Increased spontaneous activity of antibody-forming cells in the peripheral blood of patients with active SLE.

Thirty-seven patients with criteria for systemic lupus erythematosus (SLE) and 18 normal controls were studied for their spontaneous background IgM antibody plaque-forming clell number to specific chemical haptens. Active SLE patients had significantly more plaque-forming cells in their peripheral blood to a total of five chemical determinants than did patients with inactive disease or controls. This increased number of plaque forming-cells correlated with depressed serum C3 levels by Spearman rank-order analysis. The finding of elevated numberof spontaneous IgM plaque-forming cells to defined chemical haptens supports the concept that active SLE demonstrates a generalized increase in B-cell activity toward a variety of antigens.     

Increased spontaneous osteoclastogenesis from peripheral blood mononuclear cells in phenylketonuria

Phenylketonuria (PKU) is commonly complicated by a progressive bone impairment of uncertain aetiology. The therapeutic phenylalanine (Phe)-restricted diet and the possible noxious effects of high plasma Phe concentrations on bone have previously been suggested as possible determinant factors. Since osteoclasts are involved in bone reabsorption, they could play a role in determining bone damage in PKU. The reported increased excretion of bone resorption markers in PKU patients is consistent with this hypothesis. Although different diseases characterized by bone loss have been related to increased spontaneous osteoclastogenesis from peripheral blood mononuclear cells (PBMCs), to date there is no evidence of increased osteoclast formation in PKU. In this study, we compared the spontaneous osteoclastogenesis from PBMCs in 20 patients affected by PKU with that observed in age- and sex-matched healthy subjects. Phenylketonuric patients showed the number of osteoclasts to be almost double that observed in controls (159.9 ± 79.5 and 87.8 ± 44.7, respectively; p = 0.001). Moreover, a strict direct correlation between the spontaneous osteoclastogenesis in PKU patients and the mean blood Phe concentrations in the preceding year was observed (r = 0.576; p = 0.010). An imbalance between bone formation and bone resorption might explain, at least in part, the pathogenesis of bone loss in this disease. These findings could provide new insights into the biological mechanisms underlying bone damage in PKU.     

Increased immunoglobulin-secreting cells in the blood of patients with active systemic lupus erythematosus

Peripheral blood lymphoid cells actively secreting immunoglobulin G (IgG), immunoglobulin A (IgA) and immunoglobulin M (IgM) were quantitated in 24 patients with systemic lupus erythematosus (SLE) and compared with the frequency of such immunoglobulin secreting cells (IgSC) in normal controls utilizing a reverse hemolytic plaque assay. The geometric mean frequency of IgG-secreting cells in the patients with SLE was $\text{489}\text{10}{}_6$ peripheral blood mononuclear cells which was significantly higher (p < 0.005) than the mean control value of 137. The mean frequency of IgA-secreting cells in the patients with SLE was $\text{293}\text{10}{}_6$ cells which was also significantly higher (p < 0.01) than the mean of 96 in the control group. IgM-secreting cells were present in normal frequency in the patients with SLE (53 versus $\text{66}\text{10}{}^6$ in controls). The 24 patients were ranked in order of disease activity, and correlation coefficients were calculated comparing disease activity with laboratory findings including white blood cell count, erythrocyte sedimentation rate, third component of complement (C3) levels, immunoglobulin levels, anti-DNA antibody concentrations and the frequency of IgSC. Disease activity had a significant positive correlation with the serum levels of IgG (p < 0.001) and IgM (< 0.01), and the erythrocyte sedimentation rate (< 0.01), and had a significant negative correlation with serum C3 levels (p < 0.01). There was a highly significant correlation between disease activity and the level of anti-DNA antibody (r = 0.711, p < 10^?5) but the highest correlation with disease activity found was the frequency of IgG-secreting cells in the blood of the patient with SLE (r = 0.777, p < 10^?6). These data demonstrate that SLE is associated with increased numbers of IgG- and IgA-secreting cells in the peripheral blood and that increases in these parameters are closely associated with active clinical disease.     

Increased osteoclast formation and activity by peripheral blood mononuclear cells in chronic liver disease patients with osteopenia

