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.     

Alterations in levels of CD28-/CD8+ suppressor cell precursor and CD45RO+/CD4+ memory T lymphocytes in the peripheral blood of multiple sclerosis patients.

A comprehensive peripheral blood immunophenotype analysis of 16 multiple sclerosis (MS) patients was performed by three-color flow cytometric analysis, and the results were compared with those for age-matched healthy controls. The cell subsets quantified included T cells (CD3+), B cells (CD19+), NK cells (CD56+), CD4+ and CD8+ T cells, cytotoxic (CD28+) and suppressor precursor (CD28-) CD8+ T cells, CD45RA+ and CD45RO+ T cells (CD4+ and CD8+), and CD5+ T and B cells. Analysis of MS patients' peripheral blood revealed essentially normal levels of total T, B, and NK cells. In agreement with results obtained by other investigators, it was found that MS patients had an increased CD4/CD8 ratio, primarily due to a decrease in CD8+ T cells. MS patients were found to have a significantly decreased level of suppressor precursor (CD28-) CD8+ T cells compared with that of controls but to have normal levels of cytotoxic (CD28+) CD8+ T cells. These data indicate that MS patients do not have a general decrease in CD8+ T cells but that they have a specific decrease in the suppressor precursor subset only and normal levels of cytotoxic CD8+ T cells. MS patients also had a significant increase in memory (CD45RO+) CD4+ T cells and displayed a trend towards a decrease in naive (CD45RA+) T cells in the peripheral blood.     

Intrathyroidal HLA-DR-positive lymphocytes in Hashimoto's disease: increases in CD8 and Leu7 cells

The peripheral and intrathyroidal HLA-DR-positive (DR+) lymphocyte subsets that were activated in vivo in patients with Hashimoto's disease (HD) were examined by two-color flow cytometry with monoclonal antibodies against CD3, CD4, CD8, Leu7, CD19, and HLA-DR antigens. The proportions of total DR+ cells in peripheral lymphocytes and the proportions of DR+ cells in the CD3+, CD4+, and Leu7+ lymphocytes were higher in patients with HD than in normal controls. Furthermore, the proportions of total DR+ cells among intrathyroidal lymphocytes isolated from thyroid tissue of individuals with HD were higher than those in their peripheral lymphocytes. Interestingly, the proportions of DR+ cells among the CD3+, CD8+, and Leu7+ lymphocytes in the thyroid were greatly increased. These data indicate that (i) CD3+ T, especially CD4+ T helper/inducer, lymphocytes and Leu7+ NK/K cells are activated in peripheral blood in Hashimoto's disease and that (ii) CD3+ T, especially CD8+ T suppressor/cytotoxic, lymphocytes and Leu7+ NK/K cells are predominantly activated in Hashimoto's goiter, suggesting an increase of cell-mediated cytotoxicity in the thyroid in Hashimoto's disease.     

Intrathyroidal lymphocyte subsets, including unusual CD4+ CD8+ cells and CD3loTCR alpha beta lo/-CD4-CD8- cells, in autoimmune thyroid disease.

Intrathyroidal lymphocyte subsets were analysed in 13 euthyroid patients with autoimmune thyroid disease by two-colour flow cytometry and compared with subsets in peripheral blood. In both Graves' and Hashimoto's diseases, proportions of intrathyroidal CD5- B cells were higher than in peripheral blood. The numbers of such cells were correlated with serum levels of anti-thyroid microsomal antibodies. Proportions of T cells bearing alpha beta chains of T cell receptors (TCR alpha beta+ T; T alpha beta) and CD16+CD57+ natural killer (NK) cells were lower in the thyroid, but proportions of CD3hiTCR alpha beta-TCR gamma delta+ (T gamma delta) cells were not different. Proportions of CD4+Leu-8- helper T cells and CD4+CD57+ germinal centre T cells were higher and proportions of CD4+Leu-8+ suppressor-inducer T cells and CD8+CD57+ or CD8+CD11b+ suppressor T cells were lower than in the blood in both diseases. Proportions of CD5+ B cells were high in Graves' disease, and proportions of CD8+CD11b- cytotoxic T cells were high in Hashimoto's disease. Unexpectedly, CD4+CD8+ cells and CD3loTCR alpha beta lo/-CD4-CD8- cells were present in thyroid tissues of both diseases. These findings suggest that: (i) an imbalance in the numbers of regulatory T cells and of NK cells that had appeared in the thyroid resulted in the proliferation of CD5- B cells, which were related to thyroid autoantibody production; (ii) CD5+ B cells and cytotoxic T cells are important for the different pathological features in Graves' and Hashimoto's diseases, respectively; and (iii) intrathyroidal CD4+CD8+ cells and CD3loTCR alpha beta lo/-CD4-CD8- cells may be related to the pathogenesis of autoimmune thyroid disease.     

