Production of interleukin (IL)-5 and IL-10 accompanies T helper cell type 1 (Th1) cytokine responses to a major thyroid self-antigen, thyroglobulin, in health and autoimmune thyroid disease

Tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma exert detrimental effects in organ-specific autoimmune disease, while both destructive and protective roles have been demonstrated for interleukin (IL)-10, IL-4 and IL-5. We examined the production of these cytokines by peripheral blood mononuclear cells (PBMC) from patients with Hashimoto's thyroiditis (HT), Graves' disease (GD) and healthy controls, upon exposure to a thyroid self-antigen, human thyroglobulin (Tg), in the presence of autologous serum. Initially, TNF-alpha and IL-2 were produced in all three groups, accompanied by IL-10. Release of IFN-gamma, IL-4 and, notably, IL-5 ensued. Both patient groups exhibited increased TNF-alpha, IL-2, IFN-gamma and IL-10 responses, and PBMC from HT patients secreted lower amounts of IL-5 than male, but not female, controls. Enhanced TNF-alpha production by HT cells also occurred in the presence of pooled normal sera, indicating a dependency on intrinsic cellular factors. Conversely, higher production of TNF-alpha and IL-5 occurred in the presence of autologous sera than in the presence of pooled normal sera in both patient groups, indicating a dependency on serum constituents. Complement appeared to promote the production of IL-2 and particularly IL-5, the levels of which were reduced by neutralization of complement by heat- or zymosan treatment. The production of IFN-gamma and IL-2 of the three groups together correlated directly with the serum anti-Tg activity. Moreover, TNF-alpha, IFN-gamma, IL-5 and IL-10 responses were markedly inhibited by partial denaturation of Tg by boiling. We hypothesize that autoantibodies and complement may promote mixed Th1/Th2 cell cytokine responses by enhancing the uptake of autoantigens by antigen-presenting cells.     

The upregulation of TNF alpha production is not a generalised phenomenon in the elderly between their sixth and seventh decades of life

The aim of this paper was to analyse the link between the intensity of tumor necrosis factor alpha (TNF alpha) production and the health status of the elderly and to find out whether the age between sixty and seventy is a 'turning point' for the changes in the production of this cytokine. Fifty elderly volunteers (age range: 60-70), twenty-five middle-aged (age range: 36-59) and fifty young (age range: 20-35) were enrolled into the study. Their health status was graded as 'healthy' and 'almost-healthy'. The level of TNF alpha was determined by bioassay, the activity of the TNF alpha gene was analysed in non-stimulated PBMC by the RT-PCR method. The results showed that the production of TNF alpha in the 'healthy' elderly people is not upregulated until the age of sixty-seventy. The 'almost-healthy' elderly are characterised by a higher release of TNF alpha from the cultures of non- and stimulated PBMC and an activation of the TNF alpha gene in the non-stimulated PBMC. Summarising, our results indicate that the deterioration of health status is a prerequisite for an exaggerated TNF alpha production by peripheral blood mononuclear cells of people between the sixth and seventh decades of life.     

Role of podoplanin in the high interleukin‐17A secretion resulting from interactions between activated lymphocytes and psoriatic skin‐derived mesenchymal cells

