Flow cytometric analysis of peripheral T lymphocytes from patients with mycobacterial diseases]

Flow cytometric analysis was carried out on peripheral blood cells from patients with tuberculosis (TB) (n = 84) and with mycobacteriosis other than tuberculosis (MOTT) (n = 38). A whole blood-staining-hemolysis procedure was used for the preparation of samples being analyzed, and the cells were double-stained with various combinations of fluorescein isothiocyanate (FITC)- and phycoerythrin (PE)- labeled monoclonal antibodies. These procedures enabled us to obtain quite reproducible results. As patients of more than 70 years old showed apparently distinct T lymphocyte profiles compared with those less than 70 years of age, this investigation was carried out only on patients of less than 70 years old. 1) The proportion of total lymphocytes to total leukocytes was significantly low in TB- and MOTT- groups, when compared with that in the healthy control group, although the total peripheral leukocyte number was not significantly different from each other. Thus, absolute numbers of lymphocytes were decreased significantly in TB- and MOTT- patients. 2) The numbers of both T and B lymphocytes in peripheral blood decreased in patients of both groups, leaving the ratio of T/B relatively constant. 3) Both CD4+/CD8- and CD4-/CD8+ subsets of T lymphocytes decreased in TB- as well as MOTT- groups. However, the decrease in CD4+/CD8- subset was more manifest than that in CD4-/CD8+ subset. Among CD4+/CD8- subset the proportion of the Leu8+ subpopulation was slightly lower and among CD4-/CD8+ subset CD11b- subpopulation was slightly higher in both TB- and MOTT- groups than in healthy control group. 4) There was no significant difference in proportions of IL-2-receptor (p55 alpha chain) positive as well as HLA-DR positive T-lymphocytes between patient groups and healthy control group. 5) Both TB- and MOTT- groups were subdivided according to the extent of pulmonary lesion. Patients with the larger lesion showed remarkable decreases in the ratio of T lymphocytes to total peripheral leukocytes, the number of T lymphocytes, and the numbers of CD4+/CD8- and CD4-/CD8+ subsets, when compared with those with the smaller lesion. 6) Although the averages of absolute numbers of T lymphocytes, CD4+/CD8- and CD4-/CD8+ subsets were lower in patient groups than in the control group and the ratios of these to total lymphocyte counts and the ratio of CD4+ to CD8+ subsets were not significantly different between patient groups and control group, the distributions of each value of individual person were far broad in patient groups.(ABSTRACT TRUNCATED AT 400 WORDS)     

中国乙型肝炎患者外周血淋巴细胞亚群频率参考值范围

目的 调查中国乙型肝炎患者外周血淋巴细胞亚群频率参考值范围.方法 利用流式细胞术检测2846例乙型肝炎患者和117例健康人群外周血淋巴细胞亚群数值,调查我国健康人群和乙型肝炎人群的参考值范围.结果 调查了16～60岁健康人群和HBV感染相关的急性肝炎、慢性肝炎、重型肝炎和肝硬化人群外周血CD3+T淋巴细胞、CD3+CD...     

原发性胆汁性肝硬化T淋巴细胞亚群分析

目的 探讨原发性胆汁性肝硬化(PBC)患者的T淋巴细胞亚群及共刺激信号表达的特点及临床意义.方法 以未经治疗的98例PBC患者为研究组,性别、年龄匹配的健康人30名作为对照组.以流式细胞仪技术检测外周血淋巴细胞亚群以及T淋巴细胞表面的共刺激信号CD28.结果 PBC与对照组T细胞亚群差异有统计学意义:PBC组CD4+T淋巴细胞升高,CD8+T淋巴细胞下降,CD4+/CD8+比值上升(P<0.05);CD4+CD28-T细胞和CD8+CD28-T细胞明显增加(P<0.05).结论 PBC存在免疫调节功能的异常,其中CD28的表达明显减少;CD8+CD28-的细胞群可能在PBC中有一定的免疫调节作用.     

