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 activation of T lymphocytes from human immunodeficiency virus (HIV)-seropositive blood donors. I. Soluble interleukin 2 receptor (IL2R) production parallels cellular IL2R expression and DNA synthesis

We investigated the relationship of soluble interleukin 2 receptor (sIL2R) production to cellular IL2R expression and DNA synthesis by mitogen-stimulated mononuclear cells from blood donors seropositive for human immunodeficiency virus (HIV). SIL2R was measured using an enzyme-linked immunosorbent assay which employed 2 anti-IL2R monoclonal antibodies recognizing distinct IL2R epitopes. Decreased phytohemagglutinin-induced DNA synthesis and cellular IL2R expression were accompanied by decreased levels of sIL2R in cell culture supernatants. Similar findings were observed for pokeweed mitogen-induced responses. There was no detectable spontaneous secretion of sIL2R into culture supernatants by unstimulated mononuclear cells from either HIV-seropositive or control seronegative donors. These findings indicate that thein vitro T-cell activation defects which characterize HIV infection include decreased sIL2R production, as well as decreased cellular IL2R expression and DNA synthesis. Further, they show that assessment of supernatant sIL2R levels can be used as a valid, reliable assay for T-cell activation.     

The role of interleukin 2 (IL-2) and serum-soluble IL-2 receptor cells in idiopathic IgA nephropathy.

Interleukin 2 (IL-2) plays a central role in the immune response and may be involved in the derangement of cellular immunoregulation of idiopathic IgA nephropathy (IgAN). The aim of this study was to investigate the serum levels and production of IL-2 from peripheral blood mononuclear cells (PBMC) and the distribution of IL-2 receptor cells and serum-soluble IL-2 receptor cells (sIL-2R) in patients with IgAN. Twenty-four patients with IgA nephropathy and 11 healthy controls (age and sex matched) were studied during an infection-free period without signs of clinical activity at the moment of the study. Serum IL-2 concentrations did not differ between patients and controls. The supernatant levels of IL-2 taken from 24-hr cultures of PBMC stimulated with phytohemagglutinin or tumor necrosis factor increased significantly in the patients but not in the controls. The percentage of IL-2R positive cells (CD25+) was increased in patients compared with controls. Moreover, IgAN patients had increased activated CD4+ lymphocytes when compared with the controls. Serum levels of sIL-2R were significantly higher in patients than in controls. There were no correlations among renal function, serum IgA levels, and urinary findings with cellular subsets or with IL-2 levels. However, sIL-2R was higher in the subgroup of patients with episodic macrohematuria and was closely related with the presence of red blood cells in the urinary sediment. We conclude that PBMC of IgA nephropathy patients have an overproduction of IL-2 after mitogenic stimulation, an increased helper T cell activity, increased IL-2R+ cells, and elevated serum levels of sIL-2R. These alterations are present in periods of apparent clinical inactivity. Finally, sIL-2R is closely related with hematuria, providing a good marker for disease activity. Our results suggest a pivotal role of IL-2 in cellular immune responses with regard to T cell activation in patients with IgAN.     

Lactate dehydrogenase (LDH) isoenzymes and proliferative activity of lymphoid cells--an immunocytochemical study.

The regulation of cytokine production and T cell proliferation by other cytokines is mandatory in mediating inflammatory responses but the full understanding is far from complete. We have previously reported increased production of IL-2 and IL-2 receptors (IL-2R) in IgA nephropathy. The present study was undertaken to examine other cytokine production during T cell activation in IgA nephropathy. Peripheral blood mononuclear cells (PBMC) from 17 IgA nephritic patients and 14 controls were cultured with phytohaemagglutinin and phorbol myristate acetate for 48 h for maximal cytokine production. IL-2Rs and IL-4 receptors (IL-4Rs) expressed on cultured PBMC were studied by a radioimmunoassay using monoclonal antibodies against these receptors. Although the total cellular IL-2R expression and percentages of T helper and T suppressor cells did not differ between the patients and controls, there was a significant increase in activated T helper cells expressing IL-2R in patients with IgA nephropathy. The total cellular IL-4R expression was elevated in IgA nephritic patients (P less than 0.005). IL-2 production by PBMC was raised in IgA nephritic patients compared with controls (P less than 0.05) but no difference in IL-4 or IL-6 production was observed. The interferon-gamma production by PBMC was significantly increased in patients with IgA nephropathy (P less than 0.025). No correlation was observed between individual cytokine levels. Our data suggest there are selective increases in cytokine production in IgA nephropathy.     

Increased serum levels of soluble interleukin 2 receptor in patients with drug-induced allergic hepatitis.

