longitudinal study of HIV-positive and high-risk HIV-negative adolescents. Two- and three-color flow cytometry data were collected every 6 months for these subjects. We determined the effects of gender, race, and age on the following lymphocyte subset markers: total CD4⁺ cells, CD4⁺ naïve cells, CD4⁺ memory cells, all CD8⁺ cells, CD8⁺ naïve cells, CD8⁺ memory cells, CD16⁺ natural killer cells, and CD19⁺ B cells. Gender was the demographic characteristic most frequently associated with differences in lymphocyte subset measures. Females had higher total CD4⁺ cell and CD4⁺ memory cells counts and lower CD16⁺ cell counts than males. Age was associated with higher CD4⁺ memory cell counts as well as higher CD8⁺ memory cell counts. For CD19⁺ cells, there was an interaction between age and gender, with males having significantly lower CD19⁺ cell counts with increasing age, whereas there was no age effect for females. Race and/or ethnicity was associated with differences in total CD8⁺ cell counts and CD8⁺ memory cell counts, although both of these associations involved an interaction with gender.     

Flow Cytometric Analysis of Lymphocyte Subsets in the Bronchoalveolar Lavage Fluid and Peripheral Blood of Healthy Volunteers

The lymphocyte subsets in the bronchoalveolar lavage fluid (BALF) and the peripheral blood of 25 healthy volunteers were examined by analysis with a fluorescence-activated cell sorter. Comparison of the lymphocyte subsets in the BALF with those of the peripheral blood revealed much higher values for the ratios of each Leu 3a+ (CD4), Leu 3+8-, and Leu 2+15- cells, while the ratios of Leu 1+ (CD5), Leu 2a+ (CD8), Leu 7+, Leu 8+, Leu 10+, Leu 11a+ (CD16), Leu 12+, and Leu 2+15+ cells were low in the BALF. The above results indicate that the lymphocyte subsets in the BALF from healthy individuals are mainly composed of cells with surface phenotypes of helper T cells and cytotoxic T cells with virtual absence of cells carrying suppressor T and NK cell phenotypes, and with low B cell ratio. Therefore, it is assumed that the local immune mechanism of the lung is different from that of the peripheral blood.     

多发性硬化症发作期外周血和脑脊液淋巴细胞亚群的分析

为分析多发性硬化症 (MS )患者发作期淋巴细胞亚群及给予甲基强的松龙 (MP )治疗后的变化 ,流式细胞仪测定 2 6例处于复发期MS患者外周血 (PB )和脑脊液 (CSF )及 8例MS患者予MP治疗后PB淋巴细胞CD3+ 、CD4 + 、CD8+ 、CD4 5RA+ 、CD4 + /CD4 5RA+ 、CD4 + /CD2 9+ 、CD19+ 、CD5 + /CD19+ 的百分率。结果发现MS患者PB中CD8+ 、CD4 5RA+ 和CD4 + /CD4 5RA+ 百分率降低 ,CD4 + /CD2 9+ 百分率和CD4 + /CD8+ 比值升高 ;CSF中CD3+ 、CD4 + 、CD4 + /CD2 9+ 百分率和CD4 + /CD8+ 比值高于PB ;淋巴细胞亚群与临床伤残程度和距此次发作的时间无关 ;MP治疗不影响PB淋巴细胞亚群变化。表明MS患者淋巴细胞通过血脑屏障有选择性 ,淋巴细胞亚群的变化在MS发病机制中起作用     

改良微量细胞毒试验检测外周血T、B细胞总数及T细胞亚群

本文介绍一种改良微量细胞毒试验。应用抗B细胞单克隆抗体和OKT单克隆抗体系统对86例健康成人外周血中T、B细胞总数及T细胞亚群进行了检测。本法具有简便,不需特殊仪器,在白细胞计数池内计数死活细胞,结果准确等特点,结果显示B细胞、OKT_3(总T细胞),OKT_4(辅助T细胞)和OKT_8(抑制/杀伤T细胞)占外周血淋巴细胞百分率分别为20.59±0.6％;66.42±0.8％;41.65±0.8％,27.19±0.7％。并且,观察到随年龄增加,OKT_3和OKT_4细胞降低。OKT_8细胞增加,T_4/T_8比值降低。文章还讨论了老年组OKT_4细胞显著降低的意义。     