Changes in the expression of surface receptors on lymphocyte subsets in the elderly: quantitative flow cytometric analysis

The immunophenotype of circulating lymphocytes, including the intensity expression of surface receptors, changes with ageing. Until now, no results of systematic studies on age-dependent changes with respect to the expression of the major lymphocyte surface receptors in healthy elderly subjects have been reported. In order to identify age-related changes in both representation and immunophenotype of lymphocyte populations, we investigated, by means of triple-color whole-blood immunostaining and quantitative flow cytometry, the percent values and the absolute numbers, as well as the levels of surface antigen expression or antigen molecules per cell (ABC values x 10(3)), of different peripheral blood lymphocyte subsets from 23 healthy elderly subjects and 13 young donors. Naive (CD45RA+CD3+) T cells, total B cells, and CD5+ B lymphocytes are decreased (22%, 6%, 0.8% vs. 30%, 12%, 1.4%, respectively), whereas activated (HLA-DR+CD3+) and memory (CD45RO+CD3+) T cells, CD3+CD7- T lymphocytes, and lymphocytes expressing the NK marker CD56 are expanded in the elderly (2%, 53%, 13%, 6% vs. 0.8%, 45%, 8%, 8%, respectively). Moreover, T lymphocytes from elderly individuals express lower CD3 (61 +/- 10) compared to young (69 +/- 10). Considering the different T-cell populations, CD3 antigen is respectively decreased on CD45RO+ T cells (55 +/- 14 vs. 66 +/- 14) and up-regulated on CD56+ T lymphocytes (62 +/- 21 vs. 45 +/- 20). Increased CD8 expression characterizes CD3+CD7- lymphocytes (70 +/- 34 vs. 44 +/- 17) while HLA-DR on activated T cells is lower in old (39 +/- 7) than young (46 +/- 9) donors. CD7 is down-regulated both in T (22 +/- 3 vs. 28 +/- 3) and NK (48 +/- 18 vs. 71 +/- 18) cells, whereas CD2 expression, unchanged on NK cells, is up-regulated on T lymphocytes (54 +/- 10 vs. 41 +/- 8). Age-related changes in B-cell antigen expressions were also found: CD20 is increased (124 +/- 23 vs. 105 +/- 16) whereas, despite the unchanged CD5 expression of T cells, CD5 intensity on the B-cell subset co-expressing this antigen is higher in old (49 +/- 37) than in young (22 +/- 4) people. The observed changes in the expression of functionally important cellular receptors can contribute to the remodeling of immune function characteristic of the elderly. Moreover, since quantitative flow cytometry is becoming widely employed in clinical practice, our results also contribute to the assessment of specific age-dependent antigen expression changes to be considered for diagnostic approaches in the elderly.     

Increased proportion of Fas positive CD8+ cells in peripheral blood of patients with COPD

Chronic obstructive pulmonary disease (COPD) is characterised by chronic inflammation in pulmonary tissue and is also associated with systemic effects. The objective of this study was determination of lymphocyte subpopulation and the expression of Fas receptor on lymphocytes derived from peripheral blood of patients with stable COPD (n=18) and a control group: asymptomatic smokers (n=12) and non-smokers (n=12). Flow cytometry method with monoclonal antibodies was used for evaluation of lymphocyte subsets: CD4+ and CD8+ and the expression of Fas (CD95) on T lymphocytes. We found an elevated proportion of CD8+ cells in the blood of COPD patients. Proportion of Fas+ T lymphocytes was significantly higher in patients with COPD when compared with asymptomatic smokers and non-smokers (mean: 84.4% vs. 71.6% vs. 61.0% for Fas+/ CD4+ and 88.1% vs. 73.8% vs. 58.3% for Fas+/CD8+ lymphocytes). The proportion of Fas positive CD8+ cells significantly correlated with the degree of airway obstruction and hypoxemia. The significant correlations of Fas positive CD4+ and Fas positive CD8+ with smoking history expressed as pack years smoked were observed. Our observation of an elevated proportion of circulating lymphocytes bearing Fas receptor may play a role in induction of these cells' apoptosis and indicate the role of Fas/ FasL pathway in the changes in proportion of lymphocyte subpopulations in patients with COPD.     

Natural killer cell activity in multidrug-resistant pulmonary tuberculosis.

