Natural killer (NK) cells and interleukin 2 (IL-2) in Crohn disease

An analysis was made of spontaneous Natural Killer (NK) activity, the proportion of NK cells and Interleukin 2 (IL-2) production in response to a phytohemagglutinin (PHA) using isolated mononuclear peripheral blood cells from 27 patients with Crohn's disease (CD) and 27 healthy controls. The results obtained showed a decrease in NK activity in comparison with the control group (p less than 0.05), occurring mainly in patients in whom the disease was in an active phase (p less than 0.005), with no apparent relationship to the proportion of NK cells. IL-2 production was, however, similar to that of the healthy controls. A positive correlation between spontaneous NK activity and IL-2 production was evident. The origin of these findings is controversial, suggesting that other factors with a negative modulating effect may inhibit NK function.     

Serglycin expression in CD2+ and CD14+ cells from patients with various rheumatic diseases

The objective of the study was to look at the in vivo expression of serglycin in cells taken from patients with an inflammatory disease. The mRNA expression of the small proteoglycan serglycin was investigated in macrophages/monocytes and T-cells derived from the synovial fluid and blood of six patients with various rheumatic diseases and from the blood of two control subjects. Our results demonstrate higher Levels of expression in CD14+ cetts taken from patients with chronic inflammatory diseases than in control subjects. This suggests that serglycin may play a role during the inflammatory process.     

血浆白细胞介素2和白细胞介素6测定在风心病风湿活动临床诊断中的意义

目的：探讨风湿性心脏病患者白细胞介素－2和白细胞介素－6的血浆变化与风湿活动的关系。方法：随机选择25例风风病风湿活动患者及30例非风 湿活动患者,用放射免疫竞争结合法检测其血浆IL－2和IL－6水平,结果：风心病人风湿活动期血浆IL－6升高,IL－2降低,风心病非风湿活动患者血浆IL－6轻度升高,而IL－2无变化,结论：轻度IL－6升高的非风湿活动患存在隐匿风湿活动,需辅助以抗风湿治疗。     

自体骨髓单个核细胞移植治疗风湿性心脏病可行性研究

目的观察自体骨髓单个核细胞（BMMNCs）心肌内移植对风湿性心脏病患者心功能的影响,探讨其临床实用性。方法选取以左室扩大为主的风湿性心脏病患者5例,瓣膜置换手术当日采取髂骨骨髓20ml,同上法获得自体BMMNCs悬液备用。瓣膜置换完毕,于特定局部心肌内注入上述细胞悬液,分别于术前、术后1周及术后3个月行门控核素心血池显像（MGBP）检查,评价患者心功能及局部心肌舒缩能力,同时监测围手术期心电图及各项实验室指标。结果患者围手术期均未发现明显不良反应,术后3＋71反映左室收缩功能的各项指标显著升高,左室8个分区局部射血分数（rEF）亦明显升高,尤以S3～S6为著。结论自体BMMNCs心肌内移植方法简便、可靠,进一步的严格对照、多中心、随机双盲的大样本临床实验可以进行。     

IL-2 and IFN-gamma, but not IL-4 secretion by peripheral blood mononuclear cells (PBMC) are related to CD4+ T cells and clinical status in Brazilian HIV-1-infected subjects.

It has been reported that production of IL-2 and IFN-gamma, known as T-helper type 1 cytokines, by peripheral mononuclear cells (PBMC) decreases with progression of HIV infection. In contrast, IL-4 and IL-10 production, Th2 cytokine profile, increases with HIV disease progression. PBMC were evaluated from 55 HIV-infected subjects from Divis?o de Imunologia, Hospital das Clínicas, Faculdade de Medicina da Universidade de S?o Paulo, to "in vitro" cytokines production after 24 hours of stimulation with PHA. Low levels of IL-4 production in both HIV-infected patients and normal subjects, were detected. The patients with CD4+ T cell counts < 200 showed a significant decrease of IL-2 and IFN-gamma production compared to controls. Patients with higher counts of CD4+ T cells (either between 200-500 or > 500 cells/mm3) also showed decreased production of IL-2 that was not statistically significant. There was a correlation between IL-2 and IFN-gamma release with CD4+ T cells counts. HIV-1-infected individuals with CD4+ T cells > 500 cells/mm3 showed increased levels of IL-2 and IFN-gamma, than individuals with CD4+ T cells < 500 cells/mm3. In conclusion, we observed a decline of IL-2 and IFN-gamma production at advanced HIV disease. IL-4 production was not affected during HIV infection. Taken together, these findings suggest that the cytokine profile might be influenced by the HIV infection rather than the cause of disease progression.     