急性髓系白血病患者治疗前后T淋巴细胞亚群测定的临床意义

为探讨急性髓系白血病患者治疗前后T淋巴细胞亚群变化的临床意义,选择初发确诊的急性髓系白血病患者30例(初治组); 经标准方案化疗,26例完全缓解(缓解组); 健康人30名(正常对照组).应用流式细胞术检测这三组人群T淋巴细胞亚群的变化情况.结果显示,初治组CD3+T 细胞、CD4+T细胞、CD8+T细胞百分率、CD4+...     

乙型肝炎病毒感染者外周血T淋巴细胞亚群表达与其血清HBV-DNA水平的相关性分析

观察乙型肝炎病毒(HBV)感染者外周血T细胞亚群功能变化与其血清HBV-DNA水平,探讨病毒复制程度与细胞免疫功能的关系.应用流式细胞术直接荧光法检测142例HBV感染者和35名健康对照者外周血中T淋巴细胞亚群百分率,用荧光定量PCR法检测HBV感染者血清HBV-DNA.结果显示:与健康对照组比较,慢性乙型肝炎组CD4...     

急性脾破裂患者手术前后外周血淋巴细胞亚群的变化及临床意义

目的:观察急性脾破裂患者在脾切除后外周血淋巴细胞亚群水平的改变。探讨短期内脾切除术对机体免疫功能的影响。方法:利用流式细胞仪测定74例急性脾破裂患者在手术前后淋巴细胞亚群水平。结果:急性脾破裂患者手术后CD 3T淋巴细胞及CD 4T淋巴细胞较手术前升高(P<0.01);CD(16 56)细胞(即NK细胞)和CD 19B淋巴细胞较手术前降低(P<0.05);而CD 8T淋巴细胞及CD 4/CD 8较手术前无明显差异(P>0.05)。结论:脾切除术后,短期内患者的免疫功能有改变。     

低剂量辐射对不同岗位男性放射人员外周血T淋巴细胞亚群的影响

目的:探讨低剂量辐射对不同岗位男性放射人员T淋巴细胞亚群水平的影响.方法:检测168例男性放射人员CD3+T、CD3+/CD4+T、CD3+/CD8+T、CD4+/CD45RA+T、CD4+/CD45RO+T、CD8+/CD45RA+T、CD8+/CD45RO+T、CD8+/CD28+T、CD8+/CD95+T、CD8...     

Lymphocyte Subsets in The white Pulp of Human Spleen in Normal and Diseased Cases

The distribution of T and B-lymphocytes and their subsets in the white pulp of human spleens extirpated from patients with cancer of the digestive tract and those with portal hypertension was examined with the appliance of monoclonal antibodies. T-lymphocytes were distributed in the core of lymphatic sheath protuberance (Matsumoto), while the mantle zone of lymphatic nodule and germinal center were solely occupied by B-lymphocytes. On the other hand, both T and B-lymphocytes were found in the cortical zone of lymphatic sheath protuberance and outer and inner layer of lymphatic nodule. The ratio of both cells differed from one case to the other. The majority of T-lymphocytes in the core of lymphatic sheath protuberance and those localized in the light region of germinal center were helper T cells. Although the lymphatic sheaths in cases with portal hypertension were narrower than those in the controls, there was no difference in the distribution density of T cells between the two groups. Hardly any B-lymphocytes were found in the cortical zone of lymphatic sheath protuberance, while the mixture of T-lymphocytes tended to become prominent in the outer and inner layer of lymphatic nodule in those cases.     