BACKGROUND: Multidrug-resistant pulmonary tuberculosis (MDRTB), a major problem in developing countries, may result from either insufficiency of host cellular immune response or mycobacterial mechanisms which has been more intensively investigated so far. OBJECTIVES: The aim of the study was to investigate natural killer cell activity (NKA) and T lymphocyte subsets in HIV- patients with secondary MDRTB. Methods: 20 male patients with MDRTB (mean age 38 +/- 8 years), 15 nonresistant tuberculosis male patients (NRTB) (mean age 36 +/- 11 years) and 12 healthy male controls (mean age 35 +/- 8 years) were included. The percentages of CD3+, CD4+, CD8+, CD25+, CD11b+ and CD16+56+ cells were measured by flow-cytometric analysis of peripheral blood lymphocytes (PBL). NKA was evaluated using the anticandidal index method. RESULTS: The mean tuberculin response was higher in MDRTB and NRTB patients compared to controls (15.4 +/- 3.8, 15.1 +/- 3.3 and 10.9 +/- 2.8 mm, respectively; p < 0.001). There was no significant correlation between PPD response and PBL subsets or NKA. The percentages of both CD3+ and CD3+CD4+ T lymphocytes were significantly lower in MDRTB (62.4 +/- 12.1 and 33.9 +/- 9.0%) compared to NRTB (70.8 +/- 7.5 and 42.9 +/- 8.6%; p < 0.05). Patients with MDRTB had significantly lower NKA compared to NRTB and controls (30.9 +/- 11.3, 49.7 +/- 15.5 and 40.0 +/- 8.5%, respectively; p < 0.01). CONCLUSION: This reduction in NKA may suggest a role for impaired NK function in the pathogenesis of MDRTB in HIV- patients.     

Correlation between cytomegalovirus and toxoplasma gondii serology and lymphocyte phenotypes in peripheral blood and cord blood

Increased numbers of peripheral blood HNK1+ lymphocytes have been reported in transplant recipients, hemophilia patients treated with clotting factor concentrates, and HIV carriers. Our previous work has revealed a significant effect of cytomegalovirus (CMV) carrier status on HNK1+ lymphocytes. Since other antigenic stimuli may be also involved, we compared the effects of Toxoplasma gondii and CMV carrier status on lymphocyte subsets as defined by the CD3 and HNK1 markers in 288 healthy individuals. In contrast to CMV, T gondii carrier status had no significant effects on the CD3+, HNK1+ and CD3-, HNK1+ lymphocyte subsets. That result may be explained by the different relationships between these 2 microorganisms and their hosts. We also studied the effect of maternal CMV carrier status on the HNK1 expression by their offsprings' cord blood lymphocytes. None of the 100 newborns studied had serological evidence of congenital CMV infection. There were only few HNK1+ lymphocytes in the cord bloods, of which the majority was CD3-, and their proportions were not significantly influenced by maternal CMV carrier status. Since there has been probably no direct contact between maternal CMV and the newborns' immune systems, we suggest that the effect of CMV on the HNK1+ lymphocytes of its carriers, results from a direct interaction between virus or virus-infected cells and the immune system.     

Study of hematopoietic progenitor cells, hematological values and lymphocyte subsets in cord blood: application for cord blood transplantation

Hematological values, lymphocyte subsets and hematopoietic progenitor cells from normal term cord blood samples were studied, compared with normal adult blood, and analysed to determine whether a single collection of cord blood is sufficient for transplantation in adults. The parameters were assayed by automatic cells counter, flow cytometry and semisolid cell culture. All of the hematological values except RBC and MCHC were higher than in normal adult blood. Sex had an influence on RBC, Hb, Hct, Plt and reticulocyte counts. For lymphocyte subsets, all of the absolute CD3+, CD4+, CD8+ counts and T helper: suppressor ratio were higher than those of adult blood. All of the hematopoietic progenitor cells in cord blood were also higher than in adult blood. The mean volume of cord blood for each collection was 80.75 +/- 4.81 ml and the mean numbers of nucleated cells, CFU-GM and CD34+ were 13.51 +/- 0.38 x 10(8) cells, 4.33 +/- 0.66 x 10(5) colonies and 42.65 +/- 7.00 x 10(5) cells respectively. This 80 ml of cord blood would contain sufficient marrow repopulating cells for a recipient weighing about 20 kg. Recently developed technology, including ex vivo expansion may even permit transplants in adults.     

Age- and sex-related changes of the lymphocyte subsets in healthy individuals: an analysis by two-dimensional flow cytometry

The relative and absolute numbers of the peripheral blood lymphocyte (PBL) subpopulations from 156 healthy men and women of different ages (20-99 years old) were studied by the use of monoclonal antibodies (MoAbs) and two-dimensional flow cytometry. The percentage of pan-T MoAb-positive cells decreased with age, which was attributable to a relative decline in the CD8+ suppressor/cytotoxic T cells, more precisely in the CD8+ CD11- cytotoxic T cells. Those of CD4+ Leu-8- helper T cells, CD3+ HLA-DR+-activated T cells, and natural killer subsets (Leu-7+, CD16+, Leu-7+ CD16+ or Leu-7+ CD16-) increased with age. The absolute numbers of most of the lymphocyte subsets examined declined with age except that those of natural killer cell subsets and helper T cells remained unchanged. It should be noted that the PBL subsets differed markedly according to age and sex, the changes being more evident among women.     