重型乙型肝炎患者肝组织中人纤维介素基因的检测及其与临床转归关系的探讨

目的 研究人纤维介素基因或纤维介素／hfg12凝血酶原酶(简称hfg12)在重型肝炎、慢性肝炎、肝硬化患者肝组织及非配对患者外周血单个核细胞(PBMC)中的表达，分析其表达与患者临床表现和肝功能的关系，探讨hfg12检测对预测患者临床转归和预后的价值。方法 采用免疫组织化学法和免疫细胞化学法分别检测23例重型肝炎、13例慢性肝炎、14例肝硬化患者的肝组织和30例重型肝炎、10例慢性肝炎患者、10例健康对照者PBMC中的hfg12表达。应用多媒体彩色图文分析系统对hfg12的表达进行半定量分析。结果 23例重型肝炎患者肝组织中，21例可见hfg12表达，而在慢性乙型肝炎、乙型肝炎肝硬化患者肝组织中均无hfg12表达。肝组织中hfg12表达水平与血清总胆红素值呈正相关。30例重型肝炎患者中，28例外周血白细胞hfg12高表达，10例慢性乙型肝炎患者中有1例PBMC可检测到hfg12，健康对照组未检测到hfg12。PBMC中hfg12表达水平与血清总胆红素值呈正相关。结论 Hfg12的表达为重型肝炎所特有的现象，hfg12的表达与乙型肝炎患者病情的严重程度密切相关。减少和阻断其表达有可能为防治重型肝炎提供新的途径和方法。PBMC中hfg12的检测可能有助于重型肝炎的早期诊断和临床转归的判断。     

Enhanced expression of CD69 and CD25 antigen on human peripheral blood mononuclear cells by prolactin

Several clinical reports have suggested that prolactin (PRL) plays an important role in the pathogenesis of autoimmune diseases such as rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE). We have investigated the influence of PRL on immune system, by evaluating the effects of PRL on the expression of CD69 and CD25 on human peripheral blood mononuclear cells (PBMCs). Human PBMCs obtained from healthy female volunteers were incubated with phytohemagglutinin (PHA) in the presence or absence of various concentrations of PRL. The expression of CD69 and CD25 was monitored using immunofluorescence staining and flow cytometry. PRL significantly enhanced the expression of CD69 and CD25 on activated PBMCs compared with that in the absence of PRL (p<0.05, paired t-test). Increasing doses of PRL enhanced the expression of CD69 up to 2 microg/ml and CD25 up to 1 microg/ml. The enhanced expression of CD69 was observed on CD8+ T lymphocytes but not on CD4+ T lymphocytes. Our data suggest that PRL can significantly enhance the expression of CD69 and CD25 molecule on human PBMCs when induced by PHA. However, PRL would have to be at optimal concentration in order to enhance their expression.     

Clinical significance of CD45RO expression on peripheral blood mononuclear cells in HTLV-I-infected individuals

The phenotype of peripheral blood mononuclear cells (PBMC) was examined in 13 healthy volunteers, 26 HTLV-I carriers, and 58 ATL patients (22 smouldering, five chronic, 24 acute, and seven lymphoma type). The percentage of CD4⁺, CD25⁺, CD28⁺ and CD45RO⁺ cells in the PBMC of the chronic and acute type patients was significantly higher than that of the volunteers, whereas the percentage of CD8⁺ and CD45RA⁺ cells in these patients was significantly low. The histogram for CD45RO fluorescence intensity (FI) revealed two patterns: pattern A consisted of CD45RO⁺ cells with high FI (CD45RO^high) and intermediate FI (CD45RO^int). Pattern B consisted exclusively of CD45RO^high. Pattern A was evident in all volunteers. The percentage of subjects showing pattern B was increased in an order that reflected disease progression. In the patients with pattern A, the CD45RO^int cells were CD4⁺ and CD8⁻, and the FI of CD2, CD3, and Fas within the CD45RO^int cells appeared to be lower than that within the CD45RO^high cells. The acute type patients with pattern A had a significantly longer survival curve than that of these patients with pattern B. These results suggest that the presence of CD45RO^int cells may be related to protection against disease progression in HTLV-I-infected individuals.     

Cytokines and immunoglobulin in rheumatic heart disease: production by blood and tonsillar mononuclear cells

Rheumatic fever and rheumatic heart disease are considered to result from abnormal immune responses after Group A streptococcal pharyngitis. Production of interleukin 1 (IL-1), tumor necrosis factor-alpha (TNF), interleukin 2 (IL-2) and immunoglobulin (Ig) by blood and tonsillar mononuclear cells from rheumatic or healthy children was measured after stimulation in vitro by pokeweed mitogen (PWM) or the streptococcal extracellular product, blastogen A (BLA). Tonsillar cells from patients with rheumatic heart disease produced significantly less IL-1, TNF, IL-2, and Ig than control tonsillar cells. In contrast, blood mononuclear cell cultures from rheumatic children produced more TNF and IL-2 than controls. Our findings suggest that abnormal regulation of cytokine and Ig production may contribute to the pathogenesis of acute rheumatic fever and rheumatic heart disease.     

Peripheral T lymphocyte cytokine profile (IFNgamma, IL-2, IL-4) and CD30 expression/release during measles infection.