In the context of psoriasis, T helper type 17 (Th17) cells infiltrate the inflammatory site and interact with local mesenchymal cells, including skin fibroblasts. The aim of this work was to study the interactions of skin‐derived fibroblasts with peripheral blood mononuclear cells (PBMC) with a focus on the Th17 pathway and to identify a mechanism which leads to a high interleukin (IL)?17 secretion. A co‐culture system between PBMC and skin fibroblasts was developed. Healthy and patient PBMC were added to non‐lesional or lesional skin fibroblasts at a 5:1 ratio for 48 h in the presence or not of activation with phytohaemagglutinin (PHA). Monocytes were removed or not by adherence before the co‐culture. An anti‐podoplanin antibody was also used during the co‐culture. Cytokine production (IL‐8, IL‐6, IL‐1β and IL‐17) was measured by enzyme‐linked immunosorbent assay (ELISA) and cell staining (CD3, CD4, IL‐17 and podoplanin) by flow cytometry. Without T cell receptor (TCR) activation, IL‐8, IL‐6 and IL‐1β production increased in PBMC‐fibroblast co‐culture compared to PBMC alone. No additional effect was observed with TCR activation, with no difference in the Th17 cell percentage in activated‐PBMC alone or co‐cultured. Conversely, IL‐17 production was increased highly only in co‐cultures between control and patient activated‐PBMC and skin fibroblasts. Removal of monocytes decreased cytokine production, notably that of IL‐17. Addition of an anti‐podoplanin antibody decreased IL‐17 secretion by 60%. Interactions between resting PBMC and fibroblasts induce the IL‐8, IL‐6 and IL‐1β production. PBMC activation and cell interactions are critical for a high IL‐17 secretion. Podoplanin contributes largely to this massive IL‐17 secretion.     

Prediction of disease severity in neuromyelitis optica by the levels of interleukin (IL)‐6 produced during remission phase

T helper type 17 (Th17) cytokines have been implicated in the pathogenesis of neuromyelitis optica (NMO). As humanized anti‐interleukin (IL)‐6R (tocilizumab) immunoglobulin (Ig)G has been used as disease‐modifying therapy for NMO, the objective of our study was to investigate the role of endogenous IL‐6 on NMO‐derived CD4⁺ T cell behaviour. High production of IL‐6, IL‐17 and IL‐21 by CD4⁺ T‐cells was detected in NMO patients. Further, IL‐21 and IL‐6 levels were related directly to the level of neurological disabilities. The addition of anti‐IL‐6R IgG not only reduced directly the production of these cytokines, but also almost abolished the ability of activated autologous monocytes in enhancing IL‐6, IL‐17 and IL‐21 release by CD4⁺ T cells. In contrast, the production of IL‐10 was amplified in those cell cultures. Further, anti‐IL‐6R monoclonal antibodies (mAb) also potentiated the ability of glucocorticoid in reducing Th17 cytokines. Finally, the in‐vivo and in‐vitro IL‐6 levels were significantly higher among those patients who experienced clinical relapse during 2‐year follow‐up. In summary, our results suggest a deleterious role of IL‐6 in NMO by favouring, at least in part, the expansion of corticoid‐resistant Th17 cells.     

白细胞介素6基因-572C/G多态性与心肌梗塞易感性的关联研究

目的观察白细胞介素6（interleukin 6，IL6）基因-572C／G多态性在中国人群中的分布频率、与心肌梗塞（myocardial infarction，MI）易感性的关系、对MI患者冠脉病变程度的影响以及初步对该位点基因变异进行功能性分析。方法应用聚合酶链反应．限制性片段长度多态性方法对232例MI患者和260名正常对照者IL6基因-572C／G多态性进行了分析，观察了该基因多态性对冠脉病变程度及外周血单核细胞（peripheral blood mononuclear cells，PBMC）产生IL6能力的影响。结果中国汉族人群存在IL6基因-572C／G多态性；两组人群基因型、等位基因频率差异存在统计学意义，MI组CG＋GG基因型频率、G等位基因频率均显著高于对照组（P〈0．01）；基因型频率的相对风险分析发现，G等位基因携带者息MI的风险是CC基因型的1．68倍（95％CI：1．17—2．41，P〈0．01）；-572C／G多态性在单支、双支、三支冠脉病变组间的分布差异无统计学意义（P〉0．05）；G等位基因携带者氧化低密度脂蛋白刺激24h PBMC产生IL6的能力明显高于CC基因型（P〈0．05）。结论IL6基因-572G等位基因可能是中国汉族人MI的易感因子，这可能与携带该等位基因的人群存在IL6水平的高表达有关。     

A recombinant extracellular domain of the human interleukin 4 receptor inhibits the biological effects of interleukin 4 on T and B lymphocytes