Lymphocyte subpopulations in peripheral blood in primary sclerosing cholangitis

BACKGROUND/AIMS: Primary sclerosing cholangitis (PSC) is a chronic disease of autoimmunological etiology, leading to inflammation, destruction and atrophy of the bile ducts. The aim of the study was to determine peripheral lymphocyte B, T, and NK cells in PSC. METHODOLOGY: The estimation of peripheral blood lymphocytes in 17 patients (54+/-12 years old) with PSC was carried out; the control group consisted of 27 subjects (38+/-11 years). The following T lymphocyte subpopulations were determined using duo color flow cytometry: CD3+, CD4+, CD8+, CD3++HLA DR+, B cells CD19+, and NK cells CD16+ +CD56+. RESULTS: In PSC we observed doubled increase in activated T lymphocytes of CD3+ +HLA DR+ phenotype as compared to healthy subjects (7.9% vs. 2.7%, p<0.01) and NK cells (12.6% vs. 10.3%, respectively, p<0.05). There were no significant differences in the composition of CD3+, CD4+, and CD8+. In peripheral blood we noted, in patients with PSC, a decrease in B lymphocytes (11.2% vs. 12.3%, p<0.19). CONCLUSIONS: The examinations showed that activated T (HLA DR+) lymphocytes and NK cells played an important role in development of PSC.     

Immunophenotypical changes of T lymphocytes in the elderly

BACKGROUND: Substantial changes in both representation and function of T lymphocyte subsets have been reported with advancing age. However, till now, no systematic studies focused on age-dependent changes in the expression intensity of the major T lymphocyte surface receptors. OBJECTIVE: The present study was undertaken in order to establish age-related differences in lymphocyte subpopulations by simultaneously measuring three surface antigens in young and elderly people. METHOD: Peripheral blood T cell subsets from 20 healthy elderly individuals and 15 healthy young adult donors were examined by means of a quantitative three-color flow cytometry method. RESULTS: Activated (HLA-DR+) and memory (CD45RO+) T cells, CD3+CD7- T lymphocytes, and cells expressing natural killer (NK) markers (CD3-CD56+ NK cells and CD3+CD56+ T lymphocytes) were expanded, whereas T lymphocytes expressing the adhesion molecule CD62L were lower in elderly compared with young donors. In addition to alterations in the percentages of T cell subsets during senescence, several changes in the intensity expression of T cell antigens were also detected. CD3 antigen expression was downregulated on total T lymphocytes as well as on the memory T cell subset, while CD56+ T cells exhibited increased CD3 levels. Moreover, CD2 expression, unchanged on NK cells, was upregulated on T lymphocytes from elderly subjects. CD3+CD7- T cells exhibited increased expression of CD8 antigen, while the intensity expression of HLA-DR on activated T cells and CD7 on both T and NK lymphocytes was decreased. T cells from elderly subjects also exhibited higher expression of CD50 and CD62L adhesion molecules as compared with young ones. CONCLUSION: These T cell antigen expression modulations during senescence, in addition to the alteration in the frequency of the various T lymphocyte subsets, could contribute to the complex remodeling of the immune function characteristic of the elderly.     

老年患者淋巴细胞亚群及其相关因素分析

目的了解老年患者的淋巴细胞亚群情况以及与日常生活能力、心理健康、营养的关系。方法采用淋巴细胞及亚群检测、日常生活能力量表(ADL)、症状自评量表(SCL-90)、小型营养评价法(MNA)等对住院老年患者进行调查。结果老年人CD3+、CD4+、CD8+、CD19+、Lc低于对照组,CD4+/CD8+高于对照组(P<0.01),CD19+与年龄呈负相关(P<0.05);心脑血管疾病组和感染组CD8+下降(P<0.01)比CD4+更明显(P<0.05),日常生活能力与CD19+、Lc呈负相关(P<0.05),与CD56+呈正相关(P<0.01)。SCL-90与CD56+相关明显(P<0.01);MNA与CD4+/CD8+、CD56+相关(P<0.05),与CD8+呈负相关、CD19+呈正相关(P<0.01)。结论老年人细胞免疫和体液免疫功能均有所下降,且与日常生活能力、心理健康情况、营养有关。     