Osteoporosis is a common complication of chronic liver disease, and the underlying mechanisms are not understood. We aimed to determine if osteoclasts develop from osteoclast precursors in peripheral blood mononuclear cells (PBMCs) of chronic liver disease patients with osteopenia compared with controls. PBMCs were isolated and fluorescence-activated cell sorting was performed to quantify the activated T lymphocyte population and receptor activator of nuclear factor kappabeta ligand (RANKL) expression. The activated T lymphocyte populations were comparable for all 3 groups, and RANKL was not detectable. The percentage of CD14+CD11b+ cells containing osteoclast precursors was comparable between the 3 groups. To assess the formation and functional activity of osteoclasts formed from circulating mononuclear cells, PBMCs were cultured (1) without addition of cytokines, (2) with macrophage colony-stimulating factor (M-CSF), (3) with M-CSF and osteoprotegerin, and (4) with M-CSF and RANKL. The number of tartrate-resistant acid phosphatase-positive multinucleated cells and bone resorption was assessed. PBMCs from chronic liver disease patients with osteopenia formed more osteoclast-like cells, which, when cultured in the presence of M-CSF and RANKL resorbed more bone than controls. The number of osteoclast-like cells and the amount of bone resorption correlated with lumbar bone densities. Addition of M-CSF increased numbers of osteoclast-like cells formed in healthy controls; however, this was not observed in either of the chronic liver disease groups. Plasma levels of M-CSF were elevated in both patient groups compared with healthy controls. CONCLUSION: Circulating mononuclear cells from chronic liver disease patients with osteopenia have a higher capacity to become osteoclasts than healthy controls or chronic liver disease patients without osteopenia. This could partially be due to priming with higher levels of M-CSF in the circulation.     

Increased IgA-producing cells in the blood of patients with active Henoch-Sch?nlein purpura

Peripheral blood lymphocytes (PBL) producing IgA, IgM, and IgG, both spontaneously and after pokeweed mitogen stimulation in cell culture, were determined by a protein A hemolytic plaque assay in 20 children with active Henoch-Sch?nlein purpura (HSP), 42 with inactive disease, 22 normal controls of the same ages, and 18 children with upper respiratory tract infections (URTI). The geometric mean of circulating IgA-producing cells in active HSP (1,016 X divided by 1.55 cells/10(6) PBL) was increased when compared with the group with inactive disease (P less than 0.001), normal controls (P less than 0.001), and children with URTI (542 X divided by 2.03 cells/10(6) PBL, P less than 0.05). The number of circulating IgM-producing cells was slightly increased in active HSP (260 X divided by 2.65 cells/10(6) PBL) and URTI (256 X divided by 3.16 cells/10(6) PBL). Both values were higher than those found in the group with inactive disease (113 X divided by 2.26 cells/10(6) PBL, P less than 0.01). There were no significant differences among the 4 groups in the number of circulating IgG-producing cells. The number of cells producing each type of immunoglobulin after in vitro stimulation with pokeweed mitogen was similar in patients with active HSP, normal controls, and the group with inactive disease. These data demonstrate a selective increase in the number of circulating IgA-producing cells only during disease activity, and add further support to a possible pathogenic role of this immunoglobulin in HSP.     

Increased Fructose 2,6-bisphosphate in peripheral blood mononuclear cells of patients with diabetes

Fructose 2,6-bisphosphate (F2,6BP) is a powerful allosteric activator of 6-phosphofructo-1-kinase, which is the rate-limiting enzyme for glycolysis. Mitogenic stimulation of lymphocytes is related to an enhanced rate of glucose utilization and F2,6BP mediated activation of glycolysis. To determine the effect of hyperglycemia on intracellular glycolysis of lymphocytes, we measured intracellular F2,6BP content in peripheral blood mononuclear cells obtained from patients with diabetes and normal subjects. A total of 62 subjects participated in the present study. Venous blood samples were collected and peripheral blood mononuclear cells were separated by Ficoll gradients. Intracellular F2,6BP levels in peripheral blood mononuclear cells from normal control subjects were significantly lower than age-matched diabetic subjects. We observed a significant positive correlation between intracellular F2,6BP levels and long term glycemic control, as assessed by HbA1c. These data suggest that hyperglycemia increases intracellular F2,6BP in immune cells. These findings may help to clarify the impaired function in immune cells in patients with diabetes.     

Active E rosette-forming cells in the peripheral blood of patients with chronic active hepatitis

E rosette-forming cells (E-RFC) and active E-RFC were studied in the peripheral blood in patients with chronic active hepatitis (CAH), chronic persistent hepatitis (CPH), and in control subjects. A significant reduction in active E-RFC (a subpopulation of T lymphocytes considered actively involved in cell-mediated immune reactions) was found in patients with untreated CAH, both HB_sAg positive or negative. In contrast, patients with immunosuppressive-treated CAH showed an increased relative number of active E-RFC. A significant increase in active E-RFC was demonstrable in other immunosuppressivetreated chronic inflammatory disorders. In CPH active E-RFC were not different from normal controls. The number of E-RFC showed a relative and absolute decrease in both CPH and untreated CAH. These studies demonstrate an increase in active E-RFC in immunosuppressive-treated CAH and suggest that prednisone and azathioprine may have a differential effect on T lymphocyte subpopulations.This study was supported in part by a grant from the Instituto Nacional de Investigaçào Cientifica.     