Soluble interleukin 2 receptor (sIL2R) level in the serum of 10 patients with drug-induced allergic hepatitis (DIAH) were quantified with a solid-phase enzyme immunoassay using two monoclonal antibodies against the receptor. The sIL2R levels in patients with DIAH were significantly higher than in 15 normal subjects (p less than 0.01). The high sIL2R levels observed during the florid stage of DIAH returned to normal during convalescence (p less than 0.05). SIL2R from the serum of DIAH patients was found to bind to a recombinant IL2 affinity column. It seems likely that sIL2R inhibits the hyper-immune reaction that occurs in drug allergies due to bind IL2. But, we have no data that sIL2R inhibits actually the biological activity of IL2.     

Fluctuations of plasma beta 2-microglobulin, soluble interleukin 2 receptor and interferon-gamma concentrations after adoptive immunotherapy with high-dose interleukin 2 and lymphokine-activated killer cells

We report the serum levels of soluble interleukin 2 receptor (sIL2R), beta 2-microglobulin (beta 2-M) and interferon-gamma (IFN-gamma) in patients undergoing adoptive immunotherapy with rIL2 and lymphocyte-activated killer (LAK) cells. Our results indicate that rIL2 induced a marked increase of the serum concentration of these markers, although this increase varied considerably for different individuals. Parallel studies with the same patients also showed a marked rise in the number of IL2R+ lymphocytes: the IL2Rs expressed on these cells were mainly of the "low affinity" type. We suggest that evaluation of these markers may allow the monitoring of immune system activation induced by rIL2 in patients undergoing adoptive rIL2 and LAK cell immunotherapy.     

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.     

癫癎患者血清细胞因子水平的研究

目的：探讨细胞因子与癫癎发病的关系。方法：采用酶联免疫吸附法（ＥＬＩＳＡ） 对５８ 例癫癎患者及４２ 例正常对照血清可溶性白细胞介素２ 受体（ｓＩＬ２Ｒ） 及肿瘤坏死因子（ＴＮＦ） 水平进行了检测。结果：癫癎组血清ｓＩＬ２Ｒ 及ＴＮＦ 水平显著高于正常对照组,血清ｓＩＬ２Ｒ 及ＴＮＦ 水平变化与癫癎类型无关,抗癫癎药物对血清ｓＩＬ２Ｒ 及ＴＮＦ 水平无明显影响。结论：癫癎患者存在内在的细胞免疫功能紊乱,其单核—巨噬细胞及Ｔ淋巴细胞处于激活状态。     

Interleukin 2 receptor expression in renal biopsies and the diagnosis of acute allograft rejection

The aim of the study is to test the diagnostic usefulness of assessing interleukin 2 receptor (IL2R) expression in infiltrating lymphocytes in renal biopsies from patients with suspected acute renal allograft rejection and to compare the NIH-CCTT and the Banff 97 systems of classifying the histopathologic changes in acute renal allograft rejection. The expression of interleukin 2 (IL2) and IL2R, as shown immuno-histochemically, is the final step in T cell mediated acute renal allograft rejection. Renal biopsies obtained from 40 patients clinically suspected to have early acute allograft rejection were examined histologically to diagnose acute allograft rejection and classified by the two systems. Frozen sections of the biopsies were stained with specific antibody for the presence of IL2R. 31 of the 40 patients were histologically and clinically confirmed to have acute allograft rejection. There was significant correlation with this diagnosis and the demonstration of IL2R on infiltrating lymphocytes. The CCTT system of grading correlated better with the presence of IL2R and the confirmed diagnosis of acute allograft rejection. The immunohistochemical demonstration of IL2R is a useful adjunct in the evaluation of biopsies suspected to show changes of acute cellular rejection. Since IL2 expression reflects the relative proportion of activated lymphocytes in the cellular infiltrate, it is proposed that the degree of IL2 expression may reflect the response of the use of monoclonal antibodies (Humanised/Chimaerised) as anti rejection therapy.     

吸入糖皮质激素对哮喘儿童细胞免疫功能的影响

为了解糖皮质激素对免疫功能的作用，检测了哮喘发作期患儿。对经丙酸倍氯松（ＢＤＰ）吸入治疗的患儿及正常儿童外周血淋巴细胞膜ＩＬ－２受体（ｍＩＬ－２Ｒ）、超微结构、血浆游离ＩＬ－２受体（ｓＩＬ－２Ｒ）、淋巴细胞亚群及ＢＤＰ对体外培养淋巴细胞ｍＩＬ－２Ｒ的表达进行比较。结果显示，ＢＤＰ可以明显抑制ＰＨＡ刺激引起的体外培养淋巴细胞ｍＩＬ－２Ｒ的表达，引起淋巴细胞的凋亡，而吸入ＢＤＰ（２００μｇ／ｄ）的哮喘患儿（缓解期）除ｓＩＬ－２Ｒ比哮喘发作期显著降低（Ｐ＜０．０５）外，外周血淋巴细胞ｍＩＬ－２Ｒ及淋巴细胞亚群并未出现显著变化．结果提示，抑制淋巴细胞ｍＩＬ－２Ｒ表达和淋巴细胞活化可能是ＢＤＰ治疗哮喘等变态反应的重要机制之一。吸入ＢＤＰ主要是呼吸道局部作用，对全身细胞免疫指标影响不大     