Quantitative monitoring of serum hepatitis B virus DNA and blood lymphocyte subsets during combined prednisolone and interferon-alpha therapy in patients with chronic hepatitis B

Several investigators have reported a significantly reduced CD4/CD8 ratio, as defined by monoclonal antibodies, in the peripheral blood of Caucasian patients with chronic active hepatitis B (CAHB). In Asian patients with chronic hepatitis B, quantitative analyses of subpopulations of peripheral blood lymphocytes have not been able to confirm these findings. In this work, we analysed the frequency of peripheral blood lymphocyte subsets in 10 Chinese patients with histologically proven CAHB and seven healthy Chinese individuals. Four of the 10 CAHB patients received combined prednisolone/interferon-alpha2b (IFN-alpha2b) therapy. Peripheral blood samples were consecutively collected for analysis of lymphocyte subpopulations using an indirect immunofluorescence (IF) method, and hepatitis B virus (HBV) DNA was quantified by a chemiluminescent, molecular-hybridization assay. Peripheral blood mononuclear cells from seven Chinese control individuals comprised 63 +/- 3% CD3+ cells, of which 41 +/- 4% were of CD4+ and 23 +/- 2% of CD8+ subsets. The mean CD4/CD8 ratio in the healthy controls was 1.9 (95% confidence interval = 1.1-2.7). The CD4/CD8 ratios were significantly reduced (P < 0.01) in the 10 patients with chronic hepatitis B, compared with those of the controls, owing to a significant increase in the number of CD8+ cells (P < 0.005). During the treatment with prednisolone, a significant increase in the CD4/CD8 ratio was observed in all treated patients. This increase was mainly caused by a decrease in the number of CD8+ cells and was accompanied by an increase in serum HBV DNA levels, which peaked during the latter part of the prednisolone cycle. During the treatment with IFN-alpha2b, a second increase in the CD4/CD8 ratio was observed, which was caused by an increase in CD4+ cells. A marked decrease in viral load was observed, during treatment with IFN-alpha2b, in patients with HBV DNA levels below 10 000 pg ml-1. Our data indicate that the CD4/CD8 ratios in Chinese CAHB patients do not differ from those of Caucasian patients with CAHB, when analysed using similar methods for the enumeration of lymphocyte subsets. Profound effects on cellular distribution and viral replication were noted during the combined prednisolone/IFN-alpha2b therapy. Additional studies of the modulatory effect of the combined therapy on the distribution of lymphocyte subsets and cytokine profiles in relation to the therapeutic outcome of HBV infection are warranted.     

Selective depletion and activation of CD8+ lymphocytes from peripheral blood of patients with polymyalgia rheumatica and giant cell arteritis.

A prospective study of 33 patients with polymyalgia rheumatica/giant cell arteritis (PMR/GCA) was undertaken, firstly, to monitor sequentially peripheral blood CD8+ lymphocyte levels and, secondly, to assess the expression of activation markers on T lymphocyte subsets. The results indicated that there was a significant decrease in absolute numbers and relative percentages of CD8+ T lymphocytes, which returned to normal ranges after approximately 24 months' treatment, and that there was an increased percentage of CD8+ lymphocytes in PMR/GCA which express HLA class II antigens.     

BY55 monoclonal antibody delineates within human cord blood and bone marrow lymphocytes distinct cell subsets mediating cytotoxic activity.

We had previously reported, using BY55 monoclonal antibody, a cell surface 80-kDa protein restricted to human functional peripheral blood cytotoxic lymphocytes with either natural killer CD3- or cytotoxic T lymphocyte CD3+CD8+ phenotype. In the present report, we studied the cytotoxic lymphocytes in adult bone marrow and newborn cord blood as these organs are commonly used as sources of hematological stem cells for allogeneic transplantation. Our results showed that BY55 mAb labeled only 5-10% of the bone marrow lymphocytes, which included a major proportion of CD3+ CD8+ cytotoxic T lymphocytes. Interestingly, within cord blood cells, BY55+ lymphocytes represented 20-35% of the lymphocytes corresponding exclusively to a CD3- cell subset. Furthermore, we detected in cord blood no cytotoxic T lymphocyte activity but we demonstrated that the CD3-BY55+ cell subset contained the whole natural killer activity.     