BACKGROUND AND OBJECTIVE: Measles virus infection (MVI) has been reported to be characterized by an imbalanced Th(1/2)-type cytokine profile. CD30 has been proposed as a receptor preferentially associated with the Th(0/2)-type cytokine pattern. The aim of this study was therefore to define the peripheral T lymphocyte cytokine profile and to test which CD30 expression pattern it was associated with in MVI. DESIGN AND METHODS: The design of the study was a prospective evaluation with comparative analysis. The serum levels of the soluble form of CD30 (sCD30) were determined at diagnosis and at weekly intervals up to 4 weeks, using an ELISA, in 23 males (median age 19), who developed MVI while serving in the Italian army and who were admitted to the Infectious Disease Unit of the Military Hospital in Padua. In 10 of the patients at diagnosis we studied the lymphoid immunophenotype and, after non-specific ex vivo stimulation, the expression of IFNgamma, IL-2 and IL-4 by peripheral T cells using flow cytometry single cell analysis. In 3 patients such evaluations were also performed 7 weeks later. RESULTS: At diagnosis, we found (i) reduction of IFNgamma+/CD4+ T cells (p=0.048 vs controls) in the absence of substantial variation of IL-2+ and IL-4+ T cells (p=ns vs controls); (ii) expansion of CD30+/ CD4+ and CD30+/CD8+ T cell subsets (p<0.01 vs controls); (iii) high sCD30 values (median 61 U/mL; p<0.001 vs controls); (iiii) a context of lymphopenia (0. 728+/-0.292 lymph x10(9)/L). sCD30 remained elevated up to 4 weeks from MVI onset [median values 53, 49, 50, 34 U/mL after 1, 2, 3 and 4 weeks, respectively (p=ns between different time points)]. In 3 patients tested 7 weeks after diagnosis, we still observed decreased IFNgamma production by CD4+ and CD8+ T cells (p=0.05 and <0.01, respectively vs controls) and reduction of CD4+ and CD8+/IL-2+ T cells (p<0.01). INTERPRETATION AND CONCLUSIONS: MVI was characterized by featuresof inadequate Th/Tc(1) activation associated with increased circulating CD30+ T cells and elevated sCD30 levels, supporting a correlation between Th/Tc status and CD30 expression/release pattern in vivo.     

Increased expression of CD40 ligand (CD154) on CD4+ T cells as a marker of disease activity in rheumatoid arthritis

OBJECTIVES: The interaction between the activation induced surface glycoprotein CD40L (ligand) (CD154) on CD4+ T cells and its receptor CD40, which is expressed on various cell types, plays a crucial part in numerous cell mediated and humoral immune reactions that may be of pathogenetic importance in rheumatoid arthritis (RA). To further evaluate the pathogenetic role of CD40L in RA, expression of CD40L and various other T cell activation antigens as well as costimulatory molecules was investigated on CD4+ T cells in RA by flow cytometry. METHODS: Two colour flow cytometry was used to determine the percentage of CD4+ T cells expressing CD40L, CD69, CD25, HLA-DR, CD39, CD27 and CD28 in peripheral blood (PB) of 62 RA patients in comparison to 20 healthy controls (HC). Disease activity was assessed by clinical, laboratory and radiological examination. Status of clinical remission of RA was evaluated according to the ACR preliminary criteria for complete clinical remission of RA. RESULTS: CD40L was expressed on > 10% of CD4+ T cells in 29% of RA patients thus defining a CD40L(high+) patient group. Disease activity as estimated by C reactive protein, rheumatoid factor and status of clinical remission of disease (p = 0.049) was higher in this subgroup than in the RA CD40L(low+) group. Expression of CD69, CD25, and HLA-DR was significantly increased in both RA patient groups in comparison with HC. However, the percentage of CD39+ CD4+ T cells was increased only in the RA CD40L(high+) subgroup (versus HC p = 0.019, versus RA CD40L(low+) p = 0.044). Furthermore, expression of CD40L and CD39 on CD4+ T cells correlated positively as estimated by Spearman rank correlation (p<0.001). The percentage of CD4+ T cells lacking the costimulatory molecules CD27 (p = 0.002) and CD28 (p = 0.026) was increased in RA CD40L(low+) patients in comparison with HC. CONCLUSIONS: These data suggest that increased expression of CD40L on CD4+ T cells in RA indicates prolonged and increased activation of CD4+ T lymphocytes and is associated with active disease and possibly an unfavourable prognosis. Whether this phenotypically defined RA CD40L(high+) subgroup will preferentially respond to an anti-CD40L antibody treatment remains to be elucidated.     

Induction of CD94/NKG2A expression on T cells in mixed lymphocyte culture by CD14+ cells from granulocyte colony-stimulating factor-mobilized peripheral blood mononuclear cells

We report the increased expression of inhibitory natural killer (NK) cell receptor, CD94/NKG2A, for human leucocyte antigen (HLA) class I on CD8+ T cells in granulocyte colony-stimulating factor-mobilized peripheral blood mononuclear cells (G-PBMC) after mixed lymphocyte culture (MLC). The addition of purified CD14+ cells to CD14-depleted G-PBMC-induced CD94/NKG2A expression in a dose-dependent fashion; however, this enhancing effect was inhibited by the membrane between responder cells and CD14+ cells. Therefore, CD14+ cells in G-PBMC induce CD94/NKG2A expression on CD8+ T cells, which in turn appear to downregulate alloresponses after allogeneic peripheral blood stem cell transplantation.