T lymphocyte subsets and NK cell cytotoxicity in chronic hemodialysis patients. The effect of recombinant human erythropoietin (rHu-EPO) treatment.

We investigated subpopulations of T lymphocytes, NK cell number and cytotoxic activity in 14 chronic uremic patients on regular hemodialysis treatment. We observed a significantly decreased absolute lymphocyte number and percentage of CD3 cells. Relative numbers of CD16 cells were significantly elevated, but NK cell cytotoxic activity was within a normal range. Nine patients with chronic renal anemia on maintenance hemodialysis were enrolled in rHu-EPO treatment trial. The treatment was continued till the hematocrit level reached 30%. Each of the patients had corrected anemia and well-being. After 12 weeks of the treatment we observed in these patients decreases in CD3, CD4, CD8 and CD16 cell numbers and elevation of CD4/CD8 ratio. Cytotoxic activity of NK cells did not change significantly. Presented results indicate that chronic hemodialysis patients have significantly diminished lymphocyte number. rHu EPO treatment affects the T lymphocyte subsets inducing a deep decrease of CD8 and CD16 cell percentage leading to normalisation of the CD4/CD8 ratio.     

The remarkable proliferation of helper T cell subset in response to autologous thyrocytes and intrathyroidal T cells from patients with graves'' disease

We have studied cellular interactions among thyrocytes, intrathyroidal T cells and peripheral blood T cells from Graves' patients. In the autologous mixed lymphocyte reaction of Graves' patients, CD4+ cells were able to proliferate vigorously against autologous non-T cells, whereas CD8+ cells responded weakly to non-T cell stimulators. Furthermore, the proliferative response of the CD4+ 2H4+ suppressor-inducer T cell subset was increased like that of the CD4+ 2H4- helper T cell subset. In contrast to peripheral blood non-T cell stimulators, thyrocytes and intrathyroidal T cells had the ability to activate the CD4+ 2H4- helper T cell subset but were not able to cause proliferation both of CD4+ 2H4+ suppressor-inducer T cell and CD8+ suppressor/cytotoxic T cell subsets. The marked reduction in proliferative responses of CD4+ 2H4+ cells and CD8+ cells could not be attributed to a difference in kinetics or altered response to variable number of stimulator cells. On the basis of these findings, it is suggested that the concentration of the helper T cell subset may be progressively increased and suppressor circuits may be unable to be activated in thyroid tissues. These abnormalities in cellular interactions may induce the excessive production of autoantibodies.     

慢性髓系白血病患者外周血T淋巴细胞亚群和NK细胞检测的临床意义

目的 研究不同分期慢性髓系白血病患者外周血T淋巴细胞亚群、NK细胞的变化特点,以及应用伊马替尼治疗后获得完全细胞遗传学反应(complete cytogenetic reponse,CCyR)患者淋巴细胞亚群表达情况.方法 选取我院诊治40例慢性髓系白血病患者,其中急变期9例,慢性期31例.采用流式细胞术检测外周血T淋巴细胞亚群、NK细胞水平,并与正常对照组进行比较.结果 初治慢性期、急变期CML患者外周血CD3+、CD4+、CD8+T细胞百分率及CD4+/CD8+比值均低于正常对照组,且急变期CD3+、CD4+T细胞百分率及CD4 +/CD8+比值下降尤为突出(P＜0.01);初治慢性期患者NK细胞百分率与正常对照组相比无差异,而急变期患者低于正常对照组(P＜0.05).与正常组对比,伊马替尼治疗首次获得完全细胞遗传学反应患者仅CD4+T细胞百分率降低,差异具有统计学意义(P＜0.05);但获得完全细胞遗传学反应后应用伊马替尼治疗大于12月患者,CD3+、CD4+T细胞百分率及CD4 +/CD8+比值较正常对照组均有所下降(P＜0.05).与治疗前相比,治疗首次获得完全细胞遗传学反应患者CD3+、CD4+T细胞百分率升高(P＜0.05),而缓解后应用伊马替尼治疗大于12月患者T淋巴细胞亚群无改变(P＞0.05);各组的NK细胞百分比无差异(P＞0.05).初诊CML患者、急变期CD4+/CD8+的比值与BCR-ABLl/ABL1的比值呈负相关.结论 CML患者存在细胞免疫调节功能异常,且机体免疫功能与疾病分期密切相关.伊马替尼治疗初次获得完全细胞遗传学反应患者细胞免疫功能得到改善,但长期应用抑制患者细胞免疫功能.     