CD69, CD25, and HLA-DR activation antigen expression on CD3+ lymphocytes and relationship to serum TNF-alpha, IFN-gamma, and sIL-2R levels in aging

Aging is associated with changes in lymphocyte subsets and unexplained HLA-DR upregulation on T-lymphocytes. We further investigated this activation, by measuring early (CD69), middle (CD25), and late (HLA-DR) T-lymphocyte activation markers on CD3+ lymphocytes, across subjects (20-100 years) together with serum tumor necrosis factor (TNF-alpha), interferon-gamma (IFN-gamma), and soluble interleukin-2 receptor (sIL-2R). HLA-DR was present as a CD3+ HLA-DR+ subset that constituted 8% of total lymphocytes, increased twofold with age and included CD4+, CD8+, and CD45RA+ phenotypes. HLA-DR was also expressed on a CD8+ CD57+ subset. The CD3+ CD25+ subset constituted 13% of lymphocytes, fell with age but was weakly associated with the CD3+ HLA-DR+ subset especially in older subjects. A small 3-5% CD3+ CD69+ subsets showed no age effect. Serum sIL-2R, TNF-alpha, but not IFN-gamma, were associated with CD3+ HLA-DR+ lymphocytes, TNF-alpha with CD8+ CD57+ count and sIL-2R and IFN-gamma with the CD3+ CD25+/CD3+ CD4+ ratio. The study confirms age-related upregulation of HLA-DR on CD3+ lymphocytes, shows some evidence for associated upregulation of CD25 on CD3+ cells in older subjects, and links serum TNF-alpha, IFN-gamma, and sIL2-R to T-lymphocyte activation.     

慢性乙型肝炎患者外周血淋巴细胞亚群与病程相关性的研究

目的对慢性乙型肝炎轻中度、重度和肝硬化患者外周血淋巴细胞亚群的百分比和绝对细胞数进行观察,探讨慢性乙型肝炎患者外周血淋巴细胞亚群的变化与病程的关系.方法采集88例慢性乙型肝炎患者柠檬酸钠新鲜抗凝血,经流式细胞仪进行免疫分型检测.结果慢性乙型肝炎重度患者的CD3+CD4+细胞百分比显著低于轻中度患者(P＜0.05),肝硬化患者的CD3+和CD3+CD8+细胞百分比显著低于轻中度患者(P＜0.01).肝硬化患者CD3CD19+细胞百分比显著高于重度和轻中度患者(P＜0.01).CD4/CD8比例在慢性乙型肝炎轻中度、重度和肝硬化患者间无显著差异.肝硬化和重度患者淋巴细胞、CD3+、CD3+CD4+、CD3+CD8+细胞的绝对细胞数均显著低于轻中度患者(P＜0.01),且肝硬化患者CD3-CD16+56+细胞的绝对细胞数显著低于轻中度患者(P＜0.05).肝硬化患者与轻中度患者的DNA载量分布差异有显著性(P＜0.01),其高水平病毒载量的患者比例高于轻中度患者.结论慢性乙型肝炎轻中度发展为重度和肝硬化的过程中,外周血淋巴细胞亚群绝对细胞数随病情的进展显著减少,主要表现为CD3+CD4+和CD3+CD8+的T淋巴细胞亚群的百分比进行性降低.     

Assessment of peripheral blood lymphocyte subsets in idiopathic myelofibrosis

The objective of this study was to contribute to a better characterization of the immunological profile of idiopathic myelofibrosis (IM) at presentation by analysing the blood lymphocyte subsets and their possible correlations with other disease features. Absolute blood lymphocytes and lymphocyte subsets were assessed in 31 IM patients, compared with those from 34 healthy individuals, and correlated with the patients' main clinical, hematological and bone marrow histologic features. The mean lymphocyte count of the IM patients was 1.1 (SD 0.6) x 10(9)/L, versus 1.6 (SD 0.49) x 10(9)/L in controls (p = 0.0006), with 24 of the 31 patients (77.4%) showing lymphocytopenia (< 1.5 x 10(9)/L). IM patients had significantly lower counts of CD3, CD4, CD8, and CD3 -/ CD56+ cells, and significantly higher CD3 +/CD56 + lymphocyte counts. Although no significant differences were found between patients and controls with regard to CD19+/CD5+ cell counts, increased CD5 + B-cell lymphocytes were observed in three IM patients. In one of the latter patients, Ig gene rearrangement analysis of the heavy chain gene demonstrated such a subpopulation to be clonal, but the patient did not develop features of chronic lymphoid leukemia during a 5-yr follow-up. No correlation was found between the patients' blood lymphocyte counts and other disease features. We conclude that most IM patients have absolute lymphopenia, decreased T cells and increased cytotoxic T cells at diagnosis, and 10% of them show an increased CD5 + B-cell subpopulation.     