IgA肾病患者IL—2的产生及IL—2受体的表达

本文对IgA肾病(IgAN)外周血淋巴细胞白细胞介素2(IL-2)的产生、受体的表达及免疫球蛋白的产生进行了研究。结果发现:外周血淋巴细胞产生IL-2的活性明显增高,IL-2受体表达亦明显增强并伴有免疫球蛋白产生增多,提示IgAN存在着细胞免疫功能的紊乱。     

慢性乙型肝炎,肝硬化患者外周血LAK细胞活性和sIL—2R测定的意义

采用＾３Ｈ－ＴｄＲ释放法测定５１例慢性肝病患者（ＣＰＨ１０例、ＣＡＨ２３例、ＬＣ１８例）外周血ＬＡＫ细胞活性，并用酶联法测定患者血清中ｓＩＬ－２Ｒ含量；与２９例正常对照组比较，发现肝病患者ＬＡＫ活性降低，ＨＢＶＤＮＡ阳性组ＬＡＫ活性较阴性组低（Ｐ〈０．０５），ｓＩＬ－２Ｒ增高，且慢性肝病组ＬＡＫ活性与ｓＩＬ－２Ｒ水平呈负相关，说明ＬＡＫ活性与机体免疫功能状态有关，ＨＢＶ的复制和高浓度的ｓＩＬ－２Ｒ     

Multiple sclerosis: effect of myelin basic protein on interleukin 1, interleukin 2 production and interleukin 2 receptor expression in vitro.

The production of interleukin 1 (IL-1) and interleukin 2 (IL-2) by peripheral blood mononuclear cells (PBMC) stimulated with human myelin basic protein (MBP) was assessed in vitro in multiple sclerosis (MS) patients in relapse, patients with other neurological diseases (OND) and healthy subjects. Myelin basic protein significantly increased both IL-1 and IL-2 production by PBMC from MS patients during relapse when compared to OND patients or healthy controls. The most efficient concentration of MBP for the induction of IL-1 and IL-2 was 50 micrograms/ml. The optimal IL-1 production occurred after 48 h of PBMC culture and optimal IL-2 production after 72 h of PBMC culture. Anti-Tac monoclonal antibody (MoAb) was used to study IL-2 receptor expression on the same sample of PBM used for IL-2 study in MS patients in relapse. In addition IL-2 receptor expression was studied in PBMC from chronic progressive MS patients. In both MS groups IL-2 receptor expression on PBMC stimulated with MBP appeared higher than in control groups, but these differences were not statistically significant. IL-2 receptor expression on cerebrospinal fluid lymphocytes (CSF-L) either unstimulated or MBP-stimulated was, however, significantly higher in both MS groups when compared to OND patients. These results confirm the presence of activated lymphocytes in the CSF of MS patients during active stages of disease and suggest that this activation may be related to expansion of MBP specific cells.     

多发性硬化患者血清sIL-6R 水平观察

为探讨可溶性白细胞介素－６受体（ｓＩＬ－６Ｒ）在多发性硬化（ＭＳ）发生、发展中的作用，采用双抗体夹心ＥＬＩＳＡ方法，对２８例ＭＳ患者和２５例正常对照组血清ｓＩＬ－６Ｒ水平进行了测定。结果显示缓解－复发型和进展型ＭＳ患者血清ｓＩＬ－６Ｒ水平明显高于正常对照组和良性型组（Ｐ＜０．０５，Ｐ＜０．０１）；而良性型ＭＳ患者血清ｓＩＬ－６Ｒ水平与正常对照组相比，未见明显改变。缓解－复发型ＭＳ患者血清ｓＩＬ－６Ｒ水平随着病情的好转而下降。ｓＩＬ－６Ｒ水平与ＭＳ患者头颅ＭＲＩ上的病灶大小和部位无明显相关性（ｒ１＝０．１２４，Ｐ＞０．０５；ｒ２＝０．０９１，Ｐ＞０．０５）。推测ｓＩＬ－６Ｒ水平的高低可作为判断ＭＳ患者病情变化的指标之一。     

Molecular analysis for patients with IL‐12 receptor β1 deficiency

Autosomal recessive interleukin‐12 receptor β1 (IL‐12Rβ1) deficiency has been described as the most common cause of Mendelian susceptibility to mycobacterial disease (MSMD), characterized by clinical disease due to weakly virulent mycobacteria such as Bacille Calmette–Guérin (BCG) vaccines and environmental mycobacteria (EM) in children who are normally resistant to most infectious agents. Here, we report the cases of five patients with mycobacterial infection, including one with systemic lupus erythematosus (SLE). Blood samples from patients and healthy controls were activated in vitro with BCG, BCG+IL‐12, and BCG+IFN‐γ. The results showed reduced or no production of IFN‐γ after IL‐12 stimulation in all samples. IL‐12Rβ1 expression on the cell surface was negligible or absent. Genetic analysis showed five novel mutations.     