Human interleukin 4 (IL4) acts on various hematopoietic cell types through interaction with a specific cell surface receptor (IL4R), whose cDNA has been cloned. We have produced a cDNA encoding a soluble form of the extracellular domain of the human IL 4R (sIL4R) and describe here the capacity of sIL4R to antagonize the in vitro activities of IL4 on normal B and T lymphocytes. sIL4R inhibited IL4-induced proliferation of both phytohemagglutinin-preactivated peripheral blood mononuclear cells (PBMC) and anti-IgM co-stimulated tonsil B cells with similar efficiency. This inhibitory activity was specific since sIL4R did not affect IL2-dependent proliferation of these cells. sIL4R also blocked IL4-dependent induction of the low-affinity receptor for IgE on B cells and inhibited IgE production by IL4-activated PBMC. Thus, in contrast to the IL6R extracellular domain which stimulates IL6 biological activity, the IL4R extracellular domain is a powerful antagonist of its specific ligand.     

T-lymphocyte activation in IgA nephropathy: Serum-soluble interleukin 2 receptor level,interleukin 2 production,and interleukin 2 receptor expression by cultured lymphocytes

The present study was undertaken to examine the T-lymphocyte activation in IgA nephropathy. Serum-soluble interleukin 2 receptor (sIL2R) levels were studied in 29 IgA nephritic patients, 17 patients with chronic glomerulonephritis (non-IgA nephropathy), and 30 healthy controls during an infection-free period. No difference in serum sIL2R level was demonstrated among these three groups of subjects. However, the serum sIL2R levels of IgA nephritic patient rose significantly during clinical exacerbation with synpharyngitic macroscopic hematuria and the serum sIL2R levels fell when hematuria subsided. Mitogen-stimulated cellular interleukin 2 receptor (IL2R) expression, sIL2R release, and interleukin 2 (IL2) production were also examined in peripheral blood mononuclear cells (PBMC) cultured for 24–48 hr in 21 patients with IgA nephropathy, 17 patients with chronic glomerulonephritides, and 17 healthy controls. The total cellular IL2R expression and sIL2R release did not differ among these three groups of subjects. However, the individual T-cell subsets bearing IL2R were distinctly different between IgA nephritic patients and the other two groups of controls. IgA nephritic patients had increased activated CD4+ lymphocytes and reduced activated CD8+ lymphocytes. Furthermore, IL2 production in response to phytohemagglutinin and pokeweed mitogen stimulation was increased in lymphocytes from patients with IgA nephropathy. The IL2 production did not correlate with the quantities of cellular and sIL2R yet the cellular IL2R expression paralleled the sIL2R released by cultured lymphocytes. Our present study suggests that the T lymphocytes from patients with IgA nephropathy have a defect in overproduction of IL2 and increased activated T helper-cell subset upon mitogenic stimulation. Serum measurement of sIL2R could potentially be useful in monitoring the disease activity.     

Increased expression of mRNA encoding interleukin (IL)-4 and its splice variant IL-4delta2 in cells from contacts of Mycobacterium tuberculosis, in the absence of in vitro stimulation

Expression of interleukin (IL)-4 is increased in tuberculosis and thought to be detrimental. We show here that in healthy contacts there is increased expression of its naturally occurring antagonist, IL-4delta2 (IL-4delta2). We identified contacts by showing that their peripheral blood mononuclear cells (PBMC) released interferon (IFN)-gamma in response to the Mycobacterium tuberculosis-specific antigen 6 kDa early secretory antigenic target (ESAT-6). Fresh unstimulated PBMC from these contacts contained higher levels of mRNA encoding IL-4delta2 (P=0.002) than did cells from ESAT-6 negative donors (noncontacts). These data indicate that contact with M. tuberculosis induces unusual, previously unrecognized, immunological events. We tentatively hypothesize that progression to active disease might depend upon the underlying ratio of IL-4 to IL-4delta2.     