Altered cytokine expression of peripheral blood lymphocytes in polymyositis and dermatomyositis

OBJECTIVE: To investigate the intracellular and soluble cytokine levels and T cell subsets in peripheral blood of patients with active and inactive polymyositis and dermatomyositis. METHODS: The frequencies of T and B lymphocytes, T helper (Th), and T cytotoxic (Tc) cells and of interferon gamma (IFNgamma), interleukin (IL)4, and IL10 expression of CD4+ or CD8+ cells were determined by flow cytometry. The concentrations of soluble cytokines were measured with commercial enzyme linked immunosorbent assays. RESULTS: In active dermatomyositis there was a decreased percentage of T (CD3+) lymphocytes and Tc (CD8+) lymphocytes, decreased IFNgamma expression of CD4+ and CD8+ cells, but an increase in B and IL4 producing CD4+ lymphocyte frequencies. These prominent changes disappeared in the inactive stage of the disease. In polymyositis no significant change in these lymphocyte subsets or in intracellular cytokine expression could be detected in either the active or the inactive form. The frequency of IL4+/IFNgamma+ Th cells was calculated and a significantly increased Th2/Th1 frequency was found in active dermatomyositis, and a decreased frequency in inactive dermatomyositis, compared with the control population. CONCLUSIONS: There appears to be a difference between polymyositis and dermatomyositis in the level of peripheral blood lymphocytes and their intracellular cytokine content. These findings provide further evidence for a difference in the pathogenesis of polymyositis and dermatomyositis.     

Expression of activation antigens on lymphocyte surface and circulating platelet-leukocyte aggregates in ischaemic heart disease

BACKGROUND: Data from literature documented the role of the activation of circulating T lymphocytes and increased leukocyte adhesion to blood platelets in the destabilisation of an atheromatous plaque and progression of ischaemic heart disease to acute coronary syndrome. AIM: To assess whether there is an increased proportion of activated T lymphocytes and platelet-leukocyte aggregates (PLA) in the circulating blood in patients with myocardial infarction or with stable angina, and to examine whether these changes are related to the progression of clinical symptoms or coronary angiography results. METHODS: The study group consisted of 36 patients with ST-segment elevation acute myocardial infarction (STEMI), confirmed by elevated troponin T level (36 patients, 26 males, 10 females, mean age 61.8 years, range 42-78 years), 30 patients with stable angina and single-vessel disease (24 males, 6 females, mean age 58.8 years, range 43-69 years), and 20 control healthy age and gender-matched subjects. Lymphocyte activation was evaluated by the assessment of T lymphocytes CD3+ /CD69+, CD3+/HLA-DR+ and CD4+/CD154+. The PLA assessment, including platelet-granulocyte aggregates (PGA), platelet-monocyte aggregates (PMA) and platelet-lymphocyte aggregates (PlymphA) was based on the measurement of the proportion of CD45+/CD41a+ cells with the use of flow cytometry. RESULTS: The proportion of T lymphocytes CD3+/HLA-DR+, CD3+/CD69+ and CD4+/CD154 was significantly higher in patients with STEMI than in controls, and T lymphocytes CD3+/CD69+ - significantly higher in STEMI group than in both patients with angina or controls. There was no significant relationship between the proportion of activated lymphocytes and duration of anginal pain, troponin T concentration or the number of coronary vessels with critical stenosis. The proportion of PGA, PMA and PlymphA was significantly greater in STEMI patients than in patients with angina or controls. There was a significant positive correlation between the proportion of PLA and PMA, and the duration of anginal pain. Patients with stable angina had a significantly higher proportion of T lymphocytes CD3+/HLA-DR+ compared with controls. CONCLUSIONS: Circulating T lymphocyte activation is present in ischaemic heart disease. This phenomenon is more pronounced in patients with acute MI than in those with stable angina, and is not related to the degree of cardiac injury. An increased formation of platelet-leukocyte aggregates is also present in patients with acute MI.