A cell surface monoclonal antibody (H366) helps to discriminate human cytotoxic from suppressor T cells.

A study has been undertaken to differentiate T cytotoxic (Tc) and T suppressor (Ts) cell subsets using a monoclonal antibody termed H366 (mouse IgG2b) previously reported to phenotype natural killer and killer (NK/K) cells. Mononuclear cell suspensions from 14 normal subjects were depleted of H366+ cells by means of complement dependent cytotoxicity and the remaining cells were phenotyped with CD8 and CD4 monoclonal antibodies. The effects of depletion with H366 plus complement (C1) on the induction and activity of suppressor and cytotoxic T cells was also examined. The results indicate that H366 antibody recognizes in addition to NK/K cells, a population of Tc but not Ts or helper cells. Therefore, H366 antibody can be useful for obtaining Ts enriched lymphocyte subpopulations and this property may also be used for the enumeration of suppressor cells in the peripheral blood in disease states.     

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。     

腰椎间盘退变性疾病与外周血淋巴细胞亚群的关系

文题释义：淋巴细胞亚群：淋巴细胞是白细胞的一种,是机体免疫应答功能的重要细胞成分,是淋巴系统几乎全部免疫功能的主要执行者,占外周血白细胞总数的20%-40%,按其发生迁移、表面分子和功能的不同,主要分成T淋巴细胞、B淋巴细胞和自然杀伤细胞三大类。 自然杀伤细胞(natural killer cell,NK)：是机体重要的免疫细胞,不仅与抗肿瘤、抗病毒感染和免疫调节有关,而且在某些情况下参与超敏反应和自身免疫性疾病的发生。 背景：既往研究认为椎间盘退变的主要原因为遗传、衰老、营养不良和负荷史,免疫系统在椎间盘退变过程中的作用尚不清楚。 目的：观察腰椎间盘退变患者外周血淋巴细胞亚群的变化,并研究腰椎间盘退变程度与外周血各淋巴细胞亚群的关系。 方法：收集76例腰椎间盘退变性疾病患者和56例健康志愿者(对照组)的血样,用流式细胞仪检测外周血各淋巴细胞亚群,包括CD3⁺T细胞、CD4⁺T细胞、CD8⁺T细胞、CD19⁺B细胞、CD3^-CD16⁺CD56⁺自然杀伤细胞等淋巴细胞亚群的百分率,计算CD4⁺/CD8⁺比值。采用Pfirrmann分级标准评估2组腰椎间盘退变程度和分级,进一步评估外周血各淋巴细胞亚群与腰椎间盘退变程度的相关性。研究经郑州大学第一附属医院伦理审查委员会批准(伦理批号：2019-KY-285),所有受试者都签署了知情同意书。 结果与结论：①腰椎间盘退变性疾病组的腰椎间盘退变程度明显高于对照组(P < 0.05);②腰椎间盘退变性疾病组CD4⁺T细胞百分率、自然杀伤细胞百分率和CD4⁺/CD8⁺比值明显高于对照组(P < 0.05);腰椎间盘退变性疾病组CD8⁺T细胞百分率较对照组显著降低(P < 0.05);③对照组腰椎间盘退变程度与外周血各淋巴细胞亚群无相关性;腰椎间盘退变性疾病组腰椎间盘退变程度与CD4⁺ T细胞百分率、CD4⁺/CD8⁺比值、自然杀伤细胞百分率成线性正相关(r =0.412,P=0.000;r=0.715,P=0.000;r=0.494,P=0.000),与CD8⁺ T细胞百分率成线性负相关(r=-0.737,P=0.000);④结果表明,腰椎间盘发生退行性改变可能与外周血各淋巴细胞亚群改变有关,且CD4⁺T细胞增多、自然杀伤细胞增多以及CD4⁺/CD8⁺比值增高可能加速腰椎间盘退变。提示,免疫系统改变预示腰椎间盘退变发生的可能,其有望成为腰椎间盘退变性疾病的防治靶点。 ORCID: 0000-0002-8087-2356(冯阳) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Lymphocyte subset reference ranges in adult Caucasians.