外周血单个核细胞在狼疮性肾炎病人体内高效表达IL-6的观察

为了确定狼疮性肾炎（ＬＮ）病人体内是否存在内源性ＩＬ－６表达增高及其来源，作者应用ＥＬＩＳＡ法检测５８例ＬＮ病人的血清及外周血单个核细胞（ＰＢＭＣ）内ＩＬ－６蛋白的含量，用原位杂交方法结合ＩＢＡＳ２．０图像分析系统，检测其中２０例ＬＮ病人ＰＢＭＣ在未受任何刺激时ＩＬ－６ｍＲＮＡ表达强度，并分析三者之间的关系。结果表明该三者水平在ＬＮ病人体内均异常增高，活动期尤其明显，三者之间互呈直线正相关。提示ＬＮ病人ＰＢＭＣ内源性过度表达和合成ＩＬ－６是其外周血ＩＬ－６水平增高的原因之一，异常增高的循环ＩＬ－６对狼疮性全身性病理损害，尤其对肾损害具有重要的作用。此外，血清ＩＬ－６过高还提示ＬＮ处于活动期，对临床指导治疗有一定意义。     

Cytokine and chemokine levels in systemic sclerosis: relationship with cutaneous and internal organ involvement

Systemic sclerosis (SSc) is a connective tissue disorder characterized by excessive collagen deposition in the skin and internal organs. Several cytokines and chemokines have been implicated in the induction of fibrosis, but a definitive relationship between specific cytokines and organ involvement has not been established yet. Serum samples, PBMC and T cell lines (TCL) obtained from 54 patients affected by SSc and 20 healthy donors (HD) were examined by ELISA for Interferon-gamma (IFN-gamma ), interleukin (IL)-4, IL-6, IL-10, IL-18, Transforming growth factor (TGF)-beta1, Tumour necrosis factor (TNF)-alpha, sCD30, Macrophage derived chemokine (MDC), Monocyte chemoattractant protein (MCP)-1, Macrophage inflammatory protein (MIP)-1alpha and Regulated on activation normal T-cell expressed and secreted (RANTES). In all the SSc serum samples, we found significantly increased levels of IL6, TNFalpha and MCP-1 but reduced amounts of gamma-IFN and MDC. IL6, IL10, IL18, MIP-1alpha and TNFalpha measured in supernatants from PHA-stimulated PBMC and IL6, MCP-1 and RANTES in supernatants from stimulated TCL were also increased in patients. MDC was decreased in all the biological SSc sources studied. TGF-beta1, IL10, and sCD30 were produced at a significantly lower level by SSc TCL. Serum IL6 and sCD30 levels were significantly increased in dc-SSc patients compared to lc-SSc as were levels of MCP-1 produced by PBMC and IL10 from TCL. We observed a strict relationship between pulmonary fibrosis and IL10, MCP-1 (both from TCL) and serum IL6. Kidney involvement was related to serum MCP-1 levels and IL18 production from PBMC. Oesophageal involvement correlated with MDC production from PBMC and IL10 synthesis by TCL. We showed that IL-6, IL-10, MDC and MCP-1 are variably associated with internal organ involvement and allow the discrimination between limited and diffuse forms of the disease.     

In vivo interleukin 6 gene expression in the tumoral environment in multiple myeloma

Whether interleukin 6 (IL 6) is an autocrine or paracrine myeloma cell growth factor in vivo remains unresolved. To identify which cells are producing IL 6 in vivo, we have studied the IL 6 gene expression in bone marrow mononuclear cells (BMMC) of 19 patients with multiple myeloma (MM) and in peripheral blood mononuclear cells (PBMC) of 9 patients with plasma cell leukemia (PCL). We found that the IL 6 gene was transcribed by BMMC of most patients with MM (79%). Further, IL 6 mRNA was not produced by purified myeloma cells from patients with either MM (5 patients) or PCL, but by the bone marrow environment, mainly by monocytes and myeloid cells (CD13+CD15+ cells). For 2 patients with PCL, for whom PBMC and BMMC samples were available, IL 6 mRNA could be detected in BMMC but not in PBMC. Finally, no IL 6 mRNA was detected in five freshly established IL 6-dependent myeloma cell lines. The present data give a clear-cut demonstration of the paracrine origin of IL 6 in vivo in human MM.     