Lymphocyte Subsets and Mitogen Stimulation of Blood Lymphocytes in Normal Pregnancy

PROBLEM: In normal pregnancy the maternal immune system should be directed towards tolerance or suppression in order not to reject the partly foreign feto-placental unit. The aim of this investigation was to find hallmarks of systemic immunosuppression during normal pregnancy. METHODS: Five healthy primigravidae were examined during pregnancy and postpartum with flow cytometric analysis to define T and B lymphocyte subsets in peripheral blood. In addition, we studied the proliferative response of lymphocytes to mitogens or interleu-kin-2 (IL-2) alone or in combination with immunomodulating drugs or interleukin-4 (IL-4). The results were compared to healthy, non-pregnant women. RESULTS: During pregnancy and early puerperium we noted an immune balance in favour of suppression, as measured by increased numbers of T “helper/suppressor” (CD4+ CD45RA+) and “suppressor”/effector T cells (CD8+S6F1-), and decreased numbers of T “helper/inducer” (CD4+CD29+), T “helper/memory” (CD4+CD45RO+), killer/effector T cells (CD8+S6F1+), and Natural Killer cells (CD56+), as well as decreased numbers of activated lymphocytes expressing IL-2 receptor (CD25+) and T cells expressing HLA-DR (HLA-DR+CD3+). During pregnancy, lymphocyte proliferation was impaired in autologous serum with concanavalin A (ConA), phytohemagglutinin (PHA), or IL-2. A difference in proliferative response to PHA or IL-2 between cultures with AB serum and autologous serum is suggestive of an immunosuppressor factor in serum during pregnancy. Indomethacin significantly increased lymphocyte proliferation in autologous serum with ConA, indicating PGE₂ mediated suppressor activity during pregnancy. Chlorambucil and cimetidine modulated the proliferative response to ConA, indicating an alkylating agent sensitive and a histamine dependent suppressor activity during pregnancy. CONCLUSIONS: During normal pregnancy, a state of systemic suppression of the maternal immune system seems to be present.     

Blood Transit and Recirculation Kinetics of Lymphocyte Subsets in Normal Rats

Thoracic duct lymphocytes (TDL) were labelled with fluorescein isothiocyanate (FITC) and injected into normal, unanaesthetized rats with either a central venous catheter or a thoracic duct cannula. The blood transit time and the appearance in the lymph was calculated and then the percentages of B, T, T helper, and T suppressor lymphocytes were determined with monoclonal antibodies (Ox12, Ox19, W3/25, and Ox8, respectively). The blood transit time of all subsets was about 30 min. However, the percentage of B lymphocytes from 5 min after injection onwards is reduced (14.5 +/- 1.9%) compared to the injected TDL (32.7 +/- 1.7%). These cells are not in the lung vascular pool. The recovery of FITC-labelled TDL in the thoracic duct within 48 h is much higher (56.8 +/- 5.3% of the injected lymphocytes) than in previous studies using radioactive markers. B lymphocytes appear later and in a reduced number in the thoracic duct. The mean transit time is 26 h and the recovery 31.5 +/- 4.2% in contrast to T lymphocytes (18 h and 66.3 +/- 6.5%, respectively). The technique of combining FITC in vitro labelling with surface staining after completion of migration does not interfere with lymphocyte migration. It can therefore be used to study the migration of lymphocyte subsets in normal, untreated animals.     