乙型肝炎患者外周血单个核细胞端粒酶活性的研究

目的 了解乙型肝炎患者外周血单个核细胞（PBMC）端粒酶活性的表达情况。方法 通过扩增端粒重复序列（TRAP）及光度酶联免疫法，分别检测健康人及各类乙型肝炎患者PBMC的端粒酶水平。结果 各组患者PBMC在植物血凝素（PHA）刺激前均有端粒酶活性的表达，以急性型肝炎组最高，重型肝炎组最低，二者差别具有显著性（P〈0．001）。PHA刺激后与刺激前比较各组端粒酶活性均有显著性升高（P〈0．001），刺激后的端粒酶水平以重型肝炎组为最低，与其他三组比较差别具有显著性（P〈0．05）。慢性重型乙型肝炎经胸腺五肽（TP5）治疗后端粒酶活性显著增高（P〈0．05）。结论 HBV急性感染期PBMC的端粒酶水平升高；慢性感染期PBMC的端粒酶水平在体内被抑制。TP5具有免疫调节作用，能使过低的端粒酶水平趋向于正常。     

Increased proportions of peripheral blood gamma delta T cells in patients with pulmonary tuberculosis.

There is a small population of peripheral T cells bearing the gamma delta T-cell receptor, which may be involved in the defense against invading microorganisms and tumor cells. The present study was designed to evaluate the levels of gamma delta T cells in patients with pulmonary tuberculosis, bacterial pneumonia, chronic lower respiratory tract infection, lung cancer, and normal control subjects with or without old tuberculous lesion. The results showed that only patients with tuberculosis had significantly increased proportions of peripheral blood gamma delta T cells. This study suggests that the increased proportions of gamma delta T cells in tuberculosis could be related to T-cell activation by Mycobacterium tuberculosis, although it remains to be investigated which components of mycobacteria are the major ligands for gamma delta T cells.     

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.     

免疫功能受损宿主外周血单个核细胞端粒酶活性研究

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Increased production of tumor necrosis factor-alpha by peripheral blood mononuclear cells in the patients with aplastic anemia

The activity of tumor necrosis factor-alpha (TNF-alpha) in the supernatant of cultured peripheral blood mononuclear cells (PBMC) was measured in patients with aplastic anemia. It was significantly higher in patients with aplastic anemia than in normal controls, both when PBMC were unstimulated or when they were stimulated with PHA. Results from aplastic anemia patients were also significantly higher than patients who had received allogeneic bone marrow transplants. In aplastic anemia patients, the TNF-alpha value produced by PBMC upon stimulation and the platelet count were inversely correlated, as well those patients who had high TNF-alpha values tended to have lower hemoglobin and leukocyte values although this was not significant statistically. These results suggest that the increased production of TNF-alpha by PBMC plays a role in the severe suppression of hematopoiesis in aplastic anemia.     

Increased number of phagocytic cells among the peripheral blood lymphocytes of mammary tumor patients

Blood lymphocytes of 15 malignant mammary tumor patients and 16 healthy blood donors were tested for latex particle phagocytosis. The mononuclear cell fraction of breast cancer patients has been shown to contain significantly higher number of phagocytic cells (39.13 +/- 13.93%) compared to the values of healthy controls (20.34 +/- 6.96%) (p less than 0.01). It was demonstrated that the majority of phagocytic cells adhered to the glass of the hemocytometer, thus they could be distinguished morphologically from the non-adherent, non-phagocytic cells. The number of ingested latex particles per phagocytic cell was also higher in patients. This result shows that the activity of phagocytic cells was also increased in mammary tumor patients compared to controls.     

趋化因子受体CCR4在活动期类风湿关节炎患者外周血CD4+T细胞表达的研究

目的 探讨活动期类风湿关节炎(aRA)患者趋化因子受体CCR4在外周血CD4+T细胞的表达及其意义,并对该表达与血清重要细胞因子水平的相关性进行研究.方法流式细胞计数方法对12例aRA患者和10名健康对照者外周血CD4+T细胞表面CCR4的表达情况进行检测;酶联免疫吸附试验(ELISA)方法检测患者血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-10及干扰素(IFN)-γ的水平并记录患者相关临床及实验室资料.分析及评价aRA患者外周血淋巴细胞中CD4+CCR4+T细胞百分数(CD4+CCR4+T%)与TNF-α、IL-10、IFN-γ的血清水平及临床指标的相关性.结果 aRA组外周血CD4+CCR4+T%明显高于健康对照组(P＜0.01).aRA组血清TNF-α(P＜0.01)、IL-10(P＜0.01)及IFN-γ(P＜0.01)水平明显高于健康对照组.在检测的细胞因子中,aRA患者血清IL-10水平与CD4+CCR4+T%呈明显正相关关系(r＝0.66,P＜0.05).在临床指标中,CD4+CCR4+T%与aRA患者Lunsbury关节指数、血沉、C反应蛋白及血小板计数呈明显正相关关系(P＜0.05).结论 CCR4在CD4+T细胞趋化至病变关节过程中可能发挥重要作用;外周血CD4+CCR4+T%与血清IL-10水平密切相关;该百分数可以作为临床评价RA活动性的一个敏感指标.