川崎病患者外周血淋巴细胞凋亡减少的初探

目的：探讨川崎病（ＫＤ）的免疫发病机制。方法：对２６例ＫＤ患者和２０名正常儿童外周血单个核细胞（ＰＢＭＣ）经ａｎｔｉ－ＣＤ３诱导体外培养不同时间的凋亡进行计数凋亡细胞百分率和片段ＤＮＡ分析。结果：ＫＤ患者凋亡细胞百分率和片段ＤＮＡ出现时间较正常对照降低（Ｐ〈０．００１）和延迟，ＰＢＭＣ体外培养产生ＩＬ－６水平较正常对照显著升高（Ｐ〈０．００１）；加抗ＩＬ－６单抗培养或静脉注射免疫球蛋白（ＩＶＩＧ）     

Low concentrations of cytokines produced by allergen-stimulated peripheral blood mononuclear cells have potent effects on nasal polyp-derived fibroblasts

Accumulating data show that fibroblasts are important regulators in the development and maintenance of allergic airway inflammation. However, most studies so far have used individual recombinant cytokines in high concentrations, unlikely to be found in vivo. We aimed to investigate how cytokines produced by peripheral blood mononuclear cells (PBMC) affect fibroblast functions. Primary airway fibroblasts where incubated with allergen-stimulated or non-stimulated PBMC supernatants from allergic patients. The levels of cytokines in PBMC supernatants were measured and the expression of CD54, CD40 and CD106 as well as the production of eotaxin, interleukin (IL)-6 and IL-8 were assessed in fibroblasts. Although the levels of single cytokines measured in PBMC supernatants were low, a significant up-regulation of the surface molecules as well as of IL-6 and IL-8 production was found in fibroblasts cultured with allergen-stimulated PBMC supernatants as compared to non-stimulated, while the increase in eotaxin production was not significant. The evaluation of correlations between cytokines produced by PBMC and effects seen on fibroblasts did not indicate a crucial role for any single cytokine. Furthermore, the addition of comparably low concentrations of recombinant interferon (rIFN)-gamma or recombinant tumour necrosis factor (rTNF)-alpha did not induce the same effects as PBMC supernatants, the only exception being TNF-alpha as a direct inducer of CD54 expression. Our results show that synergistic mechanisms has a more important role than single mediators, highlighting important differences between in vitro experiments, where effects of individual mediators are studied, versus the actual situation in vivo.     

Heparin coating of extracorporeal circuits inhibits cytokine release from mononuclear cells during cardiac operations

PURPOSE: To evaluate whether the production of interleukin 2 (IL 2), interleukin 6 (IL 6) and interleukin 10 (IL 10) from stimulated peripheral blood mononuclear cells (PBMC) was affected by coating extracorporeal circuits in patients undergoing cardiopulmonary bypass (CPB). In addition, postoperative clinical parameters were compared between patients with heparin-coated and uncoated CPB. DESIGN: Prospective, controlled in vivo/ex vivo study. PROCEDURE: Blood samples were drawn immediately before, at the end and 24 hours after the end of CPB using either a conventional circuit (n=10) or a heparin-coated circuit (n=10) in patients undergoing CPB. Cytokine release on the supernatants of activated PBMC was detected. Cardiopulmonary parameters were measured before CPB, at ICU admission, 3 hours and 24 hours after ICU admission in both groups of patients. Statistical difference intragroups and between groups were investigated with the analysis of variance for repeated measures. RESULTS: IL 6 and IL 10 release was significantly less (p<0.05) in the heparin-coated group. No differences in clinical parameters were observed between the two groups. CONCLUSIONS: These results suggest that with the use of heparin-coated circuits there is a lower production of IL 6 and IL 10 from isolated PBMC than with uncoated circuits.     