We report here the distributions of lymphocyte populations bearing the following antigens: CD3 (T cells), CD19 (B cells), CD4 (T helper/inducer cells), CD8 (T suppressor/cytotoxic and some NK cells), and CD3-, CD16+, and/or CD56+ (NK cells). At four sites, analyses were performed on healthy, normal subjects between the ages of 18 and 70, using identical flow cytometry systems and techniques. Reference ranges (unadjusted for sex differences and age variation) are CD3 (61 to 85%), CD19 (7 to 23%), NK (6 to 29%), CD4 (28 to 58%), and CD8 (19 to 48%). The lymphocyte subpopulation distributions for all antigens were found to be similar at all sites. By combining data from all sites, it has been possible to estimate age variation and sex differences for each of these subpopulations. Age and sex associated differences are substantial for some lymphocyte subsets (CD3, CD4, NK cells), and proper accounting of these effects is essential in evaluating the individual patient, if further disease-related variation is to be accurately and consistently assessed. It appears possible to recommend reference ranges for lymphocyte population parameters applicable across national and laboratory boundaries. These ranges provide a basis for comparing results from different institutions and for combining such results on subjects and patients from several institutions, provided the methodology and equipment are identical at all sites.     

Mistletoe Lectin I-Induced Effects on Human Cytotoxic Lymphocytes. I. Synergism with Il-2 in the Induction of Enhanced Lak Cytotoxicity

This report demonstrates that in vitro activation of human cells with the β-galactoside-specific lectin from mistletoe (ML-I) or interleukin-2 (IL-2) results in different patterns of activation and function of cytotoxic cells. It is now well established that natural killer (NK) and lymphokine-activated killer (LAK) cytotoxicity is mainly mediated by resting (NK) and IL-2-activated (LAK) CD56-positive (+) cells respectively. Culture of peripheral blood lymphocytes (PBL) for 3 days with ML-I led to expansion and activation of T cells which demonstrated NK-and LAK-like cytotoxicity. T lymphocyte subset analysis revealed that in total PBL, ML-I preferentially stimulated and expanded CD8+ T cells which mediated the cytotoxic effect. Incubation of highly purified CD8+ T cells alone with ML-I did not lead to induction of cytotoxicity, which required the presence of both CD4+ and CD 14+ (monocytes) cells, suggesting that ML-I does not exert a direct effect on CD8+ T cells. Activation of PBL with both ML-I and IL-2 resulted in simultaneous induction of T and CD56+ cell-mediated NK and LAK cytotoxicity. These data suggest that treatment with ML-I and DL-2 might provide an approach to induce maximum cytotoxicity against tumors and to recruit both T and NK cells for tumor therapy.     

Differential expression of T cell antigens in normal peripheral blood lymphocytes: a quantitative analysis by flow cytometry.

AIMS: To obtain reference values of the level of expression of T cell antigens on normal lymphocyte subsets in order to disclose differences which could reflect their function or maturation stages, or both. METHODS: Peripheral blood from 15 healthy donors was processed by flow cytometry with triple colour analysis. For each sample phycoerythrin (PE) conjugated CD2, CD4, CD5, CD8, and CD56 monoclonal antibodies were combined with Cy5-R-phycoerythrin (TC) conjugated CD3 and fluorescein isothiocyanate (FITC) conjugated CD7; CD2- and CD7-PE were also combined with CD3-TC and CD4-FITC. Standard microbeads with different capacities to bind mouse immunoglobulins were used to convert the mean fluorescence intensity (MFI) values of the lymphocyte subsets identified by multiparametric flow cytometry into the number of antigen molecules per cell, measured as antibody binding capacity (ABC). RESULTS: CD4+ (helper/inducer) T cells exhibit a higher CD3 antigen expression compared with CD8+ (suppressor/ cytotoxic) T lymphocytes. Within the CD4+ T cells, the CD4+CD7- subset expressed a lower level of CD3 compared with CD4+CD7+ and CD8+CD7+ cells, and higher CD2 and CD5 expression than the main CD3+CD7+ subset. Major differences in antigen expression were also detected between CD3+ T cells and CD3-CD56+ natural killer (NK) cells: NK cells exhibited higher levels of CD7 and CD56 and lower levels of CD2 and CD5 than T cells. Significantly lower CD5 expression was also detected in the small CD5+ B lymphocyte subset compared with T cells. CONCLUSIONS: Quantitative flow cytometry with triple colour analysis may be used to detect antigen modulations in disease states and to increase the accuracy of diagnosis by comparison with findings in normal counterparts.     

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.     