大肠癌患者围手术期免疫状态变化的临床意义

目的 研究大肠癌患者围手术期的免疫状态。方法 应用ＡＰＡＡＰ夹心法ＥＬＩＳＡ和比色法分别测定大肠癌患者手术前后Ｔ细胞亚群变化,血浆可溶性白介２受体（ｓＩＬ－２Ｒ）及一氧化氮（ＮＯ）和癌组织的ＳＩＬ－２Ｒ,ＮＯ含量。结果 大谫ＣＤ３,ＣＤ４细胞、ＣＤ４／ＣＤ８比值和血浆ＮＯ水平明显低于正常,血浆ＳＩＬ－２Ｒ、ＣＤ８细胞较正常显著增高,肿瘤切除后３周左右ＣＤ３、ＣＤ４细胞、ＣＤ４／ＣＤ８和血浆术前有明     

Dissection of signals controlling T cell function and activation: H7, an inhibitor of protein kinase C, blocks induction of primary T cell proliferation by suppressing interleukin (IL)2 receptor expression without affecting IL2 production

T cell activation induced via cross-linking of the T cell receptor (TcR) stimulates hydrolysis of phosphatidylinositol to the second messengers diacylglycerol (DAG) and inositol 1,4,5-triphosphate (IP3). DAG is necessary for the activation and function of protein kinase C (PKC) which is suggested to play a key role in the cascade of signal transduction when translocated from the cytosol to the cell membrane. In this report, we investigated responses of resting vs. activated Ly-2+ and L3T4+ T lymphocytes in the presence of the PKC inhibitor H7 [1-(5-isoquinolinylsulfonyl)-2-methylpiperazine]. H7 inhibited the induction of primary T cell proliferation, while interleukin 2 (IL 2) production was fully retained. The effect of the PKC inhibitor on primary T cells depended on the type of ligand interacting with the TcR: increasing doses of concanavalin A or of immobilized anti-CD3 monoclonal antibody (mAb), but not of anti-V beta 8 or of anti-TcR alpha/beta mAb, partly overcame the blockade, indicating a differential signaling compared to the former stimuli. The blockade of T cell proliferation by H7 was not due to an inhibition of PKC translocation, but occurred even 4-8 h after T cell induction and correlated with a significant reduction of IL 2 receptor (IL 2R) expression. In contrast, the mRNA levels of IL 2R and the cellular proto-oncogenes c-fos and c-myc were not affected. On activated T cells, H7 neither blocked proliferation nor IL2R expression. Consequently, H7 dissects the signal resulting in T cell proliferation from those governing the triggering of other T cell functions, i.e. IL 2 production, during primary responses of Ly-2+ or L3T4+ murine T lymphocytes.     

活动性肺结核患者血清sIL－2R与抗PPD抗体的相关性

应用ＥＬＩＳＡ检测了正常人和肺结核患者的血清可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）和抗ＰＰＤ抗体水平。结果表明，活动性肺结核患者的血清ｓＩＬ－２Ｒ和抗ＰＰＤ抗体呈正相关（γ＝０．４３３，Ｐ＜０．００１），而且两者的水平明显高于正常对照组；抗ＰＰＤ抗体阳性的活动性肺结核患者血清中ｓＩＬ－２Ｒ浓度明显高于抗体阴性患者，提示血清ｓＩＬ－２Ｒ浓度测定可以反映肺结核患者细胞免疫功能状态。     

活动性肺结核患者血清sIL－2R与抗PPD抗体的相关性

应用ＥＬＩＳＡ检测了正常人和肺结核患者的血清可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）和抗ＰＰＤ抗体水平。结果表明，活动性肺结核患者的血清ｓＩＬ－２Ｒ和抗ＰＰＤ抗体呈正相关（γ＝０．４３３，Ｐ＜０．００１），而且两者的水平明显高于正常对照组；抗ＰＰＤ抗体阳性的活动性肺结核患者血清中ｓＩＬ－２Ｒ浓度明显高于抗体阴性患者，提示血清ｓＩＬ－２Ｒ浓度测定可以反映肺结核患者细胞免疫功能状态。