Implantation: Stimulation of human endometrial epithelial cell interleukin 6 production by interleukin 1 and placental protein 14

The effect of interleukin 1 (ILI) and placental protein 14 (PP14)on the production of interleukin 6 (IL6) by cultured human endometrialepithelial cells prepared from endometrial biopsy material obtainedat different stages in the menstrual cycle was investigated.Basal IL6 production by cells prepared from proliferative endometriumwas greater than that produced by cells prepared from secretoryendometrium (7.3 ± 0.3 and 1.1 ± 0.2ng/well/24h respectively, P < 0.001). IL1 (0.025–2.5 ng/ml) causeda dose-dependent increase in IL6 production by cells preparedfrom both proliferative and secretory endometrium, but cellsprepared from secretory endometrium responded to a lower concentrationof ILI than those prepared from proliferative endometrium. ILI-stimulatedIL6 production by epithelial cells prepared from secretory endometriumtypically reached 10 times basal values, while in cells preparedfrom proliferative endometrium stimulated levels were approximatelytwice the basal values. PP14 (1–50 µg/ml) also causeda dose-dependent increase in IL6 production by epithelial cellsprepared from secretory endometrium, but had no effect on IL6production by cells prepared from proliferative endometrium.Even in secretory cells PP14 was less effective than IL1 atstimulating IL6 production, with stimulated levels only reachingtwice the basal values. This suggests that PP14 and IL1 actvia different mechanisms in the stimulation of IL6 production.The results show that IL6 production by human endometrial epithelialcells is stimulated by other immunomodulatory peptides and thismay be part of the network of such peptides in the endometriumwhich may influence embryo implantation.     

频谱水对大鼠血浆白介素1、6和白细胞行为的影响

目的 :探讨频谱水对大鼠白介素 1、6(IL 1、IL 6)和白细胞行为的影响。方法 :静脉内注入内毒素造成炎症刺激模型 ,用生化方法检测血浆IL 1、IL 6的含量 ,用活体方法观察白细胞粘附、游出行为。结果 :实验组大鼠饮用频谱水 30、60天后 ,血浆IL 1、IL 6的含量比对照组高 ,白细胞粘附、游出的数量比对照组明显减少。对照组大鼠白细胞粘附、游出的数量较实验组明显增多 ,而血浆中IL 1、IL 6的含量比实验组低。结论 :频谱水能够提高血浆IL 1、IL 6的含量 ,显著减轻白细胞粘附及游出。     

Soluble interleukin 2 receptor release, interleukin 2 production, and interleukin 2 receptor expression in activated T-lymphocytes in vitro.

Following activation in vitro, peripheral blood mononuclear cells (PBMC) express cell-associated interleukin-2 receptors (IL2R). The present study was undertaken to define the proportion of T lymphocyte subsets that express the IL2R (CD25 antigen) upon different mitogenic stimulation. Double immunofluorescence staining with different fluorochromes, fluorescein isothiocyanate and phycoethyrin, was applied for identification of IL2R positive cells and individual lymphocyte subset. The exact percentage of individual activated lymphocyte subset bearing IL2R was enumerated by photographic counting. There was paucity of IL2R in freshly isolated, unstimulated peripheral blood, PBMC cultured without mitogen, and cultured B lymphocytes. Following pokeweed mitogen stimulation in vitro, 19% of CD4 (T-helper/inducer) lymphocytes and 14% of CD8 (T-suppressor/cytotoxic) lymphocytes expressed IL2R. Similarly, 25% of CD4 lymphocytes and 19% of CD8 lymphocytes expressed IL2R following phytohemagglutinin stimulation in vitro. Contrary to the reported data of Tac-positive cells in human lymphoid tissues, our study revealed that, upon lectin mitogen stimulation, approximately 55% of IL2R positive PBMC were CD4 lymphocytes, and 45% of them were CD8 lymphocytes. These observations imply the plausible notion that interleukin-2 mediated immune activation of T lymphocytes in PBMC is different from that in local lymphoid organs. It was also demonstrated that the release of soluble IL2R (sIL2R) and IL2 production in supernatant from cultured PBMC varied with different lectin stimulation. A significant correlation was demonstrated between the cellular and soluble IL2R but the production of IL2 from activated mononuclear cells bore no good correlation with either the cellular IL2R expression or the release of sIL2R.     