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。     

环孢素A联合沙利度胺治疗骨髓增生异常综合征对T细胞亚群影响的初步研究

为探讨环孢素A联合沙利度胺治疗骨髓增生异常综合征对T细胞亚群的影响,我们对2007年6月至2012年6月我院血液内科门诊和住院低中危MDS患者22例,在给予环孢素A联合沙利度胺治疗前后,应用流式细胞仪对于患者和健康志愿者外周血标本进行T淋巴细胞亚群检测。我们发现:CD4+T细胞在MDS患者外周血的百分比低于正常对照组(P<0.05),CD4/CD8比值与正常对照组相比有显著性差异(P<0.05)。治疗显效组患者治疗后的CD4+T细胞百分率增高(P<0.05),CD8+T细胞百分率减低(P<0.01),CD4/CD8比值在治疗前后有显著性差异(P<0.05)。在治疗前,无效组和显效组在T细胞总数百分率,CD4+T、CD8+T细胞百分率以及CD4/CD8的比值均有统计学意义(P<0.05)。由此我们认为,MDS患者存在T淋巴细胞亚群免疫失调,细胞免疫功能紊乱。CsA以及沙利度胺治疗MDS的机制可能与Th,Ts细胞比率改变有关。     

Psychological stress during exercise: Lymphocyte subset redistribution in firefighters

The purpose of this study examined the changes in heart rate (HR), catecholamines (NE, EPI) and percentages of blood lymphocyte subsets (CD3+ T cells, CD3+ CD4+ helper T cells, CD3+ CD8+ cytotoxic T cells, CD3− CD56+ NK cells, CD4/CD8 ratio, CD19+ B cells, and total lymphocytes [NK cells+ T cells+ B cells]) in firefighters exposed to a computerized firefighting strategies and tactics decision-making challenge while participating in moderate intensity exercise. Furthermore, this study also examined the possible relationships between catecholamines (NE and EPI) and blood lymphocyte subsets following combined mental and physical challenge. Ten professional male firefighters participated in two counterbalanced exercise conditions on a cycle ergometer: (1) 37 min of cycle ergometry at 60% VO_2max (exercise alone condition; EAC) and (2) 37 min of cycle ergometry at 60% VO_2max along with 20 min of a computerized firefighting strategies and tactics decision-making challenge (firefighting strategies condition; FSC). FSC elicited significantly greater HR, NE, and EPI when compared to EAC. Both EAC and FSC elicited increases in CD3− CD56+ NK cells. The percentages of CD3+ T cells, CD3+ CD4+ helper T cells, CD4/CD8 ratio, CD19+ B cells, and total lymphocytes were lower immediately following both conditions. Following dual challenge NE AUC was negatively correlated with percentage of CD19+ B cells immediately post challenge, and HR was negatively associated with the percent change in the CD4/CD8 ratio from pre to post challenge. These elevations in NE and heart rate simultaneously in response to the dual challenge suggest greater sympathetic activation that in turn would possibly explain the alteration in the distribution of lymphocyte subsets.     

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.     

Intrathyroidal accumulation of T cell phenotypes in autoimmune thyroid disease.

In this study we have correlated peripheral T cell subset phenotypes with intrathyroidal lymphocyte accumulation in patients with autoimmune thyroid disease (Graves' and Hashimoto's disease). Our study utilized euthyroid family members for one of our control groups (n = 48) thus significantly limiting familial, but not disease-specific, influences on these T cell phenotypes. Our principal new observations were found only in patients with Graves' disease. As previously reported, there was a decrease in CD8+ (suppressor/cytotoxic) T cells in the peripheral blood of patients with untreated hyperthyroid Graves' disease (n = 27) (mean +/- SEM, 19 +/- 1.1% in patients compared with 25 +/- 1.2% in controls, p = 0.03), a finding not observed in treated, euthyroid Graves' disease patients or their relatives. However, the relative number of CD8+ T cells, assessed by CD4:CD8 ratios, was increased in the intrathyroidal T cell populations (n = 10), when compared to normal and patient peripheral blood. There were no consistent changes in total CD4+ (helper) T cells in the peripheral blood of patients with treated and untreated Graves' disease but a reduction in CD4+2H4+ (suppressor-inducer) T cells was seen in patients undergoing surgery for Graves' disease (13 +/- 6.9% compared with 39 +/- 3.4%). Again, however, this T cell subset was increased within the target organ of the same patients (41 +/- 5.9%). These data point to either a selective accumulation, or a specific "homing", of certain T cell subsets within the thyroid gland of patients with Graves' disease where T cell differentiation may be strongly influenced by antithyroid drug treatment and the local immune environment.