慢性疲劳综合征患者外周血淋巴细胞亚群及CD25^＋调节性T细胞表达的研究

研究慢性疲劳综合征（CFS）患者外周血淋巴细胞亚群及CD25＋调节性T细胞的表达情况。使用流式细胞仪检测84例CFS患者（CFS组）、50例健康体检者（健康对照组）外周血淋巴细胞亚群（T细胞、CD4＋T细胞、CD8＋T细胞、B细胞、NK细胞）及CD25＋调节性T细胞的表达情况。结果显示,CFS组与健康对照组的T细胞、CD8＋T细胞百分率以及CD4＋/CD8＋比值无显著差别（P〉0.05）;而CFS组NK细胞、CD4＋T细胞及CD25＋调节性T细胞百分率显著增高,B细胞百分率显著降低（P〈0.05）。结论：慢性疲劳综合征患者外周血淋巴细胞各亚群比例异常,提示其免疫功能失衡,而CD25＋T调节性细胞可能在该病进程中有重要意义。     

多发性骨髓瘤患者外周血淋巴细胞亚群分析及与预后因子的相关性

目的 通过对多发性骨髓瘤(MM)患者外周血淋巴细胞亚群的检测以评价MM患者机体的免疫功能状态.方法 采用流式细胞术检测36例MM患者和25例健康志愿者外周血T、B淋巴细胞、NK细胞及CD4+CD25T细胞的表达.结果 与正常对照组相比,MM患者外周血的CD4、CD19细胞的表达显著下调,CD8细胞的表达显著上调,CD4/CD8比值则显著降低(P＜0.05或＜0.01);MM患者外周血的CD4+CD25T细胞占CD3+T细胞的比例明显增高(P＜0.01),且与血清中的β2-MG浓度成正相关(γ=0.56,P＜0.05).结论 MM患者体内存在淋巴细胞亚群的异常表达、CD4+CD25+Treg细胞的异常扩增,可能是MM患者体内广泛存在免疫缺陷的一个主要原因.     

Migration Pattern of Lymphocyte Subsets in the Normal Rat and the Influence of Splenic Tissue

Lymphocyte subsets leave the blood and appear in the thoracic duct of normal rats at different rates. The aim of the present study was to investigate their migration pattern through blood, spleen, bone marrow, mesenteric lymph nodes, and Peyer's patches in normal Lewis rats and to study the role of the spleen using splenectomized and spleen-transplanted animals. Fluorescein isothiocyanate (FITC)-labelled thoracic duct lymphocytes (TDL) were injected intravenously into rats and after 15 min, 1, 6, and 24 h the percentages of B, T, T helper (TH) and T-cytotoxic/suppressor (TC/S) lymphocytes in the FITC+ cells were determined in cell suspensions by means of monoclonal antibodies. B and T lymphocytes are preferentially localized in different organs, e.g. B cells in Peyer's patches and T cells in mesenteric lymph nodes. The migration of TH lymphocytes differed from that of TC/S lymphocytes in all the organs investigated. In the late phase after injection the migration of B and TH lymphocytes was influenced by the spleen, since after splenectomy the number of injected B lymphocytes increased and that of TH lymphocytes decreased in all organs investigated except the bone marrow. Splenic autotransplantation could not normalize the disturbed migration.     

原发性肺癌患者外周血PD-1淋巴细胞亚型检测与临床预后的关系研究

目的检测原发性肺癌患者外周血PD-1淋巴细胞亚型,并进行临床随访,分析其与临床预后的关系。方法采集2015年1月至2016年11月在河南省肿瘤医院呼吸内科诊治的35例可疑原发性肺癌患者的血液学标本,并进行外周血淋巴细胞亚型分析以及PD-1表达,然后进行临床随访。结果 35例患者中,31例患者最终病理确诊为原发性肺癌(腺癌23例,鳞癌3例,小细胞肺癌5例),剩余4例患者为炎症。CD3+CD8+T细胞比例数值高、CD4+/CD8+的比值低、PD-1+/PBMC比例高的患者近期疗效越好。未发现治疗前淋巴细胞亚型比例异常患者与正常患者之间存在PFS的差异。结论在本项回顾性小样本研究中,发现原发性肺癌治疗前特定淋巴细胞亚型与近期疗效相关,尤其是PD-1+PBMC比例升高的患者近期疗效更佳。需要进一步前瞻性大样本研究验证。