In vitro correction of the interleukin 2 defect of autoimmune mice

The effect of concanavalin A (Con A) and/or phorbol myristate acetate (PMA) on interleukin 2 (IL 2) production and tritiated thymidine incorporation was measured in young (6 weeks) and old (16-24 weeks) autoimmune mice by pulsing 5 X 10(6) unfractionated spleen cells with 5 micrograms of Con A and/or 5 ng of PMA for variable periods of time. The apparent deficiency in Con A-stimulated IL 2 production manifested by mice prone to the development of autoimmune disease was repaired by the addition of PMA. PMA did not enhance interleukin 1 (IL 1) secretion in autoimmune MRL-lpr mice either alone or in combination with Con A. The addition of purified IL 1 to Con A-pulsed autoimmune cells did not increase IL 2 production. Freeze-thaw experiments suggested that PMA does not promote IL 2 synthesis. Con A-pulsed cells plus PMA-pulsed cells do not syngergize. These data indicate that autoimmune-prone mice are capable of producing IL 2 and of proliferating in response to Con A provided the comitogen PMA is present. It can be argued that the failure to produce IL 2 or to respond to proliferative signals is not fundamental to the development of the disease sustained by autoimmune-prone mice.     

频谱水对大鼠血浆白介素1、6的白细胞行为的影响

目的 :探讨频谱水对大鼠白介素 1、6(IL 1、IL 6)和白细胞行为的影响。方法 :静脉内注入内毒素造成炎症刺激模型 ,用生化方法检测血浆IL 1、IL 6的含量 ,用活体方法观察白细胞粘附、游出行为。结果 :实验组大鼠饮用频谱水 30、60天后 ,血浆IL 1、IL 6的含量比对照组高 ,白细胞粘附、游出的数量比对照组明显减少。对照组大鼠白细胞粘附、游出的数量较实验组明显增多 ,而血浆中IL 1、IL 6的含量比实验组低。结论 :频谱水能够提高血浆IL 1、IL 6的含量 ,显著减轻白细胞粘附及游出。     

Multi‐parametric flow cytometric and genetic investigation of the peripheral B cell compartment in human type 1 diabetes

The appearance of circulating islet-specific autoantibodies before disease diagnosis is a hallmark of human type 1 diabetes (T1D), and suggests a role for B cells in the pathogenesis of the disease. Alterations in the peripheral B cell compartment have been reported in T1D patients; however, to date, such studies have produced conflicting results and have been limited by sample size. In this study, we have performed a detailed characterization of the B cell compartment in T1D patients (n = 45) and healthy controls (n = 46), and assessed the secretion of the anti-inflammatory cytokine interleukin (IL)-10 in purified B cells from the same donors. Overall, we found no evidence for a profound alteration of the B cell compartment or in the production of IL-10 in peripheral blood of T1D patients. We also investigated age-related changes in peripheral B cell subsets and confirmed the sharp decrease with age of transitional CD19⁺CD27⁻CD24^hiCD38^hi B cells, a subset that has recently been ascribed a putative regulatory function. Genetic analysis of the B cell compartment revealed evidence for association of the IL2–IL21 T1D locus with IL-10 production by both memory B cells (P = 6·4 × 10⁻⁴) and islet-specific CD4⁺ T cells (P = 2·9 × 10⁻³). In contrast to previous reports, we found no evidence for an alteration of the B cell compartment in healthy individuals homozygous for the non-synonymous PTPN22 Trp⁶²⁰ T1D risk allele (rs2476601; Arg⁶²⁰Trp). The IL2–IL21 association we have identified, if confirmed, suggests a novel role for B cells in T1D pathogenesis through the production of IL-10, and reinforces the importance of IL-10 production by autoreactive CD4⁺ T cells.